Biomaterial Lab 10

DEPARTMENT OF BIOMEDICAL
ENGINEERING
UNIVERSITY OF ENGINEERING AND TECHNOLOGY
LAHORE (New Campus)
Lab Report
Biomaterials
Name: ___________Haseeb-UR-Rehman____________
Roll No: __________2020-BME-10____________
Signature of Instructor:_________
Table of contents
Sr# Title Page #
1 Preparation and Micro-structure Analysis of Stainless Steel specimen 03
2 Preparation and Micro-structure Analysis of Mild Steel 10
3 To study the effect of corrosion of Stainless Steel (SS) and Mild Steel (MS) in 18
acidic media
4 To study the effect of corrosion of Stainless Steel (SS) and Mild Steel (MS) in 27
basic media
5 Immersion of Stainless Steel (SS) and Mild Steel in Ringer Solution 36
6 Preparation of Bio-ceramics Pellet Using Hydraulic Press 45
7 Densification of Bio-Ceramics Pellet by Sintering 53
8 Production and sintering of porous bio-ceramics scaffolds using naphthalene as 62
the pore former
9 To determine the hardness of metallic biomaterial using Rockwell Hardness 73

Method
10 To determine the Hardness of Metallic biomaterials using Brinell hardness 80

method
11 To determine ductility and quality of metallic samples using Erichsen Cupping 89
Index Machine
12 To determine the tensile strength of metallic samples using Universal testing machine 96
Safety Rules for Students
 Students are not allowed to enter in the lab without wearing lab coat
 Report all accidents, injuries, and breakage of glass or equipment to instructor
immediately.
 Keep pathways clear by placing extra items (books, bags, etc.) on the shelves or under the
work tables. If under the tables, make sure that these items cannot be stepped on.
 Long hair (chin-length or longer) must be tied back to avoid catching fire.
 Wear sensible clothing including footwear. Loose clothing should be secured so they do
not get caught in a flame or chemicals.
 Work quietly — know what you are doing by reading the assigned
experiment before you start to work. Pay close attention to any cautions described in the
laboratory exercises
 Do not taste or smell chemicals.
 Wear safety goggles to protect your eyes when heating substances, dissecting, etc.
 Do not attempt to change the position of glass tubing in a stopper.
 Never point a test tube being heated at another student or yourself. Never look into a test
tube while you are heating it.
 Unauthorized experiments or procedures must not be attempted.
 Keep solids out of the sink.
 Leave your work station clean and in good order before leaving the laboratory.
 Do not lean, hang over or sit on the laboratory tables.
 Do not leave your assigned laboratory station without permission of the teacher.
 Learn the location of the fire extinguisher, eye wash station, first aid kit and safety
shower.
 Fooling around or "horse play" in the laboratory is absolutely forbidden. Students found
in violation of this safety rule will be barred from participating in future labs and could
result in suspension.
 Anyone wearing acrylic nails will not be allowed to work with matches, lighted splints,
Bunsen burners, etc.
 Do not lift any solutions, glassware or other types of apparatus above eye level.
 Follow all instructions given by your teacher.
 Learn how to transport all materials and equipment safely.
 No eating or drinking in the lab at any time!
 Use of mobile phone is not allowed during lab session
[1]
General instructions for Students
 Always keep your bench CLEAN to avoid contamination. Clean surface of your bench
with 70% ethanol.
 Make your own buffers and reagents i.e. PBS, culture mediums. DO NOT USE OTHER
PEOPLE’S!!!!! You may contaminate their medium or it might be already contaminated.
 Mediums do not last forever! Always write the date that you prepared the medium and
clearly label it with your name and content.
 When you make mediums/ aliquots it is wise to use parafilm and seal the cap.
 Always clean hood with 70% ethanol before you work and when you finish! Respect the
next person who works after you. Turn on UV light in the hood when you have finished
working with highly contaminating agents.
 Stop and THINK!!!!!!! Most of the times contamination is a result of careless actions
 When you open a sterile bottle of culture medium outside the hood IT IS NOT STERILE
ANY MORE!!!
 Sterile Tips that are opened outside the hood cannot be used again inside the hood!!!
 when you are pipetting be careful not to touch the tip to any surfaces.
 Avoid pipetor contamination DO NOT EXCEED INDICATED VOLUME. If you are
using P200 NEVER use it for volumes greater than 200μl.
 IF YOU ARE NOT SURE ALWAYS ASK!!!!
 Always report spills, broken glass etc.
 Always assume that biological fluids are infectious and use GLOVES
 When you are disposing needles do not take out the cover! DISPOSE NEEDLE WITH
THE COVER!
 Check labels of reagents that you have not used before and ask your supervisor about
handling.
 Avoid filling tubes to the rim.
 Ensure that the centrifuge is properly balanced.
 Do not open the lid during or immediately after operation, attempt to stop a spinning
rotor by hand or with an object, or interfere with the interlock safety device.
 Do not forget the C/F at 4oC!!! If you do not use it anymore RAISE the temperature.
 At the end of the day leave C/F lid half-open to ensure evaporation.
[2]
Lab Assessment Rubrics (Biomaterials)
Lab #--------01--------
Name: Haseeb-UR-Rehman___________________ Reg. No.: ___2020-BME-10_____________________
Criteria Poor (=<4) Satisfactory (6) Good (8) Excellent (10) Marks
CLO1 PLO4 Set-up and The student has shown The student has shown The student has shown The student uses the
Equipment Care irresponsibility using the responsibility towards the responsibility towards equipment responsibly
(10×2) equipment and didn’t clear the cleanliness of workplace using the equipment while and clears the
leftovers at the workplace however did not follow the he didn’t care about the leftovers at the
on completion of lab work. SOP for equipment cleanliness of workplace workplace on
operation completion of lab work.
Conduction of Lacks the appropriate Demonstrates general Demonstrates good Demonstrates very good
experiment knowledge of the lab knowledge of lab knowledge of the lab knowledge of the lab
(10×2) procedures. · Often requires procedures. · Requires help procedures. Will ask procedures. Gladly helps
help from the teacher to even from teacher with some peers for help with other students to follow
complete basic procedures steps in procedures problems in lab procedures. Thoroughly
procedures. Works to and carefully follows
follow each step before each step before moving
moving on to the next step on to next step
Safety (10×2) Proper safety precautions are · Proper safety precautions Proper safety precautions Proper safety
consistently missed. Needs to are often missed. Needs to are generally used May precautions are
be reminded often during the be reminded more than once need to be reminded once consistently used.
lab during the lab during the lab Consistently thinks
ahead to ensure safety.
Will often help other
students to conduct labs
safely
Total
Formal Report illustrates inaccurate Report illustrates a limited Report illustrates an Report illustrate an
Reporting (10×1) understanding of scientific understanding of scientific accurate understanding of accurate and through
concepts underlying the lab. concepts underlying the lab. most scientific concept understanding of
Figures, graphs, tables contain Most figures, graphs, tables underlying the lab. scientific concepts
errors or are poorly constructed, OK, some still missing All figures, graphs, tables underlying the lab. All
have missing titles, captions or some important or required are correctly drawn, but figures, graphs, tables
numbers, units missing or features. Some of the results some have minor problems are correctly drawn, are
incorrect, etc. Very incomplete have been correctly or could still be improved. numbered and contain
or incorrect interpretation of interpreted and discussed; Almost all of the results titles/captions. All-
trends and comparison of data partial but incomplete have been correctly important trends and data
indicating a lack of understanding of results is interpreted and discussed, comparisons have been
understanding of results still evident only minor improvements interpreted correctly and
are needed discussed, good
understanding of results
is conveyed
Data analysis Measurements are incomplete, Measurements are Measurements are mostly Measurements are both
and inaccurate and imprecise. somewhat inaccurate and accurate. Observations are accurate and precise
interpretation Data analysis and interpretation very imprecise. Data generally complete. Work Observations are very
(10×1) work are incomplete or not analysis and interpretation is organized. Only 2 or 3 thorough and may
included. Symbols, units and work are incomplete or minor errors using recognize possible errors
significant figures are not recorded in a confusing symbols, units and in data analysis and
included way. There are 3 or more significant digits interpretation work is
minor errors using symbols, neat and
units and significant digits organized Includes
or 2 major errors appropriate symbols,
units and significant
digits
Total
CLO3 PLO9 Lab Participation was minimal OR Did the lab but did not Used time pretty well. Used time well in lab
Participation student was hostile about appear very interested. Stayed focused on the and focused attention on
(10×1) participating Focus was lost on several experiment most of the the experiment.
occasions. time.
Total
 Note: 10 % of cognitive (CLO2) will evaluate through written lab viva
Lab Instructor:
Name: Dr. Nida Iqbal Signature : _______________________
[3]
Experiment # 01
Preparation and Micro-structure Analysis of Stainless Steel specimen
 Objective
1. Metallographic Sample Preparation of Stainless Steel
2. Observation of Microstructure of Metals
 Apparatus Required
Different Grated Sand Papers.
1. Sand Paper 220C
2. Sand Paper 400C
3. Sand Paper 600C
4. Sand Paper 800C
 Raw Materials
1. Stainless Steel
 Theory
1. Metallography and Its Application:
Metallography is the study of the physical structure and components of metals, by using
microscopy. Ceramic and polymeric materials may also be prepared using metallographic
techniques, hence the terms ceramography, plastography and, collectively, materialography. It is
a scientific discipline of observing and determining the chemical and atomic structure of the grains,
constituents or inclusions in the metallic alloys.
 Mechanical Specmen Prepration:
Mechanical preparation is the most common preparation method. Successively finer abrasive
particles are used to remove material from the sample surface until the desired surface quality is
achieved. Many different machines are available for doing this grinding and polishing, and they
are able to meet different demands for quality, capacity, and reproducibility
 Systematic preparation:
This method is the easiest way to achieve the true structure. Sample preparation must therefore
pursue rules that are suitable for most materials. Different materials with similar properties
(hardness and ductility) will respond alike and thus require the same consumables during
preparation.
Applications:
 Metallography is used in almost all stages during the lifetime of the components: from the
initial materials development to inspection, production, manufacturing process control, and
even failure analysis if needed.
 Powder metallurgy techniques have been used to produce controlled porous structures,
such as porous coatings applied for dental and orthopedic surgical implants, which allow
bony tissue ingrowth within the implant surface, thereby improving fixation
[4]
Figure 1 Metallography in orthopedics Figure 2 Analysis of metals by metallography technique
 They involve nearly waste-free net-shape forming.

 They offer precise choice of chemical composition through the use of high-melting or hard-
to machine alloys to improve implant biocompatibility
 There is a lack of chemical inhomogeneity typical for cast as well as some plastic formed
materials, thus improving resistance to corrosion
 They offer porosity of appropriate pore geometry. This improves bone tissue in-growth,
thus enhancing endoprosthesis stability
 They offer the possibility to form various composite materials containing addtions that
improve biofunctionality
2. Metallurgical Microscope:
A metallurgical microscope is the type of light microscope, specialized for looking and studying
at the cross section of metals and alloys microscopically. Metallurgical Micro-scope works on the
principal of illumination. As metals are opaque substances, they must be illuminated by lightening.
Therefore, the source of light is located within the Micro-Scope tube. Image quality and resolving
power determined by the quality of the objective. Metallurgical Microscope consists of two
systems. Optical System and Illuminating System.
Features:
Metallurgical microscopes have several unique features that are not always found on a biological
or stereo microscope:
 Polarization
 Differential Interference Contrast (DIC)
 High power magnification and high resolution with reflected light illumination
 Brightfield and Darkfield options
 Reflected (and sometimes also transmitted) illumination
[5]
Figure 3 Metallurgical microscope Figure 4 Working elaboration
Optical System:
It consists of:
Eyepiece lens, which is what the observer uses to see the image
Relay system, which reflects and enlarges the image
Objective lens, which takes in the image from the sample on the stage
Illuminating System:
It is designed to prevent glare from light bouncing off the surface of the sample and consists of:
High-intensity light source
Condenser lenses
Aperture diaphragm
Plane glass reflector
Colored or polarizing filters
[6]
3. Microscopic Structure of Stainless Steel:
Stainless Steel are iron based alloys containing at least 11% Chromium. They typically contain
less than 30% of chromium and more than 50% of iron. Stainless steel obtains their stainless
characteristics because of the formation of an invisible and adherent chromium-rich oxide surface
film. The Microstructure in the annealed condition consists of ferrite grains in which fine carbides
are embedded.
Figure 5 stainless steel microscopic structures
 Experimental Procedure
1. Four to five sand papers of different gradings and stainless steel had given to us.
2. At first sand paper with 180C Grading was used to grind the stainless steel and observed
under the metallurgical microscope.
3. After that, sand paper with 400C Grading was used to grind the stainless steel and observed
4. After 400C graded sand paper, sand paper with 800C Grading was used to grind the
stainless steel and observed under the metallurgical microscope.
5. In last, sand paper with 1000C Grading was used to grind the stainless steel and observed
 Results
1st Grid Paper (220C)
Figure 2 grid paper 220
[7]
2nd Grid Paper (400C)
3rd Grid Paper (600C)
4th Grid Paper (800C)
[8]
 Discussion
For heavy sanding, to remove the bruises from the metal, a low value graded (like 180C graded
sand paper) is used. To smoother the surface, we use sand paper of high carbon graded value (for
instance 1000C graded sand paper). The sample piece is rough and have stains at microscopic level
to remove we use sand papers of different sizes. When the grit size is 200, the specimen becomes
fine piece after grinding with some water by using spray gun. The specimen becomes finer when
sand paper grit size is 400. Similarly, when we use 600 and 800 grit size sand paper the specimen
surface become more shine, polish and finer. When grit size increases the rusting and rough surface
of specimen reduces and fine piece formed. This means when the grit size of sand paper increases
the specimen become finer than before. The purpose of removing rusting is to prepare the sample
for next experiments.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
3. Wear proper covered shoes
4. Not wear jewelry and watch during the experiment
5. Wash your hands with soap and water before entering or leaving the lab.
6. 2. Wear shoes that completely cover your feet, you cannot wear sandals or other open-
toe OR open-heel shoes (clogs are ok).
7. 3. You must wear a lab coat when working in the microbiology lab. You must store
your lab coat/shirt in the lab. Your lab coat/shirt may NOT be removed from the room
during the semester.
8. 4. Long hair must be tied back in a ponytail, all hats must be removed or turned
backward.
9. 5. Accidents or injuries must be reported to your instructor as soon as possible. Know
the location of the fire blanket, fire extinguisher, shower and eyewash in case of
emergency.
10. 6. Do not eat, drink, or chew gum in the lab. Avoid hand-to-mouth activities such as
nail biting or chewing on the end of a pencil/pen.
11. 7. Cell phones are not allowed in lab under any circumstances.
12. 8. Do not put paper towels used to wipe down the bench in the bench top waste baskets.
13. 10. Disinfect your lab space at the start and end of each lab period. To disinfect your
area wipe down the table top thoroughly with laboratory disinfectant and a paper towel.
When leaving the lab, apply disinfectant to the bench area and leave to air dry (wet).
[9]
Lab #--------02--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[10]
Experiment # 02
Preparation and Micro-structure Analysis of Mild Steel
 Objective
1. Metallographic Sample Preparation of Mild Steel.
2. Observation of Microstructure of Metals.
Different Grated Sand Papers.
1. Sand Paper 180C
2. Sand Paper 400C
3. Sand Paper 800C
4. Sand Paper 1000C
 Raw Materials
1. Mild Steel
 Theory
1. Metallography and Its Application:
Metallography is the study of the physical structure and components of metals, by using
microscopy. Ceramic and polymeric materials may also be prepared using metallographic
techniques, hence the terms ceramography, plastography and, collectively, materialography. It is
a scientific discipline of observing and determining the chemical and atomic structure of the grains,
constituents or inclusions in the metallic alloys.
 Mechanical Specmen Prepration:
Mechanical preparation is the most common preparation method. Successively finer abrasive
particles are used to remove material from the sample surface until the desired surface quality is
achieved. Many different machines are available for doing this grinding and polishing, and they
are able to meet different demands for quality, capacity, and reproducibility
 Systematic preparation:
This method is the easiest way to achieve the true structure. Sample preparation must therefore
pursue rules that are suitable for most materials. Different materials with similar properties
(hardness and ductility) will respond alike and thus require the same consumables during
preparation.
Applications:
 Metallography is used in almost all stages during the lifetime of the components: from the
initial materials development to inspection, production, manufacturing process control, and
even failure analysis if needed.
 Powder metallurgy techniques have been used to produce controlled porous structures,
such as porous coatings applied for dental and orthopedic surgical implants, which allow
bony tissue ingrowth within the implant surface, thereby improving fixation
[11]
Figure 3 Metallography in orthopedics Figure 2 Analysis of metals by metallography technique
 They involve nearly waste-free net-shape forming.

 They offer precise choice of chemical composition through the use of high-melting or hard-
to machine alloys to improve implant biocompatibility
 There is a lack of chemical inhomogeneity typical for cast as well as some plastic formed
materials, thus improving resistance to corrosion
 They offer porosity of appropriate pore geometry. This improves bone tissue in-growth,
thus enhancing endoprosthesis stability
 They offer the possibility to form various composite materials containing addtions that
improve biofunctionality
2. Metallurgical Microscope:
A metallurgical microscope is the type of light microscope, specialized for looking and studying
at the cross section of metals and alloys microscopically. Metallurgical Micro-scope works on the
principal of illumination. As metals are opaque substances, they must be illuminated by lightening.
Therefore, the source of light is located within the Micro-Scope tube. Image quality and resolving
power determined by the quality of the objective. Metallurgical Microscope consists of two
systems. Optical System and Illuminating System.
Features:
Metallurgical microscopes have several unique features that are not always found on a biological
or stereo microscope:
 Polarization
 Differential Interference Contrast (DIC)
 High power magnification and high resolution with reflected light illumination
 Brightfield and Darkfield options
 Reflected (and sometimes also transmitted) illumination
[12]
Figure 3 Metallurgical microscope Figure 4 Working elaboration
Optical System:
It consists of:
Eyepiece lens, which is what the observer uses to see the image
Relay system, which reflects and enlarges the image
Objective lens, which takes in the image from the sample on the stage
Illuminating System:
It is designed to prevent glare from light bouncing off the surface of the sample and consists of:
High-intensity light source
Condenser lenses
Aperture diaphragm
Plane glass reflector
Colored or polarizing filters
[13]
Microscopic Structure of Mild Steel:
Mild Steel is a type of steel with low percentage of Carbon. The carbon content varies from piece
to piece. The range of carbon in different Mild Steel is 0.5%-0.25%. Austenitic mild steel have
a face-centered cubic (FCC) crystal structure and are composed of iron, carbon, chromium, and at
least 8% nickel. Due to their high chromium and nickel content, they are highly corrosion resistant
and non-magnetic. The microstructure is predominantly martensite but also has allotriomorphic
ferrite, widmanstätten ferrite, bainite and pearlite.
Figure 5 Microscopic Structure of SS
In mild steel, the crystalline structures within the grains have been given names such as ferrite,
austenite, martensite, or a mixture of two or more of these. The microstructure in the annealed
condition consists of ferrite grains in which fine carbides are embedded as shown in figure 2.
1. Four to five sand papers of different gradings and Mild steel had given to us.
2. At first sand paper with 180C Grading was used to grind the Mild steel and observed
3. After that, sand paper with 400C Grading was used to grind the Mild steel and
observed under the metallurgical microscope.
4. After 400C graded sand paper, sand paper with 800C Grading was used to grind the
Mild steel and observed under the metallurgical microscope.
5. In last, sand paper with 1000C Grading was used to grind the Mild steel and observed
[14]
 Results
1st Grid Paper (220C)
2nd Grid Paper (400C)
3rd Grid Paper (600C)
[15]
4th Grid Paper (800C)
 Discussion
For heavy sanding, to remove the bruises from the metal, a low value graded (like 180C graded
sand paper) is used. To smoother the surface, we use sand paper of high carbon graded value (for
instance 1000C graded sand paper). The sample piece is rough, rusty and have stains at
microscopic level to remove we use sand papers of different sizes. When the grit size is 200, the
specimen becomes fine piece after grinding with some water by using spray gun. The specimen
becomes finer when sand paper grit size is 400. Similarly, when we use 600 and 800 grit size sand
paper the specimen surface become more shine, polish and finer. When grit size increases the
rusting of specimen reduces and fine piece formed. This means when the grit size of sand paper
increases the specimen become finer than before. The purpose of removing rusting is to prepare
the sample for next experiments.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
5. Wash your hands with soap and water before entering or leaving the lab.
6. 2. Wear shoes that completely cover your feet, you cannot wear sandals or other open-
toe OR open-heel shoes (clogs are ok).
7. 3. You must wear a lab coat when working in the microbiology lab. You must store
your lab coat/shirt in the lab. Your lab coat/shirt may NOT be removed from the room
during the semester.
8. 4. Long hair must be tied back in a ponytail, all hats must be removed or turned
backward.
9. 5. Accidents or injuries must be reported to your instructor as soon as possible. Know
the location of the fire blanket, fire extinguisher, shower and eyewash in case of
emergency.
10. 6. Do not eat, drink, or chew gum in the lab. Avoid hand-to-mouth activities such as
nail biting or chewing on the end of a pencil/pen.
[16]
11. 7. Cell phones are not allowed in lab under any circumstances.
12. 8. Do not put paper towels used to wipe down the bench in the bench top waste baskets.
13. 10. Disinfect your lab space at the start and end of each lab period. To disinfect your
area wipe down the table top thoroughly with laboratory disinfectant and a paper towel.
When leaving the lab, apply disinfectant to the bench area and leave to air dry (wet).
Figure 10 safety precautions
[17]
Lab #--------03--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[18]
Experiment # 03
To study the effect of corrosion of Stainless Steel (SS) and Mild Steel (MS) in
acidic media
 Objective
1. Study the effect of the Acidic Media (like HCl) on the stainless steel
2. Study the effect of the Acidic Media (like HCl) on the mild steel
1. Beakers (4)
2. Measuring flask
3. Mild Steel
4. Stainless Steel
5. Aluminum Foil
6. Lubrication oil
 Chemicals
1. 5% HCl solution
2. 10% HCl solution
 Samples:
1. Mild Steel Sample (2)
2. Stainless Steel Sample (2)
 Theory
I. Corrosion
The degradation of a metal due to chemical interactions between it and its surroundings is known
as corrosion. When a refined metal spontaneously transforms into a more stable form, such as its
oxide, hydroxide, or sulphide state, this is called corrosion and it results in the substance degrading.
Since it interferes with the beneficial qualities of the metal, corrosion is typically a phenomenon
to be avoided. For instance, iron is recognised for its high stiffness and tensile strength (especially
alloyed with a few other elements). Iron things, however, lose their structural soundness and
become brittle and flaky when they rust. Corrosion, on the other hand, is a diffusion-controlled
process that mostly affects exposed surfaces. So, in some instances, efforts are undertaken to
decrease the activity of the exposed surface and raise the corrosion resistance of a material. It uses
techniques like chromate conversion and passivation. However, some corrosion mechanisms are
both unpredictable and not usually observable.Alternately, as it frequently involves redox reactions
between metals, corrosion might be categorised as an electrochemical process.There are different
types of corrosions
Uniform corrosion
Pitting corrosion
Crevice corrosion
Intergranular corrosion
Stress corrosion
[19]
Galvanic corrosion
Figure 4 corrosion in orthopedic implants
Types of Corrosion:
1. Uniform corrosion
 Uniform loss of metal over entire surface

 Stainless steel with pH value of less than 1 are more prone to general corrosion
2. Galvanic corrosion
 Electrochemical process where one metal corrodes preferentially compared to another

in the presence of electrolyte
3. Intergranular corrosion
 Corrosion where boundaries of crystallites are more likely to corrode than inside
surfaces
 Occurs after heating austenitic stainleUs steel at approximately 842–1562°F
4. Pitting corrosion
 Localized corrosion resulting in cavities or holes

 Occurs when stainless steel is exposed to environments containing chlorides
5. Crevice corrosion
 Localized corrosion at the crevice between two joining surfaces

 Formed between two metals or between metal and non-metal
[20]
6. Stress corrosion cracking
 Growth of crack formation in corrosive setting

 Tensile stresses in combination with corrosive environmental conditions lead to
cracking
Every metal corrodes?
Iron, zinc, and other metals ranked higher in the reactivity series corrode very easily, but metals
ranked lower in the reactivity series, such as gold, platinum, and palladium, do not corrode.
Because corrosion requires the oxidation of metals, this is how it is explained. The propensity to
being oxidized decreases dramatically with reactivity (oxidation potentials is very low).
Interestingly, while being reactive, aluminium doesn't corrode like other metals do. This is due to
the fact that aluminium already has an oxide layer on it. It is shielded from additional corrosion by
this coating of aluminium oxide.
Figure 5 corrosion in metals
II. Corrosion in mild steel

In the chemical and related industries, treating acid, alkali, and salt solutions frequently involves
the use of mild steel, one of the main building materials. Given its tendency to cause corrosion and
damage to building materials, hydrochloric acid is the most challenging of the common acids to
work with. Even at relatively low concentrations or in process solutions that contain a sizable
amount of hydrochloric acid, choosing the right materials to handle the acid alone requires extreme
caution. For the majority of common metals and alloys, this acid is exceedingly corrosive. By
using chemical (hydrogen evolution, HE, and mass loss, ML) and electrochemical
(electrochemical impedance spectroscopy, potentiodynamic polarisation), as well as
electrochemical methods at a temperature of 25 °C, the corrosion behaviour and mechanism for
mild steel in hydrochloric acid solutions, was studied. According to the chemical analysis, mild
[21]
steel corrodes in HCl solutions, and the rate of corrosion rises as the acid concentration rises. After
immersion in HCl solutions of various concentrations, microstructural experiments on mild steel
revealed general and pitting corrosion, with the latter becoming more apparent at higher levels of
HCl concentration.
III. Stainless steel in Acidic media

Stainless steels are very resistant to corrosion, yet surface damage can still happen in some
applications. Without normal cleaning and maintenance, oxidation, corrosion, rusting, or
discoloration can develop over time in severe conditions. Repeated mechanical damage also
hastens the metal's deterioration.Chromite is present in at least 10.5% of the weight of all stainless
steels. Unlike other steels, stainless steel is protected against corrosion by a layer known as the
passive layer, which is made possible by the presence of chromium. The resistance to corrosion
increases with increasing chromium concentration. When the passive layer is compromised and
there is insufficient chromium for it to regenerate, stainless steel rusts.
The passive layer is unstable due to strong acidity. . General surface corrosion may happen to
stainless steel when it is exposed to sulfuric and hydrochloric acids. The manufacture of plastics
uses hydrochloric acid, which has a high acidity. Descaling agents for eliminating limescale
contain diluted hydrochloric acid. The caustic chemical sulfuric acid can result in both secondary
thermal burns and serious chemical burns. It is applied to the refinement of oil, wastewater
treatment, and mineral processing
Figure 6 corrosion of metals under acidic environment
[22]
1. Samples (mild steel and stainless steel) were washed because they were dipped in oil to
avoid rusting
2. Then mild steel and stainless steel was weighed using weigh balance
3. Prepared the HCL solution.
4. 12 beakers were taken filled with solutions 20ml solution in each beaker.
5. Hang the sample in the beaker containing the solution, leaving the beaker's top ajar to let
air through.
6. Dry the sample and reweigh it using an electrical balance at day 1 and readings were
noted.
7. Also determined the weight loss percentage.
8. Looked at the samples' surfaces and make a comparison between the two kinds of steel.
9. Follow the same procedure for day 3 and day 7 and tracked the weight loss.
Observation
Mild Steel
Corrodent Weights Initial Day 1 Day 3 Day 7
weight
1 10% HCl W1 12.8g 13.57g
2 10% HCl W2 13.59g 13.11g
3 10% HCl W3 11.12g 10.24g
Stainless steel
4 10% HCl W1 13.73g 12.4g
5 10% HCl W2 13.53g 12.6g
6 10% HCl W3 16.87g 16.13g
Mild Steel
weight
1 20% HCl W4 12.15g 11.91g
2 20% HCl W5 11.5g 10.19g
3 20% HCl W6 13.45g 12.1g
Stainless Steel
[23]
4 20% HCl W4 16.31g 16.81g
5 20% HCl W5 12.5g 12.18g
6 20% HCl W6 13.45g 12.12g
Calculations
Percentage weight loss = (initial weight-final weight/initial weight) × 100 Percentage
Mild steel Percentage weight loss

Corrodent Weights Day 1 Day 3 Day 7
1 10% HCl W1 -6%
2 10% HCl W2 3.5%
3 10% HCl W3 7.9%
Stainless steel Percentage weight loss
4 10% HCl W1 9.6%
5 10% HCl W2 6.8%
6 10% HCl W3 4.3%
1 20% HCl W1 1.9%
2 20% HCl W2 11.73%
[24]
3 20% HCl W3 10.04%
4 20% HCl W1 0.8%
5 20% HCl W2 2.7%
6 20% HCl W3 9.8%
 Results
As mild steel has less percentage of carbon in it than stainless steel, therefore, the percentage
corrosion of mild steel is very much greater than stainless steel. Stainless steel has very high
percentage of carbon, therefore it can’t corrode easily as compare to that of the other materials
like aluminum, copper, mild steel.
Table 1 result of experiment
[25]
 Discussion
The inhibition efficiency of different different concentrations of acidic acid on mild steel was
evaluated using weight loss measurements, and the results are shown in bar graphs. In all
cases, the value increases as inhibitor concentration increases, implying an increase in the
number of molecules adsorbed on the surface of mild steel, blocking the active sites of acid
attack and protecting the metal from corrosion. Hence the outcome is weight loss. It can be
seen that increasing the acid concentration resulted in an increase in corrosion rate.
Additionally, an increase in acid concentration resulted in a significant increase in corrosion
rate. These results show that higher Cl ion concentrations destroyed the passive films that
tend to isolate the metal from the corrosive solution. The results for weight loss of stainless
steel immersed in acidic solution revealed the greatest amount of weight loss value
throughout the experiment. The weight loss decreased as the HCL concentration increased.
 Conclusion
Stainless steel has higher corrosion resistance as compare to mild steel and weight loss
decreases day by day.
 Precautions
[26]
Lab #--------04--------
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Lab Instructor:
[27]
Experiment # 04
To study the effect of corrosion of Stainless Steel (SS) and Mild Steel (MS) in
basic media
 Objective
1. Study the effect of the Basic/Alkali Media (like NaOH) on the stainless steel
2. Study the effect of the Basic/Alkali Media (like NaOH) on the mild steel
1. Beakers (4)
2. Measuring flask
3. Mild Steel
4. Stainless Steel
5. Aluminum Foil
6. Lubrication oil
 Chemicals
1. 5% NaOH solution
2. 10% NaOH solution
 Samples:
 Theory
I. Corrosion
The degradation of a metal due to chemical interactions between it and its surroundings is known
as corrosion. When a refined metal spontaneously transforms into a more stable form, such as its
oxide, hydroxide, or sulphide state, this is called corrosion and it results in the substance degrading.
Since it interferes with the beneficial qualities of the metal, corrosion is typically a phenomenon
to be avoided. For instance, iron is recognised for its high stiffness and tensile strength (especially
alloyed with a few other elements). Iron things, however, lose their structural soundness and
become brittle and flaky when they rust. Corrosion, on the other hand, is a diffusion-controlled
process that mostly affects exposed surfaces. So, in some instances, efforts are undertaken to
decrease the activity of the exposed surface and raise the corrosion resistance of a material. It uses
techniques like chromate conversion and passivation. However, some corrosion mechanisms are
both unpredictable and not usually observable.Alternately, as it frequently involves redox reactions
between metals, corrosion might be categorised as an electrochemical process.There are different
types of corrosions
Uniform corrosion
Pitting corrosion
Crevice corrosion
Intergranular corrosion
Stress corrosion
Galvanic corrosion
[28]
Figure 7 corrosion in orthopedic implants
Types of Corrosion:
1. Uniform corrosion
 Uniform loss of metal over entire surface

 Stainless steel with pH value of less than 1 are more prone to general corrosion
2. Galvanic corrosion
 Electrochemical process where one metal corrodes preferentially compared to another

in the presence of electrolyte
3. Intergranular corrosion
 Corrosion where boundaries of crystallites are more likely to corrode than inside
surfaces
 Occurs after heating austenitic stainleUs steel at approximately 842–1562°F
4. Pitting corrosion
 Localized corrosion resulting in cavities or holes

 Occurs when stainless steel is exposed to environments containing chlorides
5. Crevice corrosion
 Localized corrosion at the crevice between two joining surfaces

 Formed between two metals or between metal and non-metal
[29]
7. Stress corrosion cracking
 Growth of crack formation in corrosive setting

 Tensile stresses in combination with corrosive environmental conditions lead to
cracking
Every metal corrodes?
Iron, zinc, and other metals ranked higher in the reactivity series corrode very easily, but metals
ranked lower in the reactivity series, such as gold, platinum, and palladium, do not corrode.
Because corrosion requires the oxidation of metals, this is how it is explained. The propensity to
being oxidized decreases dramatically with reactivity (oxidation potentials is very low).
Interestingly, while being reactive, aluminium doesn't corrode like other metals do. This is due to
the fact that aluminium already has an oxide layer on it. It is shielded from additional corrosion by
this coating of aluminium oxide.
Figure 8 corrosion in metals
II. Corrosion in mild steel

In the chemical and related industries, treating acid, alkali, and salt solutions frequently involves
the use of mild steel, one of the main building materials. Given its tendency to cause corrosion and
damage to building materials, hydrochloric acid is the most challenging of the common acids to
work with. Even at relatively low concentrations or in process solutions that contain a sizable
amount of hydrochloric acid, choosing the right materials to handle the acid alone requires extreme
caution. For the majority of common metals and alloys, this acid is exceedingly corrosive. By
using chemical (hydrogen evolution, HE, and mass loss, ML) and electrochemical
(electrochemical impedance spectroscopy, potentiodynamic polarisation), as well as
electrochemical methods at a temperature of 25 °C, the corrosion behaviour and mechanism for
mild steel in hydrochloric acid solutions, was studied. According to the chemical analysis, mild
[30]
steel corrodes in HCl solutions, and the rate of corrosion rises as the acid concentration rises. After
immersion in HCl solutions of various concentrations, microstructural experiments on mild steel
revealed general and pitting corrosion, with the latter becoming more apparent at higher levels of
HCl concentration.
III. Stainless steel in Basic media

Stainless steel offers strong resistance to weak bases in both high concentrations and high-
temperature environments. However, stronger bases will cause cracking or etching corrosion.
Particular care must be taken with chloride solutions such as sodium hypochlorite.
Stainless steels are very resistant to corrosion, yet surface damage can still happen in some
applications. Without normal cleaning and maintenance, oxidation, corrosion, rusting, or
discoloration can develop over time in severe conditions. Repeated mechanical damage also
hastens the metal's deterioration.Chromite is present in at least 10.5% of the weight of all stainless
steels. Unlike other steels, stainless steel is protected against corrosion by a layer known as the
passive layer, which is made possible by the presence of chromium. The resistance to corrosion
increases with increasing chromium concentration. When the passive layer is compromised and
there is insufficient chromium for it to regenerate, stainless steel rusts
Figure 9 corrosion of metals under acidic environment
[31]
1. Samples (mild steel and stainless steel) were washed because they were dipped in oil to
avoid rusting
2. Then mild steel and stainless steel was weighed using weigh balance
3. Prepared the NaOH solution.
4. 12 beakers were taken filled with solutions 20ml solution in each beaker.
5. Hang the sample in the beaker containing the solution, leaving the beaker's top ajar to let
air through.
6. Dry the sample and reweigh it using an electrical balance at day 1 and readings were
noted.
7. Also determined the weight loss percentage.
8. Looked at the samples' surfaces and make a comparison between the two kinds of steel.
9. Follow the same procedure for day 3 and day 7 and tracked the weight loss.
Observation
Mild Steel
weight
1 5% NaOH W1 12.51g 12.83g
2 5% NaOH W2 9.94g 9.93g
3 5% NaOH W3 10.87g 10.84g
Stainless steel
4 5% NaOH W1 14.38g 13.74g
5 5% NaOH W2 17.48g 17.451g
6 5% NaOH W3 16.61 g 16.576g
Mild Steel
weight
1 10% NaOH W1 11.06g 11.0g
2 10% NaOH W2 11.34g 11.13g
3 10% NaOH W3 16.30g 16.26g
Stainless Steel
4 10% NaOH W1 7.101g 7.104g
[32]
5 10% NaOH W2 16.6g 16.58g
6 10% NaOH W3 7.9g 7.906g
Calculations

1 5%NaOH W1 -2.4%
2 5% NaOH W2 0.1%
3 5% NaOH W3 0.2%
4 5% NaOH W1 4.4%
5 5% NaOH W2 0.1%
6 5% NaOH W3 0.2%
1 10% NaOH W1 0.04%
2 10% NaOH W2 0.21%
[33]
3 10% NaOH W3 0.21%
4 10% NaOH W1 0.18%
5 10% NaOH W2 0.22%
6 10% NaOH W3 0.26%
 Results
As mild steel has less percentage of carbon in it than stainless steel, therefore, the percentage corrosion of
mild steel is greater than stainless steel. But in case of Liquid NaOH, required elevated temperatures to
corrode the metals or matrials. Stainless steel has very high percentage of carbon, therefore it can’t corrode
easily as compare to that of the other materials like aluminum, copper, mild steel.
Table 2 Result of experiment
[34]
 Discussion
The similar tendency of a decreasing weight loss rate with time is evident in weight loss
measurements taken at room temperature and exposed for up to 7 days, demonstrating a
parabolic weight loss trend. Greater material loss is observed at greater NaOH concentrations,
which is consistent with the trends found in the electrochemical tests. Iron or steel will not be
affected by sodium hydroxide because the alkaline environment prevents rust from developing.
However, if the environment is made oxidising, black magnetic oxide will develop in its place.
When combined with sequestering agents to keep the dissolved iron in solution, hot alkali
solutions like caustic soda (sodium hydroxide) can be very effective at removing rust from iron
and steel. Despite a minor difference in initial weight and the weights being measured on various
days, both steels experience weight loss, but they virtually remain the same.
 Conclusion
After exposure to concentrated caustic solutions at tem-perature, stainless steel surfaces are
covered by a black surface ﬁlm predominantly made of nickel oxide and hydroxides but mild didn’t
showed any change. Stainless steel has higher corrosion resistance as compare to mild steel and
weight loss decreases day by day.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
5. Wash your hands with soap and water before entering or leaving
the lab.
6. Wear shoes that completely cover your feet, you cannot wear
sandals or other open-toe OR open-heel shoes (clogs are ok).
7. You must wear a lab coat when working in the microbiology lab.
You must store your lab coat/shirt in the lab. Your lab coat/shirt
may NOT be removed from the room during the semester.
8. Long hair must be tied back in a ponytail, all hats must be removed
or turned backward.
9. Accidents or injuries must be reported to your instructor as soon as
possible. Know the location of the fire blanket, fire extinguisher,
shower and eyewash in case of emergency.
10. Do not eat, drink, or chew gum in the lab. Avoid hand-to-mouth
activities such as nail biting or chewing on the end of a pencil/pen.
11. Cell phones are not allowed in lab under any circumstances.
12. Do not put paper towels used to wipe down the bench in the bench top waste baskets.
[35]
Lab #--------05--------
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Lab Instructor:
[36]
Experiment # 05
Immersion of Stainless Steel (SS) and Mild Steel in Ringer Solution
 Objective
1. To study the corrosion of SS 316L in Ringer solution with variation of time.
2. To study the corrosion of Mild Steel in Ringer solution with variation of time.
1. Electrical balance
2. glass measuring cylinder
3. glass beaker
4. Water barthBeakers (4)
5. Measuring flask
6. Aluminum Foil
7. Lubrication oil
 Chemicals
1. Ringer solution
 Samples:
 Theory
I. Ringer solution:
Ringer's solution is a solution of several salts dissolved in water for the purpose of creating
an isotonic solution relative to the body fluids of an animal. Ringer's solution typically
contains sodium chloride, potassium chloride, calcium chloride and sodium bicarbonate, with the
last used to balance the pH. Other additions can include chemical fuel sources for cells,
including ATP and dextrose, as well as antibiotics and antifungals.
II. Composition:
Ringer's solution typically contains NaCl, KCl, CaCl2 and NaHCO3, sometimes with other
minerals such as MgCl2, dissolved in distilled water. The precise proportions of these vary from
species to species, particularly between marine osmoconformers and osmoregulators.
III. History:
Sydney Ringer discovered in 1882–1885 that a solution perfusing a frog's heart must contain
sodium, potassium, and calcium salts in a certain proportion if the heart is to be kept beating for
an extended period of time. Alexis Hartmann modified the use of ringer's solution in the 1930s by
adding sodium lactate to create Ringer's lactate solution.
IV. Procedure for preparation:
1. In a beaker, add 500 ml of distilled water.
2. Using distilled water, dissolve the reagents in the beaker.
3. Add water to bring the final volume to one litre.
4. Adjust the pH to 7.3-7.4.
[37]
5. Filter the solution through a 0.22-μm filter.
6. Autoclave Ringer's solution prior to use.
Figure 10 Ringer solution
Importance:
 Their sensitivity in these investigations, electrochemical methods are
often used besides gravimetric and colorometric procedures.
 These methods allow determination of parameters such as polarization resistance, pitting
potential, and breakdown potential.
 Investigations using biological systems (e.g., cell lines, organ cultures, and animal
experiments) usually apply biochemical, immunological, or histological methods.
 Sodium bicarbonated Ringer’s solution has superior effects on the acidosis and electrolyte
balance in patients with hemorrhagic shock and resulted in a lower risk of complications
than the conventional crystalloid fluid
Figure 11 method of preparation of ringer solution
[38]
Figure 12 Analysis of metals immersed in ringer solution
1. First, took the sample of Mild Steel and Stainless Steel and clean them with sand paper.
2. Weight each sample up to 4 digits by using weight balance.
3. Then took the 25 ml Ringer’s Solution and then dipped the sample into the solution and put
it into the water bath at 37 °C.
4. We take out the samples every, 3th , 4th and 7th days respectively and washed, dried and
weight them.
5. Then take the weight difference.
6. Attention need during Experiment, this time use same sample solution for each reading no
need separate beaker e.g. take the sample and dried no measure the weight change and the
again put in the same solution till 7 days 2:
7. Always check temperature of water bath should be 37C. in case of change
Observation
Sample pair 1
Ringer sample Weights Initial weight Day 3 (g) Day 5 (g) Day 7(g)
solution
(25 ml)
MS W1 16.455 16.356 - -
SS W2 7.827 7.675 - -
Sample pair 2
sample Weights Initial weight Day 3 (g) Day 5 (g) Day 7(g)
[39]
Ringer MS W1 11.143 - 11.098 -
solution
(25 ml) SS W2 7.101 - 7.099 -
Sample pair 3
Ringer sample Weights Initial weight Day 3 (g) Day 5 (g) Day 7(g)
solution
(25 ml)
MS W1 12.102 - - 12.01
SS W2 12.010 - - 12.005
Calculations
Sample pair 1
Ringer Percentage weight loss
solution Samples Day 3 Day 5 Day 7
(25 ml)
1 MS 0.012%
2 SS 0.19%
Sample pair 2
(25 ml)
1 MS 0.039%
2 SS 0.017%
[40]
Sample pair 3
(25 ml)
1 MS 0.21%
2 SS 0.039%
 Results
Ringer's solution provides lubrication on the surface of stainless steel and makes it smooth while
mild steel surface gets rough so stainless steel is better for body implants. As the composition of
ringer's solution is similar to the body fluids, thus, the rate of corrosion in mild steel and stainless
steel in ringer's solution help us to demonstrate the compatibility of steel implants in body
solution. The rate of corrosion of the steel is directly related to weight loss of the sample.
Table 3 observation made in Day 03
[41]
[42]
Table 6 calculation representing in %
 Discussion
Ringer's solution provides
lubrication on the surface of
stainless steel and makes it
smooth while mild steel
surface gets rough so
stainless steel is better for
body implants. As the
composition of ringer's
solution is similar to the
body fluids, thus, the rate of
corrosion in mild steel and
stainless steel in ringer's
solution help us to
demonstrate the
compatibility of steel
implants in body solution.
The rate of corrosion of the
steel is directly related to Figure 13 Analysis of experiment
weight loss of the sample.
[43]
The high corrosion resistance of SS is due to the formation of a thin and protective surface oxide
layer, which makes them passive. Such a characteristic is strongly related to the content of mild
steel. Microstructural and chemical-physics characteristics of SS surface, as the presence of
different phases, roughness, morphology and composition of more or less protective oxides after
welding and/or pickling-passivation treatments, etc. cannot be neglected in the research. Stainless
steels (SS) are well-known and widely used for many applications thanks to their high corrosion
resistance in several natural and industrial environments, high mechanical performances, and low
maintenance requirements. All these corrosion forms can determine severe and costly damages in
different applications.
 Conclusion
The effect of immersion of mild and stainless steel in ringer solution was observed. Mild steel
corroded as compared to stainless steel and the rate of corrosion was quicker in mild steel as
compared to stainless steel.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
the lab.
or turned backward.
[44]
Lab #--------06--------
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Lab Instructor:
[45]
Experiment # 06
Preparation of Bio-ceramics Pellet Using Hydraulic Press
 Objective
1. To make a pellet of ceramic powder by using hydraulic press.
1. Motor and pestle,
2. Vernier caliper,
3. Electrical balance,
4. Hydraulic press,
5. Die
 Material Required
1. Hydoxyapatite Powder
 Theory
I. Hydraulic Press
Hydraulic press is a machine that uses Pascal’s Principle to generate a large amount of force. It
consists of two cylinders, a smaller slave cylinder and a larger master cylinder. The slave cylinder
contains a piston that is used to apply pressure to the fluid inside the slave cylinder. This pressure
is then transferred to the master cylinder, where it is pushed by a piston.
Figure 14 Hydraulic Press
[46]
 Working Principal:
The hydraulic press relies on Pascal’s Principle, which was established by French Mathematician
Blaise Pascal in 1647-1648. Pascal’s Principle is a principle in fluid mechanics stating that the
pressure at a point has infinite direction, and thus the pressure changed at any point in a confined
incompressible liquid is transmitted throughout the fluid, such that the same change occurs
everywhere. The principle, when written mathematically, is as follows:
∆P is the hydrostatic pressure or, more simply put, the difference in pressure of two points within
a fluid column. The SI unit for pressure is in Pascals. Here, ρ is the fluid density in kilogram per
cubic meter. The term g in the above equation signifies the acceleration due to gravity (measured
in meters per second squared). ∆h is the height of the fluid above the point of measurement in the
fluid column, which is measured in meters.
The above equation can be very well understood intuitively. The change in pressure occurs due to
the change in potential energy of the liquid per unit volume of the liquid, which in turn is caused
by the acceleration due to the gravity.
 Mechanism of Hydraulic Press
Now that we are aware that the hydraulic press is based on Pascal’s Principle, we can move on to
the construction and working of the hydraulic press. The components of a hydraulic press
include cylinders, pistons, and hydraulic pipes. The working of the press is quite simple and
primitive. The system comprises two cylinders that are filled with a fluid. The fluid present inside
the two cylinders is usually oil. The fluid (the oil) is filled into the smaller cylinder, which is also
known as the slave cylinder.
A piston is inserted into the slave cylinder and pressure is applied. The pressure applied causes the
fluid to move through a pipe and into a larger cylinder. The larger cylinder is known as the master
cylinder. The pressure exerted on the master cylinder and the piston in the master cylinder pushes
the fluid back to the slave cylinder. The force applied on the fluid by the slave cylinder results in
a large force, which is experienced by the master cylinder. An industrial hydraulic press comes
along with what is known as the press plates. With the help of these press plates, the material to
be worked on is either punched or crushed into sheets.
[47]
Figure 15 Mechanism of Hydraulic Press
II. Bio Ceramics:

Bio-ceramics are a class of materials composed of biocompatible ceramics and bioglasses, with
some of the most common bio-ceramics being calcium phosphate, hydroxyapatite, polymer
composites, bioactive glasses, zirconia, titania and alumina. The materials in this class range are
often harder in structure but can either be densely packed or porous depending on the required
application. It should be noted that the ceramic-like materials bear no resemble to porcelain-type
ceramics, rather, they are materials that mimic the body’s own biomaterials (or are durable metal
oxide materials).
 Bio Ceramics Pellets:
The crystalline structure, size and morphology of the powders and the porous ceramic pellets were
investigated using advanced characterization techniques. The pellets demonstrated good
biocompatibility, including mixed cell colonisation and matrix deposition, in vivo following
surgical implantation.
 Application of Bio ceramics pellets:
Many of the applications are in assistive repair, so there are certain characteristics that bio-ceramic
materials need to exhibit for them to be used in clinical applications.
1. The most important is biocompatibility, so that they can help to prevent the body’s natural
defence system from breaking them down and they are not toxic to the cellular environment
when they are used.
2. Other properties include a high wear resistance which utilizes a materials high hardness
and a lack of plastic and elastic deformation under a load, a low friction coefficient, a high
compressional strength, a high fatigue resistance, high biological and chemical corrosion
resistance, high electrical insulating properties to prevent galvanic reactions from occurring
and the ability to synthesize a highly pure material.
[48]
3. A wide range of materials can be employed in bone regeneration and reconstruction
approaches, depending on the bone in question and the level of damage.
4. In the biomedical field ceramic materials were used as substitute materials for bone and
teeth as well as coatings for metallic implants.
5. Advanced research is going on in the molecular level modification of ceramic materials for
developing artificial transplantation materials, drug release materials as well as materials
for tissue engineering and gene therapy applications.
Figure 16 Application of bio ceramics
1. Approximately 1g of ceramic powder was measured with the help weighing balance.
2. Ceramic Powder was put in the hydraulic press.
3. Pressure was applied on the ceramic powder with the sphere-shaped metal. Pressure was
applied to about 10-30seconds.
4. After that, a dense ceramic material came out.
5. Observe the sphere-shaped ceramic material diameter and height with the help of vernier-
caliper.
6. Readings were observed.
[49]
Observation
Weight before making pallet = 1.0g
Weight after making pallet = 0.905g
Diameter of Pallet = 1.2cm
Height of pallet = 0.5cm
Calculations
Volume = 4/3(πr3) = 4/3 (π*(0.6)3) = 0.90477 cm3
Density = M/V = 0.835/0.90477 = 1.0025 g/cm3

 Results
Diameter of pallet is 1.2cm and Height of Pallet is 0.5cm.
[50]
 Discussion
Bio-ceramics are materials specially developed to use in medical and dental implants. Usually used
to replace hard tissue in the body like bone and teeth. This shape is favored for many reasons, the
main justification being that the cylindrical shape means there are only two edges to perform as
stress concentrators. This is crucial because if the initial powder lacks a binder, the following green
compact can be extremely delicate and may fracture before additional processing. A further
argument behind the selection of the pellet shape when analyzing these materials is that it can be cut to suit
most of the tests that will be applied to it. Some of the common tests that are carried out on engineering
ceramics are X-ray fluorescence (XRF) and infrared (IR) spectroscopy. A flat cylindrical sample is
normally used in these investigations. The ability to test the pellets with no need for additional
grinding or cutting is highly beneficial. There are also some important characteristics of the pellets
to consider now the reasoning behind the shape has been outlined. A pressed disc must adhere to
the following quality criteria: solidity to avoid loose particles, a homogeneous composition,
storability, and stability.
Figure 17 Procedure for designing of pellets
 Conclusion
A hard dense material formed by applying a pressure on the special ceramic powder.
[51]
 Precautions
1. Wear Gloves
2. Wear Lab Coat
the lab.
or turned backward.
[52]
Lab #--------07--------
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Lab Instructor:
[53]
Experiment # 07
Densification of Bio-Ceramics Pellet by Sintering
 Objective
1. To analyze the heating effect on Bio-ceramic Pellets
1. Motor and pestle,
2. Vernier caliper,
3. Electrical balance
4. petri dishes,
5. muffle furnace
1. Hydoxyapatite Powder
 Theory
I. Sintering Process:
Sintering is a heat treatment process where loose material is subjected to high temperature and
pressure in order to compact it into a solid piece. This is similar to when ice cubes adhere together
in a glass of water due to the temperature difference between the ice and the water, or when you
push snow together to form a compact snowball. The heat and pressure required for the sintering
process is less than the material’s melting point.
 Working principle:
Just as a material has a melting point, it will also have a desirable sintering point, at which the heat
and pressure are enough to reduce the porous spaces between the material’s particles and squeeze
loose material together into a solid lump. This use of pressure and heat takes place naturally in
mineral deposits within the Earth as well as in glacial formations. Sintering is used to increase
material properties, including thermal and electrical conductivity, material strength and integrity,
and translucency.
Figure 18 Sintering sequence by temperature ranges
[54]
 Advantages:
While the different methods and materials offer a range of benefits, there are a number of general
advantages associated with sintering:
1. Purity: Sintering offers high levels of purity and uniformity in the starting materials,
which can be maintained due to the simple fabrication process
2. Repeatable: Controlling the grain size during input allows for highly repeatable
operations
3. No Binding Contact / Inclusions: Unlike with some melting processes, sintering will not
cause binding contact between powder particles or inclusions (aka ‘stringering’)
4. Uniform Porosity: Create materials with a uniform, controlled porosity
5. Work with Difficult Materials: Sintering allows you to work with materials that cannot be
used with other technologies, such as metals with very high melting points
 Muffle Furnace sintering process for Bioceramics
A muffle furnace or muffle oven (sometimes retort furnace in historical usage is a furnace in which
the subject material is isolated from the fuel and all of the products of combustion, including gases
and flying ash. After the development of high-temperature heating elements and
widespread electrification in developed countries, new muffle furnaces quickly moved
to electric designs. CM offers our larger sintering furnaces as building blocks, with the proven
ability to customize to your particular temperature, time, and atmosphere requirements, as well as
many other customizable control options. It is the quality of our engineering and materials that set
us apart when you trust your process furnace requirements to us.
Figure 19 muffle furnace sintering process
[55]
II. Bio Ceramics:
Bio-ceramics are a class of materials composed of biocompatible ceramics and bioglasses, with
some of the most common bio-ceramics being calcium phosphate, hydroxyapatite, polymer
composites, bioactive glasses, zirconia, titania and alumina. The materials in this class range are
often harder in structure but can either be densely packed or porous depending on the required
application. It should be noted that the ceramic-like materials bear no resemble to porcelain-type
ceramics, rather, they are materials that mimic the body’s own biomaterials (or are durable metal
oxide materials).
 Dense Bio Ceramics:
The crystalline structure, size and morphology of the powders and the porous ceramic pellets were
investigated using advanced characterization techniques. The pellets demonstrated good
biocompatibility, including mixed cell colonisation and matrix deposition, in vivo following
surgical implantation.
 Application of Dense Bio ceramics:

Many of the applications are in assistive repair, so there are certain characteristics that bio-ceramic
materials need to exhibit for them to be used in clinical applications.
1. The most important is biocompatibility, so that they can help to prevent the body’s
natural defence system from breaking them down and they are not toxic to the cellular
environment when they are used.
2. Other properties include a high wear resistance which utilizes a materials high hardness
and a lack of plastic and elastic deformation under a load, a low friction coefficient, a
high compressional strength, a high fatigue resistance, high biological and chemical
corrosion resistance, high electrical insulating properties to prevent galvanic reactions
from occurring and the ability to synthesize a highly pure material.
3. A wide range of materials can be employed in bone regeneration and reconstruction
approaches, depending on the bone in question and the level of damage.
[56]
4. In the biomedical field ceramic materials were used as substitute materials for bone and
teeth as well as coatings for metallic implants.
5. Advanced research is going on in the molecular level modification of ceramic materials
for developing artificial transplantation materials, drug release materials as well as
materials for tissue engineering and gene therapy applications.
Figure 20application of dense bio-ceramics
1. Bio-ceramics pellets were prepared using Hydraulic Press.
2. The dimensions (weight, diameter and thickness) of pellet was measured using Vernier
Caliper.
3. Muffle furnace was turned ON.
4. On menu button, first program was selected and temperature was adjusted at 600OC.
RAMP 10oC and holds for 60 minutes.
5. Rate of decreasing temperature was set to decrease 50OC with RAMP 30OC.
6. After cooling down, pellet was taken out ad dimensions were measured again.
 Observation
Diameter before Heating = 1.2cm
Thickness before Heating = 0.5cm
[57]
Weight before Heating = 1g
Diameter after heating = 1.2cm
Thickness after heating = 0.4cm
Weight after heating = 0.835g
Calculations
Volume = 4/3(πr3) = 4/3 (π*(0.6)3) = 0.90477 cm3
Density = M/V = 0.835/0.90477 = 0.9229 g/cm3
 Results
After sintering, the density reduces from 1.0025 g/cm3 to 0.9229 g/cm3.
[58]
Graph:
Table 7 Graph representation of parameters
[59]
 Discussion
Sintering is the process of compacting and forming a solid mass of material by heat or pressure
without melting it to the point of liquefaction. Sintering happens as part of a manufacturing
process used with metals, ceramics, plastics, and other materials.
Bio-ceramics are materials specially developed to use in medical and dental implants. Usually
used to replace hard tissue in the body like bone and teeth. This shape is favored for many
reasons, the main justification being that the cylindrical shape means there are only two edges
to perform as stress concentrators. This is crucial because if the initial powder lacks a binder,
the following green compact can be extremely delicate and may fracture before additional
processing. A further argument behind the selection of the pellet shape when analyzing these materials
is that it can be cut to suit most of the tests that will be applied to it. Some of the common tests that are
carried out on engineering ceramics are X-ray fluorescence (XRF) and infrared (IR) spectroscopy
Figure 21 Bio-ceramics designing through muffle furnace sintering process
 Conclusion
A hard dense material formed by applying a pressure on the special ceramic powder.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
5. Wash your hands with soap and water before entering or
leaving the lab.
[60]
6. Wear shoes that completely cover your feet, you cannot wear sandals or other open-toe
OR open-heel shoes (clogs are ok).
7. You must wear a lab coat when working in the microbiology
lab. You must store your lab coat/shirt in the lab. Your lab
coat/shirt may NOT be removed from the room during the
semester.
8. Long hair must be tied back in a ponytail, all hats must be
removed or turned backward.
9. Accidents or injuries must be reported to your instructor as soon
as possible. Know the location of the fire blanket, fire
extinguisher, shower and eyewash in case of emergency.
[61]
Lab #--------08--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[62]
Experiment # 08
Production and sintering of porous bio-ceramics scaffolds using
naphthalene as the pore former
 Objective
1. Production of porous bio-ceramics scaffold using naphthalene as the pore former
2. Sintering of porous bio-ceramics scaffold
1. Mortar and Pestle
2. Weighing Balance
3. Siever
4. Hydraulic press
5. Vernier caliper
6. furnaceMotor
7. pestle
 Chemicals Required
1. polyvinyl alcohol (0.05g), Powdered HA (1g), Naphthalene (0.0g)
 Theory
I. Porous Bio-ceramics:
Porous functional graded ceramics (porous FGCs) offer immense potential to overcome the
low mechanical strengths of homogeneously porous bioceramics used as bone grafts. The
tailored manipulation of the graded pore structure including the interfaces in these materials is
of particular interest to locally control the microstructural and mechanical properties, as well
as the biological response of the potential implant. In this work, porous FGCs with integrated
interface textures were fabricated by a novel two-step transfer micro-molding technique using
alumina and hydroxyapatite feedstocks with varied amounts of spherical pore formers (0–40
Vol%) to generate well-defined porosities. Defect-free interfaces could be realized for various
porosity pairings, leading to porous FGCs with continuous and discontinuous transition of
porosity. The microstructure of three different periodic interface patterns (planar, 2D-linear
waves and 3D-Gaussian hills) was investigated by SEM and µCT and showed a shape accurate
replication of the CAD-designed model in the ceramic sample. The Young’s modulus and
flexural strength of bi-layered bending bars with 0 and 30 Vol% of pore formers were
determined and compared to homogeneous porous alumina and hydroxyapaite containing 0–
40 Vol% of pore formers. A significant reduction of the Young’s modulus was observed for
the porous FGCs, attributed to damping effects at the interface. Flexural 4-point-testing
revealed that the failure did not occur at the interface, but rather in the porous 30 Vol% layer,
proving that the interface does not represent a source of weakness in the microstructure.
 Example: Bioceramics are materials which include Alumina, Zirconia, Bioactive glass, Glass
ceramics, Hydroxyapatite, resorbable Calcium phosphates, among others.
[63]
Figure 22 classification of bio-ceramics
II. Biomedical Application of porous materials:

Different applications require a unique set of pore characteristics in the biopolymer matrix.
Various pore morphologies have different characteristics and contribute different
performances to the biopolymer matrix. Fabrication methods for bio-based porous materials
more related to the choice of material. By choosing the appropriate combination of fabrication
technique and biomaterial employment, one can obtain tunable pore characteristic to fulfill the
requirements of desired application.
Figure 23 Application of porous material in biomedical engineering
[64]
1) Porous bioceramics have been extensively investigated to satisfy the increasing
demand of bone substitute materials.
2) It mainly addressed to the development of scaffolds for bone tissue engineering and
ocular devices (orbital implants), as well as advanced therapy of bone cancer by
hyperthermia.
3) Porous biopolymer matrices are used for biomedical applications such as tissue
engineering, control drug delivery systems and wound healing.
4) It is also a popular candidate for microbiological applications such as encapsulation of
microorganism in fermentative industry, in the production of probiotic food,
biosorption and bioremediation of chemicals and as antimicrobial food packaging
material. It provides an effective way to control spoilage microorganisms and food-
borne pathogens thus enhance food safety and decrease product spoilage
5) Aside from biodegradability, biocompatibility and mechanical properties, porosity is
also an important consideration when selecting a desirable scaffold for tissue
engineering
III. Methods for fabrication of porous materials

Various processing techniques such as partial sintering, sacrificial fugitives, foaming, freeze
casting, metal injection molding, rapid prototyping, etc., and their associated parameters in
designing of porous biomaterials. Using biodegradable fibers by electrospinning, 3D
printing, Rapid prototyping, Solvent casting Particulate leaching techniques
IV. Naphthalene:
Naphthalene is a white, volatile, solid
polycyclic hydrocarbon with a strong
mothball odor. Naphthalene is obtained
from either coal tar or petroleum
distillation and is primarily used to
manufacture phthalic anhydride, but is
also used in moth repellents.
 Porogen:
Any of a mass of particles, of a specified
shape and size, used to make pores in
moulded structures used for tissue
engineering
Polymer blends can also be used as
materials to improve cell adhesion and
proliferation. Blending polymers has
indeed proved to be a useful and
economical method that is widely used to
produce new polymeric materials with
desirable properties compared to those
obtained by the copolymerization of the
corresponding monomers, which requires
complex conditions to obtain the Figure 24 Naphthalene as porogen
appropriate copolymer. Recently, the
[65]
versatility of polymer blends has enabled the implementation of new ideas in different
biomedical fields. These systems are continuing to expand and play an important role in
improving the solutions in multidisciplinary domains, notably implantable materials for
medical applications as well as investigations on bone.
V. Polyvinyl alcohol:
Polyvinyl alcohol is a water-soluble synthetic polymer obtained by polymerization of vinyl
alcohol. Polyvinyl alcohol appears as odorless white to cream-colored granules or powder.
Pure aqueous solutions are neutral or faintly acid and subject to mold growth.
 Polyvinyl alcohol as binder:
The most commonly used binder when spray-drying ceramics is probably polyvinyl alcohol
(PVA). Dispersant-binder compatibility is necessary in order to ensure optimal slurry
rheological properties. It is used in order to manufacture uniform alumina compacts with
homogeneous grain distribution and dense microstructure. As the PVA content increases, it
has been found that the density increases, but there is also less lubrication which, in turn, causes
sticking of the particles. Polyvinyl alcohol (PVA) has low toxicity, but also has the advantage
that it is already widely used as a low-cost binder in the commercial production of ceramics.
In order to increase the strength of ceramic gels, one possibility is to use another cross-linking
agent to initiate the gelation of PVA with strong chemical bonds. Several cross-linkers are
useful to form hydrogels from PVA. PVA has been widely using to fabricate porous scaffolds
for tissue engineering applications
Figure 25 Properties of PVA as boinder
[66]
1. Take 3g of Hydroxyapatite with 0.3g of PVA and 0.6g of Naphthalene using weighing
balance.
2. Mix all the chemicals and grind them completely.
3. While grinding, sieved it and separated the large sized particles to make their size
homogenous.
4. Then grind the larger particles and repeat this process until fine powder is obtained.
5. Now took approximately 1 to 1.2g of final powder and filled it in dye of hydraulic press.
6. Kept it under 50 Bar for 60 seconds.
7. Repeat the above steps for 0% and 30% Naphthlene.For 0% Naphthalene, take 0.1g
Polyvinyl alcohol and 1g powder of Hydroxyapatite and for 30% naphthalene ,take 0.3g
Polyvinyl alcohol and 3g powder of hydroxyapatite
8. After 60 seconds, release the pressure and attain the final shape of palette.
9. Apply heat treatment on palette for few hours in furnace. Heat treatment is to increase the
mechanical and compressive strength.After removing from furnace pores are formed.
 Observation
Before Sintering:
Measurement 1g(HA),0% 1g(HA),10%naphthalene 1g(HA),20%

naphthalene, 0.05g 0.05g of PVA Naphthalene, 0.05g
of PVA of PVA
Weight of pellet 0.925 g 1.052 g 1.115 g
Widht of pellet 0.50 cm 0.55 cm 0.57 cm
Length of pellet 1.22 cm 1.22 cm 1.22 cm
After Sintering:
Measurement 1g(HA),0% 1g(HA),10%naphthalene 1g(HA),20%

naphthalene, 0.05g 0.05g of PVA Naphthalene, 0.05g
of PVA of PVA
Weight of pellet 0.797 g 0.846 g 0.878 g
Widht of pellet 0.50 cm 0.57 cm 0.61 cm
[67]
Length of pellet 1.22 cm 1.22 cm 1.22 cm
 Results
Before Sintering
when sintering is not done, we make microscopic analysis of pellets which is represented
below:
Figure 26 Different Concentration of Naphthalene Figure 6 microscopic image of 0% Naphthalene
Figure 7 microscopic image of 10% Naphthalene Figure 8 microscopic image of 20% Naphthalene
[68]
After Sintering
when sintering is done, we make microscopic analysis of pellets which is represented below:
Figure 9 Different concentration of Naphthalene Figure 10 microscopic image of 0% Naphthalene
Figure 11 microscopic image of 10% Naphthalene Figure 12 microscopic image of 20% Naphthalene
[69]
Graph:
Weight:
When we compare values of weight (g) before and after sintering, we obtain following values:
Table 8 comparison of weight
Width:
When we compare values of width (cm) before and after sintering, we obtain following values:
Table 9 comparison of width
[70]
Length:
When we compare values of length (cm) before and after sintering, we obtain following values:
Table 10 Comparison of lenght
 Discussion
Porous scaffolds can be manufactured using biopolymers with specific surface-area-to-volume
ratio, crystallinity, pore size, and porosity. The three-dimensional porous scaffolds with
enhanced porosities having a homogeneous interconnected pore network are required for tissue
engineering applications. Ideal pore sizes vary for different cells and tissues. Porous
controlled-release systems should contain pores that are sufficiently large to enable diffusion
of the drug
Pore size and pore interconnectivity are more important. Usually, high total porosity is
accomplished by poor mechanical properties. Inside a porous biopolymer, there may be closed
(isolated) pores and open (connected) pores. Pore interconnectivity is important for the
accessiblity of gas, liquid, and particulate suspensions. Pore interconnectivity is defined as the
ratio of the pore volume accessible from the matrix surrounding by a sphere of known diameter,
to the total pore volume.
Sintering is the process of compacting and forming a solid mass of material by heat or pressure
without melting it to the point of liquefaction. Sintering happens as part of a manufacturing
process used with metals, ceramics, plastics, and other materials.
[71]
Figure 27 designing of Porous Bio-Ceramics
 Conclusion
We learnt about production of porous bio-ceramics scaffold using naphthalene as the pore former
and sintering of porous bio-ceramics scaffold.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
5. Wash your hands with soap and water before entering or leaving the
lab.
You must store your lab coat/shirt in the lab. Your lab coat/shirt may
NOT be removed from the room during the semester.
or turned backward.
shower and eyewash in case of emergency
[72]
Lab #--------09--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[73]
Experiment # 09
To determine the hardness of metallic biomaterial using Rockwell Hardness
Method
 Objective
1. To carry out Rockwell hardness test
2. To study different scales of Rockwell hardness test and their applications.
1. Rockwell hardness tester
2. Diamond cone indenter
3. Steel indenter
4. Micrometer travelling microscope
1. Aluminum
2. Mild Steel
3. Copper
 Theory
I. Rockwell Hardness Test:
Rockwell hardness testing is a general method for measuring the bulk hardness of metallic and
polymer materials. The Rockwell hardness test is an empirical indentation hardness test that
can provide useful information about metallic materials. This information may correlate to
tensile strength, wear resistance, ductility, and other physical characteristics of metallic
materials, and may be useful in quality control and selection of materials. Rockwell hardness is
determined using an apparatus called the Rockwell hardness tester. A standard specimen of
0.25 in. (6.35 mm) minimum thickness is used. The specimen can be either molded or cut from
a plastic sheet. The test specimen must have parallel and flat surfaces. In addition, the test
specimen must be free from sink marks, burrs, or other protrusions.
a) Rockwell hardness machine

A machine capable of performing a Rockwell hardness test and/or a Rockwell superficial
hardness test and displaying the resulting Rockwell hardness number.
b) Working Principal:
An indentation hardness test using a verified machine to force a diamond spheroconical
indenter or tungsten carbide (or steel) ball indenter, under specified conditions, into the surface
of the material under test, and to measure the difference in depth of the indentation as the force
on the indenter is increased from a specified preliminary test force to a specified total test force
and then returned to the preliminary test force.
[74]
Figure 28 working mechanism of Rockwell test
c) Classification:
The Rockwell hardness test and the Rockwell superficial hardness test. The significant
difference between the two test classifications is in the test forces that are used. For the
Rockwell hardness test, the preliminary test force is 10 kgf (98 N) and the total test forces are
60 kgf (589 N), 100 kgf (981 N), and 150 kgf (1471 N). For the Rockwell superficial hardness
test, the preliminary test force is 3 kgf (29 N) and the total test forces are 15 kgf (147 N), 30
kgf (294 N), and 45 kgf (441 N).
d) Representation of hardness values:

ockwell hardness values shall not be designated by a number alone because it is necessary to
indicate which indenter and forces have been employed in making the test. The hardness
number is followed by the symbol HR and the scale designation. Examples: 64 HRC =
Rockwell hardness number of 64 on Rockwell C scale, 81 HR30N = Rockwell superficial
hardness number of 81 on the Rockwell 30N.
[75]
e) Indentor:
 Indentor HRC:
Diamond cone with an angle of 120° at the summit and ending with a radius of 0,2 mm. Mirror
polished.
 Indentor HRB:
Tungsten carbide W ball with a hardness not less than 850 HV and a diameter of 1/16”
(1,5875mm).
Figure 29 Rockwell test with diamond indenter
1. Prepare the specimen by grinding to obtain a smooth and plain area on the surface where
hardness is to be measured
2. Place the specimen on the stage
3. Raise the specimen stage so as to bring the surface of specimen in contact with the indenter
4. Apply a small toad of 10kg by turning the capstan of machine that small power on the dial
coincides with the mark, which specifies the minor load. The purpose of minor load is to
seat the indenter into the specimen surface, eliminating the effect of any surface roughness.
5. Apply major load (140 kg for C scale 90 kg for B scale) by pushing back the crank handle.
This moves the needle on the dial anticlockwise direction
6. When needle comes to rest, pull the crank handle forward again. By doing so, the major
load is removed, but not the minor load.
7. Note the dial reading (B or C scale, depending upon major load selected). This is Rockwell
Hardness Number and can be represented as HRC and HRB for C and B scale respectively.
8. Take at least three readings for one specimen
9. Repeat the same procedure for any other specimen
[76]
 Results
Ferric materials are harder than non-ferric materials. In non-ferric materials, copper is harder than
Aluminum.
Figure 3 Rockwell scaling Figure 4 Rockwell test showing indentation for MS
Figure 5 Rockwell test showing indentation for Al Figure 6 Rockwell test showing indentation for Copper
[77]
 Observation
Hardness Values:
For Copper, hardness value is 72HRB.
For Aluminum hardness value is 55HRB.
For Steel hardness value is 42HRC
 Discussion
Rockwell testing machine is used to test the hardness level of the small sized materials. Rockwell
hardness test is used for trials that calculate indentation rigidity in metals. Most indentation
hardness tests are used to determine the measure of deformation. Rockwell is a method of hardness
testing that simply relies on the metal’s resistance to indentation using a specific load application
and a specific indenter. Rockwell hardness testing is a technique used to measure the hardness of
the small materials objects. Materials may be ferric or non-ferric materials. For ferric materials,
we use a Diamond cone depressor (HRC) and apply pressure for 10 seconds while for the non-
ferric materials, we use steel ball depressor (HRB) and apply pressure on the materials for the 30
seconds.
Figure 7 Rockwell test scrutinizing different materials
 Conclusion
We learnt about Rockwell test for different materials and scrutinize of hardness of materials.
Ferric materials are harder than non-ferric materials. In non-ferric materials, copper is harder
than Aluminum.
[78]
 Precautions
1. Wear Gloves
2. Wear Lab Coat
the lab.
6. Wear shoes that completely cover your feet, you cannot

wear sandals or other open-toe OR open-heel shoes (clogs are
ok).
[79]
Lab #--------10--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[80]
Experiment # 10
To determine the Hardness of Metallic biomaterials using Brinell hardness
method
 Objective
1. To study Brinell hardness testing machine and to determine BHN of given samples.
1. Diamond cone indenter
2. Brinell hardness tester
3. Steel indenter
4. Micrometer travelling microscope
1. Aluminum
2. Mild Steel
3. Copper
 Theory
I. Brinell Hardness Test:
The Brinell Hardness Test method is the most commonly used hardness measurement technique
in the industry. In the Brinell Hardness Testing, the hardness of a metal is determined by measuring
the permanent indentation size produced by an indenter. Harder materials will generate shallow
indentations while the softer materials will produce deeper indentations.
Figure 30 composition of Brinell hardness machine
[81]
a) Working Principal:
An indentation hardness test using a verified machine to force an indenter (tungsten carbide ball
with diameter), under specified conditions, into the surface of the material under test. The diameter
of the resulting indentation is measured after the removal of the force. The indenter is brought into
contact with the test specimen in a direction perpendicular to the surface, and the test force is
applied (Apply the test force within 1 to 8 s). The test force is held for a specified dwell time and
then removed (Maintain the fully applied test force for 10 s to 15 s). The diameter of the indentation
is measured in at least two directions perpendicular to each other. The Brinell hardness value is
derived from the mean of the diameter measurements.
Figure 31 working principal of Brinell Hardness Test
b) Representation of Hardness Values:
Brinell hardness values shall not be designated by a number alone because it is necessary to
indicate which indenter and forces have been employed in making the test. Brinell hardness
numbers shall be followed by the symbol HBW, and be supplemented by an index indicating the
test conditions in the following order: Diameter of the ball (mm) and a value representing the test
force (kgf). Examples: 450 HB10/3000 = Brinell hardness number of 450 obtained with a ball
diameter of 10mm and a test force of 3000 kgf, 150 HB2.5/62.5 = Brinell hardness number of 150
obtained with a ball diameter of 2.5mm and a test force of 62.5 kgf.
c) Sample Surface Preparation:
When necessary, the surface on which the indentation is to be made should be filed, ground,
machined or polished flat with abrasive material so that the edge of the indentation can be clearly
defined to permit the measurement of the diameter to the specified accuracy. Preparation shall be
carried out in such a way that any alteration of the surface hardness of the test surface (for example,
due to overheating or cold-working) is minimized.
[82]
d) Measurement of the Brinell Indentation:
Measure the diameter of each indentation in two directions, perpendicular (90°) to each other.
Additional measurements of the indentation diameter may also be made. The arithmetic mean of
the measurements shall be used for the calculation of the Brinell hardness number.
For tests on flat surfaces, the difference between the largest and smallest measured diameters for
the same indentation shall not exceed 1% of the indenter ball diameter unless it is specified in the
product specification, such as for an anisotropic grain structure.
When indentations are made on a curved surface, the minimum radius of curvature of the surface
shall be two and a half times the diameter of the ball. Indentations made on curved surfaces may
be slightly elliptical rather than circular in shape. The measurements of the indentation shall be
taken as the mean of the major and minor axes.
e) Indentation Spacing:
The distance between the centers of two adjacent indentations shall be at least three times the
diameter of the mean indentation. The distance from the center of any indentation to an edge of
the test piece shall be at least two and a half times the diameter of the mean indentation.
Figure 32 Indenter first tap tents for Brinell Hardness Test
f) Testing of the Ambient Temperature:
Brinell hardness tests should be carried out at an ambient temperature within the limits of 10 to
35°C (50 to 95°F). Users of the Brinell test are cautioned that the temperature of the test material
and the temperature of the hardness tester may affect the test results. Consequently, users should
ensure that the test temperature does not adversely affect the hardness measurement.
g) Brinell Hardness Scale:

The combinations of indenters and test forces define the Brinell hardness scales. The standard
Brinell hardness scales and test forces correspond to force-diameter ratios of 1, 1.25, 2.5, 5, 10 and
30.
[83]
h) Calibration:
Ddetermination of the values of the significant parameters by comparison with values indicated
by a reference instrument or by a set of reference standards.
i) Indenters:
Indenters for the Brinell hardness test shall be tungsten carbide balls of four allowed diameters (1,
2.5, 5 and 10 mm). Dust, dirt, or other foreign materials shall not be allowed to accumulate on the
indenter, as this will affect the test results.
j) Brinell hardness machine:
Equipment for Brinell hardness testing usually consists of a testing machine, which supports the
test specimen and applies an indenting force to a ball in contact with the specimen, and a system
for measuring the mean diameter of the indentation in accordance with the Brinell hardness test
principle. The design of the testing machine shall be such that no rocking or lateral movement of
the indenter or specimen occurs while the force is being applied. The design of the testing machine
shall ensure that the force to the indenter is applied smoothly and without impact forces.
Figure 33 Mechanism of Brinell Hardness testing Machine
1. Grind the surface of specimen to achieve a smooth and plane surface area
2. Place the specimen on the stage of machine
3. Raise the stage, so as to bring the surface of specimen in contact with the indenter
4. Apply load as follows
5. For hard metals (ferrous): 3000 kg
6. For softer metals (non-ferrous): 500 or 1000 kg
[84]
7. For ferrous metals, keep the load applied for a period of 10 sec and for non-ferrous
metals, keep the load applied for 30 sec
8. Release the load and remove the specimen
9. Measure the diameter of indentation in two perpendicular directions with micrometer
travelling microscope and find the mean of two values
10. Using relationship given, calculate BHN
 Observation and Calculation:

For calculations, we use the following formula.
𝐿
BHN =
(𝜋D/2) ( D−√(𝐷 2 −𝑑2 )
1. For Copper:
 L = 1000kg
 D = 10mm
 d = 4mm
 F = 76
1000
BHN = = 76.
(𝜋10/2) ( 10−√(102 −4 2 )
Figure 34: Diameter of Copper
2. For Aluminum:
 L = 1000kg
 D = 10mm
 d = 5.5mm
 F = 38.62
1000
BHN = = 38.62.
(𝜋10/2) ( D−√(102 −5.52 )
Figure 35: Diameter of Aluminum
[85]
3. Steel
 L = 3000kg
 D = 10mm
 d = 3.8mm
 F = 254.60
3000
BHN = = 254.60
(𝜋10/2) ( 10−√(102 −3.82 )
Figure 36: Depth of Steel.
 Results
For Copper Hardness is = 76 BHN

For Aluminum Hardness is = 38.62 BHN
For Steel Hardness is = 256.60 BHN
Figure 8 Brinell hardness test for MS Figure 9 Brinell hardness test for Al
[86]
Figure 10 Brinell hardness test for Copper
 Discussion
The method of Brinell testing consists of forcing a hardened steel ball, under a predetermined
pressure, into the test material. The Brinell hardness number is obtained by dividing the pressure
in kilograms spherical surface of the impression, which is expressed in square millimeters.
For non-ferric materials, we applied weight of 1000kg for 30 sec while for ferric materials, we
applied weight of 3000kg for 10 seconds.
Figure 11 Generalized Principal for rough and coarse micro-structure materials
Brinell Testing machine is used to test the hardness level of the normal sized materials. The
application of hardness testing enables you to evaluate a material's properties, such as
strength, ductility and wear resistance, and so helps you determine whether a material or
material treatment is suitable for the purpose you require. The Brinell hardness test is an
empirical indentation hardness test that can provide useful information about metallic materials.
This information may correlate to tensile strength, wear resistance, ductility, and other physical
[87]
characteristics of metallic materials, and may be useful in quality control and selection of
materials.
 Conclusion
Ferric Materials are harder than non-ferric materials. With the help of experiments, it became
clear that Copper is harder than that of Aluminum.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
the lab.
6. Wear shoes that completely cover your feet, you cannot

wear sandals or other open-toe OR open-heel shoes (clogs are
ok).
[88]
Lab #--------11--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[89]
Experiment # 11
To determine ductility and quality of metallic samples using Erichsen
Cupping Index Machine
 Objective
1. To carry out Erichsen cupping Test. Different samples (steel, brass and aluminum)
were tested under Erichsen Cupping machine and then the results were compared for
the hardness of specimens.
1. Micrometer
2. Cutter
1. Aluminum
2. Mild Steel
3. Copper
 Theory
I. Erichen Cupping Test:

The Erichsen Cupping Test (ECT) is aimed to
produce stretching conditions on thin sheets
by imposing a spherical or hemispherical
punch against a clamped specimen placed
between a die and a blank-holder. The
hemispherical tip of the punch stretches the
sample until the through-thickness failure
occurs.
Ductility can be defined as the ability of a
material to deform plastically before fracture.
Many methods can be used for measuring
the draw-ability of metal sheets, for example,
bending, scratching, coining drawing, etc.
One of them is the Erichsen cupping test for
sheet metal. Erichsen Cupping test for sheet
metal includes placing the metal sheet in the
machine and punching a cup into the sheet Figure 37 Erichen Cupping Test
until a crack appears. Then the depth of the

cup is measured and eventually, ductility is measure as a function of this depth. When we talk
about cupping testing procedure, there is a minor confusion about its important. If it is
important for testing properties of metal sheets or for testing properties of coatings? Well, for
both. Different industries use cupping test machine for assessing different quality factors. For
metal, it is done to adjudge the ductile properties. For some applications, it becomes important
that metal should bear the punch to a certain extent without getting cracked. E.g. automobiles
[90]
with metal bodies must pass this test. As vehicles are prone to dents due to external impacts,
the metal body should be ductile enough not get cracked. Again, in the automobile industry,
this cupping test is performed to check the elongation of the coating done on the body of the
vehicle. The paint should not chip off or cracked when a dent is formed due to the punch. If
coating gets removed, the metal underneath would become prone to corrosion.
a) Erichsen cupping test standards
There are two most commonly used Erichsen cupping test standards for reference
purposes. One standard is ASTM E 643-78 and the other is ISO 20482. While performing a
deep drawing test for research and standard testing purpose, it is in the best interest to follow
standard guidelines given in standards.
b) Principle
The first thing to understand while searching for Erichsen cupping test theory in principle.
The principal of the Erichsen cupping test for sheet metal is simple. Forcing a hemispherical
punch into the metallic sheet until a crack appears and at that point measuring the depth of cup
as the circular scale shows. Harder the metal, with fine grain structure, will result in lesser
depth and fine shape of a cup. On the other hand, ductile metal, with coarse grain size, results
in deeper indent with a rough cup shape.
Figure 38 Working Principal of Erichsen Cupping test
c) Construction
The principle diagram of the Erichsen cupping machine is shown below. It consists of a die
containing hollow space to move punch. A clamping system to clamp the metallic sheet
or steel sheet in between two frames of a die. A mirror is placed behind the punch and sheet to
[91]
observe crack and to know up to where the force is to apply on a punch. The punch is moved
into the plain sheet by moving the wheel. The scale is attached to the wheel to measure the
depth of indent in millimeters.
Figure 39 Specimen Specification and requirements of Erichsen Cupping test
d) Working
Working is as simple as described in the

principal. First of all, a metallic sheet is
placed between the die and is clamped.
The thickness of the metallic sheet is
important while performing the
Erichsen cupping test for sheet
metal according to international
standards. Typically the thickness of the
sheet is 1mm. when clamped, the wheel
is moved until it touches the sheet. After
that, the scale attached to the wheel is
set to zero and then a force is applied on
the sheet through cupping punch.
The punch deforms the metallic sheet.
Indentation depends upon the ductility Figure 40 Mechanism of forced cupping for Erichsen Cupping test
of material which is materials property.
Punch is moved into the sheet until a crack is observed from the mirror behind the forcing unit.
The movement of the wheel also moves the circular sliding scale. The units of scale are in mm.
at the point of crack beginning. The measurement of the scale is noticed and then the sheet is
unclamped. The measurement taken is the depth of indent which is then compared with the
values given in the literature.
[92]
1. Prepare the specimen in the form of square sheet
2. Make sure that the surface of sheet specimen is smooth
3. Measure specimen thickness up to the accuracy of 0.01 mm
4. Slightly grease the ram and die of the machine
5. Hold the sheet to be tested between cupping die and holding die socket
6. By turning the wheel clockwise, bring the holding die towards the cupping die
7. Set the hand wheel scale to zero
8. Switch on the electric lighting so as to illuminate the sheet surface being monitored
9. Disengage the clutch
10. Turn the wheel clockwise very slowly, so that ram penetrates into the specimen
11. Watch carefully the surface of specimen through the mirror on the back of specimen.
On the appearance of first incipient crack, read the drawing depth on the scale
12. By turning the wheel anti-clockwise, remove the specimen.
 Observation
A depth had produced in the plate, and after the breakdown value reached, the plate broke at
the depth.
Figure 41 Erichsen Cupping test carried out for steel
[93]
Figure 42 Erichsen Cupping test carried out for Copper
 Results
Both copper and aluminum break down when their breakdown values reached. Copper broke at
12.5mm while aluminum broke at 9.3mm. hence it proved, aluminum is more ductile than
copper.
 Discussion
Cupping test is a significant test to assess the ductile properties of the metal. In this test, a punching
force is applied to the metal sheet of thickness from 0.2mm to 2mm. A visual inspection is done
to see if the test material has developed any cracks or deformity. The force applied on the sample
is displayed by the gauge. The Erichsen cupping test is a standard method used to assess the
formability, ductility and adhesion of coatings, which induces similar strains to those experienced
during forming.
Both copper and aluminum break down when their breakdown values reached. Copper broke at
12.5mm while aluminum broke at 9.3mm. hence it proved, aluminum is more ductile than copper.
It has a wheel and cup shaped structure that produces the depth in the metals. Aluminum and
Copper are materials used in this experiment. The Erichsen cupping test is a standard method used
to assess the formability, ductility and adhesion of coatings, which induces similar strains to those
experienced during forming. The Erichsen cupping test is a standard method used to assess the
formability, ductility and adhesion of coatings, which induces similar strains to those
experienced during forming.
[94]
Figure 43 Erichsen Cupping test result comparison of Brass VS Al
 Conclusion
We learnt that Aluminum is more ductile than that of the copper. Copper break at 12.5mm while
aluminum broke at 9.3mm.
 Precautions
1. Wear Gloves
2. Wear Lab Coat
lab.
or turned backward.
[95]
Lab #--------12--------
safely
Total
is conveyed
digits
Total
occasions. time.
Total
Lab Instructor:
[96]
Experiment # 12
To determine the tensile strength of metallic samples using Universal testing
machine
 Objective
1. To perform tensile test on a given Steel Specimen and
2. To study its mechanical properties
1. UTM machine
2. Vernier caliper
1. Steel sample (28mm)
 Theory
I. Universal Testing Machine (UTM):
A Universal testing machine (UTM) is used to test the mechanical properties (tension,
compression etc.) of a given test specimen by exerting tensile, compressive or transverse
stresses. The machine has been named so because of the wide range of tests it can perform
over different kind of materials. Different tests like peel test, flexural test, tension test, bend
test, friction test, spring test etc. can be performed with the help of UTM.
Components:
A universal testing machine consists of two main parts
1. Loading Unit
2. Control Unit
1. Loading Unit
The arrangement of the test specimen and the exertion of the load is held in the loading unit.
The loading unit of a UTM consists of the following components :
 Load Frame
The load frame of a universal testing machine can be made either by single support or by
double support. The load Frame consists of a table (where the specimen is placed for the
compression test), upper crosshead, and lower crosshead.
 Upper Crosshead and Lower Crosshead
The upper crosshead is used to clamp one end of the test specimen. The lower crosshead in the
load frame is the movable crosshead whose screws can be loosened for height adjustment and
[97]
tightened. Both the crossheads have a tapered slot at the center. This slot has a pair of racked
jaws that is intended to grip and hold the tensile test specimen.
 Elongation Scale
The relative movement of the lower and upper table is measured by an elongation scale which
is provided along with the loading unit.
Figure 44 Universal Testing machine composition
2. Control Unit
The variations in the application of the load and the corresponding test result are obtained from
the control unit. The main components of the control unit in a universal testing machine are
 Hydraulic Power Unit:
This unit consists of an oil pump that provides non-pulsating oil flow into the main cylinder of
the load unit. This flow helps in the smooth application of load on the specimen. The oil pump
in a hydraulic power unit is run by an electric motor and sump.
 Load Measuring Unit
This unit has a pendulum dynamometer unit that has a small cylinder with a piston which
moves with the non-pulsating oil flow. The pendulum is connected to the piston by pivot lever.
The pivot lever deflects based on the load applied to the specimen. This deflection is converted
to the load pointer and displays as the load on the dial.
[98]
The range of load application can be adjusted by means of a knob in the load measuring unit
(0-100 kN; 0-250 kN; 0-500 kN and 0-1000 kN). The accuracy of measuring unit controls the
overall accuracy of the machine.
 Control Devices
The control devices can be electric or hydraulic. Electric control devices make use of switches
to move the crossheads and switch on/off the unit. A hydraulic control device consists of two
valves, Right Control Valve and Left Control Valve or Release Valve. A right control valve is
used to apply load on the specimen
Figure 45 Control unit with DAC system of UTM
Working Principal:
It works on the principle of elongation and deformation. These machines usually use a
hydraulic cylinder to create the force. The applied force is determined by system pressure
which can be accurately measured. The high-pressure oil pump supplies oil to the working
cylinder, and through the movement of the piston, the platen and the upper beam (the upper
jaw seat) are pushed upward to perform the tensile or compression test of the sample. The
tensile test is carried out between the upper beam of the main machine and the moving beam,
and the compression test is carried out between the platen of the main machine and the moving
beam. The adjustment of the test space is achieved by the drive mechanism (elevating motor,
sprocket, chain, etc.) driving the double screw to rotate synchronously to raise and lower the
moving beam.
[99]
Figure 46 working mechanism of UTM
1. Repeat the same procedure for any other specimen
2. First metal was selected. We select the steel sample a ferric material.
3. Sample rod was put in the UTM Machine to check the tensile strength of the rod.
4. Force was applied with the help generator. At some point, the value stops for the bit.
This was the value after which the sample broke at certain point.
5. The rod brakes at some point.
6. At this, note the value of breaking point from the machines.
7. Record the observations.
 Observation
After the application of high stress, the metal rod broke into two pieces. The break down
value depends upon the toughness and hardness of the metal rod. For 28mm metal rods, it
absorbs energy of 47kton (kilo ton). This value was the breakdown value of 28mm rod.
 Results
Here are the result represented by images of readings which has been explained aformentation.
Below are images of result after scrutinizing the procedure
[100]
Figure 47 UTM tensile testing for steel Figure 5 UTM tensile strength value for steel
Figure 6 UTM tensile testing for Al Figure 7 UTM tensile strength value for Al
[101]
 Discussion
A tensile test is performed to determine the
tensile properties of a material or
component. The test sample's deformation
is used to characterize its ductility or
brittleness as well as important
characteristics such as tensile strength, yield
point, elastic limit, percent elongation,
elastic modulus and toughness. For metals,
we check the tensile properties of the
materials using the tensile strength while for
non-metals, we use the compression
properties. In UTM Machine, we can check
by both ways either by tensile strength for
metals and by compression for non-metals.
Metal we use in this experiment is steel
sample (building steel rod). For metals, we
check the tensile properties of the materials
using the tensile strength while for non-
metals, we use the compression properties.
In UTM Machine, we can check by both
ways either by tensile strength for metals
and by compression for non-metals.
Figure 8 UTM configuration along with description of parts
 Conclusion
We learnt about 28mm metal rods absorbs 47kton (kilo ton) energy and it is the breakdown
value for the 28mm metal rod (steel rod).
 Precautions
1. Wear Gloves
2. Wear Lab Coat
lab.
or turned backward.
[102]

Biomaterial Lab 10

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DEPARTMENT OF BIOMEDICAL

UNIVERSITY OF ENGINEERING AND TECHNOLOGY

LAHORE (New Campus)

Roll No: __________2020-BME-10____________

Sr# Title Page #

1 Preparation and Micro-structure Analysis of Stainless Steel specimen 03

2 Preparation and Micro-structure Analysis of Mild Steel 10

5 Immersion of Stainless Steel (SS) and Mild Steel in Ringer Solution 36

6 Preparation of Bio-ceramics Pellet Using Hydraulic Press 45

7 Densification of Bio-Ceramics Pellet by Sintering 53

8 Production and sintering of porous bio-ceramics scaffolds using naphthalene as 62

the pore former

9 To determine the hardness of metallic biomaterial using Rockwell Hardness 73

10 To determine the Hardness of Metallic biomaterials using Brinell hardness 80

11 To determine ductility and quality of metallic samples using Erichsen Cupping 89

Name: Haseeb-UR-Rehman___________________ Reg. No.: ___2020-BME-10_____________________

 Note: 10 % of cognitive (CLO2) will evaluate through written lab viva

Name: Dr. Nida Iqbal Signature : _______________________

 They involve nearly waste-free net-shape forming.

Relay system, which reflects and enlarges the image

High-intensity light source

Plane glass reflector

Colored or polarizing filters

Figure 5 stainless steel microscopic structures

Figure 2 grid paper 220

Figure 7 grid paper 400

3rd Grid Paper (600C)

Figure 8 grid paper 600

4th Grid Paper (800C)

Figure 9 grid paper 800

Name: Haseeb-UR-Rehman___________________ Reg. No.: ___2020-BME-10_____________________

 Note: 10 % of cognitive (CLO2) will evaluate through written lab viva

Name: Dr. Nida Iqbal Signature : _______________________

 They involve nearly waste-free net-shape forming.

Relay system, which reflects and enlarges the image

High-intensity light source

Plane glass reflector

Colored or polarizing filters

Figure 5 Microscopic Structure of SS

Figure 6 grid paper 220

2nd Grid Paper (400C)

Figure 7 grid paper 400

3rd Grid Paper (600C)

Figure 8 grid paper 600

Figure 9 grid paper 800

Figure 10 safety precautions

Name: Haseeb-UR-Rehman___________________ Reg. No.: ___2020-BME-10_____________________

 Note: 10 % of cognitive (CLO2) will evaluate through written lab viva

Name: Dr. Nida Iqbal Signature : _______________________

Figure 4 corrosion in orthopedic implants

 Uniform loss of metal over entire surface

 Electrochemical process where one metal corrodes preferentially compared to another

 Localized corrosion resulting in cavities or holes

 Localized corrosion at the crevice between two joining surfaces

 Growth of crack formation in corrosive setting

Figure 5 corrosion in metals

II. Corrosion in mild steel

III. Stainless steel in Acidic media

Figure 6 corrosion of metals under acidic environment

1 10% HCl W1 12.8g 13.57g

2 10% HCl W2 13.59g 13.11g

3 10% HCl W3 11.12g 10.24g

Roll No: 2020-BME-10__

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________

Name: Haseeb-UR-Rehman_________________ Reg. No.: _2020-BME-10_____________________