Toxicology.

How can Biotechnology contribute to make things better?

An apple a day… keeps the doctor away?

A00822870 Alejandro Garibay Reyes

A01172916 Gerardo M. González Constantino

A01173120 Ma. Fernanda González Ocampo

A00825640 Carlos E. Quezada Duarte

Prof. Bertha Olivia de la Re Dávila

Monterrey, Nuevo León, November 30th, 2021.

 How can Biotechnology contribute to making things better?
A. Garibay, C. Quezada, G. González, M. González.
Toxicology by Dr. Bertha Oliva de la Re Dávila
Monterrey Institute of Technology and Higher Education
November 30th, 2021.

Abstract: Food waste is one of the most common problems in the world, as we all know, unless
recycled, our waste is commonly deposited in landfills, presumed to be the best places for it, though
this is not always the case. Scientists and recent data have proposed some mechanisms in order to
obtain biomaterials from food waste, these materials can be used as a replacement for plastic materials
or even in regenerative practices for tissue in the medical field.

Key words: food waste, biomaterials, apple waste, regenerative, antioxidants.

1. Introduction years, perform a key factor in fields such as

The handling of wastes is one of the
problems among many that concern the
toxicological and environmental departments.
As seen in class, this handling has more
negative effects than people realize, whether
e-waste, toxic metals or even medicine
dumped in water lands. This work will provide
a recycling solution of one of many wastes,
specifically the fruit waste area.
As we all know, unless recycled, our
waste is commonly deposited in landfills,
presumed to be the best places for it, though
this is not always the case. Most of our waste
if misplaced can damage the environment. The
improper management of waste can affect the
soil, water and air quality. By burning any
disposed waste and plastic materials, which is
part of the process in landfills, results in air
and environmental pollution.
Fruit waste commonly doesn’t need
the same management as inorganic waste, if
managed correctly it can bring positive effects
to the soil by composting. This still is not the
case for most of the fruits and vegetables in
the trash. Nowadays, there are a lot of benefits
obtained from organic waste, like the creation
of biomaterials from fruit waste to replace
plastic and disposable items, or like in recent
medicine. By implementing the use of this
biomaterial we could replace the use of
plastics for water bottles since eventually they
break down releasing DEHA
(diethylhydroxylamine) a carcinogen that
eventually hurts our reproductive capabilities
and causes liver dysfunction (Waste
Management, 2017).
The main objective of this project is to
present various ways where fruit waste can be
used to help the general waste problems,
mostly focusing on the biomaterials that can
be generated by fruit and the replacement of
plastics and other materials by these
biomaterials. Of course there are more positive
things that fruit waste can generate and it is
considered a new technology that we have
barely tapped into, as is the case for a lot of
biotechnology areas.
2. Objectives
a) To present a novel solution to one of
the problems seen in class.
b) To display and prove our
understanding of the subject seen
throughout the semester.
 c) To learn about waste and how it
affects negatively on the environment.
3. Background
In recent years, scientists and new
published data have shown the potential of
certain food waste in order to obtain
biomaterials that can be used in regenerative
medicine, a field in medicine which develops
methods to regrow, repair or replace damaged
or diseased cells, organs or tissues.
Before we dive into the novel biotech
approach our team wants to propose, extensive
research in this particular topic was
effectuated, thus many scientific articles and
start.ups that linked organic and plastic waste
with a new way to take advantage of these
types of materials were found, some of these
are presented in below.
a) Biodiesel waste transformed into
commodities by Yates et al (2004) [1].
b) Citrus waste transformed into p-cyme,
a pharmaceutical and fine chemical
intermediate, by Martín-Luengo et al
(2008)[2].
c) Rice production wastes used in
materials to decontaminate effluents
by Martín-Luengo et al (2010)[3].
d) Sunflower production residues
transformed into multifunctional
materials by Martín-Luengo et al
(2011)[4].
e) Beer production waste transformed
into powdered materials by Yates et al
(2008)[5].
f) Scaffolds for support of controlled
desorption of bioactive substances by
Martínez Serrano et al (2015)[6].
Nevertheless, an emerging solution
presented by Yates et al (2016) was the one
that attracted the most attention towards the
team in order to carry out the research needed
by the project, since it links the waste from (if
not the most) a well known fruit that tends to
be everyday in most of people’s houses with
medicine; the solution is presented in the next
section of this report.
4. A novel Solution: An apple a day…
keeps the doctor away? - results
4.1 The start-up
It has been found that organic residues
can derive materiales with a tissue engineering
application in biomedicine. There is a study
about the employment of food waste to
develop biomaterials that allows the
regeneration of bone and cartilage tissues for
people with osteoporosis, arthritis or
osteoarthritis, made by CTB-UPM (Centre for
Biomedical Technology at Universidad
Politécnica de Madrid)[7] in collaboration with
other institutions.
CTB-UPM study was based on “Apple
pomace” which is an abundant material with
approximately 20-30% of dried matter, mainly
used for compost or animal food. The
procedure is based on a sequential extraction
of bioactive molecules, as antioxidants or
pectin, to obtain the material with the correct
porosity and texture needed to be used as
tissue.
Apple flesh has been used to create
human tissue in the shape of an ear and
asparagus stalks to regenerate spinal cords in
the University of Ottawa[8]. The researchers
use the decellularization technique with water
and soap to remove the cells from the fruits
and veggies, the result is a naturally
vascularized cellulose scaffold which can then
be repopulated with many mammalian cells.
Also, the article that was the base for
this research was the one published by Yale et
al (2016)[9], that used apple pomace as well, a
more detailed overview of this study is
presented below.
4.1.1 The Yale Study
 In order to obtain the biomaterials for
medicine to use, three stages have to be
effectuated.
a) First stage: the antioxidants and sugars
from the AP are extracted in deionized
water, then this liquid is heated and
mixed with 1 N citric acid solution.
b) Second stage: mixture is filtered and
liquid and solids are stored at 4ºC,
then a treatment with ethanol is
applied in order to pectins to
precipitate; a centrifugation, filtration
and purification process is then
effectuated to remove traces of
monosaccharides and disaccharides.
c) Third stage: an analysis by
thermogravimetric and differential
thermal analysis (TG-DTA) is
performed, the conditions employed in
the sequential valorization of the AP
were chosen to achieve the maximum
yields of biocompatible material,
given its greater commercial value,
without forgetting the importance of
the extracts and pectins.
Also, it has to be stated that extracted
pectins were compared with the commercial
sample using TG-DTA, XRD and FTIR
analyses, other several studies are performed
in the product in order to determine the
presence of different types of elements, this
studies include: electron microscopy coupled
to microprobe analyses, X-ray diffraction
(XRD), high resolution chromatography, ion
chroma-tography (Mono and di- saccharides
Dosinos and Hamilton RCX- 30),
Finally, “Human chondrocyte
(CHON-001, ATCC) and murine osteoblast-
like (MC3T3-E1, ATCC) cell lines were
cultured on the biomaterials in DMEM
(Gibco) supplemented with 10% foetal bovine
serum, 2 mM Glutamine, 1% nonessential
amino acids and 1% penicillin- streptomycin
(basal medium) [Yale et al, 2016]”.
4.2 The improvement
According to different collected
information, this type of procedure is designed
in order to be effectuated at low temperatures
and with low toxicity solvents in order to make
the whole process a green and inexpensive
process for multi-valorisation, also this
procedure leads to an extraction of
nutraceuticals (antioxidants, carbohydrates,
and biocompatible cations) that can be later
used as food and drink additives, with the
added value of being obtained from a
sustainable source, some of the main
biological-interest substances that can be
obtained are: fructose, sorbitol, glucose,
sucrose, xylose, arabitol and arabinose.
Also, antioxidants can be obtained
from this source, as stated in baste study, “the
composition of the extracts in polyphenolic
antioxidants indicates the presence of
phloridzin, chlorogenic (5-caffeoylquinic acid)
and isomer-caffeoylquinic acid (99% of the
total) and about 1% is a mixture of epicatechin
and catechin (Yale et al, 2016)”. These
components have been proved to be beneficial
towards some diseases such as cancer,
cardiovascular dysfunctions and diabetes.
Also, these substances can be used as
nutraceutical and cosmeceutical additives.
Finally, the extracted pectin is of great
value because it is considered a safe additive
of unlimited daily consumption by the FDA[10]
(Muller-Maatsch et al, 2016) and has also an
enormous valuable towards pharmaceutical
and biomedical applications, but the process
doesn’t ends here, as he material left after
pectin production was transformed by heating
to 500 C into a material capable of acting as a
scaffold for cell growth in hard and soft tissue
engineering, being of great importance on the
medical field, and these have been proved to
be applied for use mainly in bone, tooth and
cartilage replacement therapies, being good
candidates for development of hard and soft
tissue engi- neering scaffolds.

4.3 The impact on the environment
In 2015, the world production of
apples was over 70 million metric tons. A big
part in the apple industry is when it is
converted to juice which leaves a large
quantity of apple pomace, some of it is used as
compost but otherwise the rest is just waste.
Physical apples tend to have a lot of storage
problems which if not consumed are also
perceived as just waste. This waste as given
examples in points 4.2 and 4.1 has a big area
of opportunity to be reused.
4.4 Regulatory aspects for biomaterials’
approval.
In general, biomaterials used in
medicine are regulated as medical devices.
Regulations and toxicological testing has been
a challenge due to the specificity of the
materials in the area of application. In the
United States, the Food and Drug
Administration (FDA) regulates the
manufacturing, design and marketing of
medical devices, where biomaterials are found
(Zhang, 2016). For the oversight of medical
devices, The Center for Devices and
Radiological Health (CDRH), is the branch of
the FDA that regulates all related medical
materials.
In the classification of the biomaterial,
these can be Class I, II or III, where class III is
the one that possesses the more significant
risk. For quality regulations, Quality System
Regulations (QSR) provides the rules that
manufacturers must follow, the FDA conducts
periodic inspections of manufacturers to assure
their quality systems meet the QSR standards.
4.5 Required tests
The sensitive test system for
biomaterials is really desirable for the
evaluation of possible hazardous reactions,
because of the difficulty of the obtention of the
inherent characteristics of the materials.
Extrapolating from one test system to another,
such as from animals to humans is difficult but
can be allowed, toxicologists have used the
concept of safety factors to take into account
intra- and interspecies variation. This practice
requires being able to exaggerate the
anticipated human clinical dosage in the
nonhuman test system (Hanson, 1996). Some
of the tests are: direct contact test, agar
diffusion test and elution test. In this type of
assays, the concentration gradient of toxic
chemicals can be evaluated, also, physical
trauma from movement of the specimen in the
direct contact assay is evident by patches of
missing cells interspersed with normal healthy
cells (Hanson, 1996).
5. Discussion
As seen with the presented results, this
alternative of food waste has a multi-valorized
outcome, not only because it provides
biosubtances of interest such as antioxidants
and carbohydrates, but also because it can be
extended towards one of the most important
fields out there: the health industry; with
further improvement in this type of
procedures, a before and after can be
developed in regenerative medicine practices,
since costs can be reduced by obtaining raw
material directly from an organic and very well
available source: an apple.
Also, by betting towards this type of
practices, a “zero waste” philosophy could be
created, impacting on a positive way in the
environmental field, as stated by Yale et al,
sustainable and cost effective industrial
valorization of AP into high value added
products has important economic and
environmental benefits and conversion
paths are sought to find the most suitable
one.
Most of this, if not all, has yet to be
used by big companies or establishments. One
of the messages and reasons for this work is to
remind us of the environmental benefits these
technologies can bring us. We as humankind
have a social and ethical obligation to take
care of the life on this planet, it is
understandable that the way waste is managed
today is easier and cheaper than recycling and
finding ways to reuse like this work proposes
but that is just not ideal for us as humans on
Earth. This problem goes beyond just being a
toxicological issue as it can be seen as a social
issue.
6. Conclusion
Waste contamination and waste
displacement were one of the many subjects
seen in our toxicology class. The effects these
can have are sometimes life threatening for
several species in the flora and fauna category,
even sometimes on humans. We presented one
small way to reduce some of these wastes and
showed the possible benefits these reductions
can have. As well we discussed the importance
it has in toxicology and the environmental
applications these new technologies have.
We learned various ways of reducing
waste and reusing this waste for a better
purpose. We as a team remembered the saying
of “the three Rs” that was repeatedly shown in
school classes, now in University and enrolled
in advanced courses but this paper shows still
the importance of this saying, now of course,
we know that for this saying to have a big
effect, is for companies and governments to
reduce, reuse and recycle their wastes, for a
better environment.
Our solution is not about fruit waste in
itself as the problem, but more so in plastic
wastes (and others) and using fruit waste to try
and replace some materials that would end up
in the trash and contaminate. This replacement
is what this paper is about and how the
creation of biomaterials, antioxidants and
sugars using fruit waste can try to balance the
huge existent waste on Earth.
References
[1] Yates, M., Martin-Luengo, M.A., Garcia,
G., 2014. Catalysts Prepared with
Sunflower Residues to Prepare Fuel
Additives with Biodiesel Wastes.
International Congress on
Environment (CONAMA), Madrid,
Spain.
[2] Martín-Luengo, M.A., Yates, M., Martínez
Domingo, M.J., Casal, B., Iglesias, M.,
Esteban, M., Ruiz-Hitzky, E., 2008. Synthesis
of p-cymene from limonene, a
renewable feedstock. Appl. Catal. B Environ.
81, 218e224.
[3] Martin-Luengo, M.A., Yates, M., Diaz, M.,
Saez Rojo, E., Gonzalez Gil, L., 2011.
Renewable fine chemicals from rice and citric
subproducts: Ecomaterials. Appl.
Catal. B Environ. 106, 488e493.
[4] Martin-Luengo, M.A., Yates, M., Gomez,
M., 2011c. Procedure to Obtain
Multifunc-
tional and Renewable Materials from
Sunflower Oil Production Wastes.
Patent
P201130303.
[5] Martin-Luengo, M.A., Yates, M., Ramos,
M., Saez Rojo, M., Martínez Serrano,
M.A.,
GonzalezGil,L.,2011b.Biomaterialsfrombeerm
anufacturewasteforbone
growth scaffolds. Green Chem. Lett. Rev. 4,
229e233.
[6] Martin-Luengo, M.A., Yates, M., Ramos,
M., Saez Rojo, M., Martínez Serrano,
M.A.,
GonzalezGil,L.,2011b.Biomaterialsfrombeerm
anufacturewasteforbone
growth scaffolds. Green Chem. Lett. Rev. 4,
229e233.
[7] Veronika Bátori. (2019). Fruit wastes to [10] Müller-Maatsch, J., Bencivenni, M.,
biomaterials. Noviembre 2021, from Caligiani, A., Tedeschi, T.,
Univiersity of boras Recovered from: Bruggeman, G., Bosch, M., Petrusan,
https://www.diva-portal.org/smash/get/ J., Van Droogenbroeck, B., Elst, K.,
diva2:1270941/FULLTEXT01.pdf Sforza, S., 2016. Pectin content and
composition from different food waste
[8] Duan, Yumi, et al. 2021. Apple orchard streams. Food Chem. 201, 37e45.
waste recycling and valorization of
valuable product-A review: [11] Zhang, N. (2016). In Situ Tissue
https://www.tandfonline.com/doi/full/10.1080/ Regeneration || Regulatory Aspects. ,
21655979.2021.1872905 (), 383–403.
doi:10.1016/B978-0-12-802225-2.000
[9] Yates Buxcey, M., Ramos Gómez, M., 20-9
Martín-Luengo, M. Á., Zurdo Ibañez,
V., & Martínez Serrano, A. M. (2016). [12] HANSON, STEPHEN (1996).
Multivalorization of apple pomace Biomaterials Science || Testing
towards materials and chemicals. Biomaterials. , (), 215–242.
Waste to wealth. doi:10.1016/b978-0-08-050014-0.5001
0-9