Notes Week 8 Development of Enamel and Dentin

Blue = oral epithelium; Green = mesenchyme from the NCCs; Pink = regular mesenchyme
Go thru Placode stage which is an ingavination of the oral epitehlium; following by Bud stage; then cap stage:
Cap stage: OEE = white
IEE = orange
dental papilla/odontoblasts = light brown between the orange and green is dificult to see;
dentin = dark brown, enamel = yellow

RECIPROCAL INDUCTION
The IDE forms the dental papilla which forms the odontoblasts
that go back to stimulate the IDE in reciprocal induction to form
the ameloblasts
Laid down first: dentin following by enamel
As the hard tissues are being laid down you move into bell stage.

Infolding of the dental lamina, with buds starting to come out

for primary dentition; off the dental lamina is the successional
laminae for permanent teeth

Biofabrication Ameloblasts and odontoblasts are the cells that build the hard tissues
Natural pathway: genes encode proteins that secrete matrices affected by migratory vectors that cause
mineralization leading to functional dental tissues that make up the tooth as an organ that can then perform
functions

Hierarchical • Outer hard wear-resistant brittle enamel

organization of • Inner soft tough flexible dentin
teeth • Enamel is a meshwork of rods
• Rods contain many crystallites
• Protein matrix controls mineralization
• Amelogenin is the dominant protein

Scale:
NANO - SCALE: MOLECULAR
MICRO - SCALE: CELLULAR
MACRO - SCALE: ORGAN

Function of Tooth Enamel:

structure • Wear Resistance, hardness
• Bolus penetration & mastication
• Insolubility, acid resistance

Dentin:
• Toughness, fracture resistance
• Sensation
 • Repair

Mechanical Toughness and stiffness (elastic modulus)

function: Porcelain is less tough
Dentin: more tough. Less elastic than enamel
Enamel: less tough, more easy to chip—more stiff?

DEJ Basement membrane that lies between where the IEE and mesenchymal cells that become the odontoblasts
sit
Joining enamel to Ectomesenchyme → odontoblasts (from the mesenchymal cells)
dentin Ectoderm → ameloblasts (from the IEE)
DEJ is a unique layer of connective tissue between these!
Transitional zone Then, there’s a process of reciprocal induction
between the two Odontoblasts migrated inwards FROM DEJ (towards pulp)
Ameloblasts migrate outwards FROM DEJ (towards crown)
DEJ is Gene expression → protein → matrix → mineralization
jagged/transitional
and not an abrupt Both dentin and Enamel share proteins: AmelX, Ambn, Enam, Dspp, Dmp1
interface Collagen is only in dentin during the pre-secretory phase
After initial layer of dentin then enamel is formed, there’s a change in gene expression during secretory
Change in phase:
mechanical Dentin has more Collagen + dentino-proteins (DSPP, DMP1) – COLLA2
properties as you Enamel has more enamel specific proteins (AmelX, AMBn, ENAM, Mmp20)
cross it;
After the dentin is laid down, odontoblasts stay alive; and they have odontoblastic processes that stretch out;
Ameloblasts die (senescence) so they cannot REFORM enamel

Transition Bulk enamel → specialized aprismatic enamel

(diffuses the force of damage to structure
underneath) → DEJ → specialized mantle
dentin → bulk dentin
Hardness decreases

DEJ • High surface area, scalloping

• Collagen fibers
• Gradation in properties
• Mechanical interlocking
• Intimate crystal contact
• Combination of these factors

Dentin Formation
Dentin Tough
• Soft
• Soluble
• Permeable
• Has Tubules
• Tubule Density
• Tubule size
 • Tubule occlusion
• Peritubular dentin (PTD)
• Intertubular dentin (ITD)
• The protein scaffold (of collagen) is retained
• Dentin is similar to bone! (resorption by osteoclasts)

Primary Preodontoblast differentiation: begin migrated to the IEE and attach to fibrils in the BM; IDE release signal
dentinogenesis that cause differention of odontoblasts and polarization: nucleus moves to basal area
Gap junctions develop between the odontoblasts, and also have tight junctions too!
Mineralization moves from BM inwards, just as the odontoblasts’ soma are slowly also migrating inwards

Primary dentinogeneiss
• Mantle dentin – first layer of dentin adj to DEJ; lacks odontoblastic process and tubules
• Circumpulpal dentin: rest of dentin that goes around tooth
Peritubular dentin – walls of dentinal tubules
Intertubular dentin – everything else

Collagen Key protein in dentin formation is COLLAGEN!!!!!!!!

• Basic structural unit is a triple-stranded helical molecule
• Small gaps separate heads & tails
• Adjacent molecules have 3/4 of their length overlapping, creating a mesh for mineralization
• Stabilized by covalent bonds between the N& C termini
• Contains large amounts of glycine and proline
• Contain hydroxyproline and hydroxylysine - Vitamin C
• Sequence in the pattern Gly-X-Y
• Chains are left handed helix
• but the wrapping is right-handed!
• Type I found in dentin
• 2 chains α2 chain & 1 chain α1 chain
• Osteogeneis Imperfecta, Dentinogenesis Imperfecta, Brittle Bone Disease are collagen disorders

Odontoblastic Peritubular dentin is distinguished by a dense and smooth appearance compared to the more disorganized,
processes and granular intertubular dentin.
dentin tubules Many fenestrations of the PTD.
Tubules run at different angles
Few are perpendicular to the surface
Tubules differ in size
Side-branches are present
Odontoblastic processes are present

Secondary • Continued deposition of dentin after root formation

Dentinogenesis • Much slower
• As we age the pulp canals look smaller due to this process

Reparative Dentin Formation

Tertiary • Localized dentin formation in response to potential INJURY
Dentinogenesis • Reactionary or Reparative

Reactionary dentin:
• Slow progressing, moderate sized lesions
• Primary odontoblasts will respond and form Deposition of sclerotic reactionary dentin in the
tubular area on the pulp side!!!!!

Reparative dentin:
• Aggressive, large lesions
 • Invasion of dentin
• Odontoblasts have died
• Deposition of fibrodentin or osteodentin by Newly differentiated odontoblasts
• Again, on the pulpal side!!!!!!!

Enamel Formation
Enamel formation • Two major functional stages
Secretory
Maturation

Enamel organ From basement membrane to outside the tooth: don’t quite understand the purpose of these layers yet

• Stratum intermedium
High alkaline phosphate activity
Facilitate transport of phosphate from circulation to developing enamel organ?
• Stellate reticulum
Desmosomes and gap junctions
• Outer enamel epithelium
Single layer of cuboidal cells
Protective buffer?

Secretory stage: • Ameloblasts become highly elongated and polarized

amelogenessis • Tomes processes extend into matrix
Exocytose secretory vesicles
Determine boundaries between rod and interrod
• Secretion of enamel matrix proteins (EMPs): amelogenin (AMELX), ameloblastin (AMBN), enamelin
(ENAM) thru the tomes processes
• Almost the entire enamel thickness is secreted prior to minerlization
• Gel-like matrix of EMPs, mineral, and water
• Enamel crystals form at DEJ
• Hydroxyapatite-like
• May be seeded on collagen from dentin

Amelogenin: • The dominant enamel protein

• Highly conserved
• 180 residues
• 12 - 16 isoforms
• Nanosphere structure - SELF ASSEMBLY ! – like collagen
• Protein - Protein Organization
• Protein - Crystallite Organization
• Largely removed during mineralization

Transition Phase • Ameloblasts shorten

• Tomes’ processes are lost
• Transistion into maturation phase

Maturation phase • Ameloblasts shorten

Transport ions
Maintain acid-base balance
Remove/endocytose of EMP
Apopotosis of ameloblasts
PUMPING MINERAL INTO THE TOOTH!!!!!!
Hydroxyapetite

Acidity formed in the process so has to be dealt with!

Crystalline structure is hexagonal and forms rod, then interweaving for strength. Anions like fluoride can fill
in the channels

Matrix removal Removal of amelogenin!!!!

Unlike Dentin & Bone, the proteinaceous scaffold, or matrix, is removed during enamel formation
Kallikrein is a Serine proteases!!!! Timing is everying. Problems arise if it destroys the matrix either too late or
too early.
Amelotin also important for enamel structure.
Enamel structure contributes to strength of enamel but also its softeness and susceptibility to acid attack.
Important in enamel etching.

Caries • Hard Tissue Infection

• Transmissible
• Acidogenic bacteria
• Fermentable Carbohydrate
• A Behavioral Disease
• A Dynamic Cyclic Disease Process

Tx:
• Risk Assessment
• Arrest
• Reverse
• Intervene
• Regenerate

Caries follows the enamel rod boundaries!!! Will first spread

toward DEJ then Caries spreads laterally under the DEJ.
Once caries has hit the DEJ its not remineralizable and needs to be
removed/restored.

Fluoride and • Fluoride Incorporated into the developing enamel

Enamel • F- ions substitute for OH- ions
Mineralization “Fluorohydroxyapatite”
• Enhances crystal growth rates
• Increases enamel stability
• Decreases solubility
• Fluoride Diffuses into the developed enamel from the enamel surface
• F- ions substitute or replace OH- ions lost through demineralization

Excess Flouride • Dental fluorosis or mottled enamel

• Potential decrease in activity of enamel matrix proteinases that normally would have removed the
amelogenin
• Results in weakened enamel
DMF decreases as water fluoride level increase
1ppm is good enough to decreased DMF without causing extensive fluorosis