AMELOGENESIS

Dr.I.Nanda Balan Second Year Post Graduate, Dept. of Public Health Dentistry, Ragas Dental College & Hospital, Chennai.

DEVELOPMENT OF TEETH

During development of embryo, there is formation of 3 germ layers

The primitive oral cavity is lined by stratified squamous epithelium called oral ectoderm.

Ectoderm contacts with endoderm of foregut to form the bucco pharyngeal membrane.

At about 27th day of gestation the membrane ruptures.

Connective tissue cells underlying the oral ectoderm induce overlying ectoderm to start tooth

development.

Tooth development begins during the 5th - 6th week of gestation, the first indication is the primary epithelial bands.

PRIMARY EPITHELIAL BAND

It is a continuous band of thickened epithelium formed in both upper & lower jaw.

It is a horseshoe shaped structure, which corresponds in position to the future dental arches.

Gives rise to a) Vestibular Lamina b) Dental Lamina

PRIMARY EPITHELIAL BAND

The vestibular lamina overlying the forming gums grows into the underlying gum tissue and forms the dental lamina.

Neural crest cells in the underlying mesenchyme of the gums induce the formation of the dental lamina.

PLANE OF CLEAVAGE

The dental lamina forms a C-shaped band of tissue in the gums of the upper and lower jaw.

FUNCTIONS :

1) First Phase - Act as primordium for the ectodermal portion of deciduous teeth.

2) Second Phase - Initiation of successor of deciduous teeth by successional dental lamina.

3) Third Phase -

Initiation of permanent molar.

TIME SCALE OF HUMAN TOOTH DEVELOPMENT

AGE 42-48 days 55-56 days 14 weeks DEVELOPMENT CHARACTERISTICS Dental lamina formation. Bud stage, deciduous incisor canine & molar. Bell stage for deciduous teeth, bud stage for permanent teeth. Dentin and functional ameloblasts in deciduous teeth. Dentin & functional ameloblast in permanent teeth.

18 weeks

32 weeks

DEVELOPMENTAL STAGES - BUD STAGE

Differentiation of dental lamina leads to formation of round, ovoid swelling at 10 different points corresponding to future position of deciduous teeth. These are the primordia of enamel organ. Enamel organ consists of peripherally located low columnar cells and centrally located polygonal cells.

CAP STAGE

Characterized by continuation of the ingrowth of the oral epithelium.

This stage marks the beginning of histodifferentiation and morphodifferentiation

A depression forms in the deepest part of each tooth bud and forms the cap or enamel organ.

Below this cap is a condensing mass of mesenchyme dental papilla .

The basement membrane separating the dental organ and the dental papilla becomes the future site for the dentinoenamel junction (DEJ)

Remaining mesenchyme surrounds the dental/enamel organ and condenses to form the dental sac or the dental follicle

Together the enamel organ + dental papilla + dental follicle is considered the developing tooth germ

BELL STAGE


Stage of Histodifferentiation and Morpho differentiation. The invagination of the epithelium deepens and its margins continue to grow and enamel organ assumes a bell shape. During histodifferentiation cells acquire their functional assignment. Odontoblasts are differentiated from mesenchymal cells with formation of dentin, the cells of inner dental epithelium transform into ameloblasts and enamel matrix is lead down opposite the dentin.

DENTAL ORGAN

OUTER ENAMEL EPITHELIUM: low cuboidal shape

protective At

barrier during enamel production

the end of the bell stage OEE is laid in folds. these folds the mesenchyme of dental sac

Between

forms papilla that contains capillary loops and thus provides rich nutritional supply.

CYTOPLASM CONTAINS FEW CYTOPLASMIC ORGANELLES AND SOME GLYCOGEN

INNER ENAMEL EPITHELIUM:

short,

columnar cells

differentiate

into

tall

columnar cells called ameloblasts.

separated

from the dental papilla below it by a

basement membrane.
the

IEE and OEE where they connect curved rim

of the EO = cervical loop.

GLYCOGEN STORAGE

STELLATE RETICULUM:

star-shaped cells. center of the enamel organ forms a network = reticulum synthesize and secrete GAGs - this pulls water into the EO - increasing amount of fluid in the EO forces the central cells apart - before enamel formation these cells collapse reducing the distance between ameloblasts and nutrient capillaries near OEE.

STRATUM INTERMEDIUM: Between IEE and Stellate reticulum.

inner

layer of compressed flat to cuboidal cells production of enamel

supports

AMELOGENESIS

Two step process - ameloblasts secreting organic matrix. - addition of mineral and removal of most organic matrix.

Reciprocal induction. Enamel formation is initiated in the presence of poor nutritive supply.

This is compensated by accumulation of Glycogen by IEE until OEE folds and Stellate reticulum collapse.

Stratum intermedium then contacts with ameloblasts and exhibit alkaline phosphatase activity.

ENAMEL MATRIX FORMATION

The term organic matrix is used to designate the organic component and inorganic component of the first formed enamel.

Organic component enamel protein. Inorganic protein developing enamel

ENAMELPROTEIN

Amelogenin constitutes 90% of the enamel protein on the secretory stage.

Amelogenin and enamelin plays an important role in crystal growth and organisation.

Enamelins

are

thought

to

be

aggregates

of

amelogenin breakdown products.

Serum albumin that has leaked into forming enamel. Enamelins are secreted first and are linked to the first formed enamel on the dentin surface.

Also proposed that progressive and increasing mineralization of the mineral. enamel matrix amelogenin is selectively removed while enamelin tightly bound to

HOW THESE PROTEINS ARE REMOVED ?

Pressure created by the crystal growth. Trapped proteins become low molecular weight through the proteolytic enzymes secreted by

Ameloblasts which are then squeezed out.

The remaining gets attached to the hydroxyapatite crystals.

The enamel protein is labile exhibiting quantitative and qualitative changes during the process of amelogenesis.

ELECTRON MICROSCOPY OF AMELOGEN

DIFFERENTIATION STAGE: - As IEE differentiate into Ameloblasts they elongate and their nuclei shift proximally towards stratum intermedium. - Golgi complex increases in volume and occupies central core. - amount of rough endoplasmic reticulum increase. - mitochondria clusters in proximal region. - ameloblasts becomes polarized.

SECRETORY STAGE: Synthesis rough endoplasmic reticulum condensed and

golgi apparatus

packed into membrane bound secretory granules. These granules migrate to the distal extremity of the cells and their contents are released against the newly formed mantle dentin.

SECRETORY ENDS OF AMELOBLASTS

SMOOTH AMELOBLASTS SURFACE SEPARATED FROM PREDENTIN

After this structureless enamel layer is formed, the ameloblasts migrate away from the dentin surface which permits the formation of Tomes process.

These projects into the newly formed enamel giving the junction between enamel and ameloblasts a picket fence appearance.

Once Tomes process is established, secretion of enamel protein is confined to - adjoining ameloblasts, resulting in the formation of enamel matrix wall. These walls encloses a pit into which Tomes process fits. - later fills this pit with matrix.

The walls becomes inter rod enamel and infilling becomes enamel rod.

PITS PREVIOUSLY FILLED BY TOMES PROCESS

MATURATION STAGE

Once immature enamel has been formed, the ameloblasts reduce in height, decrease in volume and organelle content.

Organelles associated with synthesis are enclosed in autophagic vacuoles and digested by lysosomal enzymes.

The remaining organelles shift to distal part of the cell.

Water and organic material are selectively removed and inorganic material is introduced.

The ameloblasts possess a ruffle border and a smooth border.

Ruffle border introduce organic material Smooth border removal of protein and water.

MINERALIZATION

Enamel protein once deposited directly in the mineralized dentin, the initial mineralization is achieved by the nucleation from the apatite crystals located in the dentin.

The mineralization occurs by displacing the matrix.

After the formation of partially mineralized enamel matrix, which involves the production of entire volume of enamel, maturation begins with

mineralization reaching 96%.

Soft x ray analysis and computer enhancement indicate mineralization of enamel occurs in 4 stages.

1st stage partial mineralization 30%. 2nd stage secondary increase in mineralization starting at surface of enamel and sweeps rapidly into deeper layers until it reach the innermost 8m layer.

3rd stage mineral rebounding from innermost layer to the enamel surface.

4th stage outer layer mineralizes rapidly and heavily becoming the most mineralized part of the enamel.

DISTURBANCE IN AMELOGENESIS DUE TO FLUORIDE

Fluorosed enamel is characterized by a retention of amelogenins in the early-maturation stage, and by the formation of a more porous enamel with a subsurface hypomineralization.

The mechanisms by which fluoride affects enamel development include specific effects on both the ameloblasts and on the developing enamel matrix.

Maturation-stage ameloblast modulation is more rapid in fluorosed enamel as compared with control enamel, and proteolytic activity in fluorosed earlymaturation enamel is reduced as compared with controls.

The severity of enamel fluorosis depends on the amount of fluoride ingested, the duration of exposure, and the stage of amelogenesis at the time of exposure.

POSSIBLE MECHANISMS

Decreased ameloblasts.

enamel

produced

by

secretary

DenBesten and Crenshaw, 1984.

Initial inhibition of matrix secretion. Bronckers et al. 1984.

Disproportionate failure to secrete amelogenins as a fraction of the total protein secreted. Eastoe and Fejerskov 1984

Fluoride damage the secretory ameloblasts, the distal (prism forming) portions of their Tomes processes are lost and come to be buried in the enamel matrix.

Walton and Eisenmann 1974.

Fluoride may alter amelogenesis through a direct effect on the extracellular enamel matrix. These effects may include alterations in the properties of the enamel matrix by fluoride-binding to specific enamel proteins, or by a direct inhibition of proteinases secreted into the extracellular matrix Crenshaw and Bawden,1984; DenBesten and Heffernan, 1989a).

Delay in the withdrawal of amelogenin in fluorosed maturation enamel as compared with control enamel Shinoda, 1983; DenBesten and Crenshaw, 1984;

The retention of amelogenins in the enamel matrix may be responsible for the hypomineralized enamel defect seen in fluorosed enamel.

Therefore, a delayed withdrawal of amelogenin may delay the growth ofenamel crystals, so that when the tooth erupts, the enamel remains incompletely mineralized.

The proteolytic activity of the maturation stage serine proteinases was less in the fluorosed enamel.

The inhibition of proteolytic activity in the presence of fluoride may be due to a reduction in ionic calcium due to fluoride binding.

Enamel overlaid by the first band of smooth-ended ameloblasts, which occurs at the transition region, had a significantly higher fluoride content than overlying the ruffle-ended ameloblasts.

Therefore, it is possible that these high concentrations of fluoride in the enamel matrix at the transition/earlymaturation stage of enamel formation could reduce the available ionic calcium, resulting in reduced proteolytic activity at this critical stage. Bawden et al. 1988

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