Griffith University Oral Biology 2 1009 DOH

Enamel
Dr. Mahmoud Bakr Lecturer in General Dental Practice B.D.S, M.D.S (Cairo University), ADC (Australia) Member of the Australian Dental Association (ADA), the Australian Biology Institute Inc. (ABI) and the Egyptian Dental Union (EDU)

Learning objectives:
After completing this lecture you should be able to: 1- Name, classify, identify and describe the structure and function of the components of Enamel. 2- Describe age related changes to Enamel and their effects. 3- By observing the histological details of cells and tissues, you should be able to use a microscope to identify different histological structures of Enamel and understand the histological processes involved in preparing slides.

 All Microscopic images are taken from the Digital Library of the Oral Biology Department (Cairo University).

INTRODUCTION
ENAMEL -Makes up the outside layer of anatomical crown of a tooth -Covers and protects the dentin of the crown -Firmly attached to the underlying dentin at the dentinoenamel junction and meets the cementum at the cementoenamel junction -The hardest and most mineralized substance of the body -Has no way to regenerate itself

PHYSICAL PROPERTEIS
1-COLOUR 2-THICKNESS

5-PERMEABILITY

3-HARDNESS

4-BRITTLNESS

1 1- COLOUR Colour:
YELLOWISH WHITE TO GRAYISH WHITE DEPENDS ON : 1- DEGREE OF
CALCIFICATION 2- HOMOGENOUSITY OF THE ENAMEL CRYSTALS.

SO: TRANSLUCENT E. YELLOWISH TEETH OPAQUE E. GREYISH TEETH

2 Thickness - THICKNESS -2
- 2 2.5 mm. at the cusp tips. - Thinning down to Feather edge at the cervical line of the tooth

2 - THICKNESS
Thin enamel is present at the bottom of the pits and fissures of the crown surface

enamel

dentin

3 HARDNESS
- ENAMEL OF THE PERMANENT TEETH IS HARDER THAN THAT OF DECIDUOUS ONES
+ IT IS THE HARDEST + CALCIFIED TISSUE IN THE BODY DUE TO: + 1- HIGH CONTENT OF THE MINERAL SALTS 2- ITS CRYSTALLINEARRANGEMENT.

3 HARDNESS
1 - IS GREATEST AT THE SURFACE AND DECREASED TOWARD DEJ. 2 - IT IS GREATER AT THE CUSPS AND INCISAL RIDGE AND DECREASES TOWARD THE CERVICAL LINE. 3- It is the hardest tissue of the body
+ + + + -

4 -4BRITTLENESS Brittleness:
-Very brittle due to large amount of minerals -Dentin, which is less mineralized and less brittle, compensates for enamel and is necessary as a support for enamel -If this supportive layer of dentin is destroyed by caries or improper cavity preparation, the unsupported enamel fractures

5- PERMEABILITY
- IT ACTS AS A SEMIPERMEABLE MEMBRANE FOR CERTAIN IONS THROUGH PORES BETWEEN THE CRYSTALS.

-PER. IS MAINLY FROM SALIVA TO OUTER LAYER OF ENAMEL, BUT LESS FROM THE PULP TO THE INNER ENAMEL LAYER ACROSS THE DENTIN.

CHEMICAL PROPERTIES
inorg organic water

INORGANIC

By weight

95-96% 1-2%

2%

ORGANIC

By volume

88-90% 10-12%

MATERIAL 96 % a-INORGANIC Inorganic material of enamel:

CALCIUM PHOSPHATE ( HYDROXY APATITE) CRYSTALS 3 Ca3 (PO4)2 . Ca (OH)2
-Most crystals are regularly hexagonal in cross section. (VERY LARGE) -They are 70 nm. in width, 25 nm. in thickness length may reach to 0.1 micron

Various ions if present during enamel formation or in the environment of fully formed enamel may replace the different ions of the molecule

. Hydroxyapatite is converted to fluoroapatite when fluoride ion replaces the hydroxyl ion 3 Ca3 (PO4)2 . Ca (F)2
Fluorapatite is 20%less soluble than hydroxyapatite it is much less susceptible to demineralization. -Fluoridation of drinking water.

HISTOLOGICAL STRUCTURE
THERE ARE TWO TYPES OF PREPARATIONS FOR HISTOLOGICAL STUDY OF HARD TISSUES :

1-DECALCIFIED SECTIONS:

2-GROUND SECTION More calcified more translucent

As Enamel contains 96% inorganic material so it cannot be seen in decalcified sections It can only be studied in Ground sections

ENAMEL ROD
The basic structural unit of enamel May be hexagonal and prism-like in cross section
Rods do not have a fixed regular geometrical outline and the term enamel rod is preferred in this text

ENAMEL ROD
LOWER CENTRAL INCISOR

1 - NUMBER

5
12

MILLIONS

UPPER FIRST MOLAR MILLIONS

2 - DIRECTION

DECIDUOUS

PERMANENT

ENAMEL ROD

3SIZE

3-4 um in diameter
Up to 2.5 mm. in length

ENAMEL ROD
4- COURSE
GNARLED ENAMEL STRUCTURELESS ENAMEL

Wavy course except near the incisal edge or cusp tips where enamel rods have a more complex course (Gnarled Enamel) to withstand masticatory forces

Gnarled Enamel

Histological structure of enamel rod

LS. TS

ENAMEL ROD
LONGITUDINAL SECTION

ENAMEL ROD
ROD

ROD SHEATH
INTERROD REGION

CROSS STRIATION

Cross section of enamel rod

Key hole pattern

Fish scales pattern

Enamel rod

ENAMEL ROD
CRYSTALS ROD SHEATH CRYSTALS INTERROD SUBSTANCE

HEAD
(occlusally)

TAIL
(Cervically)

HUNTER SCHREGERS BANDS It is an optical phenomenon can be seen in:

1-LONG.GROUND SEC. IN ENAMEL 2-OBLIQUE REFLECTED LIGHT It is due to the wavy course of Enamel rods

HUNTER SCHREGERS BANDS

Life history of ameloblasts

1-Differentiating stage (presecretory). 2- Secretory stage. 3- Transitional stage. 4- Maturative stage. 5- Protective stage. 6- Desmolytic stage

LIFE HISTORY OF THE AMELOBLASTS

6 STAGES
1-DIFFERENTIATING

DEAL WITH INNER DENTAL EPITHELIUM

LIFE HISTORY OF THE AMELOBLASTS

6 STAGES
2 SECRETORY 3 TRANSITIONAL 4- MATURATIVE

FUNCTIONS OF AMELOBLASTS

LIFE HISTORY OF THE AMELOBLASTS

6 STAGES

REDUCED DENTAL EPITHELIUM

5 PROTECTIVE 6 -- DESMOLYTIC

REDUCED DENTAL EPITHELIUM

Why life history not life cycle???? Because Ameloblasts are lost in Reduced Dental Epithelium after complete crown formation. We dont have Ameloblasts in our teeth its HISTORY. Its not a repetitive cycle.

1- Differentiating stage:
Ameloblasts differentiate from I.E.E In Early Bell Stage I.E.E cells are separated from Dental Papilla by Cell free zone I.E.E cells elongate on the expense of Cell free zone and becomes in contact with Dental Papilla Now they are called Preameloblasts Preameloblasts induce the formation of Odontoblasts by a process called Induction

1- Differentiating stage:
Before ameloblasts differentiation

INNER DENTAL EPITHELIUM BASEMENT MEMBRA CELL FREE ZONE DENTAL PAPILLA

ASG

Histological structure of Preameloblasts

As a preparatory step for Enamel matrix secretion Polarization or Reverse of Functional Polarity occurs:
Nucleus and Mitochondria move towards the proximal end of the cell RER, Golgi apparatus and centrioles move towards the distal end of the cell.

1- Differentiating stage:
Golgi apparatus

Oval nucleus
Centriol

Short columnar

Before ameloblasts differentiation

Mitochondria
ASG DISTAL END BASEMENT MEMBRANE

1- Differentiating stage:
GOLGI APP. CENTRIOL NUCLEUS

MITOCHONDRIA

DISTAL END
ASG BASEMENT MEMBRANE

PROXIMAL END PROXIMAL JUNCTIONAL COMPLEX GOLGI APP. CENTRIOL NUCLEUS

MITOCHONDRIA

DISTAL END
Basal lamina

1- Differentiating stage:
SO DURING THIS STAGE THE FOLLOWIONG OCCURE:

Centrioles
Golgi apparatus

1 - REVERSAL OF THE FUNCTIONAL POLARITY OF THE I.E.E... 2 HISTODIFFERENTIATION OF THE ODONTOBLASTS ( INDUCTION).

Mitochondria ASG

1- Differentiating stage:
INNER DENTAL EPITHELIUM (Preameloblasts)

ODONTOBLASTS ASG

2- Secretory stage:
During this stage Preameloblasts change into Ameloblasts by a process called Reciprocal Induction by signals from Odontoblasts Remember: Ameloblasts secret Enamel matrix not Enamel (only the organic component)

Histological structure of Ameloblasts (ultrastructure)

As formative (Blast) cells Ameloblasts have all the criteria of protein forming cells (Blast cells): 1- Large open face (lightly stained) nucleus. 2- Abundant (rich) cytoplasm. 3- Increase in Mitochondria 4- Increase in Golgi apparatus 5- Increase in RER 6- Increase in Alkaline Phosphatase activity 7- Presence of secretory vesicles

Tomes Process
Is a conical process that develops from the distal end of Ameloblasts during secretory stage. It is responsible for the different orientation of crystals in the Enamel rod and Interod substance.
It gives the cells Picket fence appearance Note: Tomes process not Tomes process

2-SECRETORY STAGE

2-SECRETORY STAGE
PROXIMAL TERMINAL BARS

G.A.

R.E.R.

DISTAL TERMINAL BARS TOMES PROCESS

Picket fence appearance

E. matrix

3 Transitional Stage:
1 REDUCTION IN HEIGHT.
2 DECREASE IN ITS VOLUME AND ORGANELLE CONTENT 3 WITHDRAWAL OF TOMES, PROCESS. 4-DECREASE IN OVERALL CELL NO.

BASAL LAMINA

All these changes are related to shift of function from Protein synthesis (secretory stage) to ion transport (maturative stage).

4Maturative stage
There are two types of Ameloblasts in this stage:
A- Ruffled ended Ameloblasts (80%) B- Smooth ended Ameloblasts (20%)

4Maturative stage

4 MATURATIVE STAGE
M

CYTOPLASMIC VACUOLES

AUTOPHAGIC VACUOLES

SMOOTH ENDED AMELOBLAST

RUFFLED ENDED AMELOBLAST

MATURATIVE AMELOBLAST
Leaky proximal junction
Tight proximal junction

RUFFLED ENDED

SMOOTH ENDED

Tight distal junction

Leaky distal junction

Influx of mineral ions into the present partially mineralized matrix occurs in relation to ruffled- ended cells
Withdrawal of organic matrix from the maturing enamel occur mainly in the ruffledended cells as well as through passing between the leaky distal junctions of the smooth-ended cells

5 Protective stage:
After complete Enamel matrix formation and maturation Ameloblasts become squeezed as a layer of Reduced Enamel Epithelium which protects Enamel from resorption by cells from Dental Sac.

5 PROTECTIVE STAGE
REDUCED DENTAL EPITHELIUM

PRIMARY ENAMEL CUTICLE

6 DESMOLYTIC STAGE
Reduced Enamel Epithelium (with Ameloblasts being a part of it) secrets desmolytic enzymes for elimination of Dental sac and allowing fusion between Reduced Enamel Epithelium and Oral Epithelium. This allows eruption of the tooth without bleeding. It is called Primary Enamel Cuticle which is the last product of Ameloblasts.

TOOTH ERUPTION
GINGIVAL EPITHELIUM still fusing with E.CUTICLE will wear away

REDUCED DENTAL EPITHELIUM

ENAMEL

AMELOGENESIS

ASECRETORY PHASE

B- MATURATION PHASE

THE BEGINNING OF MINERALIZATION OF THE ENAMEL MATRIX DOES NOT AWAIT THE COMPLETION OF ITS FORMATION.

A-- Secretory phase:

Beginning of secretion
End of secretion

1- Stippled material secretion:

STIPPLED MATERIAL OF ENAMEL

2- Initial rodless (aprismatic) enamel layer:

Secretory ameloblast
.The first hydroxyapatite crystals formed interdigitate with the crystals of dentin STRATUM INTERMEDIUM

AMELOBLASTS

ENAMEL MATRIX

MANTLE DENTIN

NO CLEAR CUT BAND OF ORGANIC MATRIX

INNER DENTAL EPITHELIUM

(PREAMELOBLAST)

Stippled material secretion

3-Rod (prismatic) enamel layer:

ROD

ROD

4- -Final rodless (aprismatic) enamel layer

The ameloblasts become shorter and Tomes' processes are lost
End of secretory ameloblast

Disturbances during the secretory stage of amelogenesis result in pathologically thin or hypoplastic enamel

ameloblast without Tomes process

Ameloblast with
Tomes process

Ameloblast without
Tomes process

Secretory stage

End of secretory stage

Dentin
Rodless enamel Rod (Prismatic) Enamel Rodless enamel

B-Maturation Phase
1- Primary maturation 2- Secondary maturation 3- Tertiary (post-eruptive) maturation

1- Primary maturation
The first secreted enamel matrix contains 20-30 % of the mineral content
The crystals are thin and can grow primarily in length

2- Secondary maturation

-A selective removal of enamel proteins and water occurs resulting in a change in the ratio of the proteins of the newly secreted enamel matrix and the final matrix of mature enamel

-Amelogenins and ameloblastins are removed leaving enamelins and .tuftelin in the mature enamel --In the same time the ameloblasts transport mineral ions into this partially mineralized enamel matrix which cause growth of the crystals in width and thickness The enamel crystals never fuse -

Direction of maturation

3- Tertiary maturation
After tooth eruption and exposure of enamel to saliva in the oral cavity, precipitation of certain ions on the outermost enamel layer occurs

ions

Defective maturation hypomaturative soft enamel

Hypocalcified structures of Enamel

1- Incremental lines of Enamel: a- Cross striations b- Incremental lines of Retzius c- Neonatal line 2- Enamel Tufts 3- Enamel Lamellae (Type A) 4- DEJ

INCREMENTAL LINES OF ENAMEL

They are formed as a result of the rhythmic manner of Enamel (periods of activity alternating with periods of rest) Enamel is increment formed increment by

1 SHORT INCREMENTAL LINES ( CROSS STRIATION )

They are formed as a result of the daily rest of Ameloblasts
Ameloblasts form 4 microns of Enamel then rest so the distance between cross striations is 4 microns

1 SHORT INCREMENTAL LINES

( CROSS STRIATION )

BETWEEN SEGMENT OF 2.5-6 u LENGTH, DAILY RATE OF SECRETORY ACTIVITY OF AMELOBLASTS

2 LONG INCREMENTAL LINE ( INCREMENTAL LINE OF RETZIUS ) BROWN STRIA OF RETZIUS They are formed due to the weekly rest of Ameloblasts The distance between them is 16 microns When viewed microscopically in cross-section, they appear as concentric rings. In a longitudinal section, they appear as a series of dark bands .
7 DAYS ARE NEEDED FOR THE FORMATION OF THE ENAMEL MATRIX BETWEEN TWO BANDS

3 NEONATAL LINE
It an accentuated incremental line that separates between Enamel formed before birth and that formed after birth.
It is only seen in Deciduous teeth and Permanent First Molars (Why?) As the rest of Permanent teeth are developed after birth. The quality of Prenatal Enamel is better than Postnatal Enamel (Why?) Due to constant nutrition and more protected environment inside the uterus .

3 NEONATAL LINE
Enamel of lower D formed at birth

PRENATAL ENAMEL

POSTNATAL ENAMEL

POSTNATAL ENAMEL

PRENATAL ENAMEL

ENAMEL TUFT
It appears only in T.S ground section of Enamel as a result of different crystal orientation (The function of Tomes process). It starts from DEJ and extends to 1/3 or 1/5 of the thickness of Enamel. It appears as a tuft of grass.

ENAMEL TUFT

ENAMEL TUFT

THE DENTINOENAMEL JUNCTION

It is a scalloped line between Enamel in dentin.
It is smooth sometimes in Deciduous teeth. The convexities are always towards Dentin. It is hypocalcified due to high organic content of Enamel and Dentin in this area. The bond between Enamel and Dentin is organic not mechanical.

THE DENTINOENAMEL JUNCTION

THE DENTINOENAMEL JUNCTION

ENAMEL

DENTIN

ENAMEL LAMELLAE
It extends from the outer surface of Enamel to the DEJ and may extend to Dentin.
Types of Enamel lamellae: A- Developmental (type A) B- Non Developmental (type B,C)

ENAMEL LAMELLAE

1- Developmental (true lamellae ) (Type A lamella):

Occurs during amelogenesis
Hypomineralized structures Never extend to dentin

2-Non developmental (false lamellae or cracks ) :

Unmineralized areas that occur after amelogenesis is completed
They are divided into:

i-Type B lamella A crack occurs in the enamel after its complete formation but before the tooth eruption
ii- Type C lamella A crack occurs in the enamel after the tooth eruption iii -An accidental crack during section preparation

Uncalcified structures
1- Non-Developmental lamellae
(Type C)

2- Enamel spindle

ENAMEL SPINDLES
It is actually an odontoblastic process that crosses the DEJ to reach Enamel.
It usually found under the incisal edge or cusp tips. It can be seen both in L.S. and T.S.

ENAMEL SPINDLES

ENAMEL SPINDLES
ENAMEL

ODONTOBLASTIC PROCESS

DENTIN

SURFACE STRUCTURES OF ENAMEL

1 OUTER STRUCTURELESS ENAMEL

REMEMBER: THAT THERE IS AN INNER STRUCTURELESS ENAMEL

20-70 um thick

2- PERIKYMATA

PARALLEL TO EACH OTHER AND TO CERVICAL LINE CONTINUOUS AROUND THE TOOTH. NUMEROUS AT THE CERVICAL REGION VERY FEW NEAR THE OCCLUSAL OR INCISAL EDGES

3 - ROD END MARKING

SHALLOW CERVICALLY DEEPER OCCLUSALLY

4 - CRACKS

ENAMEL LAMELLA

5 - AFIBRILLAR CEMENTUM
ENAMEL

DENTIN

CEMENTUM

1-ATTRITION
Definition. Wear rate 8 mm /year Parafunctional movements, as found in bruxism Wear polished facets Nonbacterial processes of enamel destruction 1-abrasion 2-erosion .

2- COLOUR

3- MODIFICATION IN SURFACE LAYER:

IONIC EXCHANGE BETWEEN THE ENAMEL AND THE SURROUNDING SALIVA. (TERTIARY MATURATION.)
INCREASE NITROGEN AND FLUORIDE. RESISTANCE TO DECAY IS INCREASED.

4- PERMEABILITY

Permeability of Enamel decreases by age, but remains only in the most superficial layer.
Recently Main path Old enamel

Erupted teeth
ASG

Permeability of Enamel decreases by age, but remains only in the most superficial layer.

Clinical Considerations

1- Acid-etching

A,B :itched rods C: non itched rods

Type 1 is a pattern where predominantly the enamel rods are dissolved.

Type 2 is a pattern where predominantly the interrod enamel is dissolved

Type 3 is a pattern where there is no evidence left of any enamel rods.

2-Tooth Whitening

Can be done by removal of the surface discolored layer of Enamel either : a- Mechanically (Prophylactic paste and abrasion)

b- Chemically by using bleaching agents such as:

Sodium perborate and sodium hypochlorite

3- Amelogenesis Imperfecta

It is a congenital defect that occurs during Enamel formation. It is either: a- Enamel Hypoplasia: it is a defect in the quantity of Enamel matrix. (disorder during the secretory phase of Amelogenesis) b- Enamel Hypocalcification: it is a defect in the quality of the calcified Enamel matrix. (disorder during the maturation phase of Amelogenesis)

What Enamel structures can you see in this picture?