Outline: Initiation of tooth development Stages of tooth development: Bud stage Cap stage Bell stage Advanced Bell stage Root formation Histophysiology and clinical consideration Molecular aspects of tooth development Role of genes in tooth development Tooth Type Determination
INITIATION OF TOOTH DEVELOPMENT
Early Embryogenesis Primitive oral cavity called the stomodeum, is lined stratified squamous epithelium called oral ectoderm. This oral ectoderm contacts endoderm of the foregut to form the buccopharyngeal membrane At the 27th day of gestation this membrane ruptures and the primitive oral cavity establishes a connection with the foregut
Underlying the oral ectoderm is an embryonic connective tissue that originates from the neural crest and is called ectomesenchyme
Primary Epithelium Band Around 37 th day of intrauterine life, a continuous horse shoe shaped band of thickened epithelium forms at the site of future maxillary and mandibular arches called the primary epithelial bands. They are divided into two parts: 1. Vestibular lamina 2. Dental lamina
This increase in thickness of primary epithelial band occurs because of change in orientation of mitotic spindle and cleavage plane of epithelial cell 1. Vestibular Lamina Vestibular lamina/ lip furrow band is responsible for the formation of future vestibule. The cells proliferate into the ectomesenchyme and the band enlarges It then subsequently degenerates and hollows out to form a cavity/ cleft/ vestibule between the alveolar portion of the jaws and the lips and cheeks. 2. Dental Lamina This lamina leads to development of future teeth. Dental lamina is seen as a localized proliferation of the epithelium into the ectomesenchyme. Formation of this dental lamina begins in the distal (molar) region and then proceeds towards the midline
What are neural crest cells? Neural crest cells are derived from the dorsal most region of the neural tube. These are highly pluripotent cells which migrate into the 1 st branchial arch and assume a mesenchymal phenotype. Then a series of reciprocal interactions take place with the oral epithelium leading to the formation of tooth primordial If the neural cells fail to migrate then they may lead to anodontia (absence of teeth) and microdontia (underdeveloped jaws) All tooth structures except enamel are derived from neural crest cells; enamel is ectodermal in origin
Note Tooth development (odontogenesis) takes place by specific and reciprocal epithelium mesenchymal interactions. This involves cross-talk between the epithelium derived dental lamina and the neural crest cell derived underlying ectomesenchyme Oral ectoderm gives rise to- enamel Oral ectomesenchyme gives rise to- dentin, pulp, cementum and PDL of the fully formed tooth
Tooth Development Oral Histology
BRIHASPATHI ACADEMY SUBSCRIBERS COPY NOT FOR SALE Derivatives of dental lamina: 1) Direct extension of dental lamina (lateral lamina)leads to formation of deciduous teeth 2) Lingual extension of free end of dental lamina (successional lamina) opposite to the enamel organ of each deciduous tooth gives rise to the permanent teeth or the successors of deciduous teeth 3) Distal extension (parental lamina) in dental arches provides primordia for all teeth lacking primary predecessors. e.g. permanent molars Fate of dental lamina Rudimental lamina: Total activity of dental lamina lasts for 5 years. However, the dental lamina can still be active in third molar region after it has disappeared elsewhere. After tooth germ formation, dental lamina disintegrates by autolysis. If it persists, it is called as cell rests of serre. (May give rise to eruption cysts, odontoma, or may be activated to form supernumerary teeth)
STAGES OF TOOTH DEVELOPMENT At some points along the dental lamina localized proliferation of ectodermal cells take place leading to formation of knob like structures called the enamel organ Based on the morphology and histophysiology of the enamel organ, the development of teeth can be divided into the following stages: Morphologic Stages Histophysiologic Processes Dental Lamina Initiation Bud stage Proliferation Cap stage Bell stage (early) Histodifferentiation Bell stage ( advanced) Morphodifferentiation Formation of enamel and dentin matrix Apposition
I. BUD STAGE (Initiation) The oral epithelial cells and the adjacent mesenchymal cells proliferate resulting in a bud shaped enamel organ. Histology: The enamel organ consists of peripherally located low columnar cells and centrally located polygonal cells The ectomesenchymal cells surrounding the tooth bud show condensation due to increased mitotic activity
II. CAP STAGE (Proliferation) The tooth bud continues to proliferate and exhibits unequal growth in different parts leading to cap shaped enamel organ It is characterized by a shallow invagination on the deep surface of the bud Tooth germ: enamel organ+ dental papilla + dental follicle are seen in the cap stage
Histologically, the enamel organ in the cap stage consists of: 1. Outer and Inner Enamel Epithelium The peripheral cells of the cap covering the convexity are cuboidal and are called the outer enamel epithelium (OEE). The cells lining the concavity of the cap become tall columnar and is the inner enamel epithelium (IEE) 2. Stellate Reticulum The central area of the enamel organ between the outer and inner epithelium consist of polygonal cells These cells in the centre synthesize and secrete GAGS (Glycosaminogylcans) which gives cushion like consistency, supports and protects the delicate Tooth Development Oral Histology
BRIHASPATHI ACADEMY SUBSCRIBERS COPY NOT FOR SALE enamel forming cells. As GAGs are hydrophilic in nature, water is pulled into the enamel organ. This increases the volume of extracellular compartment and cells are forced apart thus becoming star shaped and are called stellate reticulum. 3. Dental Papilla: ectomesenchymal condensation immediately subjacent to enamel organ The ectomesenchymal cells are partly covered by the invaginated portion of the inner enamel epithelium. It condenses to form the dental papilla which is the formative organ of dentin and pulp The dental papilla shows mitotic cell division along with proliferation of new blood capillaries The peripheral cells near the inner enamel epithelium increase in size and differentiate to form odontoblasts 4. Dental Sac (Dental follicle): ectomesenchymal condensation surrounding the tooth bud and the dental papilla The cells of dental sac form the cementum and periodontal ligament Blood supply for ameloblast for most of its life cycle is from the dental sac
Transient structures seen in enamel organ during cap stage:
Enamel knot / Ahrens knot: Dense cluster of non dividing epithelial cells detected first at the tip of the tooth bud Enamel cord: Vertical extension of the enamel knot between the IEE and OEE is the enamel cord Enamel septum: When the enamel cord extends to meet the OEE, it is termed as enamel septum. Enamel navel: The OEE at the point of meeting shows a small depression called enamel navel Enamel niche: An apparent structure seen in histologic sections due to funnel shaped depression of the dental lamina. It creates an impression that tooth germ has double attachment to the oral epithelium by two separate strands of dental lamina enclosing the ectomesenchyme called as enamel niche between them
III. BELL STAGE It is the 3 rd stage of enamel organ formation in which the crown form is established characterized by histodifferentiation and morphodifferentiation. As the invagination of the epithelium deepens and its margins continue to grow, the enamel organ assumes a bell shape On light microscopic examination of bell stage four different types of epithelial cells can be seen: 1. Inner Enamel Epithelium Consists of a single layer of cells that differentiate prior to amelogenesis to form tall columnar cells called ameloblasts The cells of the inner enamel epithelium exert an organizing influence on the underlying mesenchymal cells in the dental papilla, which later differentiate into odontoblasts 2. Stratum Intermedium Formed by a few layers of squamous cells between the IEE and stellate reticulum. They have a high activity of enzyme alkaline phosphatase and increased glycogen deposits which are essential to enamel formation 3. Stellate Reticulum It expands further by increase in intracellular fluid. The cells are star shaped with processes that anastomose with those of adjacent cells Significance: Enamel knot and cord act as reservoir of cells for growing enamel organ Enamel knot expresses various signalling molecules like Bmp-2, 4, 7; Fgf-4, 9; Wnt-10b, Slit- 1 and Shh FGF-4 expressed in the enamel knot, stimulates cell division in adjacent enamel epithelium and dental papilla However, enamel knot cells themselves do not express FGF-4 receptors and thus are unable to respond to the proliferative mitogenic stimuli of FGF and remain as nondividing cells Thus they regulate the epithelial folding and help in transformation from bud to cap stage They also regulate the expression of Cbfa1 gene in the mesenchyme which induces odontoblast differentiation and thus play a role in tooth morphogenesis
Tooth Development Oral Histology
BRIHASPATHI ACADEMY SUBSCRIBERS COPY NOT FOR SALE Before enamel formation begins, the stellate reticulum collapses reducing the distance between the centrally situated ameloblasts (IEE) and the nutrient blood capillaries This change begins at the height of the cusp or incisal edge and progresses cervically 4. Outer Enamel Epithelium The OEE is made of low cuboidal cells and is thrown into folds by the end of bell stage Between the folds, the mesenchyme forms papillae that contain capillary loops, thus providing a source of nutrition to the metabolically active avascular enamel organ
Zone of reflexion or cervical loop: The junction of IEE and OEE is called the zone of reflexion or cervical loop. It gives rise to the epithelial component of root formation. Changes in dental papilla: Before the inner enamel epithelium begins to produce enamel, the peripheral cells of the dental papilla differentiate into odontoblasts which are initially cuboidal and later become columnar and produce dentin. Membrana Preformativa: It is the basement membrane that separates the enamel organ and dental papilla just prior to dentin formation Changes in dental Sac: Initially it shows a circular arrangement of its fibres and resembles a capsular structure. With the development of root, the fibres of dental sac become embedded in the cementum and alveolar bone Two important events occur during bell stage Break-up of the dental lamina Inner enamel epithelium folds IV. ADVANCED BELL STAGE This stage is characterized by the commencement of mineralization and root formation
Steps in hard tissue formation (reciprocal induction): Initially, the IEE cells induce the underlying ectomesenchymal cells of dental papilla to differentiate into odontoblasts Odontoblasts lay down the first layer of dentin along the DEJ at the region of future cusps After the first layer of dentin is formed the IEE differentiate into ameloblasts ( tall columnar cells with reversal of polarity) and lay down enamel matrix over the dentin Note: Enamel does not form in the absence of dentin
The development of roots begin after enamel and dentin formation has reached the future cementoenamel junction (CEJ) The cervical portion of enamel organ gives rise to HERS which is a double layered cell sheath formed by the down growth of IEE and OEE It outlines the future root and is thus responsible for the shape, length, size and number of roots It extends around the dental pulp and forms the epithelial diaphragm which encloses primary apical formation In multirooted teeth, epithelial diaphragm exhibits differential growth, causing division of root trunk and development of multiple roots (AIIMS-89)
During expansion of pulp, the IEE cells induce the ectomesenchymal cells of the dental papilla to differentiate into odontoblasts which form dentin After the root dentin forms; the HERS cells disintegrate and move away from the surface of dentin thereby exposing it to the connective tissue Tooth Development Oral Histology
BRIHASPATHI ACADEMY SUBSCRIBERS COPY NOT FOR SALE cells of the dental sac, which differentiate into cementoblasts and deposit cementum Remnants of HERS may persist as a strand of cells found in the periodontal ligament (PDL) of erupted teeth called as Rests of Malassez The continuity of HERS may be broken or not established prior to dentin formation, leading to the development of accessory root canals
ENAMEL PEARLS If the cells of the epithelial root sheath remain adherent to the dentin surface they may differentiate into fully functioning ameloblasts and produce enamel. Such droplets of enamel are called enamel pearls and are usually found in the furcation area of roots of permanent molars
HISTOPHYSIOLOGY AND CLINICAL CONSIDERATION 1. Initiation 2. Proliferation 3. Histodifferentiation 4. Morphodifferentiation 5. Apposition 1. Initiation Specific cells within the dental lamina have the potential to develop into enamel organ or induce tooth development by epithelial mesenchymal interactions Lack of initiationAnodontia / Partial Anodontia Abnormal initiation Supernumerary teeth Initiation at abnormal locations(ectopic) Ovary (dermoid tumors or cysts) / Hypophysis
2. Proliferation It causes regular changes in the size and proportions of the developing tooth germ Disturbances has different effects depending on the time of occurrence and stage of development
3. Histodifferentiation In this stage, cells undergo definitive morphologic and functional changes, they differentiate and restrict multiplication to don their new functions During histodifferentiation, the peripheral cells of the dental papilla differentiate into odontoblasts to produce dentin which further induces IEE cells to differentiate into ameloblasts and form enamel Eg. Disturbance: Vitamin A deficiency at this stage leads to formation oosteodentin
4. Morphodifferentiation Establishment of morphologic pattern, basic form and size of future teeth takes place by differential growth at this stage
Morphogenesis is not possible without proloiferation In this stage the DEJ and CDJ are developed by the continuous deposition of enamel, dentin and cementum, giving the completed tooth its characteristic form and size Disturbances affect the form and shape of teeth: a. Supenumerary cusps/ loss of cusps b. Twining c. Peg shaped tooth d. Endocrine disturbances: hypopitutarism /hypothyroidism small crowns
5. Apposition It is the regular and rhythmic deposition of extracellular matrix of enamel and dentin Disturbances due to genetic or environmental factors leads to hypoplasia and hypocalcification
MOLECULAR ASPECTS OF TOOTH DEVELOPMENT Odontogenesis is initiated by factors resident in the first arch epithelium influencing ectomesenchyme, but with time, this potential is assumed by ectomesenchyme
Role of Homeobox genes The Homeobox gene was initially found in Drosophila melanogaster and is called as the master gene. It constitutes a large family of genes that encode the transcription factors and regulate the expression of downstream target genes A set of coded patterning homeobox genes is required to specify correct positioning and bring about the development of cephalic structures
Subfamilies of Homobox genes: Muscle segment (Msx) Distal less (Dlx) Orthodenticle (Otx) Goosecoid (Gsc) Bar class (Barx) Paired related (Prx) LIM homeobox
Tooth Development Oral Histology
BRIHASPATHI ACADEMY SUBSCRIBERS COPY NOT FOR SALE
Genes associated with tooth development Gene Function Lhx-6, Lhx-7 Lim-homeobox domain genes (Transcription factor) Earliest mesenchymal markers for tooth formation Expressed in the neural crest derived ectomesenchyme of the oral half of the first branchial arch by day 9 Fgf-8 Fibroblast growth factor 8 (Secreted protein) Expressed in the first branchial arch for induction of the Lhx genes Establishment of oral - aboral axis Determines the positions for formation of tooth germs Induction of Pax-9 gene Pax-9 Paired box homeotic gene (Transcription factor) Its expression co localizes with the exact sites where the tooth germs appear It is induced by Fgf-8 and repressed by BMP-2 & BMP- 4 Shh Sonic hedgehog (Secreted protein) Induces local epithelial cell proliferation to produce invaginations that are reminiscent of tooth buds Stimulation of epithelial cell proliferation Signaling tooth initiation Lef-1 Lymphocyte enhancer binding factor (Transcription factor) Initially expressed in dental epithelial thickenings During bud formation it shifts to being expressed in the condensing mesenchyme Gene Abbreviation Function Msx-1 Msx-2 Msh like genes (Transcription factor) Anterior teeth Msx-1 Dlx-1 Dlx-2 Distaless homologue (Transcription factor) Canines Premolars Dlx-1 Dlx-2 Barx-1 BarH1 homologue (Transcription factor) Molars
TOOTH TYPE DETERMINATION Humans have heterodont teeth (falling into 3 categories: Incisiform, caniniform and molariform) The determination of the type of tooth which will develop at a particular position is called as patterning of dentition This can be explained with the help of two hypothetical models: Field model Clone model 1. Field model (Butler 1839) It proposes that the factors responsible for tooth shape reside within the ectomesenchyme in distinct graded and overlapping fields for each tooth family This theory is supported by expression of different homeobox genes at different position
2. Clone model (Osborn 1978) It proposes that each tooth class is derived from a clone of ectomesenchymal cells programmed by epithelium to produce teeth of a given pattern. The coded pattern of homeobox gene expression in the ectomesenchyme are expressed following the epithelial signal
The specific clone induces the dental lamina to begin tooth development. At its posterior border the clone and dental lamina grow by means of the progress zone. When a clone reaches the critical size, a tooth bud is initiated at its centre. A zone of inhibition surrounds the tooth bud and the next tooth bud is not initiated until the progress zone of the clone has escaped its influence. *****