Quantitative analysis of the calcium and phosphorus content of developing and permanent human teeth.

It was the aim of this study to investigate the distribution of Ca, P and C in predentin, dentin and enamel in human tooth buds and permanent teeth by EDX element analysis. The mandible of a 16-week-old human fetus containing eight mineralizing tooth buds and three human permanent molars were fixed in formaldehyde and embedded in Technovit 9100. Serial sections of 80 microm thickness of the mandible were cut in the frontal-dorsal direction, and polarized light micrographs were taken of these sections. The permanent teeth were cut in mesio-distal direction. The sections were investigated with scanning electron microscopy and EDX element analysis with a Philips XL 30 FEG scanning microscope and an EDAX energy-dispersive X-ray system using spot measurements, EDX line-scans and element mapping. Quantitative measurements were made in predentin, mineralizing dentin adjacent to predentin, mature dentin, mineralizing enamel and young enamel of developing teeth and mature enamel of permanent teeth. In developing teeth the Ca and P content increased rapidly from outer predentin towards mineralizing dentin. In enamel prisms of developing teeth the Ca and P content increased linearly from the surface towards the enamel-dentin junction. In permanent teeth only a small layer of predentin was found. The Ca and P content in enamel and circumpulpal dentin of permanent teeth was higher than in developing teeth. The Ca/P ratio differed between predentin and dentin areas reflecting different calcium phosphate compositions, but it was the same in mineralizing and young enamel. The differences in the distribution of Ca and P reflect different mineralizing patterns of the enamel and dentin matrices.     

Morphological analysis and chemical content of natural dentin carious lesion zones

Dentin is one of the earliest bio-mineralization products to appear in the evolution of vertebrates. Dentin reactions to infection mimic earlier phylogenetic patterns, and carious lesions are divided into different zones which reflect the natural patho-morphological reaction of dentin to the carious attack. It was the aim of this study to investigate deep dentin carious lesions of human molars with combined polarization light microscopy, scanning electron microscopy and energy dispersive X-ray element analysis (EDX) for the determination of different zones of the carious lesions, their extent and the chemical content. Sixteen extracted teeth with deep dentin carious lesions were embedded in Technovit 9100 (Kulzer) and serial sections of 80 microm thickness were made. These sections were then examined with polarized light microscopy to identify the different zones of the lesions. The outlines of the zones were traced consecutively and 3D-reconstructions were made for the determination of the extent and calculation of the volumes of the different zones. From the volumes of the demineralizing dentin and the translucent zone a Dentin Demineralization Index (DDI) was calculated. Three sections of each lesion were then coated with carbon and studied with a scanning electron microscope. 3D-reconstruction of the teeth showed the rather stable translucent zone, interrupted by remnants of dead tracts, and very different volumes of demineralizing dentin. Therefore, with increasing size of the demineralizing dentin, the DDI increased. The chemical content was measured using energy dispersive X-ray analysis (EDX) in areas of intertubular dentin. The content of Ca, P, and C was significantly different in all zones. The Ca/P ratio was significantly different between sound dentin and demineralizing dentin. From the results we conclude that the mineral content of intertubular dentin of the translucent zone and demineralizing dentin is different from that of sound dentin, and the unique mineralization pattern of the translucent zone is a biological reaction to the carious attack. Because active dentin lesions exhibit many non-occluded open dentin tubules, further bacterial invasion or, in case of dentin treatment, the penetration of bonding agents towards the pulp is morphologically not prevented and therefore of clinical importance.     

Involvement of the Klotho Protein in Dentin Formation and Mineralization

Klotho‐deficient mice exhibit multiple pathological conditions resembling human aging. Our previous study showed alterations in the distribution of osteocytes and in the bone matrix synthesis in klotho‐deficient mice. Although the bone and tooth share morphological features such as mineralization processes and components of the extracellular matrix, little information is available on how klotho deletion influences tooth formation. The present study aimed to elucidate the altered histology of incisors of klotho‐deficient mice–comparing the findings with those from their wild‐type littermates, by using immunohistochemistry for alkaline phosphatase (ALP), osteopontin, and dentin matrix protein‐1 (DMP‐1), terminal deoxynucleotidyl transferase‐mediated deoxyuridinetriphosphate nick end‐labeling (TUNEL) detection for apoptosis, and electron probe microanalyzer (EPMA) analysis on calcium (Ca), phosphate (P), and magnesium (Mg). Klotho‐deficient incisors exhibited disturbed layers of odontoblasts, predentin, and dentin, resulting in an obscure dentin‐predentinal border at the labial region. Several odontoblast‐like cells without ALP activity were embedded in the labial dentin matrix, and immunopositivity for DMP‐1 and osteopontin was discernible in the matrix surrounding these embedded odontoblast‐like cells. TUNEL detection demonstrated an apoptotic reaction in the embedded odontoblast‐like cells and pulpal cells in the klotho‐deficient mice. EPMA revealed lower concentrations of Ca, P, and Mg in the klotho‐deficient dentin, except for the dentin around abnormal odontoblast‐like cells. These findings suggest the involvement of the klotho gene in dentinogenesis and its mineralization. Anat Rec, 2007. © 2008 Wiley‐Liss, Inc.     

In situ analysis of gelatinolytic activity in human dentin

Matrix metalloproteinases (MMPs) such as gelatinases are differentially expressed in human tissues. These enzymes cleave specific substrates involved in cell signaling, tissue development and remodeling and tissue breakdown. Recent evidences show that gelatinases are crucial for normal dentin development and their activity is maintained throughout the entire tooth function in the oral cavity. Due to the lack of information about the exact location and activity of gelatinases in mature human dentin, the present study was designed to examine gelatinolytic levels in sound dentin. In situ zymography using confocal microscopy was performed on both mineralized and demineralized dentin samples. Sites presenting gelatinase activity were identified throughout the entire biological tissue pursuing different gelatinolytic levels for distinct areas: predentin and dentinal tubule regions presented higher gelatinolytic activity compared to intertubular dentin. Dentin regions with higher gelatinolytic activity immunohistochemically were partially correlated with MMP-2 expression. The maintenance of gelatinolytic activity in mature dentin may have biological implications related to biomineralization of predentin and tubular/peritubular dentinal regions, as well as regulation of defensive mechanisms of the dentin-pulp complex.     

The SCPP gene family and the complexity of hard tissues in vertebrates

Diverse hard tissues constituted a tooth-like skeletal element in extinct jawless vertebrates. Today, similar tissues are found in our teeth. These tissues mineralize in the extracellular matrix and involve various macromolecules. Among these molecules are secretory calcium-binding phosphoproteins (SCPPs) coded by genes that arose by duplication. Although the repertoire of SCPPs may vary in different lineages, some SCPPs are unusually acidic and are thought to participate in the mineralization of a collagenous matrix, principally either bone or dentin. Other SCPPs are rich in Pro and Gln (P/Q) and are employed to form the tooth surface. In tetrapods, the tooth surface is usually covered with enamel which develops in a matrix comprised of P/Q-rich SCPPs. By contrast, the tooth surface tissue in teleosts is called enameloid and it forms in a dentin-like collagenous matrix. Despite the difference in their matrix, both enamel and enameloid mature into hypermineralized inorganic tissues. Notably, some P/Q-rich SCPP genes are primarily expressed at this stage and their proteins localize between the tooth surface and overlying dental epithelium. Moreover, an orthologous gene is used for maturation of these 2 different tissues. These findings suggest distinct roles of acidic and P/Q-rich SCPPs during the evolution of hard tissues. Acidic SCPPs initially regulated the mineralization of bone, dentin, or a similar ancient collagenous tissue through interaction with calcium ions. P/Q-rich SCPPs arose next and originally assembled a structure or a space that facilitated the hypermineralization of dentin or a dentin-like tissue. Subsequently, some P/Q-rich SCPPs were coopted for the mineralizing enamel matrix. More recently, however, many SCPP genes were lost in toothless birds and mammals. Thus, it appears that, in vertebrates, the phenotypic complexity of hard tissues correlates with gain and loss of SCPP genes.     

Ca/P mol ratio of cries-affected dentin structures

BACKGROUND: A new approach called "minimum intervention" has been introduced for restoration of carious lesions to preserve tooth structure. This approach suggests that perhaps caries need not always be removed completely from deeper portions of the cavity. It is, therefore, important to characterize caries-affected dentin structures, because of the potential changes in bonding quality when using different dentinal substrates. MATERIALS AND METHOD: Ninety teeth (30 teeth each group) were studied. The first group (CF) consisted of 30 caries-free teeth. The second group (CC) consisted of 30 teeth, for which caries-free dentin teeth was chemically demineralized. The third group (ND) consisted of 30 extracted human molars with coronal carious lesions. After all tooth samples were water-polished with grit #600 SiC paper, they were tested by surface contact angle measurements and the electron-probe microanalyzer to measure Ca/P mol ratio. RESULTS: Contact angles were CF = 60.07 degrees ; CC = 30.8 degrees; ND = 26.11 degrees , p<0.05. Ca/P mol ratios were as follows; CF = 1.549 (+/-0.0435); CC = 1.324 (+/-0.2305); ND = 1.568 (+/-0.0523), p<0.05. Weibull analyses for Ca/P mol ratio indicated shape parameter (m) of CF was 13.3; it was 12.8 for ND and 11.8 for CC. Above the delta point (=1.65 in Ca/P ratio), for both groups m = 3.4. CONCLUSION: Caries-affected dentin surfaces (naturally-developed and chemically created) were statistically more chemically active than caries-free dentin surface. Ca/P mol ratio of chemically created caries was less than other two groups.     

Immunocytochemistry of keratan sulfate proteoglycan and dermatan sulfate proteoglycan in porcine tooth-germ dentin

Keratan sulfate proteoglycan and dermatan sulfate proteoglycan have been reported to inhibit collagen fibrillogenesis. We investigated their distribution in order to evaluate the role of proteoglycan in dentinogenesis. Specimens of porcine tooth-germ dentin and erupted teeth were the materials on which antibodies to keratin sulfate and dermatan sulfate proteoglycan were used. Predentin was found to be positive for both antibodies and the reaction ceased in the calcification front. Uniformly thick collagen fibrils (30-70 nm in diameter) were distributed in the predentin matrix, which would become intertubular dentin in the future. Both antibodies reacted positively along these fibrils. In contrast, along the surface layer of dentin in the tooth germ and that in erupted teeth, collagen fibrils of 10-300 nm in diameter were noted occasionally in dentinal tubules whose odontoblastic processes had disappeared and these heterogeneous fibrils were negative for both antibodies. Our findings suggest that keratan sulfate proteoglycan and dermatan sulfate proteoglycan distributed in the predentin inhibit calcification of collagen fibrils in the uncalcified matrix and disappear in the calcification front. It is further suggested that keratan sulfate proteoglycan and dermatan sulfate proteoglycan distributed along collagen fibrils in the predentin matrix maintain uniform thickness, whereas collagen fibrils in dentinal tubules varied in thickness because of the absence of involvement of both proteoglycans. Therefore, keratan sulfate proteoglycan and dermatan sulfate proteoglycan were thought to be involved in both calcification and matrix formation.     

人前磨牙牙髓牙本质复合体层粘连蛋白的免疫组织化学研究

目的　研究层粘连蛋白在牙髓牙本质复合体内的表达 ,探讨其分布与功能的关系。　方法　收集不同年龄新鲜健康人前磨牙 6 0颗 ,固定 ,脱钙 ,石蜡包埋 ,层粘连蛋白免疫组织化学染色。　结果　层粘连蛋白免疫反应性 ( IR)普遍存在于牙髓、前期牙本质和成熟牙本质小管与成牙本质细胞突起之间 ,以牙髓的神经与血管周围最为丰富 ,冠髓较根髓密集。　结论　层粘连蛋白 IR不仅见于牙髓与前期牙本质 ,而且也见于成熟牙本质 ,它对支持、固定牙髓牙本质复合体各结构的正常位置、形态可能发挥了重要作用     

Postnatal Mandibular Cheek Tooth Development in the Miniature Pig Based on Two‐Dimensional and Three‐Dimensional X‐Ray Analyses

The miniature pig is a useful large laboratory animal model. Various tissues and organs of miniature pigs are similar to those of humans in terms of developmental, anatomical, immunological, and physiological characteristics. The oral and maxillofacial region of miniature pigs is often used in preclinical studies of regenerative dentistry. However, there is limited information on the dentition and tooth structure of miniature pigs. The purpose of this study was to examine the time‐course changes of dentition and tooth structure (especially the root) of the miniature pig mandibular cheek teeth through X‐ray analyses using soft X‐ray for two‐dimensional observations and micro‐CT for three‐dimensional observations. The mandibles of male Clawn strain miniature pigs (2 weeks and 3, 5, 7, 9, 11, 14, 17, and 29 months of age) were used. X‐ray analysis of the dentition of miniature pig cheek teeth showed that the eruption pattern of the miniature pig is diphyodont and that the replacement pattern is vertical. Previous definitions of deciduous and permanent teeth often varied and there has been no consensus on the number of teeth (dentition); however, we found that three molars are present in the deciduous dentition and that four premolars and three molars are present in the permanent dentition. Furthermore, we confirmed the number of tooth roots and root canals. We believe that these findings will be highly useful in future studies using miniature pig teeth. Anat Rec, 2013. © 2013 Wiley Periodicals, Inc.     

Age‐related changes in the tooth–bone interface area of acrodont dentition in the chameleon

Chameleon teeth develop as individual structures at a distance from the developing jaw bone during the pre‐hatching period and also partially during the post‐hatching period. However, in the adult, all teeth are fused together and tightly attached to the jaw bone by mineralized attachment tissue to form one functional unit. Tooth to bone as well as tooth to tooth attachments are so firm that if injury to the oral cavity occurs, several neighbouring teeth and pieces of jaw can be broken off. We analysed age‐related changes in chameleon acrodont dentition, where ankylosis represents a physiological condition, whereas in mammals, ankylosis only occurs in a pathological context. The changes in hard‐tissue morphology and mineral composition leading to this fusion were analysed. For this purpose, the lower jaws of chameleons were investigated using X‐ray micro‐computed tomography, laser‐induced breakdown spectroscopy and microprobe analysis. For a long time, the dental pulp cavity remained connected with neighbouring teeth and also to the underlying bone marrow cavity. Then, a progressive filling of the dental pulp cavity by a mineralized matrix occurred, and a complex network of non‐mineralized channels remained. The size of these unmineralized channels progressively decreased until they completely disappeared, and the dental pulp cavity was filled by a mineralized matrix over time. Moreover, the distribution of calcium, phosphorus and magnesium showed distinct patterns in the different regions of the tooth–bone interface, with a significant progression of mineralization in dentin as well as in the supporting bone. In conclusion, tooth–bone fusion in chameleons results from an enhanced production of mineralized tissue during post‐hatching development. Uncovering the developmental processes underlying these outcomes and performing comparative studies is necessary to better understand physiological ankylosis; for that purpose, the chameleon can serve as a useful model species.     

人体天然牙骨牙本质界面区元素分布变化的电子探针微区分析

采用电子探针能谱微区分析方法初步测定了天然牙牙体组织牙骨质牙本质界面区的钙（Ｃａ）、磷（Ｐ）和硫（Ｓ）三种元素的分布变化特点，以考察人体天然牙硬组织中的无机和有机成分在骨牙本质界面区的分布变化特点。实验结果表明：骨牙本质界面区的Ｃａ、Ｐ元素的分布变化波动幅度较小，在骨牙本质界面区牙骨质和牙本质中的Ｃａ、Ｐ、Ｓ元素的平均重量百分比Ｃｉ值都变化不大，提示牙体组织中的无机和有机成分在骨牙本质界面区的分布未随着牙体组织解剖结构的改变而发生明显变化。     

Comparison of Dentoalveolar Morphology in WT and P2X7R KO Mice for the Development of Biomechanical Orthodontic Models

It has been suggested that the absence of the P2X7 receptor affects long bone morphology, and that one of the cytokines dependent on its activation may also affect tooth morphology. P2X7R KO (knockout) were compared with C57B/6 WT mice (background strain) to identify differences in a maxillary molar and surrounding bone. Nineteen WT and 12 KO mouse maxillae were scanned and 3D‐reconstructed using microCT. Tooth dimensions were measured and 3D bone morphometry was conducted. A finite element model was constructed based on the results. No statistically significant differences were found in dentoalveolar characteristics between the two mouse types. A single finite element model of the tooth can be used to mechanically represent both strains. P2X7R does not have a major effect on alveolar bone or tooth morphology. The P2X7R effects are site‐specific. Anat Rec, 2009. © 2008 Wiley‐Liss, Inc.     

bmp2b and bmp4 are dispensable for zebrafish tooth development

Bone morphogenetic protein (Bmp) signaling has been shown to play important roles in tooth development at virtually all stages from initiation to hard tissue formation. The specific ligands involved in these processes have not been directly tested by loss‐of‐function experiments, however. We used morpholino antisense oligonucleotides and mutant analysis in the zebrafish to reduce or eliminate the function of bmp2b and bmp4, two ligands known to be expressed in zebrafish teeth and whose mammalian orthologs are thought to play important roles in tooth development. Surprisingly, we found that elimination of function of these two genes singly and in combination did not prevent the formation of mature, attached teeth. The mostly likely explanation for this result is functional redundancy with other Bmp ligands, which may differ between the zebrafish and the mouse. Developmental Dynamics 239:2534–2546, 2010. © 2010 Wiley‐Liss, Inc.     

Odontoblast processes of the mouse incisor are plates oriented in the direction of growth

Odontoblast processes are thin cytoplasmic projections that extend from the cell body at the periphery of the pulp toward the dentin-enamel junction. The odontoblast processes function in the secretion, assembly and mineralization of dentin during development, participate in mechanosensation, and aid in dentin repair in mature teeth. Because they are small and densely arranged, their three-dimensional organization is not well documented. To gain further insight into how odontoblast processes contribute to odontogenesis, we used serial section electron microscopy and three-dimensional reconstructions to examine these processes in the predentin region of mouse molars and incisors. In molars, the odontoblast processes are tubular with a diameter of ~1.8 μm. The odontoblast processes near the incisor tip are similarly shaped, but those midway between the tip and apex are shaped like plates. The plates are radially aligned and longitudinally oriented with respect to the growth axis of the incisor. The thickness of the plates is approximately the same as the diameter of molar odontoblast processes. The plates have an irregular edge; the average ratio of width (midway in the predentin) to thickness is 2.3 on the labial side and 3.6 on the lingual side. The plate geometry seems likely to be related to the continuous growth of the incisor and may provide a clue as to the mechanisms by which the odontoblast processes are involved in tooth development.     

TGF-beta latency-associated peptides (LAPs) in human dentin matrix and pulp

Transforming growth factor (TGF)-beta s in dentin matrix provide a pool of bioactive molecules, but association with latency-associated peptides (LAPs) may influence their activity. We investigated TGF-beta 1, -beta 2, and -beta 3 LAP expression in sound and carious human teeth. Teeth were fixed and processed immediately following extraction prior to staining with rabbit polyclonal antibodies to the TGF-beta LAPs. A soluble dentin matrix fraction was prepared from dissected human dentin and sequential extraction of pulpal ECM was performed prior to purification. Fractions were Western blotted and probed with the LAP antibodies. All three LAPs were present in odontoblasts, cells of the pulp, and predentin; however, no staining of mineralized dentin matrix was seen. Similar patterns of expression were seen in carious tissue. Expression of TGF-beta LAPs in cells and pulpal matrix of healthy and carious teeth will be important in regulation of TGF-beta activity and may modulate the tissue response to injury.     

Odontoclastic resorption at the pulpal surface of coronal dentin prior to the shedding of human deciduous teeth.

Histological and histochemical observations of more than 150 extracted human deciduous teeth revealed that, prior to shedding, odontoclastic resorption as a rule takes place at the pulpal surface of coronal dentin. We also found that this phenomenon occurs in all kinds of deciduous teeth. The process of this internal resorption of coronal dentin of deciduous teeth clearly showed time-related histological changes. During the time the roots were actively being resorbed, the pulpal tissue retained its normal structure. However, when root resorption neared completion, inflammatory cells started to gradually infiltrate into the pulp, and odontoblasts began to degenerate. After that, multinucleate odontoclasts appeared, and resorption proceeded from the predentin to the dentin. The odontoclastic activity was initially detected only on the pulpal surface at the bottom areas of the crown. It gradually spread towards the pulpal horn regions along the wall of the pulp chamber. However, this internal resorption of coronal dentin did not continue until the teeth were finally shed. After the elimination of resorption, the resorbed dentin surface was repaired by a cementum-like deposition or covered with fibrous connective tissue.     

Autoradiographic location of sensory nerve endings in dentin of monkey teeth

We have used the autoradiographic method to locate trigeminal nerve endings in monkey teeth. The nerve endings were labeled in two adult female Macaca fascicularis by 20 hours of axonal transport of radioactive protein (³H-L-proline). We found a few labeled axons in contralateral mandibular central incisors and one mandibular canine. In ipsilateral teeth, numerous myelinated and unmyelinated axons were labeled; they formed a few terminal branches in the roots but primarily branched in the crown to form the peripheral plexus of Raschkow and to terminate as free endings in the odontoblast layer, predentin, and as far as 120 μm into dentinal tubules. Electron microscopic autoradiography showed that the radioactive axonally transported protein was confined to sensory axons and endings; odontoblasts and dentin matrix were not significantly labeled. Labeled free nerve endings were closely apposed to odontoblasts in dentin but did not form distinctive junctions with them. Nerve endings were most numerous in the regular tubular dentin of the crown adjacent to the tip of the pulp horn, occurring in at least half of the dentinal tubules there. Reparative dentin was poorly innervated, even near the tip of the crown, and it had a different tubular structure and adjacent pulpal structure from the innervated dentin. Radicular dentin was not innervated in most areas but did contain a few labeled axons where the predentin was wide and the odontoblasts were columnar, as at the buccal and lingual poles of some roots. Our results show that dentinal sensory nerve endings in primate teeth can be profuse, sparse, or absent depending on the location and structure of dentin and its adjacent pulp. When dentin was innervated, the tubules were straight and contained odontoblast processes, the predentin was wide, the odontoblast cell bodies were relatively columnar, and there was an adjacent cell-free zone and pulpal nerve plexus.     

Proteoglycans in Predentin: The Last 15 Micrometers Before Mineralization

Small leucine-rich proteoglycans (SLRPs) regulate extracellular matrix organization. In order to investigate the distribution and potential functions of decorin, biglycan (BGN), and fibromodulin (3 SLRPs, potentially related to dentinogenesis), we performed light and electron immunochemistry on teeth from rats, and on wild-type and biglycan knockout mice (BGN KO). Immunohistochemical data demonstrate that chondroitin sulfate/dermatan sulfate (CS/DS) and keratan sulfate (KS) distributions displayed reverse gradients in predentin. The decrease of CS/DS labeling from the proximal to the distal predentin contrasted with the sharp decorin increase observed in the distal predentin near the predentin/dentin transition, an effect possibly attributable to the deglycosylation action of stromelysin-1. In contrast, BGN concentration was apparently constant throughout the whole predentin. Additional immunolabelings showed, for the first time, the presence of fibromodulin in predentin. Compared with the wild-type mouse, the mean diameter of collagen fibrils in the BGN KO was smaller in the proximal predentin but larger in the central and distal predentin, the metadentin was broader, and the dentin mineralization appeared altered and heterogeneous. Altogether, our data suggest an important role for BGN in dentin formation and mineralization.     

Histological and immunohistochemical studies of tissue engineered odontogenesis

The successful regeneration of complex tooth structures based on tissue-engineering principles was recently reported. The process of this regeneration, however, remains poorly characterized. In this study, we have used histochemistry to examine the regeneration process of tissue engineered teeth in order to determine the cell types that give rise to these engineered tooth structures. Porcine third molar tooth buds were dissociated into single-cell suspensions and seeded onto a biodegradable polyglycolic acid polymer scaffold. Following varying periods of growth in rat hosts, the specimens were evaluated by histology and immunohistochemistry. Aggregates of epithelial cells were first observed 4-6 weeks after implantation. These aggregates assumed three different shapes: a natural tooth germ-like shape, a circular shape, or a bilayer-bundle. Based on the structure of the stellate reticulum in the dental epithelium, the circular and bilayer-bundle aggregates could be clearly classified into two types: one with extensively developed stellate reticulum, and the other with negligible stellate reticulum. The epithelial cells in the circular aggregates differentiated into ameloblasts. The continuous bilayer bundles eventually formed the epithelial sheath, and dentin tissue was evident at the apex of these bundles. Finally, enamel-covered dentin and cementum-covered dentin formed, a process most likely mediated by epithelial-mesenchymal interaction. These results suggest that the development of these engineered teeth closely parallels that of natural odontogenesis derived from the immature epithelial and mesenchymal cells.     

Proteoglycans in predentin: the last 15 micrometers before mineralization

Small leucine-rich proteoglycans (SLRPs) regulate extracellular matrix organization. In order to investigate the distribution and potential functions of decorin, biglycan (BGN), and fibromodulin (3 SLRPs, potentially related to dentinogenesis), we performed light and electron immunochemistry on teeth from rats, and on wild-type and biglycan knockout mice (BGN KO). Immunohistochemical data demonstrate that chondroitin sulfate/dermatan sulfate (CS/DS) and keratan sulfate (KS) distributions displayed reverse gradients in predentin. The decrease of CS/DS labeling from the proximal to the distal predentin contrasted with the sharp decorin increase observed in the distal predentin near the predentin/dentin transition, an effect possibly attributable to the deglycosylation action of stromelysin-1. In contrast, BGN concentration was apparently constant throughout the whole predentin. Additional immunolabelings showed, for the first time, the presence of fibromodulin in predentin. Compared with the wild-type mouse, the mean diameter of collagen fibrils in the BGN KO was smaller in the proximal predentin but larger in the central and distal predentin, the metadentin was broader, and the dentin mineralization appeared altered and heterogeneous. Altogether, our data suggest an important role for BGN in dentin formation and mineralization.