Tooth Root Cementum Research Articles

Nanoindentation and Atomic Force Microscopy Derived Mechanical and Microgeometrical Properties of Tooth Root Cementum

In the present research, nanoindentation, atomic-force microscopy and optical microscopy were used to study the mechanical and microgeometrical parameters of tooth tissues. A nanoindentation test unit equipped with Berkovich indenter was used to determine the values of the reduced Young’s modulus and indentation hardness and both nanoindentation and atomic force microscopy using a diamond probe on a silicon cantilever were used to study microgeometrical parameters of human tooth root cementum. Three areas of cementum were studied: the cervical region near the dentine–enamel junction, the second third of the tooth root, and the apex of the tooth root. The interpretation of the results was carried out using the Oliver–Pharr method. It was established, that the mechanical properties of cementum increase from the cervical region to the central part of the root, then decrease again towards the apex of the tooth root. On the contrary, the microgeometrical characteristics of cementum practically do not demonstrate any change in the same direction. A decrease in the roughness parameters in the direction from cellular cementum to dentine was observed. Additionally, a decrease in the reduced Young’s modulus and indentation hardness of dentine in the cervical area compared to dentine in the crown part of the tooth was found using nanoindentation. The investigation of the dentine–cementum junction with high resolution revealed the interspaced collagen fiber bridges and epithelial rests of Malassez, whose sizes were studied. The parameters of the topographic features of the cementum in the vicinity of the lacunae of cementocytes were also established.

Titanium Nanosurface with a Biomimetic Physical Microenvironment to Induce Endogenous Regeneration of the Periodontium.

The periodontium supports the teeth by dentoalveolar fibrous joints that serve unique oral functions. Endogenous regeneration of the periodontium around artificial teeth (dental implants) provides a cost-effective solution for the extension of healthy life expectancy but remains a challenge in regenerative medicine. Biomimetics can create smart biomaterials that tune endogenous cells at a tissue-material interface. Here, we created a smart titanium nanosurface mimicking the surface nanotopography and micromechanical properties of the tooth root cementum (TRC), which is essential for the induction of dentoalveolar fibrous joints to regenerate the periodontium. After transplantation into the rat renal capsule, only the titanium artificial tooth with the TRC-mimetic nanosurface formed a complex dentoalveolar fibrous joint structure, with bone tissue, periodontal ligament (PDL), and TRC, in the decellularized jawbone matrix. TRC-mimetic titanium implants induce the formation of functional periodontium, even in a jawbone implantation model, which generally causes osseointegration (ankyloses). In human PDL cells, TRC analogousness in the surface mechanical microenvironment regulates matrix mineralization through bone sialoprotein expression and phosphorus metabolism, which are critical for cementogenesis. Therefore, the titanium nanosurfaces with nanotopographical and mechanical microenvironments mimicking the TRC surface induce dentoalveolar fibrous joints for periodontal regeneration by interfacial tuning of endogenous cells.

Concrescent Teeth: Extraction Implications - A Case Report

Background: Concrescent teeth belong to the group of double teeth with disturbed anatomy of shape and structure. The concrescence consists of the aggregation of root cementum of adjacent teeth. This anomaly usually concerns the upper teeth in the lateral part of the jaw. Concrescent teeth can be treated conservatively, endodontically, orthodontically and surgically. Objective: The aim of this article is to present the case of the concrescence of upper teeth, a second molar tooth with an impacted third molar, and to draw the attention of clinicians to this rare anomaly that may cause medical and legal problems. Case presentation: This article presents the case of a 53-year-old woman who reported to the dentist’s office to extract tooth 17 due to chronic periodontitis and an impacted and abnormally placed tooth 18. Result: During the extraction, concrescence of the teeth was noted. The teeth were removed, and the alveoli were surgically treated. The postoperative period was uncomplicated. Conclusion: In this paper, the authors draw the attention of clinicians to the fact that the occurrence of concrescent teeth in clinical practice is rare and that it is difficult to diagnose this anomaly on the basis of routine radiological diagnostics based on 2D images. They also point to the necessity of a thorough physical examination and a thorough assessment of teeth before extraction, the extension of imaging diagnostics by conical CT scan in justified cases, and informing the patient about possible complications. In order to avoid serious complications, the extraction of the concrescent teeth should be performed by an experienced specialist in dental or maxillofacial surgery.

Role of mitochondrial dna analyses in provenancing unknown human skeletal remains: Probabilities and limitations

Ancient degraded remains of human origin can be identified using ancient DNA analyses. The mitochondrial DNA is a useful tool to solve unsolved cases in relation to identification of an individual where nuclear DNA is too degraded or insufficient to be used for the purpose. Comparative analysis of mtDNA content in petrous bone and the tooth-root cementum have revealed that later is best substrate for studying genomic analyses of ancient human populations The dental DNA is less prone to contamination, and the preservation of ancient DNA from dental samples is easier than bones. The selection of correct decontamination method, appropriate extraction protocol provides best results and thus increases value of bones and teeth as a forensic evidence in cases where identity establishment is problematic. In this review article, the current status of the methods used in ancient DNA research, the advantages and the limitations of this archaeogenetical method have been presented.

Current Trends in In Vitro Modeling to Mimic Cellular Crosstalk in Periodontal Tissue.

Clinical evidence indicates that in physiological and therapeutic conditions a continuous remodeling of the tooth root cementum and the periodontal apparatus is required to maintain tissue strength, to prevent damage, and to secure teeth anchorage. Within the tooth's surrounding tissues, tooth root cementum and the periodontal ligament are the key regulators of a functional tissue homeostasis. While the root cementum anchors the periodontal fibers to the tooth root, the periodontal ligament itself is the key regulator of tissue resorption, the remodeling process, and mechanical signal transduction. Thus, a balanced crosstalk of both tissues is mandatory for maintaining the homeostasis of this complex system. However, the mechanobiological mechanisms that shape the remodeling process and the interaction between the tissues are largely unknown. In recent years, numerous 2D and 3D in vitro models have sought to mimic the physiological and pathophysiological conditions of periodontal tissue. They have been proposed to unravel the underlying nature of the cell-cell and the cell-extracellular matrix interactions. The present review provides an overview of recent in vitro models and relevant biomaterials used to enhance the understanding of periodontal crosstalk and aims to provide a scientific basis for advanced regenerative strategies.

Periodonto ligos. Paciento motyvacija ir papildomo gydymo galimybės

Periodonto ligos – gingivitas ir periodontitas yra uždegiminės ligos, kurios pažeidžia aplink dantį esančias atramines struktūras: dantenas, periodonto raištį, alveolės kaulą, danties šaknies cementą. Gingivitas negydomas progresuoja į periodontitą, kuris gali sukelti alveolės kaulo irimą ir danties netekimą. Tokios pasekmės blogina žmogaus gyvenimo kokybę, sukelia negalią ir socialinę atskirtį, todėl labai svarbi periodonto ligų prevencija ir laiku pradedamas gydymas. Siekiant išvengti periodonto ligų, svarbiausia yra reguliariai atlikti asmeninę burnos higieną, tačiau jos veiksmingumui svarbi paciento motyvacija. Ją galima paskatinti, taikant individualų rizikos klausimyną, kuris gerina paciento žinias apie ligą, didina tikėjimą gydymo veiksmingumu ir pasiryžimą laikytis gydymo rekomendacijų. Teigiamas poveikis pacientų motyvacijai stebimas ir po motyvacinio pokalbio, kurio metu pacientas, padedamas konsultanto, motyvuoja save teigiamiems pokyčiams. Gydant periodontitą konservatyviai, atliekama profesionali burnos higiena, tačiau, ieškant ilgalaikio poveikio, pradėtos tyrinėti ir papildomos priemonės, tokios kaip probiotikai, hialurono rūgšties aplikacijos ir maisto papildai, kurie galėtų palengvinti periodonto ligų eigą.

Effects of Intraligamentary Injection of Osteogenic-Induced Gingival Fibroblasts on Cementum Thickness in the Dog Model of Tooth Root Resorption.

Tooth root resorption is an unwanted result of orthodontic tooth movement, and it can be expressed by a reduction in cementum thickness. The aim of this experimental study was to evaluate the effect of intraligamentary injection of osteogenic-induced gingival fibroblasts (OIGF) on cellular and acellular tooth root cementum thickness in modeled orthodontic tooth movement. Six beagle dogs were used in the study. All the upper and lower third and fourth premolars were subjected to mechanical loading for 4weeks, which induced orthodontic tooth movement. Fifteen premolars were assigned to the OIGF group, which received a single OIGF injection through the periodontal ligament near the root apex (n=7 teeth), and to the control group, which received a single injection of Dulbecco's modified eagle's medium in the periapical area (n=8 teeth). The evaluation of histomorphometry was performed to assess the thicknesses of cellular and acellular cementum at the root apex and four bilateral sites distal to the apex. We found no statistically significant enhancing effects of gingival fibroblasts on either cellular or acellular cementum thicknesses when compared with the control group. We conclude that a single intraligamentary injection of OIGF does not stimulate the formation of tooth root cementum in the dog model of orthodontic tooth movement. Thus, OIGF is unlikely to prevent orthodontic-induced tooth root resorption.

The Wnt Antagonist SFRP1: A Key Regulator of Periodontal Mineral Homeostasis.

The function of mammalian periodontal tissues depends on the presence of a nonmineralized periodontal ligament (PDL) juxtaposed in between mineralized tooth anchorage tissues alveolar bone (AB) and root cementum. In the present study we have hypothesized that the Wnt antagonist secreted frizzled related protein 1 (SFRP1) is an essential regulator of periodontal tissue mineral homeostasis. Our immunoreactions and western blot data demonstrated that SFRP1 was substantially expressed higher in PDL fibroblasts than in surrounding AB progenitors and cementoblasts. SFRP1 was also detected at higher levels in PDL fibroblasts than in dental follicle (DF) cells, but the difference was less pronounced. Preferential H3K4me3 active histone mark enrichment on the SFRP1 promoter and a lack of H3K27me3 repression were most dramatic in PDL progenitors, to a lesser degree in DF cells, and not detected in AB progenitors and cementoblasts. Selective inhibition of SFRP1 using a small molecule inhibitor WAY-316606 demonstrated that SFRP1 block increased PDL cell mineralization and mineralization gene expression such as β-catenin, alkaline phosphatase, osteocalcin, collagen I, and RUNX2. The effect of SFRP1 inhibition on PDL cell mineral homeostasis was confirmed by RNA silencing. These studies also demonstrated that SFRP1 knockdown promotes PDL differentiation through histone H3K4me3-mediated activation of RUNX2 and SP7. Finally, when SFRP1 inhibition and silencing studies were performed using AB progenitors instead of PDL progenitors, there was little effect on mineralized state control and gene expression, with the exception of osteocalcin, which was dramatically upregulated upon SFRP1 silencing. Together, the results of our study document the highly specific role of the Wnt inhibitor SFRP1 in maintaining the nonmineralized state of PDL progenitors.

Tissue-specific mineralization defects in the periodontium of the Hyp mouse model of X-linked hypophosphatemia

X-linked hypophosphatemia (XLH) is a dento-osseous disorder caused by inactivating mutations in the PHEX gene, leading to renal phosphate wasting and hypophosphatemia, and impaired mineralization of bones and teeth. In the oral cavity, recent reports suggest a higher susceptibility of XLH patients to periodontitis, where patients present with impaired tooth cementum – a bone-like tissue involved in tooth attachment to the jaw bones and post-eruption tooth positioning – and a higher frequency of intrabony defects. In the present study, the pathobiology of alveolar bone and tooth cementum was investigated in the Hyp mouse, the murine analog of XLH. PHEX deficiency in XLH/Hyp dramatically alters the periodontal phenotype, with hypoplasia of tooth root cementum associated with a lack of periodontal ligament attachment and the presence of an immature apatitic mineral phase of all periodontal mineralized tissues. Challenging the Hyp periodontium in two surgical experimental models – ligature-induced periodontal breakdown and repair, and a model of tooth movement adaptation inducing cementum formation – we show that bone and cementum formation, and their healing, are altered. Bone and cementum mineralization appear similarly disturbed, where hypomineralized pericellular matrix surrounds cells, and where the protein osteopontin (OPN, a mineralization inhibitor) accumulates in a tissue-specific manner, most notably in the perilacunar matrix surrounding osteocytes. Although the pathobiology is different between XLH/Hyp bone and cementum, our results show a major XLH phenotype in oral mineralized tissues consistent with variations in patient susceptibility to periodontal disorders.

Hertwig's epithelial root sheath cells contribute to formation of periodontal ligament through epithelial-mesenchymal transition by TGF-β.

In tooth root development, periodontal ligament (PDL) and cementum are formed by the coordination with the fragmentation of Hertwig's epithelial root sheath (HERS) and the differentiation of dental follicle mesenchymal cells. However, the function of the dental epithelial cells after HERS fragmentation in the PDL is not fully understood. Here, we found that TGF-β regulated HERS fragmentation via epithelial-mesenchymal transition (EMT), and the fragmented epithelial cells differentiated into PDL fibroblastic cells with expressing of PDL extracellular matrix (ECM). In the histochemical analysis, TGF-β was expressed in odontoblast layer adjacent of HERS during root development. Periostin expression was detected around fragmented epithelial cells on the root surface, but not in HERS. In the experiment using an established mouse HERS cell line (HERS01a), TGF-β1 treatment decreased E-cadherin and relatively increased N-cadherin expression. TGF-β1 treatment in HERS01a induced further expression of important ECM proteins for acellular cementum and PDL development such as fibronectin and periostin. Taken together, activation of TGF-βsignaling induces HERS fragmentation through EMT and the fragmented HERS cells contribute to formation of PDL and acellular cementum through periostin and fibronectin expression.

Mineralization defects in cementum and craniofacial bone from loss of bone sialoprotein

Bone sialoprotein (BSP) is a multifunctional extracellular matrix protein found in mineralized tissues, including bone, cartilage, tooth root cementum (both acellular and cellular types), and dentin. In order to define the role BSP plays in the process of biomineralization of these tissues, we analyzed cementogenesis, dentinogenesis, and osteogenesis (intramembranous and endochondral) in craniofacial bone in Bsp null mice and wild-type (WT) controls over a developmental period (1–60days post natal; dpn) by histology, immunohistochemistry, undecalcified histochemistry, microcomputed tomography (microCT), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and quantitative PCR (qPCR). Regions of intramembranous ossification in the alveolus, mandible, and calvaria presented delayed mineralization and osteoid accumulation, assessed by von Kossa and Goldner's trichrome stains at 1 and 14dpn. Moreover, Bsp−/− mice featured increased cranial suture size at the early time point, 1dpn. Immunostaining and PCR demonstrated that osteoblast markers, osterix, alkaline phosphatase, and osteopontin were unchanged in Bsp null mandibles compared to WT. Bsp−/− mouse molars featured a lack of functional acellular cementum formation by histology, SEM, and TEM, and subsequent loss of Sharpey's collagen fiber insertion into the tooth root structure. Bsp−/− mouse alveolar and mandibular bone featured equivalent or fewer osteoclasts at early ages (1 and 14dpn), however, increased RANKL immunostaining and mRNA, and significantly increased number of osteoclast-like cells (2–5 fold) were found at later ages (26 and 60dpn), corresponding to periodontal breakdown and severe alveolar bone resorption observed following molar teeth entering occlusion. Dentin formation was unperturbed in Bsp−/− mouse molars, with no delay in mineralization, no alteration in dentin dimensions, and no differences in odontoblast markers analyzed. No defects were identified in endochondral ossification in the cranial base, and craniofacial morphology was unaffected in Bsp−/− mice. These analyses confirm a critical role for BSP in processes of cementogenesis and intramembranous ossification of craniofacial bone, whereas endochondral ossification in the cranial base was minimally affected and dentinogenesis was normal in Bsp−/− molar teeth. Dissimilar effects of loss of BSP on mineralization of dental and craniofacial tissues suggest local differences in the role of BSP and/or yet to be defined interactions with site-specific factors.

Age estimation using annulations in root cementum of human teeth: A comparison between longitudinal and cross sections.

Background:Age estimation is an important factor in the identification of an individual in forensic science. Research indicates that cemental annulations may be used more reliably than other morphological or histological traits of human skeleton for age estimation.Materials and Methods:Twenty-five teeth were sectioned longitudinally, and twenty-five teeth were cross-sectioned at the mid portion of the root. Sections were ground, mounted and viewed under a bright light microscope. The area selected for counting was photographed under ×10 objective, magnified 5 times; cemental lines were counted and added to the eruption age of that patient, to obtain the chronological age. The statistical software SAS 9.2, SPSS 15.0, Stata 10.1, MedCalc 9.0.1, Systat 12.0 and R environment ver.2.11.1 were used for the analysis of the data.Results:The P value comparing actual age and calculated age using longitudinal sections is moderately significant and the P value comparing actual age and calculated age in the age group of <30 years is significant.Interpretation and Conclusion:The middle third of tooth root was most suitable to count annulations. The cross sections are easier to count but longitudinal sections give more appropriate results on age estimation. Though the procedure predicts under assessment of age in the younger age group and over assessment of age in the older age group, it provides a close estimate of the actual age of an individual. It can be correlated with other age estimation methods for better reliability.

Tooth root dentin mineralization defects in a mouse model of hypophosphatasia.

Tissue-nonspecific alkaline phosphatase (TNAP) is expressed in mineralizing tissues and functions to reduce pyrophosphate (PP(i) ), a potent inhibitor of mineralization. Loss of TNAP function causes hypophosphatasia (HPP), a heritable disorder marked by increased PP(i) , resulting in rickets and osteomalacia. Tooth root cementum defects are well described in both HPP patients and in Alpl(-/-) mice, a model for infantile HPP. In Alpl(-/-) mice, dentin mineralization is specifically delayed in the root; however, reports from human HPP patients are variable and inconsistent regarding dentin defects. In the current study, we aimed to define the molecular basis for changes in dentinogenesis observed in Alpl(-/-) mice. TNAP was found to be highly expressed by mature odontoblasts, and Alpl(-/-) molar and incisor roots featured defective dentin mineralization, ranging from a mild delay to severely disturbed root dentinogenesis. Lack of mantle dentin mineralization was associated with disordered and dysmorphic odontoblasts having disrupted expression of marker genes osteocalcin and dentin sialophosphoprotein. The formation of, initiation of mineralization within, and rupture of matrix vesicles in Alpl(-/-) dentin matrix was not affected. Osteopontin (OPN), an inhibitor of mineralization that contributes to the skeletal pathology in Alpl(-/-) mice, was present in the generally unmineralized Alpl(-/-) mantle dentin at ruptured mineralizing matrix vesicles, as detected by immunohistochemistry and by immunogold labeling. However, ablating the OPN-encoding Spp1 gene in Alpl(-/-) mice was insufficient to rescue the dentin mineralization defect. Administration of bioengineered mineral-targeting human TNAP (ENB-0040) to Alpl(-/-) mice corrected defective dentin mineralization in the molar roots. These studies reveal that TNAP participates in root dentin formation and confirm that reduction of PP(i) during dentinogenesis is necessary for odontoblast differentiation, dentin matrix secretion, and mineralization. Furthermore, these results elucidate developmental mechanisms underlying dentin pathology in HPP patients, and begin to explain the reported variability in the dentin/pulp complex pathology in these patients.

Methods for studying tooth root cementum by light microscopy

The tooth root cementum is a thin, mineralized tissue covering the root dentin that is present primarily as acellular cementum on the cervical root and cellular cementum covering the apical root. While cementum shares many properties in common with bone and dentin, it is a unique mineralized tissue and acellular cementum is critical for attachment of the tooth to the surrounding periodontal ligament (PDL). Resources for methodologies for hard tissues often overlook cementum and approaches that may be of value for studying this tissue. To address this issue, this report offers detailed methodology, as well as comparisons of several histological and immunohistochemical stains available for imaging the cementum–PDL complex by light microscopy. Notably, the infrequently used Alcian blue stain with nuclear fast red counterstain provided utility in imaging cementum in mouse, porcine and human teeth. While no truly unique extracellular matrix markers have been identified to differentiate cementum from the other hard tissues, immunohistochemistry for detection of bone sialoprotein (BSP), osteopontin (OPN), and dentin matrix protein 1 (DMP1) is a reliable approach for studying both acellular and cellular cementum and providing insight into developmental biology of these tissues. Histological and immunohistochemical approaches provide insight on developmental biology of cementum.

Hypoplastic root cementum and premature loss of primary teeth in Coffin–Lowry syndrome: a case report

Coffin-Lowry syndrome (CLS) is a rare genetic disorder. The syndrome presents with psychomotor retardation, short stature, skeletal deformations, digit abnormalities, and distinctive facial features. Oral and dental findings in CLS are common and they include thick prominent lips, high palate, midline lingual furrow, hypodontia, microdontia, delayed eruption, and early tooth loss. Only one earlier case suggesting hypoplastic root cementum as cause for primary loss of teeth in CLS has been published. This case describes a 3-year-old boy with premature loss of primary incisors without preceding root resorption. In addition to the dental findings, the boy had several general signs and symptoms and the dental findings together with the other characteristics led to the clinical diagnosis of CLS, which later was genetically verified. Histological analysis of an extracted primary incisor showed hypoplastic root cementum. Hypoplastic root cementum may explain early tooth loss in CLS. As early loss of primary teeth is rare, especially when there is no previous root resorption, the individual is likely to seek dental care. Thus, the dentist may play an important role in assisting in the diagnosing of CLS.

The Progressive Ankylosis Protein Regulates Cementum Apposition and Extracellular Matrix Composition

Background/Aims: Tooth root cementum is sensitive to modulation of inorganic pyrophosphate (PP_i), an inhibitor of hydroxyapatite precipitation. Factors increasing PP_i include progressive ankylosis protein (ANK) and ectonucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) while tissue nonspecific alkaline phosphatase hydrolyzes PP_i. Studies here aimed to define the role of ANK in root and cementum by analyzing tooth development in Ank knock-out (KO) mice versus wild type. Materials and Methods: Periodontal development in KO versus control mice was analyzed by histology, histomorphometry, immunohistochemistry, in situ hybridization, electron microscopy, and nanoindentation. Cementoblast cultures were used in vitro to provide mechanistic underpinnings for PP_i modulation of cell function. Results: Over the course of root development, Ank KO cervical cementum became 8- to 12-fold thicker than control cervical cementum. Periodontal ligament width was maintained and other dentoalveolar tissues, including apical cementum, were unaltered. Cervical cementum uncharacteristically included numerous cells, from rapid cementogenesis. Ank KO increased osteopontin and dentin matrix protein 1 gene and protein expression, and markedly increased NPP1 protein expression in cementoblasts but not in other cell types. Conditional ablation of Ank in joints and periodontia confirmed a local role for ANK in cementogenesis. In vitro studies employing cementoblasts indicated that Ank and Enpp1 mRNA levels increased in step with mineral nodule formation, supporting a role for these factors in regulation of cementum matrix mineralization. Conclusion: ANK, by modulating local PP_i, controls cervical cementum apposition and extracellular matrix. Loss of ANK created a local environment conducive to rapid cementogenesis; therefore, approaches modulating PP_i in periodontal tissues have potential to promote cementum regeneration.

Incremental lines in root cementum of human teeth: An approach to their role in age estimation using polarizing microscopy

Age estimation is an important factor in the identification of an individual in forensic science. The hard tissues of the human dentition are able to resist decay and degradation long after other tissues are lost. This resistance has made teeth useful indicators for age calculation. Recent research indicates that tooth cementum annulations (TCA) may be used more reliably than any other morphological or histological traits of the adult skeleton for age estimation. The purpose of this study was to examine the correlation between age and the number of incremental lines in human dental cementum and to ascertain the best method of studying cementum with respect to different forms of microscopy. Thirty nonrestorable teeth were extracted from 20 people, and longitudinal ground section of each tooth was prepared. Photomicrographs of the area at the junction of apical and middle third of the root under light and polarized microscope were taken. The cementum was composed of multiple light and dark bands that were counted on the photomicrograph with the help of image analysis software and added to the average eruption time of individual tooth. The predicted age of the individual was thus obtained. Results showed a significant correlation between the predicted age and actual chronological age of the individual. These data indicate that quantitation of cementum annuli is a moderately reliable means for age estimation in humans and polarizing microscopy is a better means of visualizing incremental lines of cementum compared to light microscopy.

Cementum analysis in cleidocranial dysostosis

Cleidocranial dysostosis (CCD) is a skeletal disorder associated with dental anomalies such as failure or delayed eruption of permanent teeth and multiple impacted supernumerary or permanent teeth. Absence of cellular cementum at the root apex is presumed to be one of the factors for failure or delayed eruption. The aim of this study was to analyze the root cementum of supernumerary(S) and permanent teeth (P) of a CCD patient and to compare the findings with those of normal individuals with special emphasis on delayed eruption. Ground sections of 12 extracted teeth (9S + 3P) of a CCD patient and 12 teeth of normal individuals (5S + 7P) were taken for the study. The sections were longitudinal and 50 microm thick. Root characteristics were recorded with the aid of a light microscope. The absence of apical cellular cementum, increased root resorption and increased percentage of the gap type of cemento-enamel (C-E) junctions were significant findings noted both in the supernumerary and permanent teeth of the CCD patient. The presumption of cellular cementum is essential for tooth eruption is not supported by the findings of the present study. Delayed exception could be attributed to various other factors like mechanical obstruction, reduced serum alkaline phosphatase levels etc. Also, the significance of the increased percentage of the gap type of C-E junctions and its role in tooth eruption has to be further evaluated.

Targeting and Immobilization of Bioactive Peptides on Dentin Matrix

The regeneration of structurally/functionally competent tooth root cementum is a critical step for the successful restoration of periodontal attachment. In this study, we tested whether a poly-glutamic acid-rich domain and glutamine-containing transglutaminase substrate can be used to target biologically active peptides to the mineralized root matrix and to bind such peptides covalently to the organic matrix. As a biologically active model molecule, the integrin-binding motif, RGD, was used. The effects of immobilization of such synthetic peptides to the dentin matrix on cementoblastic adhesion in vitro and cementogenesis in vivo were studied. In vitro, cementoblastic adhesion improved significantly when the dentin surface contained covalently bound peptides. In vivo, this bound peptide significantly increased cementum formation compared with that attained in control conditions. Transglutaminase-catalyzed covalent binding of bioactive peptides targeted to mineralized collagenous dentin matrix via the poly-glutamate domain can be readily achieved. This approach offers potential for clinical use in periodontal regeneration.

Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo

The dental follicle is a mesenchymal tissue that surrounds the developing tooth germ. During tooth root formation, periodontal components, viz., cementum, periodontal ligament (PDL), and alveolar bone, are created by dental follicle progenitors. Here, we report the presence of PDL progenitors in mouse dental follicle (MDF) cells. MDF cells were obtained from mouse incisor tooth germs and immortalized by the expression of a mutant human papilloma virus type 16 E6 gene lacking the PDZ-domain-binding motif. MDF cells expressing the mutant E6 gene (MDF( E6-EGFP ) cells) had an extended life span, beyond 150 population doublings (PD). In contrast, normal MDF cells failed to proliferate beyond 10 PD. MDF( E6-EGFP ) cells expressed tendon/ligament phenotype-related genes such as Scleraxis (Scx), growth and differentiation factor-5, EphA4, Six-1, and type I collagen. In addition, the expression of periostin was observed. To elucidate the differentiation capacity of MDF( E6-EGFP ) cells in vivo, the cells were transplanted into severe combined immunodeficiency mice. At 4 weeks, MDF( E6-EGFP ) cell transplants had the capacity to generate a PDL-like tissue that expressed periostin, Scx, and type XII collagen and the fibrillar assembly of type I collagen. Our findings suggest that MDF( E6-EGFP ) cells can act as PDL progenitors, and that these cells may be a useful research tool for studying PDL formation and for developing regeneration therapies.