Dental Follicle Stem Cells Promote Periodontal Regeneration through Periostin-Mediated Macrophage Infiltration and Reprogramming in an Inflammatory Microenvironment.

Dental follicle stem cells (DFSCs) have been verified to promote periodontal regeneration in an inflammatory microenvironment. When coping with inflammatory stimulation, DFSCs highly express periostin, a bioactive molecule closely related to periodontal homeostasis. It is worth exploring whether and how periostin plays a role in the promotion of periodontal regeneration by DFSCs. By tracking the fate of DFSCs, it was found that DFSCs significantly contributed to periodontal regeneration in rat periodontal defects while they had a low survival rate. They highly expressed periostin and improved the immune microenvironment in the defect area, especially via the recruitment and reprogramming of macrophages. Silencing periostin attenuated the effects of DFSCs in promoting periodontal regeneration and regulating macrophages. Recombinant human periostin (rhPeriostin) could not only directly promote macrophage reprogramming through the integrin αM/phosphorylated extracellular signal-regulated kinase (p-Erk)/Erk signaling pathway, but it also exhibited the potential to promote periodontal regeneration in rats when loaded in a collagen matrix. These results indicated that periostin is actively involved in the process by which DFSCs promote periodontal regeneration through the regulation of macrophages and is a promising molecular agent to promote periodontal regeneration. This study provides new insight into the mechanism by which DFSCs promote periodontal regeneration and suggests a new approach for periodontal regeneration therapy.

Effects of Cellular Senescence on Dental Follicle Cells.

The dental follicle is part of the tooth germ, and isolated stem cells from this tissue (dental follicle cells; DFCs) are considered, for example, for regenerative medicine and immunotherapies. However somatic stem cells can also improve pharmaceutical research. Cell proliferation is limited by the induction of senescence, which, while reducing the therapeutic potential of DFCs for cell therapy, can also be used to study aging processes at the cellular level that can be used to test anti-aging pharmaceuticals. Unfortunately, very little is known about cellular senescence in DFCs. This review presents current knowledge about cellular senescence in DFCs.

Effects of aging on DNA hydroxymethylation and methylation in human dental follicles.

OBJECTIVE: Despite the high frequency of impacted teeth and increased frequency of lesions in dental follicles (DF) with aging, DF age-changes remain unclear. We compared the global methylation and hydroxymethylation profiles in DF in relation to age. DESIGN: DF associated with impacted lower third molars were obtained from 59 individuals. Global DNA methylation (5mC content) and hydroxymethylation (5hmC) were evaluated by ELISA. We tested the correlation between 5mC and 5hmC content, and the correlation of each with patients' age. The differences in age, 5mC, and 5hmC in DF from men/women, and location (left/right mandible) was tested. RESULTS: The mean age of the 59 individuals was 19.56 ±â€¯3.92, ranging from 13 to 31 years, and most were women (n = 39). 5hmC content and age up to 19 years were inversely correlated (Spearman's correlation coefficient=-0.552, p = 0.0003, n = 38). There was no relationship between 5hmC and 5mC content. There was no difference in the medians of age (p = 0.25), 5hmC (p = 0.33) and 5mC (p = 0.86) between men/women, nor in the medians of age (p = 0.39), 5hmC (p = 0.99) and 5mC (p = 0.22) between the left/right side of the tooth extraction. CONCLUSION: An inverse correlation between 5hmC and age was established, with no correlation between 5mC and 5hmC content in DF. The biological meaning of such a decrease of global DNA hydroxymethylation with age in DF remains to be clarified.

Stem cells from human exfoliated deciduous teeth as an alternative cell source in bio-root regeneration.

A stem cell-mediated bioengineered tooth root (bio-root) has proven to be a prospective tool for the treatment of tooth loss. As shown in our previous studies, dental follicle cells (DFCs) are suitable seeding cells for the construction of bio-roots. However, the DFCs which can only be obtained from unerupted tooth germ are restricted. Stem cells from human exfoliated deciduous teeth (SHEDs), which are harvested much more easily through a minimally invasive procedure, may be used as an alternative seeding cell. In this case, we compared the odontogenic characteristics of DFCs and SHEDs in bio-root regeneration. Methods: The biological characteristics of SHEDs and DFCs were determined in vitro. The cells were then induced to secrete abundant extracellular matrix (ECM) and form macroscopic cell sheets. We combined the cell sheets with treated dentin matrix (TDM) for subcutaneous transplantation into nude mice and orthotopic jaw bone implantation in Sprague-Dawley rats to further verify their regenerative potential. Results: DFCs exhibited a higher proliferation rate and stronger osteogenesis and adipogenesis capacities, while SHEDs displayed increased migration ability and excellent neurogenic potential. Both dental follicle cell sheets (DFCSs) and sheets of stem cells from human exfoliated deciduous teeth (SHEDSs) expressed not only ECM proteins but also osteogenic and odontogenic proteins. Importantly, similar to DFCSs/TDM, SHEDSs/TDM also successfully achieved the in vivo regeneration of the periodontal tissues, which consist of periodontal ligament fibers, blood vessels and new born alveolar bone. Conclusions: Both SHEDs and DFCs possessed a similar odontogenic differentiation capacity in vivo, and SHEDs were regarded as a prospective seeding cell for use in bio-root regeneration in the future.

Recent Advances of Useful Cell Sources in the Periodontal Regeneration.

BACKGROUND: Periodontitis is an inflammatory disease that can result in destruction of the tooth attachment apparatus. Therefore, periodontal tissue regeneration is currently an important focus of research in the field. Approaches using stem cells and reprogrammed cells, such as induced pluripotent stem cells (iPSCs) or trans-differentiated cells, represent the cutting edge in periodontal regeneration, and have led to many trials for their clinical application. Objectives and Results: In this review, we consider all available stem cell sources, methods to obtain the cells, their capability to differentiate into the desired cells, and the extent of their utilization in periodontal regeneration. In addition, we introduce the new concepts of using iPSCs and transdifferentiated cells for periodontal regeneration. Finally, we discuss the promise of tissue engineering for improving cell therapy outcomes for periodontal regeneration. CONCLUSIONS: Despite their limitations, iPSCs and trans-differentiated cells may be promising cell sources for periodontal tissue regeneration. Further collaborative investigation is required for the effective and safe application of these cells in combination with tissue engineering elements, like scaffolds and biosignals.

Isolation, Characterization and MicroRNA-based Genetic Modification of Human Dental Follicle Stem Cells.

To date, several stem cell types at different developmental stages are in the focus for the treatment of degenerative diseases. Yet, certain aspects, such as initial massive cell death and low therapeutic effects, impaired their broad clinical translation. Genetic engineering of stem cells prior to transplantation emerged as a promising method to optimize therapeutic stem cell effects. However, safe and efficient gene delivery systems are still lacking. Therefore, the development of suitable methods may provide an approach to resolve current challenges in stem cell-based therapies. The present protocol describes the extraction and characterization of human dental follicle stem cells (hDFSCs) as well as their non-viral genetic modification. The postnatal dental follicle unveiled as a promising and easily accessible source for harvesting adult multipotent stem cells possessing high proliferation potential. The described isolation procedure presents a simple and reliable method to harvest hDFSCs from impacted wisdom teeth. Also this protocol comprises methods to define stem cell characteristics of isolated cells. For genetic engineering of hDFSCs, an optimized cationic lipid-based transfection strategy is presented enabling highly efficient microRNA introduction without causing cytotoxic effects. MicroRNAs are suitable candidates for transient cell manipulation, as these small translational regulators control the fate and behavior of stem cells without the hazard of stable genome integration. Thus, this protocol represents a safe and efficient procedure for engineering of hDFSCs that may become important for optimizing their therapeutic efficacy.

Gene expression profiles in dental follicles from patients with impacted canines.

Animal studies suggest that the dental follicle (DF) plays a major role in tooth eruption. However, the role of the DF during tooth impaction and related root resorptions in adjacent teeth is not clear. The hypothesis for the present study is that expression of regulatory factors involved in the bone remodelling process necessary for tooth eruption may differ between dental follicles from teeth with different clinical situations. We have analysed the gene expression profiles in the DF obtained from impacted canines, with (N = 3) or without (N = 5) signs of root resorption, and from control teeth (normal erupting teeth, mesiodens) (N = 3). DF from 11 patients (mean age: 13 years) obtains at the time of surgical exposure of the tooth. Due to the surgical time point, all teeth were in a late developmental stage. Gene expression related to osteoblast activation/bone formation, osteoclast recruitment and activation was analysed by RTqPCR. Genes related to bone formation (RUNX2, OSX, ALP, OCN, CX43) were highly expressed in all the samples, but osteoclast recruitment/activation markers (OPG, RANKL, MCP-1, CSF-1) were negligible. No apparent patterns or significant differences in gene expression were found between impacted canines, with or without signs of root resorption, or when compared to control teeth. Our results suggest the DF regulation of osteoclastic activity is limited in the late pre-emergent stage of tooth development, irrespective if the tooth is normally erupting or impacted. We suggest that the follicle may have an important regulatory function for alveolar bone formation in the final eruption process and CX43-gap junction communication could be an important signalling pathway.

Dental Tissue-Derived Mesenchymal Stem Cells: Applications in Tissue Engineering.

In recent years, mesenchymal stem cells (MSCs) derived from dental tissue have gained in popularity for tissue-engineering and regenerative medicine applications. The highly proliferative and self-renewing population of dental stem cells has the neural crest as their origin. This expands their applicability for regeneration of tissues from both ectochyme and mesenchymal origin. Ease of tissue harvest, high initial yield of cells, low population-doubling time, plasticity, multipotential capabilities, and immunomodulatory properties make them a suitable candidate for various therapeutic strategies. Furthermore, dental tissue-derived cells can be transformed into induced pluripotent stem cells to customize cell-based regenerative approaches. However, there is currently a lack of exhaustive comparative profiles of these dental tissues and their regenerative applications. We thereby present a comprehensive compilation of morphofunctional analyses and tissue-engineering applications of MSCs that are derived from tooth germ, exfoliated deciduous teeth, periodontal ligament, gingiva, dental pulp, alveolar bone, dental follicle, and apical papilla. Immunoregulatory properties of dental stem cells provide potential for both autologous and allogenic tissue-engineering approaches. In vitro and animal studies show promise for using dental stem cells in regenerative medicine. Eventually, the orchestration of clinical trials will require systematic monitoring of spontaneous in vitro transformations and complications associated with graft versus host response as well as a thorough understanding of underlying anabolic mechanisms.

RUNX2 Mutation Impairs 1α,25-Dihydroxyvitamin D3 mediated Osteoclastogenesis in Dental Follicle Cells.

Cleidocranial dysplasia (CCD), a skeletal disorder characterized by delayed permanent tooth eruption and other dental abnormalities, is caused by heterozygous RUNX2 mutations. As an osteoblast-specific transcription factor, RUNX2 plays a role in bone remodeling, tooth formation and tooth eruption. To investigate the crosstalk between RUNX2 and 1α,25-dihydroxyvitamin D3 (1α,25-(OH)2D3) in human dental follicle cells (hDFCs) during osteoclast formation, we established a co-culture system of hDFCs from CCD patient and healthy donors with peripheral blood mononuclear cells (PBMCs). Expression of the osteoclast-associated genes and the number of TRAP(+) cells were reduced in CCD hDFCs, indicating its suppressed osteoclast-inductive ability, which was reflected by the downregulated RANKL/OPG ratio. In addition, 1α,25-(OH)2D3-stimulation elevated the expression of osteoclast-related genes, as well as RANKL mRNA levels and RANKL/OPG ratios in control hDFCs. Conversely, RUNX2 mutation abolished this 1α,25-(OH)2D3-induced RANKL gene activation and osteoclast formation in CCD hDFCs. Therefore, RUNX2 haploinsufficiency impairs dental follicle-induced osteoclast formation capacity through RANKL/OPG signaling, which may be partially responsible for delayed permanent tooth eruption in CCD patients. Furthermore, this abnormality was not rescued by 1α,25-(OH)2D3 application because 1α,25-(OH)2D3-induced RANKL activation in hDFCs is mediated principally via the RUNX2-dependent pathway.

Orthodontic movement in deciduous teeth.

Deciduous teeth exfoliate as a result of apoptosis induced by cementoblasts, a process that reveals the mineralized portion of the root while attracting clasts. Root resorption in deciduous teeth is slow due to lack of mediators necessary to speed it up; however, it accelerates and spreads in one single direction whenever a permanent tooth pericoronal follicle, rich in epithelial growth factor (EGF), or other bone resorption mediators come near. The latter are responsible for bone resorption during eruption, and deciduous teeth root resorption and exfoliation. Should deciduous teeth be subjected to orthodontic movement or anchorage, mediators local levels will increase. Thus, one should be fully aware that root resorption in deciduous teeth will speed up and exfoliation will early occur. Treatment planning involving deciduous teeth orthodontic movement and/or anchorage should consider: Are clinical benefits relevant enough as to be worth the risk of undergoing early inconvenient root resorption?

Orthodontic movement in deciduous teeth

Deciduous teeth exfoliate as a result of apoptosis induced by cementoblasts, a process that reveals the mineralized portion of the root while attracting clasts. Root resorption in deciduous teeth is slow due to lack of mediators necessary to speed it up; however, it accelerates and spreads in one single direction whenever a permanent tooth pericoronal follicle, rich in epithelial growth factor (EGF), or other bone resorption mediators come near. The latter are responsible for bone resorption during eruption, and deciduous teeth root resorption and exfoliation. Should deciduous teeth be subjected to orthodontic movement or anchorage, mediators local levels will increase. Thus, one should be fully aware that root resorption in deciduous teeth will speed up and exfoliation will early occur. Treatment planning involving deciduous teeth orthodontic movement and/or anchorage should consider: Are clinical benefits relevant enough as to be worth the risk of undergoing early inconvenient root resorption?.

O dente decíduo é esfoliado graças à apoptose em seus cementoblastos, que desnuda a parte mineralizada da raiz e atrai os clastos. A rizólise é lenta, pois faltam mediadores em quantidade para acelerar o processo, mas ela se acelera e unidireciona quando se aproxima um folículo pericoronário de dente permanente rico em EGF e outros mediadores da reabsorção óssea - os responsáveis pelas reabsorções óssea na erupção e dentária decídua na rizólise e esfoliação. Se houver movimentação ortodôntica ou ancoragem em dentes decíduos, aumenta-se, também, o nível local desses mesmos mediadores, devendo-se estar bem consciente de que haverá uma aceleração da rizólise e, em decorrência, uma antecipação de sua esfoliação. No planejamento de casos em que dentes decíduos estejam envolvidos na movimentação ortodôntica e/ou ancoragem, deve-se ponderar: o benefício clínico para o paciente será relevante, a ponto de valer o risco de uma rizólise abreviada e inconveniente?.

A therapeutic strategy for spinal cord defect: human dental follicle cells combined with aligned PCL/PLGA electrospun material.

Stem cell implantation has been utilized for the repair of spinal cord injury; however, it shows unsatisfactory performance in repairing large scale lesion of an organ. We hypothesized that dental follicle cells (DFCs), which possess multipotential capability, could reconstruct spinal cord defect (SCD) in combination with biomaterials. In the present study, mesenchymal and neurogenic lineage characteristics of human DFCs (hDFCs) were identified. Aligned electrospun PCL/PLGA material (AEM) was fabricated and it would not lead to cytotoxic reaction; furthermore, hDFCs could stretch along the oriented fibers and proliferate efficiently on AEM. Subsequently, hDFCs seeded AEM was transplanted to restore the defect in rat spinal cord. Functional observation was performed but results showed no statistical significance. The following histologic analyses proved that AEM allowed nerve fibers to pass through, and implanted hDFCs could express oligodendrogenic lineage maker Olig2 in vivo which was able to contribute to remyelination. Therefore, we concluded that hDFCs can be a candidate resource in neural regeneration. Aligned electrospun fibers can support spinal cord structure and induce cell/tissue polarity. This strategy can be considered as alternative proposals for the SCD regeneration studies.

Functional tooth restoration by next-generation bio-hybrid implant as a bio-hybrid artificial organ replacement therapy.

Bio-hybrid artificial organs are an attractive concept to restore organ function through precise biological cooperation with surrounding tissues in vivo. However, in bio-hybrid artificial organs, an artificial organ with fibrous connective tissues, including muscles, tendons and ligaments, has not been developed. Here, we have enveloped with embryonic dental follicle tissue around a HA-coated dental implant, and transplanted into the lower first molar region of a murine tooth-loss model. We successfully developed a novel fibrous connected tooth implant using a HA-coated dental implant and dental follicle stem cells as a bio-hybrid organ. This bio-hybrid implant restored physiological functions, including bone remodelling, regeneration of severe bone-defect and responsiveness to noxious stimuli, through regeneration with periodontal tissues, such as periodontal ligament and cementum. Thus, this study represents the potential for a next-generation bio-hybrid implant for tooth loss as a future bio-hybrid artificial organ replacement therapy.

Tooth eruption results from bone remodelling driven by bite forces sensed by soft tissue dental follicles: a finite element analysis.

Intermittent tongue, lip and cheek forces influence precise tooth position, so we here examine the possibility that tissue remodelling driven by functional bite-force-induced jaw-strain accounts for tooth eruption. Notably, although a separate true 'eruptive force' is widely assumed, there is little direct evidence for such a force. We constructed a three dimensional finite element model from axial computerized tomography of an 8 year old child mandible containing 12 erupted and 8 unerupted teeth. Tissues modelled included: cortical bone, cancellous bone, soft tissue dental follicle, periodontal ligament, enamel, dentine, pulp and articular cartilage. Strain and hydrostatic stress during incisive and unilateral molar bite force were modelled, with force applied via medial and lateral pterygoid, temporalis, masseter and digastric muscles. Strain was maximal in the soft tissue follicle as opposed to surrounding bone, consistent with follicle as an effective mechanosensor. Initial numerical analysis of dental follicle soft tissue overlying crowns and beneath the roots of unerupted teeth was of volume and hydrostatic stress. To numerically evaluate biological significance of differing hydrostatic stress levels normalized for variable finite element volume, 'biological response units' in Nmm were defined and calculated by multiplication of hydrostatic stress and volume for each finite element. Graphical representations revealed similar overall responses for individual teeth regardless if incisive or right molar bite force was studied. There was general compression in the soft tissues over crowns of most unerupted teeth, and general tension in the soft tissues beneath roots. Not conforming to this pattern were the unerupted second molars, which do not erupt at this developmental stage. Data support a new hypothesis for tooth eruption, in which the follicular soft tissues detect bite-force-induced bone-strain, and direct bone remodelling at the inner surface of the surrounding bony crypt, with the effect of enabling tooth eruption into the mouth.

Inferior alveolar nerve block and third-molar agenesis: a retrospective clinical study.

BACKGROUND: Children often receive inferior alveolar nerve blocks (IANBs) when their third molars are just beginning to develop. The location of the third-molar follicle is close to where the needle penetrates during an IANB. The authors examined the possible association between IANBs and missing third molars. METHODS: The authors examined 439 potential sites of third-molar development for evidence of third-molar follicles on panoramic radiographs of randomly selected children 7 years and older. The authors conducted a statistical comparison of the incidence of missing third-molar follicles in a control group of children who had no history of receiving IANBs with children in a test group who had a definitive history of receiving IANBs by means of generalized estimating equations. RESULTS: The authors found a statistically significant greater incidence of missing third-molar follicles in mandibular quadrants that had a definitive history of receiving IANBs compared with mandibular quadrants that had no history of receiving IANB. CONCLUSION: IANBs administered to young children when the third-molar tooth bud is immature may stop third-molar development. Owing to the significant clinical implications, further research is needed to verify these results. PRACTICAL IMPLICATIONS: Dentists inadvertently may be stopping the development of third molars when administering IANBs to children.

Dental follicle cells and cementoblasts induce apoptosis of ameloblast-lineage and Hertwig's epithelial root sheath/epithelial rests of Malassez cells through the Fas-Fas ligand pathway.

Hertwig's epithelial root sheath (HERS), epithelial rests of Malassez (ERM) cells, and reduced ameloblasts undergo apoptosis during tooth development. This study examined the effects of dental follicle cells and cementoblasts on the apoptosis of ameloblast-lineage and HERS/ERM cells derived from the enamel organ. We also elucidated the induction pathways and identified the apoptotic pathway involved in this process. Here, we showed terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick-end labeling (TUNEL)-positive HERS cells and reduced ameloblasts near dental follicle cells during tooth development. Co-culturing ameloblast-lineage cell line (ALC) ameloblasts and HERS/ERM cells with either dental follicle cells or OCCM-30 cementoblasts markedly enhanced the apoptosis of ameloblasts and HERS/ERM cells compared with cells cultured alone. However, dental follicle cells and cementoblasts did not modulate the apoptotic responses of co-cultured non-odontogenic MCF10A or KB cells. When ameloblasts + HERS and cementoblasts + dental follicle cells were co-cultured, the expression of Fas ligand (FasL) increased in cementoblasts + dental follicle cells, while the expression of Fas increased in ameloblasts + HERS. Interestingly, recombinant FasL induced ameloblast apoptosis while the cementoblast-induced ameloblast apoptosis was suppressed by the Fas/FasL antagonist Kp7-6. These results suggest that during tooth development, dental follicle cells and cementoblasts induce apoptosis of ameloblast-lineage and HERS/ERM cells through the Fas-FasL pathway, but do not induce the apoptosis of non-odontogenic epithelial cells.

Requirement of alveolar bone formation for eruption of rat molars.

Tooth eruption is a localized event that requires a dental follicle (DF) to regulate the resorption of alveolar bone to form an eruption pathway. During the intra-osseous phase of eruption, the tooth moves through this pathway. The mechanism or motive force that propels the tooth through this pathway is controversial but many studies have shown that alveolar bone growth at the base of the crypt occurs during eruption. To determine if this bone growth (osteogenesis) was causal, experiments were designed in which the expression of an osteogenic gene in the DF, bone morphogenetic protein-6 (Bmp6), was inhibited by injection of the first mandibular molar of the rat with a small interfering RNA (siRNA) targeted against Bmp6. The injection was followed by electroporation to promote uptake of the siRNA. In 45 first molars injected, eruption was either delayed or completely inhibited (seven molars). In the impacted molars, an eruption pathway formed but bone growth at the base of the crypt was greatly reduced compared with the erupted first-molar controls. These studies show that alveolar bone growth at the base of the crypt is required for tooth eruption and that Bmp6 may be essential for promoting this growth.

Influence of ADAM28 on biological characteristics of human dental follicle cells.

OBJECTIVES: The aim of this study was to investigate the effects of a disintegrin and metalloproteinase 28 (ADAM28) on the biological characteristics of human dental follicle cells (HDFCs) and possible action mechanism. METHODS: Eukaryotic expression plasmid containing ADAM28 coding region and ADAM28 antisense oligodeoxynucleotides (AS-ODN) with FITC labelling were constructed and synthesised by gene clone and recombination. Then we respectively transfected them into HDFCs by Lipofectamine 2000 system and detected their effects on proliferation, differentiation and apoptosis of HDFCs by MTT assay, cell cycle detection, ALP activity and Annexin V-FITC/PI analysis. Finally we observed the effects of ADAM28 AS-ODN on HDFCs expressing extracellular matrix (ECM) proteins by immunocytochemical staining. RESULTS: ADAM28 eukaryotic plasmid was constructed and identified successfully, and could be correctly translated and expressed in HDFCs, furthermore overexpression of ADAM28 promoted the HDFCs proliferation and inhibited specific differentiation of HDFCs, while inhibition of ADAM28 exerted the opposite effects and induced apoptosis. Moreover ADAM28 could significantly inhibit the secretion of OPN and type III collagen of HDFCs. CONCLUSIONS: ADAM28 might actively participate in the network regulation which associates HDFCs proliferation, differentiation, apoptosis with matrix mineralisation during tooth development by interacting with multiple signal molecules.

Fate of HERS during tooth root development.

Tooth root development begins after the completion of crown formation in mammals. Previous studies have shown that Hertwig's epithelial root sheath (HERS) plays an important role in root development, but the fate of HERS has remained unknown. In order to investigate the morphological fate and analyze the dynamic movement of HERS cells in vivo, we generated K14-Cre;R26R mice. HERS cells are detectable on the surface of the root throughout root formation and do not disappear. Most of the HERS cells are attached to the surface of the cementum, and others separate to become the epithelial rest of Malassez. HERS cells secrete extracellular matrix components onto the surface of the dentin before dental follicle cells penetrate the HERS network to contact dentin. HERS cells also participate in the cementum development and may differentiate into cementocytes. During root development, the HERS is not interrupted, and instead the HERS cells continue to communicate with each other through the network structure. Furthermore, HERS cells interact with cranial neural crest derived mesenchyme to guide root development. Taken together, the network of HERS cells is crucial for tooth root development.

Cellular and molecular basis of tooth eruption.

OBJECTIVES: Tooth eruption requires the presence of a dental follicle (DF), alveolar bone resorption for an eruption pathway, and alveolar bone formation at the base of the bony crypt. The objectives of our investigations have been to determine how the DF regulates both the osteoclastogenesis and osteogenesis needed for eruption. MATERIAL AND METHODS: Multiple experimental methods have been employed. RESULTS: The DF regulates osteoclastogenesis and osteogenesis by regulating the expression of critical genes in both a chronological and spatial fashion. In the rat 1st mandibular molar there is a major burst of osteoclastogenesis at day 3 postnatally and a minor burst at day 10. At day 3, the DF maximally expresses colony-stimulating factor-1 (CSF-1) to down-regulate the expression of osteoprotegerin (OPG) such that osteoclastogenesis can occur. At day 10, the minor burst of osteoclastogenesis is promoted by upregulation of vascular endothelial growth factor (VEGF) and RANKL in the DF. Spatially, the bone resorption is in the coronal portion of the bony crypt and genes such as RANKL are expressed more in the coronal region of the DF than in its basal one-half. For osteogenesis, bone formation begins at day 3 at the base of the bony crypt and maximal growth is at days 9-14. Osteo-inductive genes such as bone morphogenetic protein-2 (BMP-2) appear to promote this and are expressed more in the basal half of the DF than in the coronal. Conclusion - The osteoclastogenesis and osteogenesis needed for eruption are regulated by differential gene expression in the DF both chronologically and spatially.