Evolution, development, and regeneration of tooth-like epithelial appendages in sharks.

Sharks and their relatives are typically covered in highly specialized epithelial appendages embedded in the skin called dermal denticles; ancient tooth-like units (odontodes) composed of dentine and enamel-like tissues. These 'skin teeth' are remarkably similar to oral teeth of vertebrates and share comparable morphological and genetic signatures. Here we review the histological and morphological data from embryonic sharks to uncover characters that unite all tooth-like elements (odontodes), including teeth and skin denticles in sharks. In addition, we review the differences between the skin and oral odontodes that reflect their varied capacity for renewal. Our observations have begun to decipher the developmental and genetic shifts that separate these seemingly similar dental units, including elements of the regenerative nature in both oral teeth and the emerging skin denticles from the small-spotted catshark (Scyliorhinus canicula) and other chondrichthyan models. Ultimately, we ask what defines a tooth at both the molecular and morphological level. These insights aim to help us understand how nature makes, replaces and evolves a vast array of odontodes.

Six1 Regulates Mouse Incisor Development by Promoting Dlx1/2/5 Expression.

Tooth development is a complex process orchestrated by intricate gene regulatory networks, involving both odontogenic epithelium and ectomesenchyme. Six1, a pivotal transcription factor (TF), is involved in the development of the lower incisor. However, its precise role during incisor development and the molecular mechanisms underpinning its regulatory functions remain poorly understood. This study employs Six1 deletion mouse models to elucidate the critical regulatory role of Six1 in governing dental mesenchyme development. By performing single-cell RNA sequencing, we constructed a comprehensive transcriptome atlas of tooth germ development from the bud to bell stage. Our analyses suggest that the dental follicle and the dental papilla (DP) are differentiated from dental ectomesenchyme (DEM) and identify the key TFs underlying these distinct states. Notably, we show that Dlx1, Dlx2, and Dlx5 (Dlx1/2/5) may function as the key TFs that promote the formation of DP. We further show that the deletion of Six1 perturbs dental mesenchyme development by impeding the transitions from DEM to DP states. Importantly, SIX1 directly binds to the promoters of Dlx1/2/5 to promote their co-expression, which subsequently leads to widespread epigenetic and transcriptional remodeling. In summary, our findings unveil Six1's indispensable role in incisor development, offering key insights into TF-driven regulatory networks that govern dental mesenchyme cell fate transitions during tooth development.

Testing the patterning cascade model of cusp development in Macaca fascicularis mandibular molars.

OBJECTIVE: Molar crown configuration plays an important role in systematics, and functional and comparative morphology. In particular, the number of cusps on primate molars is often used to identify fossil species and infer their phylogenetic relationships. However, this variability deserves renewed consideration as a number of studies now highlight important developmental mechanisms that may be responsible for the presence of molar cusps in some mammalian taxa. Experimental studies of rodent molars suggest that cusps form under a morphodynamic, patterning cascade model of development (PCM) that involve the iterative formation of enamel knots. This model posits that the size, shape and location of the first-forming cusps determines the presence and positioning of later-forming cusps. DESIGN: Here we test whether variation in accessory cusp presence in 13 Macaca fascicularis mandibular second molars (M2s) is consistent with predictions of the PCM. Using micro-CT, we imaged these M2s and employed geometric morphometrics to examine whether shape variation in the enamel-dentine junction (EDJ) correlates with accessory cusp presence. RESULTS: We find that accessory cusp patterning in macaque M2s is broadly consistent with the PCM. Molars with accessory cusps were larger in size and possessed shorter relative cusp heights compared to molars without accessory cusps. Peripheral cusp formation was also associated with more centrally positioned primary cusps, as predicted by the PCM. CONCLUSIONS: While these results demonstrate that a patterning cascade model is broadly appropriate for interpreting cusp variation in Macaca fascicularis molars, it does not explain all manifestations of accessory cusp expression in this sample.

Secular changes in dental development of Korean children aged 4 to 16 years over a 10-year period.

This study evaluated 10-year secular changes in dental maturity and dental development among Korean children. A retrospective analysis of panoramic radiograph samples from Korean children (4-16 years old) taken in 2010 and 2020 was conducted. The 2010 group consisted of 3491 radiographs (1970 boys and 1521 girls), and the 2020 group included 5133 radiographs (2825 boys and 2308 girls). Using Demirjian's method, dental maturity scores and dental developmental stages were assessed. For intra-observer reliability, Weighted Cohen's kappa was used, and Mann-Whitney U tests were performed to compare the 2020 and 2010 groups. A slight acceleration in dental maturity was observed in both boys and girls, with the difference being more noticeable in boys at an earlier age. Statistically significant differences were noted at ages 4, 5 and 7 for boys, and at age 6 for girls. Despite these differences, the individual dental development stages of 2020 and 2010 showed inconsistent trends with limited differences. Generally, girls demonstrate more advanced dental maturity than boys. A slight acceleration in Korean children's dental maturity was observed over a 10-year period when comparing the 2020 groups to the 2010 groups.

Resilience of the replacing dentition in adult reptiles.

The dentition is critical to animal survival and teeth are present in modern vertebrates including teleost fish, sharks, amphibians, mammals and reptiles. The developmental processes that give rise to teeth are not just preserved through evolution but also share high level of similarity with the embryogenesis of other ectodermal organs. In this review we go beyond the embryonic phase of tooth development to life-long tooth replacement. We will address the origins of successional teeth, the location of putative tissue-resident stem cells, how de novo tooth formation continues throughout life and how teeth are shed in a spatially and temporally controlled manner. We review the evidence that the dental epithelium, which is the earliest recognizable dental structure in the reptilian dentition, serves as a putative niche for tissue-resident epithelial stem cells and recent molecular findings from transcriptomics carried out in reptilian dentitions. We discuss how odontoclasts resorb the primary tooth allowing eruption of the successional tooth. The reptiles, particularly lizards, are emerging as some of the most accessible animals to study tooth replacement which has relevance to evolution of the dentition and human dental disorders.

Spatiotemporal regulation of dental pulpal innervation in the rat.

The dental pulp is a highly innervated tissue transmitting pain-related sensations in the tooth. Consequently, understanding the intricacies of its innervation mechanism in odontogenesis is crucial for gaining insights into dental pain and developing dental pain-modulating agents. This study examined neuroregulatory molecules such as neurotrophic factors (nerve growth factor [NGF], brain-derived neurotrophic factor [BDNF], neurotrophin-4 [NTF-4], and neurturin [NRTN]) and neuroinhibitory factors (slit2, ephrin isoforms and netrin-1) in developing rat teeth with follicles. NGF, BDNF and NRTN transcriptions showed time-dependent upregulation, particularly during the root formation stage. In contrast, NTF-4 mRNA was highly expressed at the cap stage, but became downregulated over time. Slit2 and ephrin-B2 expression was distinct at the cap stage and then downregulated in a time-dependent manner. Ephrin-A5 and netrin-1 expression did not significantly change. Immunofluorescence analysis revealed a robust expression of both ephrin-B2 and slit2 in the outer and inner dental epithelia of the enamel organ, a non-neurogenic tissue, during the cap stage of 3rd molar germs. In contrast, BDNF was predominantly localized in dental papilla cells and odontoblasts during the root formation stage. These results suggest that neuroregulatory molecules, such as BDNF, slit2 and ephrin-B2, may be important in identifying therapeutic targets for modulating dental pulp pain.

The ectopic mandibular canines can start tooth formation in three different locations: a case series study based on single orthopantomograms from 47 individuals.

INTRODUCTION: A former study on orthopantomograms from young children with abnormal dental development (not canine ectopia) demonstrated that the tooth bud of the mandibular canine, compared to a stable longitudinal canine axis, could be located normally, anteriorly or posteriorly, with close relation to the first premolar. AIM: The aim of the present study is to analyse on orthopantomograms if the canine axis can demonstrate where the ectopic mandibular canine started tooth formation. MATERIALS: The material consists of orthopantomograms with ectopic mandibular canines and presence of primary mandibular canines from 47 cases (29 cases 9-21 years old and 18 cases with unknown ages). The primary canines demonstrated from minor apical resorption to more severe apical resorption. METHODS: Based on canine maturity, location of the canine axes and the interrelationships between the roots of the permanent canine and first premolar, the location from where the canine started tooth formation was determined. Canine maturity. Maturity stage below half root length and maturity stage above half root length revealed that 11 ectopic canines had less than half root length and 36 cases more than half root length. Canine axes. The canine axis, through the length of the primary canines Ax, is inserted on drawings of the orthopantomograms using the tracing programme Inkscape®. Interrelationship between roots. By visual inspection, the distance between the canine and first premolar was designated close distance, normal distance and extended distance. RESULTS: The results are divided into 3 groups. Group 1: The initial site of the permanent ectopic canine is located within the canine axis (6 cases). Group 2: The initial site of the permanent ectopic canine is located posterior to the canine axis (36 cases). Group 3: The initial site of the permanent ectopic canine is located anterior to the canine axis (5 cases). CONCLUSION: The study explained that the canine axis could divide cases of ectopic canines into three groups according to the location from where tooth formation starts. For getting closer to the pattern of the ectopic canine eruption, it is necessary to analyse series of orthopantomograms taken from the same individual over several years.

Histone-modifying enzymes: Roles in odontogenesis and beyond.

OBJECTIVES: Odontogenesis, an intricate process initiated by epithelium-mesenchyme interaction, is meticulously regulated by a cascade of regulatory mechanisms. Epigenetic modifications, especially histone modification, have been found to exhibit spatiotemporal specificity during tooth development. However, the expression patterns and roles of enzymes associated with histone modifications have yet to be systematically explored in odontogenesis. This review aims to summarize the histone-modifying enzymes in odontogenesis and their regulation mechanism during tooth development and provide the potential theoretical basis for the clinical management and intervention of dental developmental diseases. SUBJECTS AND METHODS: This study conducted a systematic search across PubMed and Web of Science databases, utilizing the keywords "odontogenesis," "histone modification," and "enzyme" for pertinent articles. RESULTS: No doubt histone modification contributes extensively to odontogenesis regulation, and the disturbances in histone modifications can derange the odontogenesis process. CONCLUSION: Further studies are warranted to elucidate these roles and their potential downstream effects, positioning histone modifications as a pivotal focal point for unraveling the intricacies of tooth development and regeneration.

Roles of SP/KLF transcription factors in odontoblast differentiation: From development to diseases.

OBJECTIVES: A zinc-finger transcription factor family comprising specificity proteins (SPs) and Krüppel-like factor proteins (KLFs) plays an important role in dentin development and regeneration. However, a systematic regulatory network involving SPs/KLFs in odontoblast differentiation has not yet been described. This review examined the expression patterns of SP/KLF gene family members and their current known functions and mechanisms in odontoblast differentiation, and discussed prospective research directions for further exploration of mechanisms involving the SP/KLF gene family in dentin development. MATERIALS AND METHODS: Relevant literature on SP/KLF gene family members and dentin development was acquired from PubMed and Web of Science. RESULTS: We discuss the expression patterns, functions, and related mechanisms of eight members of the SP/KLF gene family in dentin development and genetic disorders with dental problems. We also summarize current knowledge about their complementary or synergistic actions. Finally, we propose future research directions for investigating the mechanisms of dentin development. CONCLUSIONS: The SP/KLF gene family plays a vital role in tooth development. Studying the complex complementary or synergistic interactions between SPs/KLFs is helpful for understanding the process of odontoblast differentiation. Applications of single-cell and spatial multi-omics may provide a more complete investigation of the mechanism involved in dentin development.

Fibroblast growth factor signalling regulates the development of tooth root.

Fibroblast growth factor (FGF) signalling plays a crucial role in the morphogenesis of multiple tissues including teeth. While the role of the signal has been studied in tooth crown development, little is known about root development. Of several FGF ligands involved in hard tissue formation, we suggest that FGF18 regulates the development of murine tooth roots. We implanted FGF18-soaked heparin beads into the lower first molar tooth buds at postnatal day 6 (P6), followed by transplantation under the kidney capsule. After 3 weeks, FGF18 significantly facilitated root elongation and periodontal tissue formation compared to the control. In situ hybridisation showed that Fgf18 transcripts were initially localised in the dental pulp along Hertwig's epithelial root sheath at P6 and P10 and subsequently in the dental follicle cells at P14. Fgf receptors were expressed in various dental tissues during these stages. In vitro analysis using the dental pulp stem cells revealed that FGF18 inhibited cell proliferation and decreased expression levels of osteogenic markers, Runx2, Alpl and Sp7. Consistently, after 1 week of kidney capsule transplantation, FGF18 application did not induce the expression of Sp7 and Bsp, but upregulated Periostin in the apical region of dental mesenchyme in the grafted molar. These findings suggest that FGF18 facilitates molar root development by regulating the calcification of periodontal tissues.

Control radiográfico de trasplante dentario: reporte de caso / Radiographic control of dental transplantation: a case report

El trasplante dentario es una opción terapéutica para reemplazar un órgano dental perdido, causado por un proceso carioso extenso, agenesia, trauma-tismos o iatrogenias. Este procedimiento quirúrgico traslada un órgano dental íntegro desde un alveolo donante hacia su lecho receptor; para lo cual debe poseer ciertas características que permitan tener un pronóstico favorable a largo plazo. El presente estudio describe la evolución de un trasplante dental autólogo realizado hace 14 años a una paciente que acudió a la consulta para valoración del órgano den-tal 4.7, el que presentó un pronóstico desfavorable, por lo cual se realizó exodoncia y trasplante inme-diato del diente vital 4.8 al alveolo del órgano dental 4.7. Tras la planificación quirúrgica se procedió con la intervención conservando la vitalidad pulpar del diente a ser trasplantado, se realizó control clínico y radiográfico a los 15 días, 30 días, 6 meses, 1 año, 5 años y 14 años, en el que se observó conservación del paquete vasculonervioso y ligamento periodontal del órgano dental; a su vez se pudo evidenciar rizo-génesis en el diente trasplantado y un aumento de la altura del proceso alveolar, mediante mediciones realizadas en Auto CAD 2023 (AU)

Tooth transplantation is a therapeutic option to re-place a lost dental organ, caused by an extensive carious process, agenesis, trauma or iatrogenesis. This surgical procedure transfers a complete den-tal organ from a donor alveolus to its recipient bed; for which it must have certain characteristics that allow it to have a favorable long-term prognosis. The present study describes the evolution of an autolo-gous dental transplant carried out 14 years ago to a female patient who attended the consultation for evaluation of the dental organ 4.7, the same one that presented an unfavorable prognosis, for which an extraction and immediate transplantation of the 4.8 vital tooth was performed to the alveolus of the den-tal organ 4.7. After surgical planning, the intervention was carried out preserving the pulpal vitality of the tooth to be transplanted; clinical and radiographic control was performed at 15 days, 30 days, 6 months, 1 year, 5 years and 14 years, in which preservation of the vascular-nervous bundle and periodontal liga-ment of the dental organ was observed; in turn, rhizo-genesis in the transplanted tooth and an increase in the height of the alveolar process could be evidenced, through measurements made in Auto CAD 2023 (AU)

The Role of Histone Acetylation Modification in Dental Tissue-Derived Mesenchymal Stem Cells and Odontogenesis.

Odontogenesis is a complex physiological process that is based on dental tissue-derived mesenchymal stem cells (MSCs). Dental tissue-derived MSCs are the stem cell populations isolated and characterized from different parts of the oral cavity, and are considered as promising candidates for stem cell-based therapy. During odontogenesis, epigenetic factors can influence the proliferation, differentiation, or apoptosis of dental tissue-derived MSCs. As one of the epigenetic modifications, histone acetylation modification is critical for the proper regulation of many biological processes, including transcriptional regulation of cell cycle progression and cell fate. In odontogenesis, histone acetylation and deacetylation play crucial roles in odontogenic differentiation of dental tissue-derived MSCs. In this review, we aim to outline the general features of acetylation modification and describe their roles in odontogenic differentiation of dental tissue-derived MSCs, as well as their future implications in the field of novel regenerative therapies for the dentine-pulp complex.

Acetylsalicylic-acid (ASA) regulation of osteo/odontogenic differentiation and proliferation of human dental pulp stem cells (DPSCs) in vitro.

OBJECTIVE: The study aimed to investigate acetylsalicylic acid (ASA) effects on osteo/odontogenic differentiation and proliferation of dental pulp stem cells (DPSCs) in vitro and the potential involvement of adenosine monophosphate-activated protein kinase (AMPK) pathway in these processes. DESIGN: DPSCs were isolated from third molars pulp tissues of five patients and grown in osteogenic medium alone or supplemented with ASA. Expression of DPSCs markers was tested by flow-cytometry. Cytotoxicity of ASA at concentrations of 10, 50 and 100 µg/ml was tested by MTT and NR assays. Osteo/odontogenic differentiation was analyzed via alizarin red staining and ALP activity. Quantitative PCR (qPCR) was used for osteo/odontogenic markers' (DSPP, BMP2, BMP4, BSP, OCN and RUNX2) and c-Myc expression analysis. AMPK inhibition of ASA-induced osteo/odontogenesis was tested by qPCR of selected markers (DSPP, OCN and RUNX2). RESULTS: Cytotoxicity assays showed that only the highest ASA dose decreased cell viability (89.1 %). The smallest concentration of ASA applied on DPSCs resulted in a remarkable enhancement of osteo/odontogenic differentiation, as judged by increased mineralized nodules' formation, ALP activity and gene expression of analyzed markers (increase between 2 and 30 folds), compared to untreated cells. ASA also increased DPSCs proliferation. Interestingly, AMPK inhibition per se upregulated DSPP, OCN and RUNX2; the gene upregulation was higher when ASA treatment was also included. c-Myc expression level decreased in cultures treated with ASA, indicating undergoing differentiation processes. CONCLUSIONS: Low concentrations of ASA (corresponding to the standard use in cardiovascular patients), were shown to stimulate osteo/odontogenic differentiation of dental pulp stem cells.

Diphyodont tooth replacement of Brasilodon-A Late Triassic eucynodont that challenges the time of origin of mammals.

Two sets of teeth (diphyodonty) characterise extant mammals but not reptiles, as they generate many replacement sets (polyphyodonty). The transition in long-extinct species from many sets to only two has to date only been reported in Jurassic eucynodonts. Specimens of the Late Triassic brasilodontid eucynodont Brasilodon have provided anatomical and histological data from three lower jaws of different growth stages. These reveal ordered and timed replacement of deciduous by adult teeth. Therefore, this diphyodont dentition, as contemporary of the oldest known dinosaurs, shows that Brasilodon falls within a range of wide variations of typically mammalian, diphyodont dental patterns. Importantly, these three lower jaws represent distinct ontogenetic stages that reveal classic features for timed control of replacement, by the generation of only one replacement set of teeth. This data shows that the primary premolars reveal a temporal replacement pattern, importantly from directly below each tooth, by controlled regulation of tooth resorption and regeneration. The complexity of the adult prismatic enamel structure with a conspicuous intra-structural Schmelzmuster array suggests that, as in the case of extant mammals, this extinct species would have probably sustained higher metabolic rates than reptiles. Furthermore, in modern mammals, diphyodonty and prismatic enamel are inextricably linked, anatomically and physiologically, to a set of other traits including placentation, endothermy, fur, lactation and even parental care. Our analysis of the osteodental anatomy of Brasilodon pushes back the origin of diphyodonty and consequently, its related biological traits to the Norian (225.42 ± 0.37 myr), and around 25 myr after the End-Permian mass extinction event.

Dentin Particulate for Bone Regeneration: An In Vitro Study.

The aim of this in vitro study was to investigate the commitment and behavior of dental pulp stem cells (DPSCs) seeded onto two different grafting materials, human dentin particulate (DP) and deproteinized bovine bone matrix (BG), with those cultured in the absence of supplements. Gene expression analyses along with epigenetic and morphological tests were carried out to examine odontogenic and osteogenic differentiation and cell proliferation. Compressive testing of the grafting materials seeded with DPSCs was performed as well. DPSC differentiation into odontoblast-like cells was identified from the upregulation of odontogenic markers (DSPP and MSX) and osteogenic markers (RUNX2, alkaline phosphatase, osteonectin, osteocalcin, collagen type I, bmp2, smad5/8). Epigenetic tests confirmed the presence of miRNAs involved in odontogenic or osteogenic commitment of DPSCs cultured for up to 21 days on DP. Compressive strength values obtained from extracellular matrix (ECM) synthesized by DPSCs showed a trend of being higher when seeded onto DP than onto BG. High expression of VEGF factor, which is related to angiogenesis, and of dentin sialoprotein was observed only in the presence of DP. Morphological analyses confirmed the typical phenotype of adult odontoblasts. In conclusion, the odontogenic and osteogenic commitment of DPSCs and their respective functions can be achieved on DP, which enables exceptional dentin and bone regeneration.

Dental anatomy and replacement patterns in the early Permian stem amniote, Seymouria.

The exquisite preservation of maxillary and mandibular fragments of Seymouria has allowed us to examine for the first time in detail the dental anatomy and patterns of development in this stem amniote. The results obtained through histological examination show that Seymouria has pleurodont implantation with ankylosis of the tooth to the labial side of the jawbone. The dentary and maxillary teeth exhibit similar dental characteristics, such as the attachment bone (alveolar bone) and cementum rising above the jawbone on the base of the tooth, and smooth carinae extending lingually toward the tooth apex. Additionally, the clear presence of plicidentine, infolding of dentine into the pulp cavity, was found within the tooth root extending into the tooth crown. Lastly, the tooth replacement pattern is alternating, illustrating that Seymouria retains the classic primitive condition for tetrapods, a pattern that is continued in amniotes. Our results provide an important basis for comparison with other stem amniotes and with amniotes.

Long non-coding RNA IGFBP7-AS1 promotes odontogenic differentiation of stem cells from human exfoliated deciduous teeth through autophagy: An in vitro study.

OBJECTIVE: In the present study, we aimed to investigate whether long non-coding RNA (lncRNA) insulin-like growth factor binding protein 7-antisense 1 (IGFBP7-AS1) regulates the odonto-differentiation of stem cells from human exfoliated deciduous teeth (SHED) and its underlying mechanism. DESIGN: Real-time polymerase chain reaction (PCR) and correlation analysis were used to determine the expression of IGFBP7-AS1 during odontogenesis. Alkaline phosphate staining, alizarin red S staining, and real-time PCR in vitro were performed to investigate the effects of IGFBP7-AS1 during odontogenesis. Western blot and immunostaining (with or without chloroquine treatment) were applied to detect the expression of the autophagy-related markers, microtubule-associated proteins 1A/1B light chain 3B (LC3B) and p62. The autophagy inhibitor 3-methyladenine was used to further clarify the effect of autophagy in odonto-differentiation as promoted by IGFBP7-AS1. RESULTS: The expression of lncRNA IGFBP7-AS1 is significantly upregulated during odonto-differentiation of SHED and promotes odontogenesis of SHED in vitro. IGFBP7-AS1 promotes autophagy during odontogenesis. CONCLUSIONS: IGFBP7-AS1 elicits odontogenic differentiation of SHED through autophagy. Furthermore, IGFBP7-AS1 shows promise as a gene target in the regeneration of dental hard tissue and dental-pulp complex.

Cytokine co-stimulation effect on odontogenic differentiation of stem cells.

OBJECTIVE: The study aims to evaluate the effect of bone morphogenetic protein-2 (BMP-2) and transforming growth factor-beta 1 (TGF-ß1) co-stimulation on odontogenic differentiation of human dental pulp stem cells (hDPSCs). MATERIALS AND METHODS: The viability/proliferation of hDPSCs treated with BMP-2 (group B), TGF-ß1 (group T), or BMP-2/TGF-ß1 (group BT) were evaluated. The experiments on odontogenic differentiation were done for 14 days. The following subgroups were added to investigate the effect of co-stimulation with different timing: subgroup B1, TGF-ß1 co-stimulation in the first week; subgroup B2, TGF-ß1 co-stimulation in the second week; subgroup T1, BMP-2 co-stimulation in the first week; and subgroup T2, BMP-2 co-stimulation in the second week. The mineralization was assessed using alizarin red staining. The expression of following genes was assessed using quantitative real-time polymerase chain reaction: dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP1), osteopontin (OPN), and alkaline phosphatase. RESULTS: All groups showed viability similar to the control group (P > .05). The greater mineralization was detected in B groups on day 14. The expressions of DSPP, DMP-1, and OPN increased on day 14 (P < .05). In the combination groups, the higher expressions of DSPP and DMP-1 were observed in subgroups B1 and B2 than groups B and T (P < .05). CONCLUSIONS: BMP-2 was the key in odontogenic differentiation of hDPSCs, which was further enhanced by co-stimulation with TGF-ß1. Continuous stimulation with TGFß-1 did not improve the differentiation of hDPSCs. CLINICAL RELEVANCE: Combined use of the BMP-2 and TGFß-1 at the specific sequence can provide a tissue engineering approach for the future guided dentin regeneration.

Odontogenic Effect of Icariin on the Human Dental Pulp Cells.

Background and Objectives: Human dental pulp cells (HDPCs) can be used for dentin regeneration due to its odontogenic differentiation property. Icariin can induce osteogenic differentiation of stem cells. However, its potential to induce odontogenic differentiation of HDPCs remains unclear. Thus, the aim of this study was to evaluate the capacity of icariin to induce odontogenic differentiation of HDPCs and investigate the underlying molecular mechanism. Materials and Methods: Cell viability assay was used to detect the cytotoxicity of icariin to HDPCs. Effect of icariin on HDPCs chemotaxis was measured by scratch migration assay. The mineralized and odontogenic differentiation of HDPCs was assessed by alkaline phosphatase (ALP) staining, alizarin red S (ARS) staining, real-time PCR, and Western blot of dentin matrix protein 1 (DMP 1) and dentin sialophosphoprotein (DSPP). In addition, Mitogen-activated protein kinase (MAPK) signaling pathway of icariin-induced biomineralization was investigated by Western blot. Results: Cells treated with icariin at all concentrations tested maintained viability, indicating that icariin was biocompatible. Icariin accelerated HDPCs chemotaxis (p < 0.05). Expression levels of related odontogenic markers were increased in the presence of icariin (p < 0.05). Icariin-induced odontogenic differentiation occurred via activation of the MAPK signaling pathway. Furthermore, MAPK inhibitors suppressed expression levels of DSPP and DMP 1 protein, ALP activity, and mineralization of HDPCs. Conclusions: Icariin can upregulate odontogenic differentiation of HDPCs by triggering the MAPK signaling pathway.