Drug Repurposing in Dentistry; towards Application of Small Molecules in Dentin Repair.

One of the main goals of dentistry is the natural preservation of the tooth structure following damage. This is particularly implicated in deep dental cavities affecting dentin and pulp, where odontoblast survival is jeopardized. This activates pulp stem cells and differentiation of new odontoblast-like cells, accompanied by increased Wnt signaling. Our group has shown that delivery of small molecule inhibitors of GSK3 stimulates Wnt/ß-catenin signaling in the tooth cavity with pulp exposure and results in effective promotion of dentin repair. Small molecules are a good therapeutic option due to their ability to pass across cell membranes and reach target. Here, we investigate a range of non-GSK3 target small molecules that are currently used for treatment of various medical conditions based on other kinase inhibitory properties. We analyzed the ability of these drugs to stimulate Wnt signaling activity by off-target inhibition of GSK3. Our results show that a c-Met inhibitor, has the ability to stimulate Wnt/ß-catenin pathway in dental pulp cells in vitro at low concentrations. This work is an example of drug repurposing for dentistry and suggests a candidate drug to be tested in vivo for natural dentin repair. This approach bypasses the high level of economical and time investment that are usually required in novel drug discoveries.

Natural Bioactive Epigallocatechin-Gallate Promote Bond Strength and Differentiation of Odontoblast-like Cells.

The (-)-Epigallocatechin-gallate (EGCG) metabolite is a natural polyphenol derived from green tea and is associated with antioxidant, biocompatible, and anti-inflammatory effects. OBJECTIVE: To evaluate the effects of EGCG to promote the odontoblast-like cells differentiated from human dental pulp stem cells (hDPSCs); the antimicrobial effects on Escherichia coli, Streptococcus mutans, and Staphylococcus aureus; and improve the adhesion on enamel and dentin by shear bond strength (SBS) and the adhesive remnant index (ARI). MATERIAL AND METHODS: hDSPCs were isolated from pulp tissue and immunologically characterized. EEGC dose-response viability was calculated by MTT assay. Odontoblast-like cells were differentiated from hDPSCs and tested for mineral deposition activity by alizarin red, Von Kossa, and collagen/vimentin staining. Antimicrobial assays were performed in the microdilution test. Demineralization of enamel and dentin in teeth was performed, and the adhesion was conducted by incorporating EGCG in an adhesive system and testing with SBS-ARI. The data were analyzed with normalized Shapiro-Wilks test and ANOVA post hoc Tukey test. RESULTS: The hDPSCs were positive to CD105, CD90, and vimentin and negative to CD34. EGCG (3.12 µg/mL) accelerated the differentiation of odontoblast-like cells. Streptococcus mutans exhibited the highest susceptibility < Staphylococcus aureus < Escherichia coli. EGCG increased (p < 0.05) the dentin adhesion, and cohesive failure was the most frequent. CONCLUSION: (-)-Epigallocatechin-gallate is nontoxic, promotes differentiation into odontoblast-like cells, possesses an antibacterial effect, and increases dentin adhesion.

Wnt signaling in dental pulp homeostasis and dentin regeneration.

OBJECTIVE: Wnt signaling is crucial in the physiological and pathological processes of dental pulp tissues. The present study described the effects of Wnt signaling in dental pulp homeostasis and regeneration. DESIGN: Publications in Pubmed and Scopus database were searched, and a narrative review was performed. The roles of Wnt signaling in dental pulp tissue were reviewed and discussed. RESULT: In vitro and in vivo evidence have confirmed the involvement of Wnt signaling in tooth development, dental pulp homeostasis, and physiological processes in dental pulp responses. Manipulating Wnt signaling components generates beneficial effects on pulp healing, dentin repair, and epigenetic regulation related to stemness maintenance, implying that Wnt signaling is a potential therapeutic target for future clinical dental applications. Additionally, an overview of the epigenetic control of dental pulp stem cells by Wnt signaling is provided. CONCLUSION: This review provides basic knowledge on Wnt signaling and outlines its functions in dental pulp tissues, focusing on their potential as therapeutic treatments by targeting the Wnt signaling pathway.

Dentin Matrix Metalloproteinases: A Futuristic Approach Toward Dentin Repair and Regeneration.

Matrix metalloproteinases (MMPs) have been linked to modulating healing during the production of tertiary dentin, as well as the liberation of physiologically active molecules and the control of developmental processes. Although efforts to protect dentin have mostly centered on preventing these proteases from doing their jobs, their role is actually much more intricate and crucial for dentin healing than anticipated. The role of MMPs as bioactive dentin matrix components involved in dentin production, repair, and regeneration is examined in the current review. The mechanical characteristics of dentin, especially those of reparative and reactionary dentin, and the established functions of MMPs in dentin production are given particular attention. Because they are essential parts of the dentin matrix, MMPs should be regarded as leading applicants for dentin regeneration.

The Role of GH/IGF Axis in Dento-Alveolar Complex from Development to Aging and Therapeutics: A Narrative Review.

The GH/IGF axis is a major regulator of bone formation and resorption and is essential to the achievement of normal skeleton growth and homeostasis. Beyond its key role in bone physiology, the GH/IGF axis has also major pleiotropic endocrine and autocrine/paracrine effects on mineralized tissues throughout life. This article aims to review the literature on GH, IGFs, IGF binding proteins, and their respective receptors in dental tissues, both epithelium (enamel) and mesenchyme (dentin, pulp, and tooth-supporting periodontium). The present review re-examines and refines the expression of the elements of the GH/IGF axis in oral tissues and their in vivo and in vitro mechanisms of action in different mineralizing cell types of the dento-alveolar complex including ameloblasts, odontoblasts, pulp cells, cementoblasts, periodontal ligament cells, and jaw osteoblasts focusing on cell-specific activities. Together, these data emphasize the determinant role of the GH/IGF axis in physiological and pathological development, morphometry, and aging of the teeth, the periodontium, and oral bones in humans, rodents, and other vertebrates. These advancements in oral biology have elicited an enormous interest among investigators to translate the fundamental discoveries on the GH/IGF axis into innovative strategies for targeted oral tissue therapies with local treatments, associated or not with materials, for orthodontics and the repair and regeneration of the dento-alveolar complex and oral bones.

DMP1-derived peptides promote remineralization of human dentin.

Remineralization of dentin during dental caries is of considerable clinical interest. Dentin matrix protein 1 (DMP1) is a non-collagenous calcium-binding protein that plays a critical role in biomineralization. In the present study, we tested if peptides derived from DMP1 can be used for dentin remineralization. Peptide pA (pA, MW = 1.726 kDa) and peptide pB (pB, MW = 2.185), containing common collagen-binding domains and unique calcium-binding domains, were synthesized by solid-phase chemistry. An extreme caries lesion scenario was created by collagenase digestion, and the biomineral-nucleating potential of these peptides was ascertained when coated on collagenase-treated dentin matrix and control, native human dentin matrix under physiological levels of calcium and phosphate. Scanning electron microscopy analysis suggests that peptide pB was an effective nucleator when compared with pA. However, a 1:4 ratio of pA to pB was determined to be ideal for dentin remineralization, based on hydroxyapatite (HA) morphology and calcium/phosphorus ratios. Interestingly, HA was nucleated on collagenase-challenged dentin with as little as 20 min of 1:4 peptide incubation. Electron diffraction confirmed the presence of large HA crystals that produced a diffraction pattern indicative of a rod-like crystal structure. These findings suggest that DMP1-derived peptides may be useful to modulate mineral deposition and subsequent formation of HA when exposed to physiological concentrations of calcium and phosphate.

Pulp stem cells: implication in reparative dentin formation.

Many dental pulp stem cells are neural crest derivatives essential for lifelong maintenance of tooth functions and homeostasis as well as tooth repair. These cells may be directly implicated in the healing process or indirectly involved in cell-to-cell diffusion of paracrine messages to resident (pulpoblasts) or nonresident cells (migrating mesenchymal cells). The identity of the pulp progenitors and the mechanisms sustaining their regenerative capacity remain largely unknown. Taking advantage of the A4 cell line, a multipotent stem cell derived from the molar pulp of mouse embryo, we investigated the capacity of these pulp-derived precursors to induce in vivo the formation of a reparative dentin-like structure upon implantation within the pulp of a rodent incisor or a first maxillary molar after surgical exposure. One month after the pulp injury alone, a nonmineralized fibrous matrix filled the mesial part of the coronal pulp chamber. Upon A4 cell implantation, a mineralized osteodentin was formed in the implantation site without affecting the structure and vitality of the residual pulp in the central and distal parts of the pulp chamber. These results show that dental pulp stem cells can induce the formation of reparative dentin and therefore constitute a useful tool for pulp therapies. Finally, reparative dentin was also built up when A4 progenitors were performed by alginate beads, suggesting that alginate is a suitable carrier for cell implantation in teeth.

Effects of WNT10A on proliferation and differentiation of human dental pulp cells.

INTRODUCTION: Wingless-type MMTV integration site family, member 10A (WNT10A) plays crucial roles in odontogenesis. The aim of this study was to investigate the effects of WNT10A on human dental pulp cells (DPCs), which contain a mixed population of cells, including stem and progenitor cells, and participate in dentin repair or dentin-pulp regeneration. METHODS: Healthy human premolars extracted for orthodontic reasons were used as a study model. The expression of WNT10A protein in dental pulp was determined by immunohistochemistry. The messenger RNA expression of WNT10A and Wnt-related genes was analyzed by semiquantitative reverse-transcription polymerase chain reaction. DPCs were enzymatically separated from pulp tissues, cultured, and passaged. The biological effects of WNT10A on DPCs were investigated using recombinant lentivirus encoding WNT10A complementary DNA. WNT10A-induced changes in DPC proliferation were assessed by methyltetrazolium assay and flow cytometry. In order to determine the effects of WNT10A on DPC differentiation, the activity of alkaline phosphatase (ALP), an early marker of odontoblastic differentiation, was assessed using an ALP activity assay kit, and the expression levels of odontoblast-specific genes, including DSPP, DMP1, ALP, and COL1A1, were detected by quantitative polymerase chain reaction and Western blot. RESULTS: WNT10A protein was clearly identified in the cytoplasm of DPCs. Semiquantitative reverse-transcription polymerase chain reaction indicated the expression of WNT10A and Wnt-related genes in pulp tissues as well as in passaging DPCs. Lentiviral overexpression of WNT10A enhanced proliferation of DPCs and down-regulated ALP activity and the expression of odontoblast-specific genes. CONCLUSIONS: WNT10A promotes the proliferation of DPCs and negatively regulates their odontoblastic differentiation.