Investigation of the early apical release from endodontic hydrogels: A 3D printed model.

AIM: Regenerative Endodontic Procedures (REPs) using new materials such as hydrogels aim to replace current endodontic treatments, but numerous limitations are to overcome. Apical release was little explored in previous studies, especially regarding hydrogels that incorporate molecules, such as growth factors and antibiotics. Apical release is a key mechanism in achieving regeneration, as it could regulate disinfection or cell colonization. Few models exist for apical release, limiting the transfer of these devices from bench to bedside. This study aims to design a simple and standardized model to identify parameters that influence the early apical release kinetic of molecules from endodontic hydrogels. METHODOLOGY: Endodontic Release Inserts (ERI) were designed to mimic the situation of an immature incisor using three different diameters (Ø 0.5 to 2 mm) and to allow the study of the early release from a hydrogel in a 96-well plate. ERI was produced with a 3D printing machine. The kinetic release was investigated using 2 fluorescent, hydrophobic (BDP-500) and hydrophilic (Fluorescein) molecules, in different hydrogels (fibrin and agarose) and in various media (PBS or serum). The release kinetics were estimated by measuring the fluorescence at different time points (1 to 24 h). RESULTS: ERI use made it possible to report that apical diameters increase from 500 to 1000 µm was associated with an increase in release from 4.02 ± 1.63% to 11.53 ± 2.38% over 24 h. It also allowed us to report that bottom solution composition change from PBS to human serum was associated with an increase in the release of fatty acid molecules, whilst a decrease in the hydrogel concentration was associated with a variation in release kinetics. Moreover, nano-encapsulation of a molecule was associated with a decreased release over the first 24 h from 5.25 to 0%. CONCLUSION: ERI use enables investigation of the parameters influencing release kinetics from endodontic hydrogels. Further investigations are necessary to evaluate the interaction of these parameters with each other, in animal models and clinic.

Next generation antibacterial strategies for regenerative endodontic procedures: A scoping review.

BACKGROUND: The clinical results following regenerative endodontic procedures (REPs) vary according to numerous parameters, including the presence of bacteria. This limitation reduces the indications for REPs and calls for the development of next generation antibacterial strategies (NGAS) providing alternatives to current antibacterial strategies (CAS) such as double or triple antibiotic paste (DAP/TAP) and (Ca(OH)2). OBJECTIVES: The present scoping review aims to describe the current trends regarding the use of such strategies and highlight future perspectives. METHODS: Four databases (PUBMed, Cochrane, ClinicalTrials and Science Direct) were searched until 1st May 2023. RESULTS: A total of 918 records were identified, 133 were screened and assessed for eligibility, and 87 articles were included. The findings show that (1) clinical studies are only available for CAS, (2) although next generation strategies are the most studied approach since 2017, they are all at the pre-clinical stage, (3) most of the next generation strategies use galenic forms which offer cell support and colonization and which simultaneously contain antibacterial molecules as alternatives to CAS and to antibiotics in general, (4) standardization is required for future research, specifically regarding the bacterial strains studied, the use of biofilm studies and the cellular behaviour assessments. CONCLUSION: Although NGAS are promising strategies to improve REPs in the context of infection, the current evidence is mostly limited to pre-clinical studies. Further methodological improvement is required to allow relevant comparisons between studies and to reduce the time from bench to bedside.

Bioactive Endodontic Hydrogels: From Parameters to Personalized Medicine.

Regenerative endodontic procedures (REPs) aim at recreating dental pulp tissue using biomaterials such as hydrogels. Their bioactivity is mostly related to the nature of biomolecules or chemical compounds that compose the endodontic hydrogel. However, many other parameters, such as hydrogel concentration, bioactive molecules solubility, and apex size, were reported to influence the reciprocal host-biomaterial relationship and hydrogel behavior. The lack of knowledge regarding these various parameters, which should be considered, leads to the inability to predict the clinical outcome and suggests that the biological activity of endodontic hydrogel is impossible to anticipate and could hinder the bench-to-bedside transition. We describe, in this review, that most of these parameters could be identified, described, and studied. A second part of the review lists some challenges and perspectives, including development of future mathematical models that are able to explain, and eventually predict, the bioactivity of endodontic hydrogel used in a clinical setting.

A critical analysis of research methods and experimental models to study pulpitis.

Pulpitis is the inflammatory response of the dental pulp to a tooth insult, whether it is microbial, chemical, or physical in origin. It is traditionally referred to as reversible or irreversible, a classification for therapeutic purposes that determines the capability of the pulp to heal. Recently, new knowledge about dental pulp physiopathology led to orientate therapeutics towards more frequent preservation of pulp vitality. However, full adoption of these vital pulp therapies by dental practitioners will be achieved only following better understanding of cell and tissue mechanisms involved in pulpitis. The current narrative review aimed to discuss the contribution of the most significant experimental models developed to study pulpitis. Traditionally, in vitro two (2D)- or three (3D)-dimensional cell cultures or in vivo animal models were used to analyse the pulp response to pulpitis inducers at cell, tissue or organ level. In vitro, 2D cell cultures were mainly used to decipher the specific roles of key actors of pulp inflammation such as bacterial by-products, pro-inflammatory cytokines, odontoblasts or pulp stem cells. However, these simple models did not reproduce the 3D organisation of the pulp tissue and, with rare exceptions, did not consider interactions between resident cell types. In vitro, tissue/organ-based models were developed to better reflect the complexity of the pulp structure. Their major disadvantage is that they did not allow the analysis of blood supply and innervation participation. On the contrary, in vivo models have allowed researchers to identify key immune, vascular and nervous actors of pulpitis and to understand their function and interplay in the inflamed pulp. However, inflammation was mainly induced by iatrogenic dentine drilling associated with simple pulp exposure to the oral environment or stimulation by individual bacterial by-products for short periods. Clearly, these models did not reflect the long and progressive development of dental caries. Lastly, the substantial diversity of the existing models makes experimental data extrapolation to the clinical situation complicated. Therefore, improvement in the design and standardisation of future models, for example by using novel molecular biomarkers, databased models and artificial intelligence, will be an essential step in building an incremental knowledge of pulpitis in the future.

Manufacturing of an immediate removable partial denture with an intraoral scanner and CAD-CAM technology: a case report.

BACKGROUND: Incisor loss constitutes a strong aesthetic and psychologic traumatism for the patient and it remains a challenging situation for the dental practitioner because of the necessity to rapidly replace the lacking tooth. Various therapeutic procedures have been proposed to replace the incisor concerned, for example by using a removable partial denture. However, the manufacturing of such a denture with classical procedures is often subject to processing errors and inaccuracies. The computer-aided design and computer-aided manufacturing (CAD-CAM) technology could represent a good alternative, but it is currently difficult because of the lack of dental softwares able to design easily immediate removable partial dentures. CASE PRESENTATION: A 30-year- patient complained about pain caused by a horizontally and vertically mobile maxillary right central incisor. After all options were presented, extraction of the traumatized incisor was decided due to its very poor prognosis, and the patient selected the realization of a removable denture for economic reasons. The present paper proposes an innovative procedure for immediate removable denture, based on the use of an intraoral scanner, CAD with two different softwares used sequentially, and CAM with a 5-axis machine. CONCLUSIONS: We show in this report that associating an intraoral scanner and CAD-CAM technology can be extended to immediate dentures manufacturing, which could be a valuable procedure for dental practitioners and laboratories, and also for patients.

Dental Pulp Defence and Repair Mechanisms in Dental Caries.

Dental caries is a chronic infectious disease resulting from the penetration of oral bacteria into the enamel and dentin. Microorganisms subsequently trigger inflammatory responses in the dental pulp. These events can lead to pulp healing if the infection is not too severe following the removal of diseased enamel and dentin tissues and clinical restoration of the tooth. However, chronic inflammation often persists in the pulp despite treatment, inducing permanent loss of normal tissue and reducing innate repair capacities. For complete tooth healing the formation of a reactionary/reparative dentin barrier to distance and protect the pulp from infectious agents and restorative materials is required. Clinical and in vitro experimental data clearly indicate that dentin barrier formation only occurs when pulp inflammation and infection are minimised, thus enabling reestablishment of tissue homeostasis and health. Therefore, promoting the resolution of pulp inflammation may provide a valuable therapeutic opportunity to ensure the sustainability of dental treatments. This paper focusses on key cellular and molecular mechanisms involved in pulp responses to bacteria and in the pulpal transition between caries-induced inflammation and dentinogenic-based repair. We report, using selected examples, different strategies potentially used by odontoblasts and specialized immune cells to combat dentin-invading bacteria in vivo.

Dental pulp mesenchymal stem cells-response to fibrin hydrogel reveals ITGA2 and MMPs expression.

Regenerative endodontic procedures (REP) aim at reestablishing tooth vitality by replacing the irreversibly damaged dental pulp removed by the dental practitioner with a new functional one. The current treatment of advanced caries relies on the replacement of the inflamed or necrosed dental pulp with an inert filling material. This leads to a functional but non-vital tooth, which lacks the ability to sense dental tissue damage, and to protect from further bacterial attack. Therapeutic strategies inspired by tissue engineering called REP propose to regenerate a fully functional dental pulp directly in the canal space. Promising results were obtained using dental pulp mesenchymal stem cells (DP-MSCs) in combination with bio-inspired artificial and temporary 3D hydrogels made of extracellular matrix molecules such as collagen and fibrin biomacromolecules. However, the uncontrolled mechanisms of DP regeneration from DP-MSCs in 3D biomacromolecules fail to regenerate a fully functional DP and can induce fibrotic scarring or mineralized tissue formation to a non-negligible extent. The lack of knowledge regarding the early molecular mechanisms initiated by DP-MSCs seeded in ECM-made hydrogels is a scientific lock for REP. In this study, we investigated the early DP-MSC-response in a 3D fibrin hydrogel. DP-MSCs isolated from human third molars were cultured for 24 h in the fibrin hydrogel. The differential transcript levels of extracellular and cell surface genes were screened with 84-gene PCR array. Out of the 84 genes screened, 9 were found to be overexpressed, including those coding for the integrin alpha 2 subunit, the collagenase MMP1 and stromelysins MMP3, MMP10 and MMP12. Over-expression of ITGA2 was confirmed by RT-qPCR. The expression of alpha 2 integrin subunit protein was assessed over time by immunoblot and immunofluorescence staining. The increase in the transcript level of MMP1, MMP3, MM10 and MMP12 was confirmed by RT-qPCR. The overexpression of MMP1 and 3 at the protein level was assessed by immunoblot. MMP3 expression by DP-MSCs was observed by immunofluorescence staining. This work demonstrates overexpression of ITGA2 and of MMP1, 3, 10 and 12 by DP-MSCs cultured in a fibrin hydrogel. The main preliminary extracellular and cell surface response of the DP-MSCs to fibrin hydrogel seems to rely on a ITGA2/MMP3 axis. Further investigations are needed to precisely decipher the role of this axis in dental pulp tissue building. Nevertheless, this work identifies extracellular and cell surface molecules that could be potential checkpoints to be targeted to guide proper dental pulp tissue regeneration.

Real-time simulation of the transplanted tooth using model order reduction.

The biomechanics of transplanted teeth remain poorly understood due to a lack of models. In this context, finite element (FE) analysis has been used to evaluate the influence of occlusal morphology and root form on the biomechanical behavior of the transplanted tooth, but the construction of a FE model is extremely time-consuming. Model order reduction (MOR) techniques have been used in the medical field to reduce computing time, and the present study aimed to develop a reduced model of a transplanted tooth using the higher-order proper generalized decomposition method. The FE model of a previous study was used to learn von Mises root stress, and axial and lateral forces were used to simulate different occlusions between 75 and 175N. The error of the reduced model varied between 0.1% and 5.9% according to the subdomain, and was the highest for the highest lateral forces. The time for the FE simulation varied between 2.3 and 7.2 h. In comparison, the reduced model was built in 17s and interpolation of new results took approximately 2.10-2s. The use of MOR reduced the time for delivering the root stresses by a mean 5.9 h. The biomechanical behavior of a transplanted tooth simulated by FE models was accurately captured with a significant decrease of computing time. Future studies could include using jaw tracking devices for clinical use and the development of more realistic real-time simulations of tooth autotransplantation surgery.

Surgical and prosthetic treatment in an elderly patient affected by unilateral idiopathic gingival fibromatosis: a case report.

OBJECTIVES: The aim was to present the diagnosis and treatment of a case of unilateral idiopathic gingival fibromatosis (IGF) in a geriatric patient. BACKGROUND: IGF is a rare condition characterised by an enlargement of the attached and marginal gingivae with no obvious association to any causative factor. Gingival overgrowth causes inaesthetic changes and clinical symptoms such as speech disturbances, tooth movement, and occlusal problems. MATERIALS AND METHODS: A 65-year-old female presented localised enlargement of mandibular gingiva, malpositioning of anterior mandibular teeth, and difficulty in speech and mastication. IGF also causes numerous aesthetic and psychological problems. Treatment consisted of multiple extractions, surgical therapy, and early prosthetic rehabilitation to restore function and appearance and to prevent recurrence. RESULTS: Excellent aesthetic result and psychological benefit were achieved, and no fibromatosis recurrence was detected after 1 year following surgery. CONCLUSION: Gingival resection of the tissue excess and early oral rehabilitation with removable dentures have so far greatly improved patient's quality of life.

Multifactorial Analysis of Endodontic Microsurgery Using Finite Element Models.

BACKGROUND: The present study aimed to classify the relative contributions of four biomechanical factors-the root-end filling material, the apical preparation, the root resection length, and the bone height-on the root stresses of the resected premolar. METHODS: A design of experiments approach based on a defined subset of factor combinations was conducted to calculate the influence of each factor and their interactions. Sixteen finite element models were created and analyzed using the von Mises stress criterion. The robustness of the design of experiments was evaluated with nine supplementary models. RESULTS: The current study showed that the factors preparation and bone height had a high influence on root stresses. However, it also revealed that nearly half of the biomechanical impact was missed without considering interactions between factors, particularly between resection and preparation. CONCLUSIONS: Design of experiments appears to be a valuable strategy to classify the contributions of biomechanical factors related to endodontics. Imagining all possible interactions and their clinical impact is difficult and can require relying on one's own experience. This study proposed a statistical method to quantify the mechanical risk when planning apicoectomy. A perspective could be to integrate the equation defined herein in future software to support decision-making.

Validated Finite Element Models of Premolars: A Scoping Review.

Finite element (FE) models are widely used to investigate the biomechanics of reconstructed premolars. However, parameter identification is a complex step because experimental validation cannot always be conducted. The aim of this study was to collect the experimentally validated FE models of premolars, extract their parameters, and discuss trends. A systematic review was performed following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Records were identified in three electronic databases (MEDLINE [PubMed], Scopus, The Cochrane Library) by two independent reviewers. Twenty-seven parameters dealing with failure criteria, model construction, material laws, boundary conditions, and model validation were extracted from the included articles. From 1306 records, 214 were selected for eligibility and entirely read. Among them, 19 studies were included. A heterogeneity was observed for several parameters associated with failure criteria and model construction. Elasticity, linearity, and isotropy were more often chosen for dental and periodontal tissues with a Young's modulus mostly set at 18-18.6 GPa for dentine. Loading was mainly simulated by an axial force, and FE models were mostly validated by in vitro tests evaluating tooth strains, but different conditions about experiment type, sample size, and tooth status (intact or restored) were reported. In conclusion, material laws identified herein could be applied to future premolar FE models. However, further investigations such as sensitivity analysis are required for several parameters to clarify their indication.

Odontoblasts in the dental pulp immune response.

Recent studies have demonstrated that human dental pulp cells sense pathogens and elicit innate and/or adaptive immunity. Particular attention has been paid to odontoblasts that are situated at the pulp-dentin interface and constitute the first line of defense to cariogenic bacteria entering dentin after enamel disruption. In this review, recent in vitro and in vivo data suggesting that odontoblasts initiate immune/inflammatory events within the dental pulp in response to cariogenic bacteria are discussed. These data include sensing of pathogens by Toll-like receptors (TLRs), production of chemokines upon cell stimulation with microbial by-products and induction of dendritic cell migration. Additional results presented here reveal that all TLR genes are expressed in the healthy human dental pulp that is thus well equipped to combat pathogens entering the tissue. Seventeen chemokine genes including CXCL12, CCL2, CXCL9, CX3CL1, CCL8, CXCL10, CCL16, CCL5, CXCL2, CCL4, CXCL11 and CCL3, and 9 chemokine receptor genes including CXCR4, CCR1, CCR5, CX3CR1, CCR10 and CXCR3, are also expressed in pulp. TLR2, CCL2 and CXCL1 are upregulated in odontoblasts both under caries lesions and upon stimulation with pathogen by-products. These molecules thus appear as preferential targets for the design of therapeutic agents able to reduce the immune/inflammatory response to cariogenic bacteria and favor pulp healing.

Comparison of Two Innovative Strategies Using Augmented Reality for Communication in Aesthetic Dentistry: A Pilot Study.

During dental prosthetic rehabilitation, communication and conception are achieved using rigorous methodologies such as smile design protocols. The aim of the present pilot study was to compare two innovative strategies that used augmented reality for communication in dentistry. These strategies enable the user to instantly try a virtual smile proposition by taking a set of pictures from different points of view or by using the iPad as an enhanced mirror. Sixth-year dental students (n=18, women = 13, men = 5, mean age = 23.8) were included in this pilot study and were asked to answer a 5-question questionnaire studying the user experience using a visual analog scale (VAS). Answers were converted into a numerical result ranging from 0 to 100 for statistical analysis. Participants were not able to report a difference between the two strategies in terms of handling of the device (p=0.45), quality of the reconstruction (p=0.73), and fluidity of the software (p=0.67). Even if the participants' experience with the enhanced mirror was more often reported as immersive and more likely to be integrated in a daily dental office practice, no significant increase was reported (p=0.15 and p=0.07). Further investigations are required to evaluate time and cost savings in daily practice. Software accuracy is also a major point to investigate in order to go further in clinical applications.

Phenotypic Identification of Dental Pulp Mesenchymal Stem/Stromal Cells Subpopulations with Multiparametric Flow Cytometry.

Dental pulp (DP) is a specialized, highly vascularized, and innervated connective tissue mainly composed of undifferentiated mesenchymal cells, fibroblasts, and highly differentiated dentin-forming odontoblasts. Undifferentiated mesenchymal cells include stem/stromal cell populations usually called dental pulp mesenchymal stem cells (DP-MSCs) which differ in their self-renewal properties, lineage commitment, and differentiation capabilities. Analysis of surface antigens has been largely used to precisely identify these DP-MSC populations. However, a major difficulty is that these antigens are actually not specific for MSCs. Most of the markers used are indeed shared by other cell populations such as progenitor cells, mature fibroblasts, and/or perivascular cells. Accordingly, the detection of only one of these markers in a cell population is clearly insufficient to determine its stemness. Recent data reported that multiparametric flow cytometry, by allowing for the detection of several molecules on the surface of one single cell, is a powerful tool to elucidate the phenotype of a cell population both in vivo and in vitro. So far, DP-MSC populations have been characterized mainly based on the isolated expression of molecules known to be expressed by stem cells, such as Stro-1 antigen, melanoma cell adhesion molecule MCAM/CD146, low-affinity nerve growth factor receptor p75NTR/CD271, and the mesenchymal stem cell antigen MSCA-1. Using multiparametric flow cytometry, we recently showed that human DP-MSCs are indeed phenotypically heterogeneous and form several populations.The present paper describes the multiparametric flow cytometry protocol we routinely use for characterizing DP-MSCs. The description includes the design of the antibody panel and explains the selection of the different parameters related to the data quality control.

Design and characterization of a chitosan-enriched fibrin hydrogel for human dental pulp regeneration.

OBJECTIVE: Regenerating a functional dental pulp in the pulpectomized root canal has been recently proposed as a novel therapeutic strategy in dentistry. To reach this goal, designing an appropriate scaffold able to prevent the growth of residual endodontic bacteria, while supporting dental pulp tissue neoformation, is needed. Our aim was to create an innovative cellularized fibrin hydrogel supplemented with chitosan to confer this hydrogel antibacterial property. METHODS: Several fibrin-chitosan formulations were first screened by rheological analyses, and the most appropriate for clinical use was then studied in terms of microstructure (by scanning electron microscopy), antimicrobial effect (analysis of Enterococcus fæcalis growth), dental pulp-mesenchymal stem/stromal cell (DP-MSC) viability and spreading after 7 days of culture (LiveDead® test), DP-MSC ultrastructure and extracellular matrix deposition (transmission electron microscopy), and DP-MSC proliferation and collagen production (RT-qPCR and immunohistochemistry). RESULTS: A formulation associating 10mg/mL fibrinogen and 0.5% (w/w), 40% degree of acetylation, medium molar mass chitosan was found to be relevant in order to forming a fibrin-chitosan hydrogel at cytocompatible pH (# 7.2). Comparative analysis of fibrin-alone and fibrin-chitosan hydrogels revealed a potent antibacterial effect of the chitosan in the fibrin network, and similar DP-MSC viability, fibroblast-like morphology, proliferation rate and type I/III collagen production capacity. SIGNIFICANCE: These results indicate that incorporating chitosan within a fibrin hydrogel would be beneficial to promote human DP tissue neoformation thanks to chitosan antibacterial effect and the absence of significant detrimental effect of chitosan on dental pulp cell morphology, viability, proliferation and collagenous matrix production.

A clinical and histological study of dental defects in a 10-year-old girl with pseudoxanthoma elasticum and amelogenesis imperfecta.

BACKGROUND: The prominent dental feature of a 10-year-old girl was severely hypoplastic enamel in permanent teeth. CASE REPORT: Severe dental defects were detected in a 10-year-old female patient affected by pseudoxanthoma elasticum and amelogenesis imperfecta. An orthopantomographic examination revealed a reduction of enamel thickness on the crown of all erupted and unerupted teeth, agenesis of the maxillary right second premolar, delayed eruption of mandibular first premolars, and the presence of large calcifications in all tooth pulp chambers. A detailed histological analysis of permanent mandibular first molars showed that pulp calcifications presented a concentric laminate organization and merged to almost completely obliterate the pulp chamber. Osteodentine was visible all along the pulpal surface of the radicular dentine. Broad resorption areas were present in the outermost dentine at both coronal and radicular levels. Radicular resorption areas presented a typical rectangular form and were filled with acellular cementum. Cementum thickness was highly increased on the root surface. Apposition of cellular cementum-like tissue was also observed on the coronal dentine surface. CONCLUSION: Before treating patients affected by amelogenesis imperfecta and/or pseudoxanthoma elasticum, paediatric dentists should be aware of the presence of pulp calcifications that add to the complexity of endodontic procedures.

Multi-Fiber-Reinforced Composites for the Coronoradicular Reconstruction of Premolar Teeth: A Finite Element Analysis.

A coronoradicular reconstruction (CRR) has conventionally used a metallic inlay core (MIC) or a single-fiber-reinforced composite (sFRC) but extensive dentin removal can lead to root fracture. We propose herein a multi-fiber-reinforced composite (mFRC) based on a bundle of thin flexible fibers that can be adapted to the root anatomy without removing additional dentin. The aim of this study was to compare the mechanical behavior of the root reconstructed with mFRC, MIC, or sFRC using a finite element analysis (FEA). Models with or without a ferrule effect were created using Autodesk© software and divided into four parts: root, post, bonding composite or cement, and zirconia crown. For both models, extreme stress values (ESV), stress distribution, and risk of fracture were calculated for an oblique force (45°) of 100 N applied to the top of the buccal cusp. Results indicated that mFRC and mFRCG present a lower risk of fracture of the root and of the CRR without ferrule and thus could be valuable alternatives for premolar CRR. Further studies are necessary to evaluate the clinical success of these CRR.

Expression of the TGF-beta/BMP inhibitor EVI1 in human dental pulp cells.

Members of the TGF-beta/BMP family of growth factors induce odontoblast differentiation and reparative dentin synthesis, and their use has been proposed to stimulate pulp healing during dental therapeutics in human. However, factors that modulate TGF-beta and/or BMP signalling during odontoblast differentiation and physiology remain largely unknown. To identify them, we compared expression profiles of TGF-beta/BMP-related genes in pulp fibroblast- and odontoblast-like cells cultured from human dental pulp explants using cDNA gene arrays. We evidenced that the gene encoding ecotropic viral integration site-1 (EVI1), a transcription factor that inhibits TGF-beta/BMP signalling, was under-expressed in odontoblast-like cells. This result was verified by real-time PCR and, at the protein level, by immunohistochemistry. In vivo, real-time PCR analysis revealed that EVI1 was expressed in the dental pulp, at a level similar to brain, but lower than in lung, kidney or trachea. The protein was localized in dental pulp samples in pulp core and subodontoblast cells. Staining intensity progressively decreased from the radicular to the coronal pulp where EVI1 staining was almost undetectable in odontoblasts. Our data suggest that fine regulation of the EVI1 level in the human dental pulp might be important in the TGF-beta/BMP-induced modulation of dental pulp cell kinetics and/or odontoblast differentiation.

Current challenges in human tooth revitalization.

Tooth vitality and health are related to the presence of a living connective tissue, the dental pulp (DP), in the center of the dental organ. The DP contains the tooth immune defence system that is activated against invading oral cariogenic bacteria during the caries process and the tissue repair/regeneration machinery involved following microorganisms' eradication. However, penetration of oral bacteria into the DP often leads to complete tissue destruction and colonization of the endodontic space by microorganisms. Classical endodontic therapies consist of disinfecting then sealing the endodontic space with a gutta percha-based material. However, re-infections of the endodontic space by oral bacteria can occur, owing to the lack of tightness of the material. Recent findings suggest that regenerating a fully functional pulp tissue may be an ideal therapeutic solution to maintain a tooth defence system that will detect and help manage future injuries. The objective of this paper was to explain the different revascularization and regeneration strategies that have been proposed to reconstitute a living DP tissue and to discuss the main challenges that have to be resolved to improve these therapeutic strategies.