Chitosan-based endodontic irrigation solutions and TGF-ß1 treatment: Creating the most favourable environment for the survival and proliferation of stem cells of the apical papilla in vitro.

BACKGROUND: The dental pulp's environment is essential for the regulation of mesenchymal stem cells' homeostasis and thus, it is of great importance to evaluate the materials used in regenerative procedures. AIM: To assess in vitro (i) the effect of chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse®, 17% EDTA, 10% citric acid and 2.5% NaOCl on DSCS viability; (ii) the effect of different concentrations of TGF-ß1 on DCSC proliferation; and (iii) whether treatment with TGF-ß1 following exposure to the different irrigation solutions could compensate for their negative effects. METHODOLOGY: (i) DSCS were treated with three dilutions (1:10, 1:100 and 1:1000) of the six irrigation solutions prepared in DMEM for 10 and 60 min to assess the effect on viability. (ii) The effect of different concentrations (0, 1, 5 and 10 ng/mL) of TGF-ß1 on DCSC proliferation was assessed at 1, 3 and 7 days. (iii) The proliferative effect of TGF-ß1 following 10-min exposure to 1:10 dilution of each irrigation solution was also tested. We used MTT assay to assess viability and proliferation. We performed statistical analysis using Prism software. RESULTS: (i) The different endodontic irrigation solutions tested showed a significant effect on cell viability (p ≤ .0001). Significant interactions between the endodontic irrigation solutions and their dilutions were also found for all parameters (p ≤ .0001). Chitosan nanoparticles and 0.2% chitosan irrigation solution were the least cytotoxic to DSCS whilst 2.5% NaOCl was the most cytotoxic followed by 17% EDTA. (ii) TGF-ß1 at concentrations of 1 and 5 ng/mL resulted in significantly higher proliferation compared to the control group. (iii) Exposure to 17% EDTA or 2.5% NaOCl for 10 min was sufficient to make DSCS cells refractory to the proliferative effects of TGF-ß1. DSCS groups treated with TGF-ß1 following exposure to chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse® and 10% CA demonstrated significantly higher proliferation compared to non-TGF-ß1-treated groups (p ≤ .0001, p ≤ .0001, p ≤ .0001 and p = .01 respectively). CONCLUSIONS: The current study offers data that can be implemented to improve the outcome of regenerative endodontic procedures by using less toxic irrigation solutions and adding TGF-ß1 to the treatment protocol.

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.

Biomimetic pulp scaffolds prepared from extracellular matrix derived from stem cells from human exfoliated deciduous teeth promote pulp-dentine complex regeneration.

AIM: To evaluate the role of biomimetic pulp scaffolds derived from the extracellular matrix derived of stem cells from human exfoliated deciduous teeth (SHED-ECM-PS) in promoting pulp-dentine complex regeneration. METHODOLOGY: SHED-ECM-PS was prepared through cell aggregation and decellularization techniques. Histological and immunofluorescence analyses, scanning electron microscopy, and DNA quantification assays were used to characterize the SHED-ECM-PS. Additionally, a tooth slice implantation model was established to evaluate the effects of SHED-ECM-PS on regeneration of the pulp-dentine complex in vivo. Extraction medium for SHED-ECM-PS was prepared, and its effect on bone marrow mesenchymal stem cells (BMMSCs) was assessed in vitro. Cell counting kit-8 and Ki-67 staining assays were performed to determine cell proliferation. The rate of apoptosis was evaluated by flow cytometry. Wound healing and transwell assays were conducted to evaluate cell migration. Alizarin red S staining was performed to examine mineralized nodule formation. Western blotting was used to detect the expression of osteogenic and odontogenic markers. The results were analysed using an independent two-tailed Student's t-test. p < .05 was considered statistically significant. RESULTS: SHED-ECM-PS was successfully constructed, exhibiting a striped dental pulp-like shape devoid of nuclear structures or DNA components, and rich in fibronectin, collagen I, DMP1 and DSPP. Notably, SHED-ECM-PS showed no impact on the proliferation or apoptosis of BMMSCs. Histological analysis revealed that dental pulp fibroblasts formed an interwoven mesh in the root canal, and angiogenesis was observed in the SHED-ECM-PS group. Moreover, a continuous, newly formed tubular dentine layer with polarized odontoblast-like cells was observed along the inner wall of the root canal. SHED-ECM-PS promoted the migration, polar alignment and mineralized nodule formation of BMMSCs and specifically elevated the expression levels of odontogenic markers, but not osteogenic markers, compared with the control group in vitro. CONCLUSION: SHED-ECM-PS exhibited no cytotoxicity and promoted pulp-dentine complex regeneration in vivo as well as cell migration and odontogenic differentiation of BMMSCs in vitro. These findings provide evidence that SHED-ECM-PS, as a novel biological scaffold, has the potential to improve the outcomes of REPs.

In vitro and in vivo evaluation of iRoot BP Plus as a coronal sealing material for regenerative endodontic procedures.

OBJECTIVES: To investigate in vitro effects of a nanoparticle bioceramic material, iRoot BP Plus, on stem cells from apical papilla (SCAP) and in vivo capacity to induce pulp-dentin complex formation. MATERIALS AND METHODS: The sealing ability of iRoot BP Plus was measured via scanning electron microscopy (SEM). SCAP were isolated and treated in vitro by iRoot BP Plus conditioned medium, with mineral trioxide aggregate (MTA) conditioned medium and regular medium used as controls, respectively. Cell proliferation was assessed by BrdU labeling and MTT assay and cell migration was evaluated with wound healing and transwell assays. Osteo/odontogenic potential was evaluated by Alizarin red S staining and qPCR. Pulp-dentin complex formation in vivo was assessed by a tooth slice subcutaneous implantation model. RESULTS: iRoot BP Plus was more tightly bonded with the dentin. There was no difference in SCAP proliferation between iRoot BP Plus and control groups (P > 0.05). iRoot BP Plus had a greater capacity to elevated cell migration (P < 0.05) and osteo/odontogenic marker expression and mineralization nodule formation of SCAP compared with MTA groups (P < 0.05). Furthermore, the new continuous dentine layer and pulp-like tissue was observed in the iRoot BP Plus group in vivo. CONCLUSIONS: iRoot BP Plus showed excellent sealing ability, promoted the migration and osteo/odontogenesis of SCAP and induced pulp-dentin complex formation without affecting the cell proliferation, which indicated iRoot BP Plus was a promising coronal sealing material in REPs. CLINICAL RELEVANCE: The coronal sealing materials play crucial roles for the outcomes of REPs. This study showed that iRoot BP Plus has good coronal sealing and promote pulp-dentin complex formation compared with MTA, providing experimental evidences for the clinical application of iRoot BP Plus as a promising coronal seal material in REPs.

Outcome of regenerative endodontic procedures in delayed replanted immature permanent teeth with apical periodontitis: A retrospective study.

BACKGROUND: Regenerative endodontic procedures (REPs) is effective for treating young permanent teeth with pulp necrosis. However, its efficacy on delayed replanted avulsed teeth is unclear. AIM: This retrospective study aimed to assess the efficacy of REPs in treating delayed replanted immature permanent teeth with apical periodontitis. DESIGN: Avulsed teeth receiving REPs were systematically screened based on predetermined criteria. This study assessed the REP outcomes, postoperative periodontal healing, and overall treatment efficacy. Samples were grouped by REP outcomes and root development stage, with Fisher's exact tests used to compare outcomes among different groups. RESULTS: Among the included 17 teeth, 47.1% exhibited successful REPs and periodontal healing. Another 47.1%, due to replacement resorption or REP failure, were categorized as tooth survival. Healing of periapical lesions was observed in 88.2% of the cases, but only 41.2% demonstrated continued root development. Although differences were not significant (p = 0.05), teeth with continued root development had a higher rate of functional healing (85.7%) compared to those without (30%). CONCLUSION: Within the limitations of this study, REPs presented reliable outcomes for treating delayed replanted immature permanent teeth with apical periodontitis mainly in periapical lesion healing. Teeth with continued root development after REPs exhibited a higher rate of functional healing. Further investigation is required to explore potential synergies between REP outcomes and periodontal healing.

Does the use of different scaffolds have an impact on the therapeutic efficacy of regenerative endodontic procedures? A systematic evaluation and meta-analysis.

BACKGROUND: In the regenerative endodontic procedures, scaffolds could influence the prognosis of affected teeth. Currently, there is controversy regarding the postoperative evaluation of various scaffolds for pulp regeneration. The objective of this study was to access whether other scaffolds, used alone or in combination with blood clot (BC), are more effective than BC in regenerative endodontic procedures. METHODS: We systematically search the PubMed, the Cochrane Central Register of Controlled Trials (CENTRAL), Embase, and Google Scholar databases. Randomized controlled trials examining the use of BC and other scaffold materials in the regenerative endodontic procedures were included. A random effects model was used for the meta-analysis. The GRADE method was used to determine the quality of the evidence. RESULTS: We screened 168 RCTs related to young permanent tooth pulp necrosis through electronic and manual retrieval. A total of 28 RCTs were related to regenerative endodontic procedures. Ultimately, 12 articles met the inclusion criteria and were included in the relevant meta-analysis. Only 2 studies were assessed to have a low risk of bias. High quality evidence indicated that there was no statistically significant difference in the success rate between the two groups (RR=0.99, 95% CI=0.96 to 1.03; 434 participants, 12 studies); low-quality evidence indicated that there was no statistically significant difference in the increase in root length or root canal wall thickness between the two groups. Medium quality evidence indicated that there was no statistically significant difference in pulp vitality testing between the two groups. CONCLUSIONS: For clinical regenerative endodontic procedures, the most commonly used scaffolds include BC, PRP, and PRF. All the different scaffolds had fairly high clinical success rates, and the difference was not significant. For regenerative endodontic procedures involving young permanent teeth with pulp necrosis, clinical practitioners could choose a reasonable scaffold considering the conditions of the equipment and patients.

Impact of using XP-endo finisher and nanobubble water during EDTA dentin conditioning on TGF-ß1 release in regenerative endodontic procedures.

INTRODUCTION: Transforming Growth Factor-Beta 1 (TGF-ß1) plays a crucial role in the success of Regenerative Endodontic Procedures (REPs) as they directly impact the proliferation and differentiation of stem cells. TGF-ß1 is released by conditioning of the dentin matrix using 17% EDTA. EDTA was found to have deleterious effects on dentin especially in immature teeth with fragile dentin walls. Decreasing the irrigation time was reported to decrease these effects. Accordingly, enhancement and activation of the EDTA solution to maintain its efficiency in TGF-ß1 release from dentin and thus compensating the reduction in irrigation time was employed. EDTA solution was enhanced by adding Nanobubble (NB) water which contains oxygen filled cavities less than 200 nm in diameter. Additionally, EDTA was activated with XP-endo Finisher rotary file. The aim of this study was to assess the impact of NB enhancement and/or XP-endo Finisher activation of the EDTA solution on the TGF-ß1 release from dentin. METHODS: Fifty standardized root segments with open apex were allocated to two main groups according to whether EDTA was enhanced with NB water or not, and within each group whether XP-endo Finisher activation was used or not in addition to a Negative Control group. The concentration of the released TGF-ß1 in the root canal was measured using enzyme-linked immunosorbent assay (ELISA). The statistical analysis was done using the Shapiro- Wilk, Kolmogorov Smirnov, ANOVA and Post-hoc Tukey tests. RESULTS: All groups released a considerable amount of TGF-ß1 with the highest values in the EDTA/NB/XP group, followed by EDTA/NB, EDTA/DW/XP, EDTA/DW and Negative Control groups respectively. CONCLUSIONS: The results of this study suggest that NBs can promote the success of REPs since it revealed a significant increase in the TGF-ß1 release following its use in the enhancement of the EDTA solution. A comparable effect was obtained by XP-endo finisher activation of the EDTA solution. The combined use of NBs and XP-endo Finisher can be a promising addition in REPs. Accordingly, Enhancement and activation of the EDTA solution may compensate decreasing the EDTA irrigation time attempted to avoid the deleterious effect of EDTA on dentin.

Comparative Evaluation of Platelet-rich Fibrin and Treated Dentin Matrix in Regenerative Endodontic Treatment of Nonvital Immature Permanent Teeth: A Randomized Clinical Trial.

AIM: Clinical and radiographic evaluation of the efficacy of platelet-rich fibrin (PRF) and treated dentin matrix (TDM) in regenerative endodontic treatment and periapical healing of nonvital immature permanent teeth with chronic apical periodontitis. MATERIALS AND METHODS: Twenty-four children aged between 7 and 11 years, each presenting with a nonvital immature permanent upper central incisor, were selected. They were randomly allocated into two groups (n = 12), group I (PRF) and group II (TDM). Baseline clinical findings were recorded, and preoperative cone-beam computed tomography (CBCT) was taken. Follow-up was done clinically for 15 months at 3-month intervals (3, 6, 9, 12, and 15 months), and CBCT was taken at the end of the 15-month follow-up. Root length, apical diameter, radiographic root area (RRA), and size of the periapical lesion were quantitively assessed at the end of follow-up period and compared to the preoperative CBCT. RESULTS: Clinical success was 100% in both groups by the end of the follow-up period. Radiographically, after a 15-month follow-up, there was a significant increase in root length and RRA, and there was also a significant reduction in apical diameter and lesion size within each group (p < 0.05). However, there was no statistically significant difference between both groups regarding the mean percentage of increase in root length and mean percentage of reduction of apical diameter (p > 0.05). On the other hand, PRF showed more increase in RRA and more reduction in lesion size, with a statistically significant difference between both groups (p < 0.05). CONCLUSION: Both PRF and TDM were clinically successful. Platelet-rich fibrin showed better radiographic outcomes and periapical healing. CLINICAL SIGNIFICANCE: Platelet-rich fibrin is a viable scaffold to aid further root development and resolution of periapical lesions of nonvital immature permanent teeth. Further studies with different forms of TDM are needed to assess the efficacy of TDM in regenerative endodontic treatment of nonvital immature permanent teeth. How to cite this article: Asal MA, Elkalla IH, Awad SM, et al. Comparative Evaluation of Platelet-rich Fibrin and Treated Dentin Matrix in Regenerative Endodontic Treatment of Nonvital Immature Permanent Teeth: A Randomized Clinical Trial. J Contemp Dent Pract 2024;25(6):563-574.

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.

The capability of regenerative endodontics to promote the complete root formation in immature permanent teeth with different initial pre-operative conditions.

OBJECTIVES: To describe the potential possibility of complete root formation after regenerative endodontic procedures (REPs) in immature permanent teeth with different initial pre-operative conditions. MATERIALS AND METHODS: Children who underwent REPs in the Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing, China, from January 2013 to December 2022 were enrolled. Whether the tooth achieved complete root formation was determined using post-operative periapical radiography. The potential possibility of complete root formation after REPs in immature permanent teeth with different initial conditions is discussed. RESULTS: A total of 428 teeth from 401 patients were included, of which 258 (60.2%) achieved complete root formation. In cases that achieved complete root formation, the most serious type of trauma was avulsion, the teeth with the worst initial root development were less than one-third of the development (stages 6-7), and the longest duration of the presence of apical lesions in the tooth before the first visit was more than 180 days. There is a significant difference in the root formation of teeth with different initial developmental stages and diagnoses. CONCLUSIONS: The active period of continued root formation was 5-21 months post-operatively. The higher the initial stage of root development and the shorter the duration of the presence of apical lesions in the tooth before the first visit, the greater the possibility of root formation. However, complete root formation may still be expected after REPs in severely injured immature permanent teeth and teeth with poor initial pre-operative root development conditions (less than stage 7) at the first visit. This is also true for teeth with long-term and large-scale apical lesions.

Regenerative endodontic treatment using autologous blood from alveolar bone for mature permanent premolar with apical periodontitis: a case report.

OBJECTIVE: The formation of blood clot, achieved through periapical bleeding or the use of peripheral venous blood, platelet-rich plasma (PRP) or platelet-rich fibrin (PRF), has been widely utilized in regenerative endodontic procedures (REPs). Except for inducing periapical bleeding, the scaffolds obtained from PRP, PRF, and venous blood are not derived from oral and maxillofacial tissues. In this case report, autologous blood clots from alveolar bone were described, which involved puncturing the maxillary terminal alveolar bone. The purpose of this case report was to assess the efficacy of the alveolar-derived blood clot in regenerative endodontic procedures in a mature permanent premolar. METHODS: A mature premolar had a fractured central cusp, one of the developmental aberrations of tooth, without a proper treatment, resulting in chronic periapical lesion. To address this condition, REPs were employed, utilizing blood obtained through aspiration from the maxillary terminal alveolar bone aspiration. RESULT: Follow-up examinations of tooth #29 revealed the absence of clinical symptoms, a progressive resolution of a periapical lesion area and thickening of the root canal walls. CONCLUSION: This case demonstrated that blood clot obtained through paracentesis of the maxillary terminal alveolar bone could be successfully applied in REPs and has potential to manage periapical lesions. However, further and more clinical trials are required to verify the feasibility of the alveolar-derived blood clot in REPs and compare the outcome of alveolar-derived blood clot with currently used biological scaffolds. CLINICAL RELEVANCE: In comparison to PRP or PRF from peripheral venous blood, blood clots obtained via paracentesis of the maxillary terminal alveolar bone could be administrated by dentists, eliminating the need for reliance on professional nurses. The use of blood clots from alveolar bone could simplify the REPs in cases where periapical blood was insufficient to brim the root canals.

Treatment outcomes of regenerative endodontic procedures in nonvital mature permanent teeth: a retrospective study.

OBJECTIVES: This retrospective study was undertaken to clinically and radiographically evaluate the long-term outcomes of regenerative endodontic procedures (REPs) for nonvital mature permanent teeth, to analyze predictors influencing treatment outcomes. METHODS: Nonvital mature permanent teeth treated by REPs with a minimum follow-up period of 6 months were included from 2015 to 2017. Treatment outcomes were categorized as success and failure. The periapical status and lesion healing were assessed in terms of the periapical index (PAI) and the percentage changes in periapical radiolucency (PARL) area. The clinical and radiographic outcomes of REPs were assessed by Mann-Whitney test at different follow-up period. Kaplan-Meier curves and Univariate Cox regression analysis were conducted to assess the success and identify potential predictors affecting outcomes, respectively. RESULTS: A total of 37 mature teeth with an average follow-up of 4.3 years satisfied the criteria, and 89.2% of the teeth had a successful outcome. Significant differences in PAI scores were found between each period with respect to the baseline (p < .05). Among different periods, there was a significant difference between intervals of 3-6 months and 7-12 months (p = .039) and no significant difference between each interval of more than 12 months (p > .05). Eighty-seven percent of teeth with preoperative PARL presented completely healed. REPs significantly decreased the PARL area at the interval of 7-12 months compared to 3-6 months (p = .025), with no significant difference between each interval of more than 12 months (p > .05). No significant predictor was found for the success of outcome (p > .05). Thirteen teeth (35.1%) regained pulp sensibility, and 40.5% of the teeth exhibited intracanal calcification. CONCLUSIONS: Within the limitations of this study, REPs provided a high long-term success rate and promoted the resolution of PARL as a biologically-based alternative treatment option for nonvital mature teeth. CLINICAL RELEVANCE: REPs provide a high long-term success rate and promoted healing of apical periodontitis comparable with reported outcomes for root canal therapy of mature teeth.

Implications of root, pulp chamber, and canal anatomy on pulpotomy and revitalization procedures.

OBJECTIVES: This review aims to discuss the implications of anatomy of the root, pulp chamber, and canals on pulpotomy and revitalization procedures (RPs) as treatment alternatives to root canal treatment procedures. METHODS: This narrative review was undertaken to address two main questions - why remove vital pulp tissue in teeth with complex canal anatomy when it can be preserved? And why replace the necrotic pulp in teeth with mature roots with a synthetic material when we can revitalize? This review also aims to discuss anatomical challenges with pulpotomy and revitalization procedures. RESULTS: Maintaining the vitality of the pulp via partial or full pulpotomy procedures avoids the multiple potential challenges faced by clinicians during root canal treatment. However, carrying out pulpotomy procedures requires a meticulous understanding of the pulp chamber anatomy, which varies from tooth to tooth. Literature shows an increased interest in the application of RPs in teeth with mature roots; however, to date, the relation between the complexity of the root canal system and outcomes of RPs in necrotic multi-rooted teeth with mature roots is unclear and requires further robust comparative research and long-term follow-up. CONCLUSIONS: Whenever indicated, pulpotomy procedures are viable treatment options for vital teeth with mature roots; however, comparative, adequately powered studies with long-term follow-up are needed as a priority in this area. RPs show promising outcomes for necrotic teeth with mature roots that warrant more evidence in different tooth types with long-term follow-ups.  CLINICAL RELEVANCE: Clinicians should be aware of the pulp chamber anatomy, which is subject to morphological changes by age or as a defensive mechanism against microbial irritation, before practicing partial and full pulpotomy procedures. RP is a promising treatment option for teeth with immature roots, but more evidence is needed for its applications in teeth with mature roots. A universal consensus and considerably more robust evidence are needed for the standardization of RPs in teeth with mature roots.

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.

Evaluation of the characteristics of root canal calcification after regenerative endodontic procedures: A retrospective cohort study over 3 years.

AIM: This study aimed to evaluate the characteristics of root canal calcification after regenerative endodontic procedures (REPs) during long-term follow-up. DESIGN: Data of children who underwent REPs and were followed up for >3 years in the Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing, China, from January 2013 to January 2019, were collected. All the patients were treated by the protocol of REPs based on the American Association of Endodontists (AAE) protocol. A total of 91 teeth of 54 boys and 37 girls (average age 10.4 ± 1.9 years) with follow-up duration >3 years were included. The follow-up duration ranged from 36 to 92 months (average, 53.2 ± 13.4 months). The prevalence, contributing factors, and long-term prognoses of root canal calcification after REPs are discussed. Independent t-test and χ2 test were used for statistical analysis. RESULTS: The incidence of root canal calcification was 78% (71/91). The use of calcium hydroxide paste was significantly correlated with the occurrence of root canal calcification (p < .05). Some teeth showed aggravation of calcification with time; however, not all teeth showed calcification after longer follow-up duration. CONCLUSIONS: Teeth treated with REPs had a relatively high probability of root canal calcification detection during the long-term follow-up. The occurrence of calcification is related to the use of calcium hydroxide paste but does not affect the long-term prognosis of teeth.

Treatment outcomes of immature permanent necrotic evaginated teeth: A retrospective study comparing regenerative endodontic procedures with apexification.

BACKGROUND: Few studies have compared the outcomes of regenerative endodontic procedures (REPs) and calcium hydroxide apexification focusing on necrotic teeth with dens evaginatus. AIM: To qualitatively and quantitatively compare the treatment outcomes of REPs and calcium hydroxide apexification in teeth with dens evaginatus. DESIGN: Immature permanent necrotic evaginated teeth treated with REPs or calcium hydroxide apexification for a follow-up period of at least 12 months were included. Tooth success and survival rates were analyzed. Changes in radiographic root length, apical diameter, and radiographic root area (RRA) were quantified. Prognostic factors that might influence RRA were identified via multivariate linear regression analysis. RESULTS: A total of 112 teeth (50 REP cases and 62 apexification cases) with a median follow-up period of 26.5 months were included. Regenerative endodontic procedures and calcium hydroxide apexification exhibited similar satisfactory success and survival rates (p > .05). Additionally, 88 teeth were quantitatively analyzed. The REP group presented a significantly greater percentage increase in RRA and less decrease in apical diameter than the calcium hydroxide apexification group (p < .05). Teeth treated with REPs and with Stages 7 and 8 of root development showed a better gain in RRA (p < .05). CONCLUSION: While REP and calcium hydroxide apexification had similar success and survival rates, teeth with REPs showed an increase in RRA, indicating that REP is the preferred choice.

Analysis of the achievement of primary and secondary goals and influencing factors in single-rooted immature permanent teeth after regenerative endodontic procedures: a retrospective study.

OBJECTIVE: This study explored the achievement of primary and secondary goals and factors influencing their achievement in regenerative endodontic procedures (REPs) for immature permanent teeth. METHODS: Dental records of all patients who received REPs for immature permanent teeth at the Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology between January 2012 and January 2023 were retrieved. The evaluation of the primary goal was based on medical and radiographic records. The achievement of the primary goal was defined as the absence of clinical signs and symptoms, such as pain, swelling, or sinus tract and the absence of periapical radiolucency, as assessed on postoperative periapical radiographs. The achievement of secondary goal represented increased root wall thickness and/or increased root length, that is, continued root development. Periapical radiographs before and after treatment were used to evaluate the achievement of the secondary goal. The secondary goal was required to be achieved alongside the achievement of primary goal. RESULTS: A total of 436 teeth (136 anterior and 300 posterior teeth) were included in this study, 96.1% of which demonstrated achievement of the primary goal. Signs of failure (17 teeth) included crown fracture and uncontrolled and recurrent periapical lesions. In addition, 77.8% of teeth demonstrated achievement of the secondary goal, and more than half of the teeth exhibited a complete root development. Evaluation factors included patients' age, sex, tooth type, etiology, preoperative periapical lesion, duration of clinical signs and symptoms, follow-up period, and stage of root development. The achievement of the primary and secondary goals were significantly related to age and tooth type (p < 0.05). CONCLUSIONS: Children with a younger initial visit age are more likely to achieve primary and secondary goals. Additionally, posterior teeth had an advantage over anterior teeth in achieving primary and secondary goals.

Regenerative endodontic procedures for the treatment of necrotic mature teeth: A preliminary randomized clinical trial.

AIM: This preliminary randomized, prospective, controlled trial aimed to compare the clinical and radiographic outcomes of two regenerative endodontic procedures (REPs), revitalization and a platelet-rich fibrin (PRF)-based technique, in the treatment of mature permanent teeth with necrotic pulps. METHODOLOGY: The trial has been reported according to the Preferred Reporting Items for Randomized Trials in Endodontics 2020 guidelines. The study protocol was registered at the clinical trial registry (ClinicalTrials.gov) with identifier number NCT04158232. Twenty patients with mature necrotic anterior teeth with large periapical lesions were randomly allocated into two groups (n = 10): group I, treated with revitalization with the blood clot (BC) technique and group II, treated with a PRF-based technique. The follow-up was for 12 months. Periradicular healing was assessed using standardized radiographs taken at baseline, and at 6 and 12 months after treatment. An electric pulp tester was used to assess whether pulp sensibility had been regained during the follow-up period. Statistical analysis was conducted using Mann-Whitney test and Wilcoxon test for non-parametric data. For parametric data, repeated measures analysis of variance was used. The significance level was set at p ≤ .05. RESULTS: There was a significant increase in periradicular healing in both groups at 6 and 12 months, compared to that at baseline, with no significant difference between the studied groups after 12 months (p = .143). There was a significant difference between the tooth sensibility readings at baseline, 6-month and 12-month follow-up timepoints (p < .001). CONCLUSIONS: The findings of this preliminary trial indicate the potential for using REPs, such as revitalization or PRF-based techniques, as treatment options for mature teeth with necrotic pulps. A higher level of evidence obtained through adequately powered clinical trials and longer follow-up periods are required to conclusively validate the different outcomes of REPs.

Improvement of apical papilla cell attachment after erbium, chromium-doped yttrium, scandium, gallium, and garnet laser application: a study in an ex vivo immature tooth model.

The purposes of this study were (1) to investigate the direct effect of an Er,Cr:YSGG laser on human apical papilla cell (APC) proliferation and mineralization and (2) to examine the effect of Er,Cr:YSGG laser, when applied to an ex vivo immature tooth model, on APC attachment. An Er,Cr:YSGG laser at various power outputs (0.1, 0.5, and 1 W) was used at different positions (2, 5, or 8 mm from the cells) to irradiate cultured APCs. APC proliferation and mineralization were assessed at various intervals. For the cell attachment evaluation, ex vivo tooth models containing dentin samples were irrigated with either EDTA or normal saline solution (NSS) and supplemented with laser activation. Fibronectin-positive-staining cells were counted and analyzed. The number of APCs was significantly greater when power outputs of 0.1 W and 0.5 W were used than when 1 W was used (P < 0.05). The close contact of laser application, at 2 and 5 mm, exerted a negative effect on cell proliferation at 24 and 48 h. The application at 8 mm did not show the deterioration effect. APC mineralization was reduced after laser irradiation, regardless of the power and the tip positioning, at 21 days. APC attachment in all laser-activated groups was significantly greater than in the groups without laser. The use of Er,Cr:YSGG laser significantly promoted APC attachment on the root canal dentin.

Leptin in Dental Pulp and Periapical Tissues: A Narrative Review.

Leptin is a non-glycosylated 16 kDa protein synthesized mainly in adipose cells. The main function of leptin is to regulate energy homeostasis and weight control in a central manner. There is increasing evidence that leptin also has systemic effects, acting as a link between innate and acquired immune responses. The expression of leptin and its receptor in human dental pulp and periradicular tissues have already been described, as well as several stimulatory effects of leptin protein expression in dental and periodontal tissues. The aim of this paper was to review and to compile the reported scientific literature on the role and effects of leptin in the dental pulp and periapical tissues. Twelve articles accomplished the inclusion criteria, and a comprehensive narrative review was carried out. Review of the available scientific literature concluded that leptin has the following effects on pulpal and periapical physiology: 1) Stimulates odontogenic differentiation of dental pulp stem cells (DPSCs), 2) Increases the expression of dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1), odontoblastic proteins involved in odontoblastic differentiation and dentin mineralization, 3) Stimulates vascular endothelial growth factor (VEGF) expression in human dental pulp tissue and primary cultured cells of human dental pulp (hDPCs), 4) Stimulates angiogenesis in rat dental pulp cells, and 5) Induces the expression of interleucinas 6 and 8 in human periodontal ligament cells (hPDLCs). There is evidence which suggests that leptin is implicated in the dentin mineralization process and in pulpal and periapical inflammatory and reparative responses.