Effect of Vehicles and Irrigation Solutions and Methods on Removal of Double Antibiotic Paste from Root Canals: Insights from High-performance Liquid Chromatography Analysis.

INTRODUCTION: In regenerative endodontics, eradicating antibiotic residues from root canals is imperative, given their detrimental effects on human apical papilla stem cells. Previous antibiotic removal studies lacked precision in identifying types and quantities of residual antibiotics. High-performance liquid chromatography (HPLC) enhances sensitivity and specificity, enabling accurate detection and quantification of residual drugs. Using HPLC analysis, this study explored the influence of vehicles and irrigation solutions and methods on double antibiotic paste (DAP) removal from root canals. METHODS: Two DAP formulations, each containing 5 mg/mL ciprofloxacin and metronidazole, were created using distinct vehicles: macrogol and propylene glycol (MP) or hydroxypropyl methylcellulose (HPMC). Subsequently, 5 µL of DAP was applied to 200 simulated immature teeth with open apices (n = 100 per formulation) and cultured for 28 days at 37°C. Samples were then divided into 11 groups (n = 20 per group), and canals were irrigated with 17% ethylenediaminetetraacetic acid or 10% citric acid, employing a positive pressure syringe or passive ultrasonic irrigation. The irrigation solution and dentin sample from each tooth were evaluated via HPLC for ciprofloxacin and metronidazole quantification. RESULTS: Citric acid exhibited significantly superior efficacy in antibiotic removal from root canals, with no observable effect of irrigation methods on drug removal. The HPMC-based DAP formulation significantly enhanced ciprofloxacin removal compared with MP-based DAP. CONCLUSIONS: For antibiotic paste removal from root canals, citric acid is effective, and HPMC is a preferable vehicle over MP. Overall, HPLC is a valuable method for detecting, removing, and quantifying residual antibiotics in root canals.

GelMA-based hydrogel biomaterial scaffold: A versatile platform for regenerative endodontics.

Endodontic therapy, while generally successful, is primarily limited to mature teeth, hence the pressing need to explore regenerative approaches. Gelatin methacryloyl (GelMA) hydrogels have emerged as pivotal biomaterials, promising a bright future for dental pulp regeneration. Despite advancements in tissue engineering and biomaterials, achieving true pulp tissue regeneration remains a formidable task. GelMA stands out for its injectability, rapid gelation, and excellent biocompatibility, serving as the cornerstone of scaffold materials. In the pursuit of dental pulp regeneration, GelMA holds significant potential, facilitating the delivery of stem cells, growth factors, and other vital substances crucial for tissue repair. Presently, in the field of dental pulp regeneration, researchers have been diligently utilizing GelMA hydrogels as engineering scaffolds to transport various effective substances to promote pulp regeneration. However, existing research is relatively scattered and lacks comprehensive reviews and summaries. Therefore, the primary objective of this article is to elucidate the application of GelMA hydrogels as regenerative scaffolds in this field, thereby providing clear direction for future researchers. Additionally, this article provides a comprehensive discussion on the synthesis, characterization, and application of GelMA hydrogels in root canal therapy regeneration. Furthermore, it offers new application strategies and profound insights into future challenges, such as optimizing GelMA formulations to mimic the complex microenvironment of pulp tissue and enhancing its integration with host tissues.

The Effect of Glycyrrhizin on the Viability and Proliferation of Dental Pulp Stem Cells Compared to Intracanal Medicaments.

AIM: To study the effect of glycyrrhizin (GA) on the viability and proliferation of dental pulp stem cells (DPSCs) compared with intracanal medicaments. MATERIALS AND METHODS: Third molars of an adult donor were used to obtain the DPSCs. Flow cytometry was utilized to conduct phenotypic analysis for DPSCs. The methyl-thiazol tetrazolium (MTT) test was used to detect the cell viability. Cell proliferation assay was conducted at distinct time intervals: 3, 5, and 7 days. RESULTS: The flow cytometry analysis verified the positive expression of mesenchymal cell surface antigen molecules (CD73, CD90, and CD105) and the absence of hematological markers (CD14, CD34, and CD45) in the DPSCs. The cells that treated with concentrations more than 0.5 mg/mL of Ca(OH2) and triple antibiotic paste (TAP) gave significant decrease in viability in comparison to the untreated cells (p < 0.05). Also, the cells treated with concentrations 50 and 25 µM of GA showed no significant difference compared with the untreated cells (p > 0.05), while concentrations 12.5 and 6.25 µM expressed a significant increase in viability compared with the untreated cells (p < 0.05). At 7 days, cells treated with the three different concentrations of GA (12.5, 25, and 50 µM) demonstrated a significant increase in cell density compared with Ca(OH)2 and TAP-treated cells (p < 0.05). CONCLUSION: Based upon the potential of GA on DPSCs proliferation compared with Ca(OH)2 and TAP, It is conceivable to acknowledge that GA could be used as an intracanal medicaments for revascularization process of necrotic immature teeth. CLINICAL SIGNIFICANCE: This study emphasizes the significance of assessing alternative root canal medicaments and their impact on the proliferation and viability of DPSCs. The results regarding GA, specifically its impact on the viability and growth of DPSCs, provide essential understanding for its potential application as an intracanal medicine. This study adds to the continuous endeavors in identifying safer and more efficient intracanal therapies, which are essential for improving patient outcomes in endodontic operations. How to cite this article: Alrashidi MA, Badawi MF, Elbeltagy MG, et al. The Effect of Glycyrrhizin on the Viability and Proliferation of Dental Pulp Stem Cells Compared to Intracanal Medicaments. J Contemp Dent Pract 2024;25(3):267-275.

Access to Care Considerations for the Endodontic Treatment of Immature Permanent Teeth: A National Survey of Pediatric Dentists and Endodontists.

INTRODUCTION: This study aimed to investigate access to care and financial considerations associated with the endodontic treatment of immature permanent teeth. METHODS: Surveys were distributed to endodontists (n= 2,457) and pediatric dentists (n= 3,974) in the United States. Data were analyzed using X2 analysis and logistic regression. The level of significance was set to 0.05. RESULTS: The response rate was 13% (n=840). Respondent specialist groups were similar by age and years since specialty residency completion, but significantly different with regard to primary practice setting (e.g. private practice, Federally Qualified Health Center, hospital…), (p=0.001). The majority (91%) of respondents reported participation with dental insurance. Pediatric dentists (69%) were significantly more likely than endodontists (17%) to participate with public-payer dental insurance (p<0.001). The majority of respondents (82%) indicated that patients reported economic factors (time or money) as a barrier to accessing endodontic treatment. Pediatric dentists were significantly more likely to consider economic factors when treatment planning (p<0.001). Pediatric dentists were more likely than endodontists to have the opinion that endodontic procedures for treatment of necrotic immature permanent teeth should cost less than root canal therapy (apexification, p<0.001; regenerative endodontic procedures, p=0.002). Pediatric dentists (33%) reported encountering barriers when attempting to refer their patients to an endodontist. Inability to find an endodontist that participates with dental insurance was the most frequently cited barrier. CONCLUSIONS: Limited clinician participation with dental insurance and gaps in insurance coverage for endodontic procedures appear to contribute to access to care barriers for pediatric patients.

Micro-computed tomography (micro-CT) evaluation of root canal morphology in immature maxillary third molars.

The endodontic treatment of immature permanent teeth with necrotic pulp is a significant clinical challenge. The success of regenerative endodontic procedure is highly dependent on disinfection of the root canal and an accurate anatomical knowledge of the root canal. The aim of this study was to use micro-computed tomography (micro-CT) analysis to investigate the configuration of root canals in the upper permanent third maxillary molars with incomplete root development in their coronal, apical and middle third portions. Thirty immature third permanent maxillary molars were scanned using a micro-CT system. Then, we measured the diameters and areas of the root canal in the coronal, middle and apical third of the roots. The ratio between the long and short diameter of each root canal was then calculated and the canals were divided into several groups: round, oval, long oval, flat and irregular. The round configuration was not observed in the distobuccal and mesiobuccal roots in any of their anatomical regions. Oval and long oval canals predominated in the distobuccal root. The greatest variations were observed in the mesiobuccal root, with the ribbon-shaped canal predominating in the middle region and an irregular shape in the apical region. In the coronal region of the palatal canal, the round configuration predominated; in the middle third, we observed an almost equivalent distribution between round and oval configurations; apically, the oval shape predominated. In conclusion, we observed significant complexity and variation in the morphology and configuration of root canals in immature permanent molars, thus generating additional obstacles for the success of regenerative endodontics.

Laser Applications in Regenerative Endodontics: A Review.

Introduction: Developing regenerative endodontic treatment (RET) is an exciting approach to managing immature permanent teeth with pulp necrosis. RET is usually performed in two clinical steps: disinfection (first step) and tissue engineering (second step). Recently, laser therapy has entered the field of RET. This study aimed to provide an overview of the literature that employed laser therapy for root regeneration. Methods: A comprehensive search was performed on four databases, including PubMed, Web of Science, Scopus, and Google Scholar. The searched keywords were laser, regenerative endodontics, immature permanent teeth, and dental pulp necrosis, and related English-published articles were included up to October 2023. Results: Thirteen studies utilized a laser for RET. In the first step of RET, both high-power and low-level lasers (through photodynamic therapy [PDT]) may be applied for canal disinfection. In contrast, regenerative procedures in the second step of RET are just accelerated by low-power lasers (biostimulation). The literature does not support the benefit of laser-assisted irrigation in improving the clinical success of RET. There is some evidence that laser-assisted disinfection with a diode laser may provide comparable results to triple antibiotic paste in reducing bacterial counts in root canals while providing slightly better clinical and radiographic outcomes. PDT may be an effective and suitable adjunct to conventional disinfection methods in immature, necrotic teeth. Conclusion: Low-power lasers may be beneficial tools for improving the results of regenerative endodontics through chemical disinfection in the first step (PDT) or by biostimulation in the second step of RET.

Revascularization of a Previously Treated Mandibular First Molar with Open Apices: A Case Report.

In this case report, the regeneration method was used to treat the previously treated first molar, which presented with symptoms of pain during chewing and secondary acute apical periodontitis. Due to the importance of preserving the patient's permanent teeth in order to maintain optimal space and function, the regeneration treatment was performed in two visits and according to the American Association of Endodontists (AAE) guidelines. The continuation of tooth root development, healing of periapical radiolucency and positive response to sensibility test (cold test) were observed during follow-up radiographs at intervals of 6 months, 1 year & 2 year.

Use of Endodontic Procedures in Young Permanent Teeth.

Background: Regenerative endodontic procedures (REPs) have emerged as a promising treatment option for young permanent teeth with pulp necrosis, offering the potential for tissue repair and preservation. Materials and Methods: A retrospective analysis was conducted on a cohort of 30 patients aged 8 to 16 years with pulp necrosis in young permanent teeth. The patients underwent REPs, including disinfection, triple antibiotic paste application, and a coronal barrier. Clinical and radiographic data were collected at baseline and follow-up appointments at 6, 12, and 24 months. Radiographs were analyzed for root lengthening, apical closure, and resolution of periapical lesions. Results: The mean increase in root length after 24 months was 3.42 mm (SD ± 1.12 mm), and 90% of cases demonstrated complete apical closure. The overall success rate, defined as the absence of clinical symptoms and radiographic evidence of pathology, was 80. Conclusion: REPs show promising outcomes in young permanent teeth with pulp necrosis, promoting root development, and apical closure.

Long-term Success of Regenerative Endodontic Procedures.

Background: Regenerative endodontic procedures (REPs) have emerged as a transformative approach to treating immature permanent teeth with necrotic pulp tissue. Materials and Methods: A prospective study was conducted, enrolling 100 patients with immature permanent teeth requiring REPs. All procedures were performed by a single experienced endodontist following established protocols. Patients were followed up for a minimum of 5 years' post-treatment. Clinical examinations, radiographic assessments, and patient-reported outcomes were recorded at regular intervals. Data were analyzed using statistical methods to determine the success rates, complications, and factors influencing long-term outcomes. Results: The results of this original research reveal a significant and sustained success rate for REPs. After a minimum follow-up period of 5 years, an arbitrary value of 92% for tooth survival was achieved. Radiographic assessments demonstrated consistent healing of apical lesions, and continued root development was observed in the majority of cases. Patient-reported outcomes indicated a high level of satisfaction with the procedure. Complications such as crown discoloration and tooth fracture occurred in a minority of cases but were effectively managed without compromising the overall success of REPs. Conclusion: This original research provides strong evidence for the long-term success of REPs in the treatment of immature permanent teeth with necrotic pulp tissue. The high tooth survival rate, continued root development, and patient satisfaction support the efficacy of REPs as a reliable treatment option.

Enhancing Antibiotic Efficacy in Regenerative Endodontics by Improving Biofilm Susceptibility.

INTRODUCTION: Various strategies have been researched to enhance the susceptibility of biofilms, given their tolerance to antibiotics. This study evaluated the effect of the anti-microbial peptide nisin in association with antibiotics used in regenerative endodontics, exploring different treatment times and biofilm growth conditions. METHODS: A mixture of 10 bacterial species was cultivated on dentin specimens anaerobically for 21 days. Biofilms were treated with 1 mL of high-purity nisin Z (nisin ZP, 200 µg/mL) and a triple antibiotic mixture (TAP: ciprofloxacin + metronidazole + minocycline, 5 mg/mL), alone or in combination. The effectiveness of antimicrobial agents was assessed after 1 and 7 days. During the 7-day period, biofilms were treated under 2 conditions: a single dose in a nutrient-depleted setting (ie, no replenishment of growth medium) and multiple doses in a nutrient-rich environment (ie, renewal of medium and antimicrobial agents every 48 h). After treatments, biofilm cells were dispersed, and total colony-forming units were counted. RESULTS: After 1 d-treatment, nisin ZP + TAP resulted in 2-log cell reduction compared to TAP alone (P < .05). After 7 d-treatment with a single dose, nisin ZP + TAP and TAP reduced bacteria to nonculturable levels (P < .05), whereas repeated antimicrobial doses did not eliminate bacteria in a nutrient-rich environment. No bacterial reduction was observed with nisin ZP alone in any treatment time. CONCLUSIONS: The additional use of nisin improved the TAP activity only after a short exposure time. Longer exposure to TAP or nisin + TAP in a nutrient-deprived environment effectively eliminated biofilms.

Influence of extracellular matrix scaffolds on histological outcomes of regenerative endodontics in experimental animal models: a systematic review.

BACKGROUND: Decellularized extracellular matrix (dECM) from several tissue sources has been proposed as a promising alternative to conventional scaffolds used in regenerative endodontic procedures (REPs). This systematic review aimed to evaluate the histological outcomes of studies utilizing dECM-derived scaffolds for REPs and to analyse the contributing factors that might influence the nature of regenerated tissues. METHODS: The PRISMA 2020 guidelines were used. A search of articles published until April 2024 was conducted in Google Scholar, Scopus, PubMed and Web of Science databases. Additional records were manually searched in major endodontic journals. Original articles including histological results of dECM in REPs and in-vivo studies were included while reviews, in-vitro studies and clinical trials were excluded. The quality assessment of the included studies was analysed using the ARRIVE guidelines. Risk of Bias assessment was done using the (SYRCLE) risk of bias tool. RESULTS: Out of the 387 studies obtained, 17 studies were included for analysis. In most studies, when used as scaffolds with or without exogenous cells, dECM showed the potential to enhance angiogenesis, dentinogenesis and to regenerate pulp-like and dentin-like tissues. However, the included studies showed heterogeneity of decellularization methods, animal models, scaffold source, form and delivery, as well as high risk of bias and average quality of evidence. DISCUSSION: Decellularized ECM-derived scaffolds could offer a potential off-the-shelf scaffold for dentin-pulp regeneration in REPs. However, due to the methodological heterogeneity and the average quality of the studies included in this review, the overall effectiveness of decellularized ECM-derived scaffolds is still unclear. More standardized preclinical research is needed as well as well-constructed clinical trials to prove the efficacy of these scaffolds for clinical translation. OTHER: The protocol was registered in PROSPERO database #CRD42023433026. This review was funded by the Science, Technology and Innovation Funding Authority (STDF) under grant number (44426).

Effect of erbium yttrium aluminium garnet laser dentin conditioning on dental pulp stem cells viability.

Objective: This study aims to investigate the effect of dentin conditioning by subablative Er:YAG (erbium-doped yttrium aluminium garnet) laser on dental pulp stem cells (DPSCs) viability. Methods: For this in-vitro experimental study, root fragments were longitudinally hemisected after decoronation of single-rooted extracted teeth and preparation of root canals. Prepared samples were randomly assigned to 2 experimental groups (n = 17) as follows; 1) laser conditioning: irradiation with Er:YAG laser beams (2940 nm, 50 mJ per pulse, 20 Hz) 2) Chemical conditioning: 1.5% NaOCl, followed by phosphate-buffered saline (PBS), 17% EDTA, followed by PBS as a final rinse. The samples were ultraviolet-sterilized, and DPSCs were seeded on the samples. MTT assay was performed after 1, 4 and 7 days of incubation to assess the cell viability (n = 5/group per day). Also, after 7 days, two samples of each group underwent SEM (scanning electron microscope) analysis. Statistical analysis was done using independent t-test, one-way ANOVA and two-way ANOVA at a significance level of 0.05. Results: Laser irradiated samples exhibited significantly higher cell viability of DPSCs on days 4 (p < 0.0001) and 7 (p < 0.0001), unlike day 1 (p = 0.131). SEM photomicrographs revealed that Er:YAG laser performed much better smear layer removal and created surface irregularities. Several different cell morphologies were observable on the laser-treated samples, which cells with cytoplasmic extensions being the most frequent. Conclusions: Dentin conditioning by Er:YAG laser enhances DPSCs viability and can be a valuable modality for conditioning dentin to perform regenerative endodontic procedures. Further clinical studies are suggested.

Ortho-Endo Management of Traumatically Intruded Immature Incisors: A Case Report of Novel Interventional Approach With 20 Months Follow-Up.

The case report describes the successful interdisciplinary management of a nine-year-old girl who suffered from traumatic intrusion and rotation of her immature teeth with 12 mm apical displacement of teeth 11 and 21 and 10 mm of teeth 12 and 22. It emphasizes the importance of a comprehensive and contemporary approach, which includes orthodontic intervention and regenerative endodontic procedures (REP). The report follows the CARE (case report) guidelines. The patient was initially observed for four weeks to see if there was potential for spontaneous re-eruption. After eight weeks of orthodontic intervention, the rotation was corrected. However, negative pulp sensitivity and external inflammatory (infection-related) resorption (EIR) occurred, which led to REP using injectable platelet-rich fibrin (i-PRF). Follow-ups over 20 months showed the favorable outcomes of the treatment. A long-term perspective is essential in understanding the outcomes and potential complications associated with traumatic dental injuries (TDI) in children. The case report highlights the importance of collaboration between orthodontists and pediatric dentists, among other specialists. It provides valuable insights into the complexities of managing TDI in children and highlights contemporary regenerative strategies as viable solutions.

Isolation and Culture of Primary Human Dental Pulp Cells-A Description of Technical and Methodological Steps to Maximise Predictability and Yield.

The dental pulp has critical functions in tooth development as well as an ongoing role in promoting and maintaining the vitality of teeth. In particular, its regenerative ability allows dental tissues to be restored following damage caused by traumatic injury or caries. Regenerative endodontic procedures aim to utilise these processes to stimulate dental pulp repair in a minimally invasive manner and reduce the need for more invasive procedures such as root canal treatment. Dental pulp is a source of dental pulp cells (DPCs), which has a subpopulation of dental pulp stem cells (DPSCs), which are attractive for use in regenerative medicine due to their high proliferation rate, ability to differentiate into multiple cell types, and their preserved vitality following cryopreservation. The development of next-generation clinical therapeutics that maximise the potential of dental pulp relies on strong empirical evidence arising from in vitro experimentation. Here, we describe a modified method for the efficient isolation of primary human DPCs from sound third molar teeth for culture using an explant outgrowth method on basement membrane-coated flasks, as well as using high-resolution macro-photography to illustrate the methods. Critically, steps are taken to minimise potential physical and mechanical trauma to the cells and maximise yield. Human DPCs cultured using this method can be further expanded in cell culture flasks to facilitate their use in various in vitro experimental procedures.

Characterization of cells in blood evoked from periapical tissues in immature teeth with pulp necrosis and their potential for autologous cell therapy in Regenerative Endodontics.

OBJECTIVE: The objectives of this study were to isolate, characterize progenitor cells from blood in the root canals of necrotic immature permanent teeth evoked from periapical tissues and evaluate the applicable potential of these isolated cells in Regenerative Endodontics. DESIGN: Ten necrotic immature permanent teeth from seven patients were included. Evoked bleeding from periapical tissues was induced after chemical instrumentation of the root canals. Cells were isolated from the canal blood and evaluated for cell surface marker expression, multilineage differentiation potential, proliferation ability, and target protein expression. Cell sheets formed from these cells were transferred into human root segments, and then transplanted into nude mice. Histological examination was performed after eight weeks. Data analysis was conducted using one-way ANOVA followed by Tukey's post-hoc comparison, considering p < 0.05 as statistically significant. RESULTS: The isolated cells exhibited characteristics typical of fibroblastic cells with colony-forming efficiency, and displayed Ki67 positivity and robust proliferation. Flow cytometry data demonstrated that at passage 3, these cells were positive for CD73, CD90, CD105, CD146, and negative for CD34 and CD45. Vimentin expression indicated a mesenchymal origin. Under differentiation media specific differentiation media, the cells demonstrated osteogenic, adipogenic, and chondrogenic differentiation potential. Subcutaneous root canals with cell sheets of isolated cells in nude mice showed the formation of pulp-like tissues. CONCLUSIONS: This study confirmed the presence of progenitor cells in root canals following evoked bleeding from periapical tissues of necrotic immature teeth. Isolated cells exhibited similar immunophenotype and regenerative potential with dental mesenchymal stromal cells in regenerative endodontic therapy.

Dental Pulp Cell Transplantation Combined with Regenerative Endodontic Procedures Promotes Dentin Matrix Formation in Mature Mouse Molars.

Regenerative endodontic procedures (REPs) are promising for dental pulp tissue regeneration; however, their application in permanent teeth remains challenging. We assessed the potential combination of an REP and local dental pulp cell (DPC) transplantation in the mature molars of C57BL/6 mice with (REP + DPC group) or without (REP group) transplantation of DPCs from green fluorescent protein (GFP) transgenic mice. After 4 weeks, the regenerated tissue was evaluated by micro-computed tomography and histological analyses to detect odontoblasts, vasculogenesis, and neurogenesis. DPCs were assessed for mesenchymal and pluripotency markers. Four weeks after the REP, the molars showed no signs of periapical lesions, and both the REP and REP + DPC groups exhibited a pulp-like tissue composed of a cellular matrix with vessels surrounded by an eosin-stained acellular matrix that resembled hard tissue. However, the REP + DPC group had a broader cellular matrix and uniquely contained odontoblast-like cells co-expressing GFP. Vasculogenesis and neurogenesis were detected in both groups, with the former being more prominent in the REP + DPC group. Overall, the REP was achieved in mature mouse molars and DPC transplantation improved the outcomes by inducing the formation of odontoblast-like cells and greater vasculogenesis.

Osteogenic differentiation and proliferation of apical papilla stem cells using nanoparticles of Neo MTA and bioactive glass.

Objective: The aim of this study was to evaluate the osteogenic differentiation ability and proliferation of apical papilla stem cells using nanoparticles of Neo MTA and bioactive glass. Methods: Neo MTA and bioactive glass 45S5 nanoparticles were prepared and characterized using a transmission electron microscope and X-ray diffraction. Apical papilla stem cells were harvested from freshly-extracted fully-impacted wisdom teeth, cultured, and characterized using flow cytometric analysis. Tested nanomaterials were mixed and samples were classified into four equal groups as follows; Negative control group: SCAP with Dulbecco's modified eagle's medium, Positive control group: SCAP with inductive media, First experimental group: Neo MTA nanoparticles with SCAP, Second experimental group: Bioactive glass nanoparticles with SCAP. Osteoblastic differentiation was assessed using an alkaline phosphatase assay and RANKL expression using specific polyclonal antibodies by fluorescence microscope. The proliferation of SCAP was assessed using cell count and viability of Trypan Blue in addition to an MTT assay. Results: Isolated SCAP showed a non-hematopoietic origin. Neo MTA showed the highest ALP concentration followed by bioactive glass nanoparticles, and negative control. Bioactive glass nanoparticles showed the highest H score for RANKL protein expression followed by Neo MTA, and negative control. Bioactive glass nanoparticles showed the highest viable cell count. Conclusions: SCAP isolation is achievable from extracted fully impacted immature third molars. Both tested nanobiomaterials have the ability to induce osteogenic differentiation and proliferation of SCAP.

Allogeneic Bone Marrow Mesenchymal Stromal Cell Transplantation Induces Dentin Pulp Complex-like Formation in Immature Teeth with Pulp Necrosis and Apical Periodontitis.

INTRODUCTION: Dental pulp regeneration is challenging in endodontics. Cellular therapy is an alternative approach to induce dental pulp regeneration. Mesenchymal stromal cells (MSCs) have the capacity to induce dental pulp-like tissue formation. In this study, we evaluated the capacity of allogeneic bone marrow MSCs (BM-MSCs) to regenerate pulp following necrosis and apical periodontitis in children's permanent immature apex teeth. METHODS: Patients aged 8 to 12 years with pulp necrosis and apical periodontitis were evaluated. The study included 15 teeth (13 incisors and 2 molars) from 14 patients (8 boys and 6 girls). Radiographic evaluation showed periapical radiolucency and immature apex teeth. There was no response to cold or electric pulp testing. The root canal of each tooth was cleaned, shaped, and Ca(OH)2 used as an interappointment medication. Cryopreserved allogeneic BM-MSCs were thawed, expanded, incorporated into preclotted platelet-rich plasma, and implanted into the tooth's pulp cavity. They were sealed with bioceramic cement and composite. Sensibility, apical foramen, calcium deposits within the root canal, and resolution of periapical lesions were evaluated in each tooth over the following 12 months. RESULTS: Based on 9 variables established for dental pulp-like tissue regeneration, all MSC-treated teeth showed evidence of successful regeneration. Clinical and radiographic evaluation of the treated teeth showed periapical lesion healing, sensitivity to cold and electricity, decreased width of the apical foramen, and mineralization within the canal space. CONCLUSIONS: Transplantation of allogeneic MSCs induces the formation of dental pulp-like tissue in permanent immature apex teeth with pulp necrosis and apical periodontitis. Implant of MSCs constitutes a potential therapy in regenerative endodontics in pediatric dentistry. Future studies incorporating a larger sample size may confirm these results.

NOTUM plays a bidirectionally modulatory role in the odontoblastic differentiation of human stem cells from the apical papilla through the WNT/ß-catenin signaling pathway.

OBJECTIVE: Notum is a secreted deacylase, which is crucial for tooth dentin development in mice. This study aimed to investigate the effect of NOTUM on the odontoblastic differentiation of human stem cells from the apical papilla (hSCAPs), to reveal the potential value of NOTUM in pulp-dentin complex regeneration. DESIGN: The expression pattern of NOTUM in human tooth germs and during in vitro odontoblastic differentiation of hSCAPs was evaluated by immunohistochemical staining, and quantitative polymerase chain reaction, respectively. To manipulate the extracellular NOTUM level, ABC99 or small interfering RNA was used to down-regulate it, while recombinant NOTUM protein was added to up-regulate it. The effects of changing NOTUM level on the odontoblastic differentiation of hSCAPs and its interaction with the WNT/ß-catenin signaling pathway were studied using alkaline phosphatase staining, alizarin red staining, quantitative polymerase chain reaction, and western blot. RESULTS: NOTUM was observed in the apical papilla of human tooth germs. During in vitro odontoblastic differentiation of hSCAPs, NOTUM expression initially increased, while the WNT/ß-catenin pathway was activated. Downregulation of NOTUM hindered odontoblastic differentiation. Recombinant NOTUM protein had varying effects on odontoblastic differentiation depending on exposure duration. Continuous addition of the protein inhibited both odontoblastic differentiation and the WNT/ß-catenin pathway. However, applying the protein solely in the first 3 days enhanced odontoblastic differentiation and up-regulated the WNT/ß-catenin pathway. CONCLUSION: NOTUM demonstrated a bidirectional impact on in vitro odontoblastic differentiation of hSCAPs, potentially mediated by the WNT/ß-catenin pathway. These findings suggest its promising potential for pulp-dentin complex regeneration.

Two-Dimensional Radiographs and Cone-beam Computed Tomography Assessment of Concentrated Growth Factor and Platelet-Rich Fibrin Scaffolds in Regenerative Endodontic Treatment of Immature Incisors with Periapical Radiolucency: A Randomized Clinical Trial.

INTRODUCTION: The primary aim of this study was to compare the radiographic changes of immature incisors with periapical radiolucency after treatment with platelet-rich fibrin (PRF) and concentrated growth factor (CGF) platelet concentrate scaffolds as well as assessment of the clinical success rate over 12 months. The secondary aim was to monitor the radiographic changes in terms of reduction of periapical lesion diameter (PALD), root dentine thickness (RDT), root length (RL), and apical foramen width (AFW). The tertiary aim was to assess and pulp responses, after 12 months. METHODS: Fifty six children with seventy necrotic, single-rooted maxillary incisors with periapical radiolucency were treated with either CGF or PRF scaffolds (35 teeth per group). Two patients with 4 teeth (2 teeth in each group) failed to attain the follow-up recalls. Radiographic changes in terms of reduction of PALD, RDT, RL, and AFW were monitored using a 2-dimensional (2D) radiograph and cone-beam computed tomography (CBCT) scan. The clinical performance of teeth receiving both scaffolds was assessed after 6 and 12 months. Categorical and continuous data were analyzed using the chi-square test and the t test, respectively. The time and group effects on the means of different radiographic dimensions were tested using the general linear model. Bland-Altman plots were used to assess the level of agreement between the 2D radiographs and CBCT. The level of significance was defined at 0.05 and a 95% confidence interval. RESULTS: The means of PALD and RL showed significant enhancement in the CGF group compared to the PRF group (P < .05). While the difference between the 2 scaffolds in terms of RDT and AFW was not significant (P > .05). The findings of the 2D radiograph and CBCT were consistent. Clinically, both scaffold success rates were similar (93.9%) over the follow-up intervals. The influence of study independent variables had no significant effect on the success of the regenerative endodontic procedures outcome (P > .05). There was no significant difference in the positive pulp responses to the thermal and electric pulp tests after one year of treatment (P > .05). CONCLUSIONS: According to the short-term follow-up, PRF and CGF were successful in treating immature teeth with periapical radiolucency by regenerative endodontics. Both scaffold systems induced periapical healing and root lengthening with significant superiority of CGF.