Regenerative Endodontics Research Articles

Quantitative assessment of transforming growth factor-β1 release from dentin matrix upon conditioning with ethylene diamine tetra-acetate, doxycycline hydrochloride, and propolis: An in vitro study.

Transforming growth factor-beta 1 (TGF-β1) is a key morphogen in regenerative endodontics that plays a central role in regulating cellular functions. Various chelating agents have been shown to release this growth factor upon conditioning. The objective of the study was to evaluate TGF-β1 release from the dentin matrix upon conditioning with ethylene diamine tetra-acetate (EDTA), doxycycline hydrochloride, and propolis. Forty-two human 3rd molar teeth were collected and coronal portion of the teeth was sectioned to obtain dentin blocks with dimensions 2 mm × 2 mm × 2 mm. The blocks were then randomly divided into three groups depending on the conditioning agent used; Group 1: 17% EDTA, Group 2: doxycycline hydrochloride (100 mg/mL), and Group 3: propolis (250 μg/mL). Conditioned blocks were placed in 0.5 mL of phosphate buffered saline and incubated for 1 week for quantification. Highest TGF-β1 release was noted for propolis (0.21 ng/mL), followed by doxycycline hydrochloride (0.18 ng/mL) and 17% EDTA (0.14 ng/mL). Doxycycline hydrochloride and propolis significantly enhanced the release of TGF-β1 from the dentin matrix compared to EDTA (P < 0.05). No significant difference was observed between doxycycline hydrochloride and propolis (P > 0.05). Doxycycline and propolis can be used as effective alternatives to EDTA during regenerative endodontic procedures.

Platelet-rich fibrin used in regenerative endodontics and dentistry: Current uses, limitations, and future recommendations for application

Numerous strategies, including pulp implantation, revascularization, and postnatal stem cell treatments, have been developed by regenerative endodontics. Revascularization has been effectively used in clinical settings recently, giving dentists amazing outcomes. When administered alone or in conjunction with a bone transplant, platelet-rich fibrin (PRF) encourages bone development and vascularization. There are many uses for PRF in regenerative endodontics, including as a tool for mending iatrogenic pulpal floor perforations and for revascularizing developing permanent teeth with necrotic pulps. It serves as a matrix for the ingrowth of tissue. It was seen that the dentinal walls thickened with time, the roots lengthened, the periapical pathology regressed, and the apex closed. The specific mechanism of action of PRF for dental pulp rejuvenation, including both vitro and in vivo, necessitates further study.

Evaluation of postoperative pain and healing following regenerative endodontics using platelet-rich plasma versus conventional endodontic treatment in necrotic mature mandibular molars with chronic periapical periodontitis. A randomized clinical trial.

To evaluate the post-operative pain and the healing of necrotic mature permanent mandibular molar teeth with peri-apical periodontitis after conventional endodontic treatment versus after platelet-rich plasma (PRP) revascularization. The protocol of this randomized clinical trial was registered at www. gov with identification number NCT03350841. Twenty-eight patients were included in the study after confirming the diagnosis clinically and radiographically. In the first visit, mandibular molar teeth of both groups were mechanically prepared. Double antibiotic paste was prepared and injected then the cavity that was sealed with glass ionomer. At the second visit, the patients were randomized either to control group where standard endodontic treatment was completed by lateral condensation technique or assigned to intervention group where PRP revascularization technique. The degree of spontaneous pain was assessed using numerical rating scale (NRS) pre-operatively, then post-operatively after 6, 12 h and daily for 5 days. Patients were given a placebo to be administrated in case of pain. An analgesic (ibuprofen 400mg) was prescribed in case of persistent pain. Clinical and radiographic healing was assessed after 6 and 12 months. All demographic, baseline and outcome data were collected and statistically analysed. Regarding the post revascularization/obturation pain, there was no statistically significant difference between severity of pain in the two groups at all time intervals except after 12 h, revascularization group showed statistically significantly higher prevalence of no pain than endodontic treatment group. Regarding the healing; the periapical lesions decrease in size significantly from the pre-operative lesion size in both groups without significant difference between the both groups. Within the limitation of this study, PRP revascularization could be an alternative treatment to root canal treatment but further randomized clinical trials with standardized techniques and long follow up periods are recommended for more reliable results.

Assessment of the Clinical Outcomes of Single Visit Regenerative Endodontic Procedure In Treating Necrotic Mature Teeth with Apical Periodontitis Using Biological Irrigating Solution

Aim: to observe the effect of propolis and its nano counterpart when used as final irrigation materials and compare them to the conventionally used EDTA and saline on the clinical outcomes of necrotic mature anterior teeth with apical periodontitis after treating them using regenerative endodontic procedure.&#x0D; Methodology: 84 patients who fulfilled the inclusion criteria were recruited for the study. Access cavity was prepared, rubber dam isolation was done. Mechanical preparation was performed up to #60 using K-files. Final irrigation procedure was done as follows:&#x0D; Group A (Control ): Final irrigation was done using saline (3ml/min) followed by 17% EDTA solution (20 mL/canal, 5 min).&#x0D; Group B (comparator 1): Final irrigation was done using propolis[1] irrigation (20 mL/canal, 5 min).&#x0D; Group C (comparator 2): Final irrigation was done using Nano-propolis[2] irrigation (20 mL/canal, 5 min).&#x0D; Bleeding was then induced, PRF was placed in the canals, followed by Collacote[3] and Bio-dentine[4] placement. Patients were recalled after 6 and 12 months. Teeth were checked for sensibility using electric pulp tester and thermal tests. All data was analysed using Kruskal Wallis test.&#x0D; Results: All of the 84 patients (100%) within the 3 groups showed both clinical success and survival. Using Kruskal Wallis, there was no statistically significant difference among the three groups (p=1). Two teeth in the nano propolis group responded positively to thermal test at 12 months follow up. None of the teeth in the three tested groups (100%) responded to the&#x0D; electric pulp test at 12 months. There was nostatistically significant difference between the three groups(p=1)&#x0D; Conclusion: within the limitation of the study, single visit regenerative endodontics can be used to treat necrotic mature teeth with apical periodontitis and a periapical lesion.&#x0D; Key words: Regenerative endodontics, single visit regeneration, propolis, Nano-propolis, Platelet-rich fibrin&#x0D; &#x0D; [1] Propolis: Imtinan/Cairo, Egypt&#x0D; [2] Nanopropolis: NanoTech/Cairo/Egypt&#x0D; [3] Collacote: Collacote/Zimmer/Indiana /USA&#x0D; [4] Biodentine: Acrostone/Cairo/Egypt&#x0D;

Clinical usage of dental stem cells and their derived extracellular vesicles.

Stem cell-based therapies remain at the forefront of tissue engineering and regenerative medicine because stem cells are a unique cell source with enormous potential to treat incurable diseases and even extend lifespans. The search for the best stem cell candidates continues to evolve and in recent years, dental stem cells have received significant attention due to their easy accessibility, high plasticity, and multipotential properties. Dental stem cells have been the subject of extensive research in both animal models and human clinical trials over the past two decades, and have demonstrated significant potential in ocular therapy, bone tissue engineering, and, of course, therapeutic applications in dentistry such as regenerative endodontics and periodontal tissue regeneration. These new sources of cells may be advantageous for cellular therapy and the advancement of regenerative medicine strategies, such as allogeneic transplantation or therapy with extracellular vesicles (EVs), which are functional nanoscale membrane vesicles produced by cells. This chapter discusses the accumulating research findings on cell-based regenerative therapy utilizing dental stem cells and their derived EVs, which could be a viable tool for the treatment of a variety of diseases and hence extremely valuable to mankind in the long run.

Gelatin methacryloyl hydrogel as an injectable scaffold with multi-therapeutic effects to promote antimicrobial disinfection and angiogenesis for regenerative endodontics.

Regenerative endodontics represents a paradigm shift in dental pulp therapy for necrotic young permanent teeth. However, there are still challenges associated with attaining maximum root canal disinfection while supporting angiogenesis and preserving resident stem cells viability and differentiation capacity. Here, we developed a hydrogel system by incorporating antibiotic-eluting fiber-based microparticles in gelatin methacryloyl (GelMA) hydrogel to gather antimicrobial and angiogenic properties while prompting minimum cell toxicity. Minocycline (MINO) or clindamycin (CLIN) was introduced into a polymer solution and electrospun into fibers, which were further cryomilled to attain MINO- or CLIN-eluting fibrous microparticles. To obtain hydrogels with multi-therapeutic effects, MINO- or CLIN-eluting microparticles were suspended in GelMA at distinct concentrations. The engineered hydrogels demonstrated antibiotic-dependent swelling and degradability while inhibiting bacterial growth with minimum toxicity in dental-derived stem cells. Notably, compared to MINO, CLIN hydrogels enhanced the formation of capillary-like networks of endothelial cells in vitro and the presence of widespread vascularization with functioning blood vessels in vivo. Our data shed new light onto the clinical potential of antibiotic-eluting gelatin methacryloyl hydrogel as an injectable scaffold with multi-therapeutic effects to promote antimicrobial disinfection and angiogenesis for regenerative endodontics.

Regenerative endodontics: repair or true regeneration? A narrative review

Regenerative endodontics therapy (RET) is a pioneering approach in dentistry that aims to restore damaged dental pulp and promote tissue regeneration in non-vital teeth. It belongs to the broader field of regenerative medicine, where stem cells, scaffolds, and bioactive molecules are central components. The goal of RET is to achieve true tissue regeneration, fully restoring both the structure and function of damaged tissues, in contrast to repair, which only restores basic functionality through scar formation or tissue remodeling. Histological studies conducted in various research models, ranging from animal studies to human case studies, have provided valuable insights into the nature of tissue regeneration in RET. However, the quality of regenerated tissue can vary significantly among studies. Some research reports have described the presence of fibrous or fibrovascular tissue with limited functionality, resembling repair rather than true regeneration. This variation can be attributed to the impact of disinfection protocols on stem cell viability and differentiation potential. In conclusion, RET has the potential for both repair and true regeneration processes, with outcomes influenced by factors such as disinfection methods and the source of stem cells. Despite the remaining challenges, promising results suggest that RET could revolutionize dental pulp regeneration, especially when approaches that utilize the body\'s stem cells are optimized for clinical use. Further research and protocol refinement is essential to fully unlock the regenerative potential of this exciting field.

Injectable decellularized dental pulp matrix-functionalized hydrogel microspheres for endodontic regeneration.

The sufficient imitation of tissue structures and components represents an effective and promising approach for tissue engineering and regenerative medicine applications. Dental pulp disease is one of the most common oral diseases, although functional pulp regeneration remains challenging. Herein, we propose a strategy that employs hydrogel microspheres incorporated with decellularized dental pulp matrix-derived bioactive factors to simulate a pulp-specific three-dimensional (3D) microenvironment. The dental pulp microenvironment-specific microspheres constructed by this regenerative strategy exhibited favorable plasticity, biocompatibility, and biological performances. Human dental pulp stem cells (hDPSCs) cultured on the constructed microspheres exhibited enhanced pulp-formation ability in vitro. Furthermore, the hDPSCs-microcarriers achieved the regeneration of pulp-like tissue and new dentin in a semi-orthotopic model in vivo. Mechanistically, the decellularized pulp matrix-derived bioactive factors mediated the multi-directional differentiation of hDPSCs to regenerate the pulp tissue by eliciting the secretion of crucial bioactive cues. Our findings demonstrated that a 3D dental pulp-specific microenvironment facilitated by hydrogel microspheres and dental pulp-specific bioactive factors regenerated the pulp-dentin complex and could be served as a promising treatment option for dental pulp disease. STATEMENT OF SIGNIFICANCE: Injectable bioscaffolds are increasingly used for regenerative endodontic treatment. Despite their success related to their ability to load stem cells, bioactive factors, and injectability, conventional bulk bioscaffolds have drawbacks such as ischemic necrosis in the central region. Various studies have shown that ischemic necrosis in the central region can be corrected by injectable hydrogel microspheres. Unfortunately, pristine microspheres or microspheres without dental pulp-specific bioactive factor would oftentimes fail to regulate stem cells fates in dental pulp multi-directional differentiation. Our present study reported the biofabrication of dental pulp-derived decellularized matrix functionalized gelatin microspheres, which contained dental pulp-specific bioactive factors and have the potential application in endodontic regeneration.

Newer prospects of regenerative endodontics: A comprehensive and updated review of literature

Newer prospects of regenerative endodontics: A comprehensive and updated review of literature

METTL3-Mediated lncSNHG7 m6A Modification in the Osteogenic/Odontogenic Differentiation of Human Dental Stem Cells

Background: Human dental pulp stem cells (hDPSCs) play an important role in endodontic regeneration. N6-methyladenosine (m6A) is the most common RNA modification, and noncoding RNAs have also been demonstrated to have regulatory roles in the expression of m6A regulatory proteins. However, the study on m6A modification in hDPSCs has not yet been conducted. Methods: Single base site PCR (MazF) was used to detect the m6A modification site of lncSNHG7 before and after mineralization of hDPSCs to screen the target m6A modification protein, and bioinformatics analysis was used to analyze the related pathways rich in lncSNHG7. After knockdown and overexpression of lncSNHG7 and METTL3, the osteogenic/odontogenic ability was detected. After METTL3 knockdown, the m6A modification level and its expression of lncSNHG7 were detected by MazF, and their binding was confirmed. Finally, the effects of lncSNHG7 and METTL3 on the Wnt/β-catenin pathway were detected. Results: MazF experiments revealed that lncSNHG7 had a m6A modification before and after mineralization of hDPSCs, and the occurrence site was 2081. METTL3 was most significantly upregulated after mineralization of hDPSCs. Knockdown/ overexpression of lncSNHG7 and METTL3 inhibited/promoted the osteogenic/odontogenic differentiation of hDPSCs. The m6A modification and expression of lncSNHG7 were both regulated by METTL3. Subsequently, lncSNHG7 and METTL3 were found to regulate the Wnt/β-catenin signaling pathway. Conclusion: These results revealed that METTL3 can activate the Wnt/β-catenin signaling pathway by regulating the m6A modification and expression of lncSNHG7 in hDPSCs to enhance the osteogenic/odontogenic differentiation of hDPSCs. Our study provides new insight into stem cell-based tissue engineering.

Towards a New Concept of Regenerative Endodontics Based on Mesenchymal Stem Cell-Derived Secretomes Products.

The teeth, made up of hard and soft tissues, represent complex functioning structures of the oral cavity, which are frequently affected by processes that cause structural damage that can lead to their loss. Currently, replacement therapy such as endodontics or implants, restore structural defects but do not perform any biological function, such as restoring blood and nerve supplies. In the search for alternatives to regenerate the dental pulp, two alternative regenerative endodontic procedures (REP) have been proposed: (I) cell-free REP (based in revascularization and homing induction to remaining dental pulp stem cells (DPSC) and even stem cells from apical papilla (SCAP) and (II) cell-based REP (with exogenous cell transplantation). Regarding the last topic, we show several limitations with these procedures and therefore, we propose a novel regenerative approach in order to revitalize the pulp and thus restore homeostatic functions to the dentin-pulp complex. Due to their multifactorial biological effects, the use of mesenchymal stem cells (MSC)-derived secretome from non-dental sources could be considered as inducers of DPSC and SCAP to completely regenerate the dental pulp. In partial pulp damage, appropriate stimulate DPSC by MSC-derived secretome could contribute to formation and also to restore the vasculature and nerves of the dental pulp.

Nanofibrous scaffolds for regenerative endodontics treatment.

Untreated dental caries, tooth trauma and dental anatomical variations such as dens invaginatus can result in pulpitis. However, standard root canal therapy cannot treat immature permanent teeth due to an open apical foramen and thin dentinal walls. Thus, regenerative endodontics treatment (RET) following a disinfection step with pulp regeneration has been developed. Pulp connective-tissue, dentin formation, revascularization and reinnervation can occur in this procedure which should be supplemented with intelligent biomaterials to improve repeatability and support well-coordinated regeneration. Furthermore, nanofibrous scaffolds, as one of the most commonly used materials, show promise. The purpose of this article is to highlight the advantages of nanofibrous scaffolds and discuss the future modification and application of them.

Research progress of odontogenic extracellular vesicles in regeneration of dental pulp.

It is well understood that maintaining viable pulp is critical for tooth retention. This review focused on cell-free therapy based on extracellular vesicles (EVs), a novel minimally invasive treatment strategy for endodontic restoration. This study was conducted by searching mainstream electronic databases such as Web of Science and PubMed for relevant studies on the therapeutic role of odontogenic EVs in pulp healing published in the last five years. We selected 89 relevant articles and discovered that dental stem cells (DSCs) derived EVs (DSC-EVs) have become a research hotspot in oral regenerative medicine, with significant advantages over cell transplantation in terms of low immunogenicity, ease of isolation, preservation, and management. Here, we introduce in detail the therapeutic effects of DSC-EVs for pulp restoration from three perspectives: excellent odontogenic properties, clinical applications, and possible molecular mechanisms. This article contributes a new viewpoint to the field of regenerative endodontics.

Biomimetic Approaches in Clinical Endodontics.

In the last few decades, biomimetic concepts have been widely adopted in various biomedical fields, including clinical dentistry. Endodontics is an important sub-branch of dentistry which deals with the different conditions of pulp to prevent tooth loss. Traditionally, common procedures, namely pulp capping, root canal treatment, apexification, and apexigonesis, have been considered for the treatment of different pulp conditions using selected materials. However, clinically to regenerate dental pulp, tissue engineering has been advocated as a feasible approach. Currently, new trends are emerging in terms of regenerative endodontics which have led to the replacement of diseased and non-vital teeth into the functional and healthy dentine-pulp complex. Root- canal therapy is the standard management option when dental pulp is damaged irreversibly. This treatment modality involves soft-tissue removal and then filling that gap through the obturation technique with a synthetic material. The formation of tubular dentine and pulp-like tissue formation occurs when stem cells are transplanted into the root canal with an appropriate scaffold material. To sum up tissue engineering approach includes three components: (1) scaffold, (2) differentiation, growth, and factors, and (3) the recruitment of stem cells within the pulp or from the periapical region. The aim of this paper is to thoroughly review and discuss various pulp-regenerative approaches and materials used in regenerative endodontics which may highlight the current trends and future research prospects in this particular area.

Pulp tissue regeneration in the treatment of immature premolars accompanied by infection: A case report

Background: Young permanent teeth are one of the challenges in performing endodontic treatment. Revascularization is a good yet simple treatment option for pulp tissue regeneration to stimulate periapical lesion healing, continue root growth and restore pulp tissue function. Purpose: This case report describes management of pulp tissue regeneration in teeth with infection. Case: Female, 16 years old, came complaining of pain in the left lower back tooth accompanied by recurrent swelling. Radiograph showed an open apex with apical lesion on tooth 35. The diagnosis was pulpal necrosis accompanied by a chronic apical abscess. Case management: The treatment plan was revascularization and direct composite restoration. After accessing the cavity, the teeth were prepared, irrigated, and medicated with TAP, which was repeated for 2 visits. On the next visit, after the root canal was cleaned, over instrumentation was performed for bleeding and Collaplug was placed over the blood clot, covered with MTA, and composite resin for coronal sealing. Follow up was carried out for up to 9 months and was asymptomatic. Radiographs show healing of the lesion accompanied by closure of the root canal. Conclusion: Regenerative endodontics has been shown the successful in stimulating root canal closure to treat immature teeth

Investigating the residual effect of silver nanoparticles gel as an intra-canal medicament on dental pulp stromal cells

BackgroundThe aim of this study was to evaluate the indirect effects of residual silver nanoparticles (AgNPs) gel on human dental pulp stromal cells (DPSCs).MethodsNinety-five dentin discs (4x4x1 mm) were prepared from freshly extracted human single-rooted teeth following institutional ethical approval and informed consent. Samples were cleaned, autoclaved, and treated with: 1.5%NaOCl, Saline and 17% EDTA then randomly assigned to 5 groups that received 50 μl of one of the following treatments: 0.01%AgNPs, 0.015%AgNPs, 0.02%AgNPs, Calcium hydroxide (Ca (OH)2) or no treatment for 1 week. Discs were washed with Saline and 17%EDTA then seeded with DPSCs and incubated for 3 and 7 days. At 24 hours unattached cells were collected and counted. At each time point cytotoxicity (LDH assay), cell viability (live/dead staining and confocal microscopy) and cell proliferation (WST1 assay) were assessed. All experiments were repeated a minimum of 3 times using DPSCs isolated from 3 different donors for each time point assessed (n = 9/group). Statistical analysis was done using One-Way ANOVA followed by Tukey’s test and Kruskal Wallis followed by post-hoc comparisons with significance set at p ≤ 0.05.ResultsAfter 24 hours, the percentage of DPSCs attachment ranged between 92.66% ±4.54 and 95.08% ±1.44 with no significant difference between groups (P = 0.126). Cell viability was ≥92% at 24 hours for all groups. However this percentage dropped to less than 60% at 3 days then started to rise again at 7 days. There was no significant difference in cytotoxicity between different groups at all time points except for 0.01%AgNPs group which had the highest cytotoxicity. DPSCs proliferation increased significantly from 3 to 7 days in all groups except for Ca (OH)2 which showed lower proliferation rates at both 3 (45.89%) and 7 days (79.25%).ConclusionDentin discs treated for 7 days with concentrations of AgNPs gel (0.01–0.02%) allowed more than 90% DPSCs cell attachment after 24 hours. The cytotoxicity and proliferation of DPSCs in response to AgNPs gel were comparable to those with calcium hydroxide. This suggests that AgNPs gel may represent a promising future candidate for clinical use in regenerative endodontics. However, its effects may be concentration-dependent warranting further investigation.

Effectiveness of Teeth Whitening after Regenerative Endodontics Procedures: An In Vitro Study.

Discolouration resulting from regenerative endodontic procedures may have a negative impact on the quality of life of treated patients; therefore, it is recommended to minimize this risk by selecting appropriate scaffolds and barrier materials, and if discolouration occurs, the use of a simple, cost-effective and minimally invasive technique, such as whitening, should be considered. This in vitro study aimed to evaluate tooth discolouration after two- and single-visit regenerative endodontic procedures and the effectiveness of subsequent whitening procedures with carbamide peroxide. Two hundred bovine incisors were included in this study and divided into twenty groups based on the tested material combinations. Two groups were control groups, one with saline and the other group with blood. In the 12 groups, the experiment was designed to be consistent with the two-visit regenerative endodontic procedures. Triple antibiotic paste or calcium hydroxide were placed in the root canal, and then scaffolds (e.g., blood or platelet-rich fibrin) and barrier materials (Biodentine, OrthoMTA or MTA Repair HP) were applied after rinsing the disinfectant pastes. In the six groups that corresponded to the single-visit regenerative endodontic procedure, the use of a disinfectant paste was omitted. Subsequently, the specimens were bleached twice with carbamide peroxide at a 7-day interval. Colour change measurements were performed using a spectrophotometer (VITA Easyshade Compact 5.0, VITA Zahnfabrik, Bad Säckingen, Germany). Statistical analysis was performed with the Kruskal−Wallis H test, the independent t-test and t-test for related samples. Tooth discolouration was noticed after two- and single-visit regenerative endodontic procedures, except for the platelet-rich fibrin+MTA Repair HP group. After the first and second whitening procedures, all of the tested two- and single-visit regenerative endodontic procedures groups showed a change in the colour of the crown, which was noticeable to the naked eye (∆E > 3.3). When analysing the ∆E value between the first and second bleaching procedures, no changes in the colour of teeth were visually noticed in the calcium hydroxide and platelet-rich fibrin +MTA Repair HP groups (∆E < 3.3). Single-visit regenerative endodontic procedures are suggested if possible; however, if two-visit regenerative endodontic procedures are performed, it is recommended to use calcium hydroxide as the disinfectant paste because of the lower staining potential. In the context of discolouration, platelet-rich fibrin is advisable for use as a scaffold. The whitening procedure is worth considering, but does not guarantee a return to the original tooth colour, especially when triple antibiotic paste is used.

Micro-RNA Profiling in Dental Pulp Cell Cultures.

Recently, the central role of microRNAs (miRNAs) and epigenetic modifiers in biological and pathological processes, such as stem cell differentiation and inflammation, has stimulated interest. In particular, their influence in dental pulp stem cell (DPSC) differentiation has been highlighted as an exciting avenue of research in the field of Regenerative Endodontics. Although specific miRNAs have been shown to be altered in expression during dental pulp mineralization and repair processes, their interaction with epigenetic modifiers, such as histone deacetylase inhibitors (HDACi) or DNA methyltransferase inhibitors (DNMTi), has not been explored. Currently available next-generation sequencing (NGS) technologies offer the potential to explore in detail the miRNA expression profile of cells, and to investigate the effects of pharmacological inhibitors such as epigenetic modifiers on this profile. This chapter describes the experimental methods required to induce mineralization of DPCs in the presence and absence of epigenetic modifiers and analyze the resulting miRNA expression profiles using RNA sequencing (RNAseq), with a focus on bioinformatic analysis.

Effect of chitosan irrigant solutions on the release of bioactive proteins from root dentin.

To identify the effect of two chitosan solutions on the release of root dentin matrix proteins and to describe the chemical changes observed following conditioning with chelating agents. The release of dentin sialoprotein (DSP), transforming growth factor-beta 1 (TGF-β1), vascular endothelial growth factor (VEGF), and platelet-derived growth factor-BB (PDGF-BB) with different chelating agents, including ethylenediaminetetraacetic acid (EDTA), chitosan solution (CS), and nanoparticulate chitosan (CSnp), was investigated. DSP was quantified using an enzyme-linked immunosorbent assay (ELISA). TGF-β1, VEGF, and PDGF-BB were quantified using a cytokine bead panel (CBA). Raman spectroscopy was performed to identify surface chemical changes. Statistical analysis was performed using Kruskal-Wallis test with Mann-Whitney-Wilcoxon rank-sum test (p < 0.05). TGF-β1, VEGF, and DSP solubilized in all irrigants tested. CSnp showed the highest concentration of DSP. PDGF-BB did not exceed the detection limits. Raman spectroscopy revealed a decrease in the phosphate and carbonate peaks, representing the chelating effect of EDTA, CS, and CSnp. Additionally, CSnp showed the greatest preservation of the amide I and III content. Proteins can be released from dentin via EDTA, CS, and CSnp conditioning. Raman spectroscopic revealed changes in the inorganic content of the root dentin after chelation. Furthermore, use of CSnp facilitated a preservation of the organic content. Chelation allows the release of proteins, justifying the use of chelating agents in regenerative endodontics. The chitosan-dentin matrix interaction also promotes the protection of the organic content as an additional benefit to its protein releasing effect.

Tooth Bleaching of Discolorations Caused by Hydraulic Cements in Regenerative Endodontic Treatment: A 3-Year In Vitro Study.

This study aimed to evaluate the color change caused by hydraulic cements after 3 years in vitro by simulating their use in regenerative endodontic treatment (RET) and to quantify the color change after external bleaching with 40% hydrogen peroxide at 1, 6, and 12 months of follow-up. Fifty teeth were treated simulating RET. Samples were distributed according to the hydraulic cement to be used (n = 10 per group): negative control (no cement), ProRoot-MTA, MM-MTA, TotalFill BC-RRM, or Biodentine. Three years after RET, two sessions of external bleaching with Opalescence Boost were performed. The color was measured in the cervical and incisal halves of the teeth at different time points: baseline, 3 years after performing RET, and after 1, 6, and 12 months after bleaching. The ΔL, Δa, and Δb were determined. A generalized linear model was used to compare color considering group and time. The ΔEab and the ΔE00 were calculated and the acceptability in color change was determined. Three years after RET, a reduction in lightness (negative ΔL values) was found in all groups. These values significantly increased 1 month after bleaching in all groups (p &lt; 0.05) and reversed at 6 months. One month after bleaching, ΔE00 values (color difference tolerance (CIEDE2000)) ranged from very good (&gt;3.6 ≤ 5.4) to excellent (&gt;5.4). One year after bleaching, the color reverted to values similar to those found 3 years after RET. All groups became darker after RET. The color recovered and even improved compared with the baseline measurement after one month of bleaching but did not remain stable over time.