Regenerative endodontics model in immature infected rat molars using two step protocol

Aim: to develop a model for regenerative endodontics using newly-weaned Wistar rats immature molars with pulp necrosis to histologically describe the evolution of apical tissues following treatment with a bi-antibiotic paste, induced bloodclot formation and MTA. Methods: Ten 25-day-old female Wistar rats were divided into an initial control group (Ci) and two experimental groups in which pulp necrosis was experimentally induced on the left mandibular first molar by exposing the pulp chamber and leaving it open to the oral environment. One of the experimental groups was left untreated (E1) while the other was submitted to a protocol of regenerative endodontics 10 days thereafter (E2). Fifteen days after placement of a bi-antibiotic paste, bleeding was induced into the root canal space and MTA was placed upon. Animals were euthanized 30 days later. Right mandibular first molars served as an 80-day-old final control group (Cf). Each hemimandible was histologically processed to analyse parameters associated with root development. Statistical analysis was carried by means of ANOVA; p values below 0.05 were considered statistically significant. Results: baseline (i.e. 25-days old) mean root length and apical diameter of the distal root canal were 1.84±0.25 and 0.38±0.02mm respectively. Following the regenerative endodontic protocol, cells lining the walls of the root canal and significant increase to both length (2.37±0.22mm) and diameter (0.32±0.03 mm) were observed. Conclusions: newly-weaned Wistar rats serve as a suitable model to evaluate regenerative endodontic protocols. However, further research is needed in order to disclose the nature of the cells and/or cell mediators involved.

Biocompatibility of a HA/β-TCP/C Scaffold as a Pulp-Capping Agent for Vital Pulp Treatment: An In Vivo Study in Rat Molars

Bioceramic materials possess desirable biological properties, highlighting their non-reactivity and osteoconductivity. Their use has been extended in vital pulp treatment. The purpose of this study was to evaluate and compare the effects of beta-tricalcium phosphate (β-TCP), hydroxyapatite (HA), and collagen (C) scaffold with mineral trioxide aggregate (MTA) on the vital pulp of rat molars. Thirty-two molars of Sprague–Dawley rats underwent direct pulp capping with β-TCP/HA/C (n = 16) and MTA (n = 16). After 30 days, the following parameters were evaluated in the tested samples: the degree of pulp inflammation and pulp vitality, the presence of reparative dentin, the homogeneity of the odontoblastic layer, and the presence of pulp fibrosis. No statistically significant differences were observed between HA/β-TCP/C and MTA in terms of the degree of inflammation (p = 0.124). Significant differences were found in reparative dentin formation between the treatment groups (p = 0.0005). Dentin bridge formation was observed in the MTA-treated group. The local action of HA/β-TCP/C is similar to that of MTA when used as an agent for pulp vital treatment in terms of absence of inflammation and maintenance of pulp vitality, although there are significant differences between both materials regarding the formation of dentin bridges.

Impact of remnant healthy pulp and apical tissue on outcomes after simulated regenerative endodontic procedure in rat molars

When regenerative endodontic procedures (REPs) are performed on immature teeth diagnosed with pulp necrosis and apical periodontitis, various healing patterns occur. Furthermore, infected immature teeth with endodontic disorders often exhibit some remnant pulp and apical tissue. Therefore, this study investigated the impact of remnant healthy or fully functional pulp and apical tissue on healing patterns after REPs. Simulated REPs were performed on non-infected immature rat molars with different amounts of remnant pulp and apical tissue. Healing patterns in these teeth were assessed after 28 days. Teeth with 0.81–0.91 mm of remnant pulp healed with pulp-like tissue, dentin, and osteodentin-like dentin-associated mineralized tissue (OSD-DAMT); teeth with 0.60–0.63 mm of remnant pulp healed with pulp-like tissue and OSD-DAMT; teeth with 0.13–0.43 mm of remnant pulp healed with periodontal ligament (PDL)-like tissue, OSD-DAMT, and cementum-like dentin-associated mineralized tissue (CEM-DAMT); and teeth with disorganization of pulp and apical tissues at 0.15–0.38 mm beyond the root apex healed with PDL-like tissue, CEM-DAMT, and intracanal bone (IB). Loss of Hertwig’s epithelial root sheath was observed with IB formation. These results showed that four distinct healing patterns occurred after REPs, depending on the preoperative amount of remnant healthy pulp and apical tissue.

Translation Approach for Dentine Regeneration Using GSK-3 Antagonists

The canonical Wnt/β-catenin signaling pathway is crucial for reparative dentinogenesis following tooth damage, and the modulation of this pathway affects the rate and extent of reparative dentine formation in damaged mice molars by triggering the natural process of dentinogenesis. Pharmacological stimulation of Wnt/β-catenin signaling activity by small-molecule GSK-3 inhibitor drugs following pulp exposure in mouse molars results in reparative dentinogenesis. The creation of similar but larger lesions in rat molars shows that the adenosine triphosphate (ATP)–competitive GSK-3 inhibitor, CHIR99021 (CHIR), and the ATP noncompetitive inhibitor, Tideglusib (TG), can equally enhance reparative dentine formation to fully repair an area of dentine damage up to 10 times larger, mimicking the size of small lesions in humans. To assess the chemical composition of this newly formed dentine and to compare its structure with surrounding native dentine and alveolar bone, Raman microspectroscopy analysis is used. We show that the newly formed dentine comprises equal carbonate to phosphate ratios and mineral to matrix ratios to that of native dentine, both being significantly different from bone. For an effective dentine repair, the activity of the drugs needs to be restricted to the region of damage. To investigate the range of drug-induced Wnt-activity within the dental pulp, RNA of short-term induced (24-h) molars is extracted from separated roots and crowns, and quantitative Axin2 expression is assayed. We show that the activation of Wnt/β-catenin signaling is highly restricted to pulp cells in the immediate location of the damage in the coronal pulp tissue with no drug action detected in the root pulp. These results provide further evidence that this simple method of enhancement of natural reparative dentinogenesis has the potential to be translated into a clinical direct capping approach.