Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration.

OBJECTIVES: To evaluate the effect of SHED-CM on the proliferation, differentiation, migration ability, cell death, gene expression and production of VEGF of HUVEC in vitro and in a rodent orthotopic dental pulp regeneration. METHODS: Three culture media [M199, DMEM/Ham's F12 and DMEM/Ham's F12 conditioned by SHEDs] were used as experimental groups. SHED-CM was prepared maintaining confluent cells in culture without serum for 3 days. The proliferation and cell death marker of HUVECs were assessed using flow cytometry. The capacity of formation of vascular-like structures was analyzed in cells grown over Matrigel® in hypoxic condition. HUVECs migration was followed using the scratch test. VEGF-A expression in HUVECs was assessed using real time RT-qPCR; and VEGF synthesis with ELISA test. SHED-CM was also applied in rodent ortotopic model of dental pulp regeneration in rats. The formed tissue was submitted to histological and immunohistochemical analyses. RESULTS: SHED-CM promoted significantly lower expression of 7AAD in HUVECs; whereas the expression of the Ki67 was similar in all groups. The vascular-like structures were observed in all groups. Migration of SHED-CM group was faster than DMEM/Ham's F12. SHED-CM induced similar expression of VEGF-A than M199, and higher than DMEM/Ham's F12. SHED-CM induced significantly higher VEGF synthesis than other media. SHED-CM induced formation of a vascularized connective tissue inside the root canal. CONCLUSION: The study showed that SHEDs release angiogenic and cytoprotective factors, which are of great importance for tissue engineering. CLINICAL SIGNIFICANCE: SHED-CM could be an option to the use of stem cells in tissue engineering.

Bioactive glass plus laser phototherapy as promise candidates for dentine hypersensitivity treatment.

Treatments for dentine hypersensitivity (DH) may produce positive effects, though do not have lasting results. We investigated the reparative potential of stem cells derived from deciduous teeth (SHEDs) in response to components delivered from substances used in the treatment of the DH, associated or not to laser phototherapy (LPT), to stimulate dentine formation. SHEDs were submitted to substances delivered from a laboratorial P-rich bioactive glass [57SiO2 -26CaO-17P2 O5 (wt %)] or a commercially available desensitizer (Gluma® Desensitizer), associated (or not) to LPT (InGAlP diode laser, 660 nm, 0.028 cm2 , 20 mW, 5 J/cm2 , 7 s, contact mode). Biomaterial characterization was performed by X-ray diffraction, scanning electron microscopy and the particle size was evaluated by dynamic light scattering. SHEDs proliferation and differentiation were analyzed by MTT and Alizarin Red staining, respectively. The conditioned media used in these tests were evaluated regarding their pH and the ionic concentration changes due to ions leached from the bioactive glass (BG). BG majority presented a non-crystalline solid structure and mixed particle sizes characterized by the agglomeration of nanoparticles. Cultures treated with BG alone or in association to LPT showed improved cell growth in relation to Gluma® (p < 0.05). Gluma® was cytotoxic in all tested conditions, regardless irradiated or not. BG associated to LPT induced intense mineral matrix formation. In conclusion, BG releases ionic dissolution products able to promote SHEDs differentiation. BG associated to LPT improves SHEDs proliferation and differentiation in vitro, and may be a promise therapeutic approach for the DH treatment. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 107-116, 2017.

Photobiomodulation of Dental Derived Mesenchymal Stem Cells: A Systematic Review.

OBJECTIVE: This study aimed to conduct a systematic review of the literature published from 2000 to August 2015, to investigate the effect of photobiomodulation (PBM) therapy on dentoalveolar-derived mesenchymal stem cells (ddMSCs), assessing whether a clear conclusion can be reached from the data presented. BACKGROUND DATA: Systematic reviews provide the best evidence on the effectiveness of a procedure and permit investigation of factors that may influence the performance of a method. To the best of our knowledge, no previous systematic review has evaluated the effects of PBM only on ddMSCs. METHODS: The search was conducted in PubMed /MEDLINE®, Scopus and Web of Science databases, and reported according to the Preferred Reporting Items for Systematic Reviews and Metaanalyses (PRISMA Statement). Original research articles investigating the effects of PBM therapy on ddMSCs, published from 2000 to August 2015, were retrieved and used for this review according to the following eligibility criteria: evaluating PBM therapy, assessing stem cells of dentoalveolar origin, published in English, dealing with cells characterized as stem cells, and using light that did not need external chromophores. RESULTS: From the initial 3467 potentially relevant articles identified, 6 were excluded because they were duplicates, and 3453 were considered ineligible based on the inclusion criteria. Therefore, eight articles remained, and these were fully analyzed in order to closely check exclusion criteria items. Only one of them was excluded because the cultured cells studied were not characterized as stem cells. Finally, seven articles served as the basis for this systematic review. CONCLUSIONS: PBM therapy has no deleterious effects on ddMSCs. Although no other clear conclusion was obtained because of the scarce number of publications, the results of these studies are pointing to an important tendency of PBM therapy to improve ddMSCs' viability and proliferation.

Cytotoxicity evaluation of a copaiba oil-based root canal sealer compared to three commonly used sealers in endodontics.

BACKGROUND: The constant development of new root canal sealers has allowed the solution of a large number of clinical cases in endodontics, however, cytotoxicity of such sealers must be tested before their validation as filling materials. The aim of this study was to evaluate the cytotoxic effect of a new Copaiba oil-based root canal sealer (Biosealer [BS]) on osteoblast-like Osteo-1 cells. MATERIALS AND METHODS: The experimental groups were formed according to the culture medium conditioned with the tested sealers, as follows: Control group (CG) (culture medium without conditioning); Sealer 26 (S26) - culture medium + S26; Endofill (EF) - culture medium + EF; AH Plus (AHP) - culture medium + AHP; and BS - culture medium + BS (Copaiba oil-based sealer). The conditioned culture medium was placed in contact with 2 × 10(4) cells cultivated on 60 mm diameter Petri dishes for 24 h. Then, hemocytometer count was performed to evaluate cellular viability, using Trypan Blue assay. The normal distribution of data was tested by the Kolmogorov-Smirnov test and the values obtained for cellular viability were statistically analyzed (1-way ANOVA, Tukey's test - P < 0.05), with a significance level of 5%. RESULTS: S26, EF and AHP presented decreased cellular viability considerably, with statistical significance compared with CG (P < 0.05). BS maintained cellular viability similar to CG (P > 0.05). CONCLUSION: The Copaiba oil-based root canal sealer presented promising results in terms of cytotoxicity which indicated its usefulness as a root canal sealer.