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Regenerative endodontic procedures rely on the delivery of mesenchymal stem cells into the root canal and on the effect of local growth factors from the dentin and blood clot. The aim of this study was to assess the effect of dentin conditioning with ethylenediamine tetraacetic acid (EDTA) and diode lasers with different wavelengths (808 nm and 980 nm) on the expression of odontoblast-like cell markers. Forty dentin cylinders were divided into four groups according to the irrigation protocol: EDTA, EDTA + 808 nm diode laser, EDTA + 980 nm diode laser, and phosphate-buffered saline as the control group. Dental pulp stem cells were seeded into the previously conditioned cylinders and incubated for 14 days. The quantitative real-time polymerase chain reaction was used to evaluate the expression of dentin sialophosphoprotein (DSPP), dentin morphoprotein-1 (DMP-1), and transforming growth factor-beta 1 (TGF-ß1). Data analysis was performed using the Kruskal-Wallis test. The activation of EDTA with 980 nm and 808 nm diode lasers resulted in lower DSPP and DMP-1 expression than that for EDTA alone (p < 0.05 and p < 0.01, respectively). The expression of TGF was similar among all groups. The highest level of expression of odontoblast-like differentiation markers was observed with EDTA alone. However, the use of an 808 nm diode laser during EDTA irrigation reduced the expression of odontoblastic differentiation markers.

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INTRODUCTION: Dental pulp stem cells (DPSC) are very attractive in regenerative medicine. In this study, we focused on the characterization of the functional properties of mesenchymal stem cells derived from DPSCs. Currently, it is unknown whether inflammatory conditions present in an inflamed dental pulp tissue could alter the immunomodulatory properties of DPSCs. This study aimed to evaluate the immunomodulatory capacity in vitro of DPSCs derived from healthy and inflamed dental pulp. METHODS: DPSCs from 10 healthy and inflamed dental pulps (irreversible pulpitis) were characterized according to the minimal criteria of the International Society for Cell Therapy, proliferation, differential potential, and colony-forming units. Furthermore, the immunomodulatory capacity of DPSCs was tested on the proliferation of T lymphocytes by flow cytometry and the in vitro enzyme activity of indoleamine 2, 3-dioxygenase. RESULTS: There were no significant differences in the DPSC characteristics and properties such as immunophenotype, tridifferentiation, colony-forming units, and proliferation of the DPSCs derived from normal and inflamed pulp tissue. Furthermore, there were significant differences in the immunomodulatory capacity of DPSCs obtained from human healthy dental pulp and with the diagnosis of irreversible pulpitis. CONCLUSIONS: Our results showed that DPSCs isolated from inflamed dental pulp showed typical characteristics of MSCs and diminished immunosuppressive capacity in vitro in comparison with MSCs derived from healthy dental pulp. Further investigation in vivo is needed to clarify the mechanism of this diminished immunosuppressive capacity.

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INTRODUCTION: Regenerative endodontic procedures have emerged as a new treatment. The aim of this case report was to describe a regenerative autologous cellular therapy using mesenchymal stem cells from inflamed dental pulp and leukocyte platelet-rich fibrin (L-PRF) in a mature tooth. METHODS: A healthy 50-year-old man consulting for spontaneous dental pain was referred for endodontic treatment in tooth #28, which was diagnosed with symptomatic irreversible pulpitis. Inflamed dental pulp was extracted and transported to a good manufacturing practice laboratory for the isolation and culture of dental pulp stem cells (DPSCs). L-PRF was obtained from the patient's blood and was introduced into the instrumented and disinfected root canal, and expanded DPSCs were inoculated into the clot. The cervical part of the root canal was sealed with Biodentine (Septodont, Saint-Maur-des-Fosses, France) and a composite resin. RESULTS: Follow-up examinations were performed 6 months and 3 years later. The examinations included periapical radiographs (to measure the periapical index [PAI]), cone-beam computed tomographic (CBCT) imaging, sensitivity, and vitality tests. Clinical evaluations revealed normal responses to percussion and palpation tests. The tooth had a delayed response to cold, and the electric pulp test was responsive. The PAI and CBCT imaging revealed that the periapical area remained normal with a PAI score of 1 and a CBCT PAI score of 0. The vitality test performed indicated low blood perfusion units. CONCLUSIONS: This case study reveals the potential use of a patient's own DPSCs and L-PRF as an alternative procedure for the treatment of pulpitis in mature permanent teeth. It also paves the way for the design of personalized cell-based clinical trials in regenerative endodontics.

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La ingeniería tisular se plantea como tratamiento ideal para la regeneración de tejidos con la utilización de andamiajes, células madres y factores de crecimiento. Las células madres de origen gingival plantean ventajas de obtención, mientras que el OPLA 3D permite cultivos de alta densidad celular. El objetivo de esta investigación fue evaluar la biocompatibilidad de células madres de origen gingival en OPLA. Las células se obtuvieron de tejido gingival y fueron caracterizadas fenotípica y funcionalmente. La biocompatibilidad se evaluó mediante la proliferación celular, prueba de viabilidad con azul tripán y diferenciación celular a linaje condrogénico y osteogénico. El recultivo del constructo se utilizó para evaluar la capacidad de transporte. Las células al interior del OPLA se visualizaron mediante cortes teñidos con H-E. Las células madres mesenquimales en OPLA proliferaron, 80% de confluencia a la cuarta semana. La viabilidad celular en OPLA fue de 83,32%. En el recultivo, las células comienzan a proliferar a la semana. El OPLA permite la diferenciación celular a linaje condrogénico y osteogénico. Se observan células al interior del OPLA, permite la proliferación, viabilidad y diferenciación celular. El OPLA podría ser utilizado como andamiaje celular para la ingeniería de tejidos.

Tissue engineering arises as the ideal treatment for tissue regeneration with the use of scaffolds, stem cells and growth factors. Stem cells derived from gingival tissue present benefits in its objection. 3D OPLA allow high cell density cultures. The objective of this study was to evaluate the biocompatibility of gingival stem cells in OPLA. Cells were obtained from gingival tissue and were characterized phenotypically and functionally. The biocompatibility was evaluated through cell proliferation, viability test with trypan blue and cell differentiation to chondrogenic and osteogenic lineage. Recultivation of the construct was used to evaluate transportability. Cells inside OPLA were visualized by stained sections with H&E. Mesenchymal stem cells proliferated in OPLA, 80% confluence at the fourth week. Cell viability in OPLA was 83.32%. In recultivation, cells start proliferating in a week. OPLA allows cell differentiation to chondrogenic and osteogenic lineage. Cells were observed within OPLA. In conclusion OPLA allows proliferation, viability and cell differentiation. OPLA could be used as scaffolds for cells in tissue engineering.

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