Avaliação de meios de armazenagem e substâncias moduladoras na preservação da viabilidade e inibição clástica

Abstract

Studies have proposed the use of different solutions for storage of avulsed teeth, such as milk, HBSS, and recently the coconut water. However, it would be interesting the association of these storage media with some immunomodulatory substance that be able to control the inflammatory response after late replantation, and improve the prognosis in such cases. Thus, the general objective of the present study was to evaluate the effect of storage media and modulatory substances on preservation viability and inhibit the osteoclast activity, by in vitro and in vivo studies. This study involved five specific objectives: Objective 1: evaluate the effect of storage media on viability of human gingival fibroblasts (HGF) by multiparametric cytotoxic assay; and, on the extern superficial of bovine root dentine by FTIR analysis. Objective 2: evaluate the the periodontal healing process quantitatively and qualitatively of dog teeth replanted after storage in two different formulations of powdered coconut water by histomorphometric analysis. Objective 3: evaluate the effect of lectin (ScLL), stimulated with LPS, on the viability, proliferation and release of IL-10 in HGF Objective 4: evaluate the effect of 15d-PGJ2 and ScLL in HGF LPS-stimulated on viability, and release of IL-6 and TGFβ-1. Objective 5: evaluate the viability of ScLL in peripheral blood mononuclear cells (PBMC) stimulated with LPS and in murine macrophage RAW 264.7, as well as their effects on the osteoclastogenesis in vitro, and on the release of TNF-α and NO. Based on the results of these objectives is possible to conclude that: 1) the soy milk presented higher viability when compared with HBSS and whole milk, whereas the solutions behaved similarly on the dentine composition. Clinically the time at 1 h is a crucial factor in the prognosis of dental replantation; 2) In relation the powdered coconut water, ACP-404 I formulation was able to preserve the normal periodontium and limit the level of surface resorption. Additionally, this study demonstrated that the adjustment of the pH of coconut water is essential to maintain the normal periodontium; 3) The ScLL concentrations tested as possible immunomodulator agent were not cytotoxic, and had not effect on the cell proliferation and release of IL-10 parameters; 4) In the others analysis assay, the concentration of ScLL 5 μg/ml did not interfere in the cell viability. In relation to release of IL-6 parameter, ScLL 2 μg/ml and 15d-PGJ2 1 μg/ml demonstrated the lowest levels of this inflammatory cytokine, and all the tested substances had no effect on the release of TGFβ-1; 5) Finally, ScLL in the concentration 5 μg/ml supressed the in vitro osteoclastogenesis, without interfering in the PBMC and murine osteoclast precursors viability, and no presented effect on the release of TNF-α and NO. Therefore, it is the first step to evaluate the use of ScLL as immunomodulatory agent to prevent the resorption in avulsed teeth, and in the future their association with the powdered coconut water, which may open up new perspectives innovation and transferring of technology with the generation of the new products for treating avulsed teeth.