Desenvolvimento e avaliação "In vitro" de membranas compósitas de quitosana, álcool polivinílico, polietileno glicol e policaprolactona triol com adição de nanopartículas de vidro bioativo para regeneração periodontal

Abstract

Several diseases affect the integrity of periodontium causing the destruction of the connective tissue and cells. The biologic goal of periodontal regeneration is restoration of the periodontium to its original form and function. Tissue engineering using human periodontal ligament cells (HPDL) cultured on composite membranes polymer and bioactive glass (BG) is a possible strategy. It is known that the effect of BG can be enhanced in the nanoscale. Preparations of membranes based on chitosan (CHI) with other biomaterials such as polyvinyl alcohol (PVA), polyethylene glycol (PEG), polycaprolactone triol (PCL-T) and bioactive glass nanoparticles (BGNP) may be promising regarding mechanical properties, cell proliferation and particularly cell adhesion. For this purpose, in this work, HPDL cells were extracted of teeth with orthodontic indication for extraction. It was observed the expression of periostin, osteocalcin (OC), alkaline phosphatase (ALP), collagen I (COL I ), F-Spondin and RUNX2 by RT-PCR. In addition, the cells expressed stem cell markers CD105 and CD166. In a second stage, composite membranes CHI / PVA / PEG with the addition of 0%, 5% and 10% of VBNP were developed. The samples were characterized by Scanning Electron Microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), diffraction of x-ray (XRD), mechanical tests, contact angle, degradation assay and bioactivity test "in vitro". Samples with the addition of 5% and 10% VBNP showed a significant increase in strength and decrease in contact angle. Also, there was a significant increase in cell viability, cell proliferation and ALP activity in HPDL cells when in contact with composite membranes containing VBNP. The results described above indicate that it is possible to establish a primary cell culture from human periodontal ligament cells and mesenchymal stem cells probably exist in this culture. Moreover, they suggest that the introduction of VBNP added interesting characteristics in composite membranes creating a suitable environment for the culture and proliferation of cells HPDL, which is a promising step for the development of improved biomaterials for periodontal regeneration.