Effect of Cell Sheet of human dental pulp stem cell, associated or not with photobiomodulation therapy, on the regeneration of critical size bone defects in calvaria of rats with Type 2 Diabetes Mellitus
Diabetes mellitus (DM) compromises bone repair, generating the need for new therapeutic strategies. One is based on Cell Sheets (CSs) cultures that preserve cell-cell contact, structural organization, and stem cell phenotype (SCs). Photobiomodulation therapy (PBMT) also induces beneficial effects that favor cell therapy. The objective of this study is to evaluate the effect of the use of CSs of human dental pulp SCs, associated or not to PBMT, on the regeneration of critical size bone defects in the calvaria of diabetic rat (ZDF). The study will have two steps: one in vitro (obtaining the CSs) and other in vivo. In 72 diabetic rats, standardized critical size bone defects will be performed on the parietal bone, which will then be treated according to the following experimental groups (n = 12 per group): GI: Negative control: lesion with no additional treatment; GII: Positive Control: Placement of commercial swine collagen membrane (BioGideTM); GIII: Placement of BioGideTM followed by application of PBMT; GIV: placement of CS; GV: Placement of CS followed by PBMT and GVI: lesion with no additional treatment followed by PBMT. PBMT will be performed with infrared laser (780 nm, 40 mW, 0.04 cm2 beam area, 1 W/cm2, 3s, 3J/cm2, 0.12J per point) immediately, 48 and 96 hours after surgery. Four and eight weeks after surgery, the skull bone will be dissected, fixed and submitted to the following analysis: micro-computed tomography (MicroCT), histological (HE), histochemistry (Picro Sirius and TRAP) and immunohistochemistry (cell tracking-BrDU and markers of the Osteogenesis). The data will be treated statistically (pd0.05).
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