de Oliveira Lana Sarita de Souza, de Araújo Aurigena Antunes, de Araújo Júnior Raimundo Fernandes, Barboza Carlos Augusto Galvão, Borges Boniek Castillo Dutra, da Silva José Sandro Pereira
Department of Dentistry, Post-Graduation Program in Public Health, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
Department of Biophysics and Pharmacology, Post Graduation Program in Public Health/Post Graduation Program in Pharmaceutical Science, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
Int J Exp Pathol. 2017 Apr;98(2):75-85. doi: 10.1111/iep.12226. Epub 2017 May 29.
The aim of this study was to evaluate the effect of collagen sponge scaffold (CSS) implantation associated with low-level laser therapy (LLLT) on repairing bone defects. A single 5-mm cranial defect was surgically created in forty Wistar rats, which then received one of the following four interventions (n = 10 per group): no treatment (G0); bone defect implanted with collagen sponge scaffold (CSS) alone (G1); defect treated with low-level laser therapy (LLLT) (wavelength 780 nm; total energy density 120 J/cm ; power 50 mW) alone (G2); and CSS associated with LLLT treatment (G3). After surgery, animals in each group were euthanized at 21 days and 30 days (n = 5 per euthanasia time group). Bone formation was monitored by X-ray imaging analysis. Biopsies were collected and processed for histological analysis and immunohistochemical evaluation of transforming growth factor-beta (TGF-β), fibroblast growth factor-2 (FGF-2), osteoprotegerin (OPG) and receptor activator of nuclear factor ƙ (RANK). Osteocalcin (OCN) was detected by immunofluorescence analysis. Compared to the G0 group, defects in the 30-day G3 group exhibited increased bone formation, both by increase in radiopaque areas (P < 0.01) and by histomorphometric analysis (P < 0.001). The histopathological analysis showed a decreased number of inflammatory cells (P < 0.001). The combined CCS + LLLT (G3) treatment also resulted in the most intense immunostaining for OPG, RANK, FGF-2 and TGF-β, and the most intense and diffuse OCN immunofluorescent labelling at 30 days postsurgery (G3 vs. G0 group, P < 0.05). Therefore, the use of CCS associated with LLLT could offer a synergistic advantage in improving the healing of bone fractures.
本研究旨在评估胶原海绵支架（CSS）植入联合低强度激光治疗（LLLT）对修复骨缺损的效果。在40只Wistar大鼠中通过手术制造一个单一的5毫米颅骨缺损，然后将其分为以下四种干预措施之一（每组n = 10）：不治疗（G0）；仅植入胶原海绵支架（CSS）的骨缺损（G1）；仅用低强度激光治疗（LLLT）（波长780 nm；总能量密度120 J/cm²；功率50 mW）治疗的缺损（G2）；以及CSS联合LLLT治疗（G3）。手术后，每组动物在21天和30天时实施安乐死（每个安乐死时间组n = 5）。通过X射线成像分析监测骨形成情况。收集活检样本并进行处理，以进行组织学分析以及对转化生长因子-β（TGF-β）、成纤维细胞生长因子-2（FGF-2）、骨保护素（OPG）和核因子κB受体激活剂（RANK）进行免疫组织化学评估。通过免疫荧光分析检测骨钙素（OCN）。与G0组相比，30天的G3组缺损处骨形成增加，这在不透射线区域增加（P < 0.01）以及组织形态计量分析中均有体现（P < 0.001）。组织病理学分析显示炎症细胞数量减少（P < 0.001）。联合CCS + LLLT（G3）治疗还导致术后30天时OPG、RANK、FGF-2和TGF-β的免疫染色最强，以及OCN免疫荧光标记最强且最弥散（G3组与G0组相比，P < 0.05）。因此，使用CSS联合LLLT在改善骨折愈合方面可能具有协同优势。
