Ma Shu-Yuan, Feng Zhi-Qiang, Lai Ren-Fa, Yin Zhong-Da
First Affiliated Hospital of Ji'nan University, Guangzhou, Guangdong-510630, China.
First Affiliated Hospital of Ji'nan University, Guangzhou, Guangdong-510630, China.
Asian Pac J Trop Med. 2015 Jan;8(1):53-9. doi: 10.1016/S1995-7645(14)60187-5.
To investigate the synergistic effect and mechanism of the combined application of recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF).
24 KM male mice were randomly divided into 6 groups with 4 mice in each group, namely, Group A (control group), Group B (only treated with collagen), Group C (treated with 2 ng bFGF+collagen), Group D (treated with 4 μ g rhBMP-2+collagen), Group E (treated with 4 μ g rhBMP-2+2 ng bFGF+collagen) and Group F (treated with 4 μ g rhBMP-2+4 ng bFGF+collagen). The composites were implanted into the intermuscular septum of hind legs mice; whereas in control group, intermuscular septum of mice was separated and no implantation was performed. General observation, detection of concentration of calcium content, micro computed tomography (Micro-CT), three-dimensional reconstruction scan, measurement of bone mineral density (BMD), bone volume fraction (BVF) and trabecular thickness (Tb.Th), as well as histological observation with HE staining and ALP and CD34 immumohistochemical staining were performed.
Ectopic osteogenesis was found in Groups D, E and F mice. The difference in concentration of calcium contents was statistically significant between Groups D and E (P<0.05), but insignificant between Groups E and F (P>0.05). Micro-CT and three-dimensional reconstruction revealed continuous newborn bone substance in external surface of ectopic bone formation, and the center of bone formation did not show obvious substantial filling by bone substance. The differences in BMD, BVF and Tb.Th were statistically significant between Groups D and E or F (P<0.01 or <0.05). HE staining showed that in Groups D, E and F, newborn bone substance was mainly located at the edge of ectopic bone formation, and the bone formation in Groups E and F was better than that in Group D. ALP and CD34 immumohistochemical staining revealed the positive expression mainly at the edge of ectopic bone formation, and area of positive expression in Groups E and F was larger than that in Groups D.
rhBMP-2 possesses the capacity to induce ectopic osteogenesis independently, but bFGF does not have this ability; the combined application of rhBMP-2 and bFGF can enhance the synergetic effect on inducing ectopic osteogenesis.
探讨重组人骨形态发生蛋白-2（rhBMP-2）与碱性成纤维细胞生长因子（bFGF）联合应用的协同效应及机制。
将24只雄性KM小鼠随机分为6组，每组4只，即A组（对照组）、B组（仅用胶原蛋白处理）、C组（用2 ng bFGF+胶原蛋白处理）、D组（用4 μg rhBMP-2+胶原蛋白处理）、E组（用4 μg rhBMP-2+2 ng bFGF+胶原蛋白处理）和F组（用4 μg rhBMP-2+4 ng bFGF+胶原蛋白处理）。将复合材料植入小鼠后腿肌间隔；而对照组分离小鼠肌间隔但不进行植入。进行一般观察、钙含量浓度检测、微型计算机断层扫描（Micro-CT）、三维重建扫描、骨密度（BMD）、骨体积分数（BVF）和骨小梁厚度（Tb.Th）测量，以及HE染色、碱性磷酸酶（ALP）和CD34免疫组织化学染色的组织学观察。
D、E、F组小鼠出现异位成骨。D组和E组钙含量浓度差异有统计学意义（P<0.05），但E组和F组差异无统计学意义（P>0.05）。Micro-CT和三维重建显示异位骨形成外表面有连续的新生骨质，骨形成中心未见明显的骨质实质性填充。D组与E组或F组之间的BMD、BVF和Tb.Th差异有统计学意义（P<0.01或<0.05）。HE染色显示，D、E、F组新生骨质主要位于异位骨形成边缘，E组和F组的骨形成优于D组。ALP和CD34免疫组织化学染色显示阳性表达主要位于异位骨形成边缘，E组和F组的阳性表达面积大于D组。
rhBMP-2具有独立诱导异位成骨的能力，而bFGF没有这种能力；rhBMP-2与bFGF联合应用可增强诱导异位成骨的协同效应。
