论文部分内容阅读
目的:构建UHMWPE颗粒诱导的小鼠颅盖骨骨溶解模型以及小鼠卵巢切除骨质疏松模型,通过动物模型证实P60PLAD蛋白在体内具有破骨细胞抑制特性和骨保护作用。方法:用8周C57BL/6雄性小鼠实验。在小鼠颅盖骨沿正中线做一矢状切口,暴露0.5 cm×0.5 cm范围,保证骨膜完整性,将直径在0.1-10μm的超高分子量聚乙烯(Ultra High Molecular Weight Polyethylene)颗粒20μg均匀撒在骨膜,构建小鼠颅盖骨骨溶解模型,在局部注射PBS或者P60PLAD蛋白溶液。2周后处死,将颅骨分离进行Micro-CT检查比较骨溶解区域变化。手术切除10周雌性小鼠卵巢构建小鼠去卵巢骨质疏松模型,腹腔注射PBS或P60PLAD蛋白溶液。12周后处死后分离股骨,经4%多聚甲醛固定后行Micro-CT检查比较骨密度等参数。结果:超高分子量聚乙烯(Ultra High Molecular Weight Polyethylene)颗粒诱导的骨溶解模型中,加入P60PLAD蛋白后,颅骨局部骨溶解区域较对照组减少,骨量相关参数与对照组相比有统计学意义。在小鼠卵巢切除骨质疏松模型中,注射了P60PLAD蛋白的实验组,其骨密度(BMD)等与骨量相关参数较对照组有明显增加。结论:P60PLAD蛋白可有效抑制UHMWPE颗粒诱导的小鼠颅盖骨骨吸收模型局部的骨吸收,抑制局部破骨细胞的骨吸收作用,并可以部分逆转小鼠卵巢切除骨质疏松模型的骨质疏松,提高骨密度,在破骨细胞抑制活性和骨保护方面具有良好临床应用前景。
OBJECTIVE: To construct a mouse model of calvarial osteolysis induced by UHMWPE particles and a model of ovariectomized osteoporosis in mice. The animal model of osteoclasts and osteoprotegerin in vivo was established. Methods: Eight weeks of C57BL / 6 male mice were used. A sagittal incision was made along the midline of the calvaria of the mouse to expose the range of 0.5 cm × 0.5 cm to ensure the periosteal integrity. The Ultra High Molecular Weight Polyethylene particles with a diameter of 0.1-10 μm were evenly distributed In the periosteum, a mouse calvarial osteolysis model was constructed and a PBS or P60PLAD protein solution was injected locally. After 2 weeks, the skull was sacrificed and the skull separated for micro-CT examination to compare the change of osteolysis area. The ovariectomized mice were ovariectomized for 10 weeks to establish ovariectomized osteoporosis model. Intraperitoneal injection of PBS or P60PLAD protein solution. After 12 weeks of sacrifice, femur was isolated and fixed with 4% paraformaldehyde. Micro-CT was used to compare the parameters such as bone mineral density. RESULTS: In the osteolysis model induced by Ultra High Molecular Weight Polyethylene (PEG) particles, the local osteolysis area of skull was decreased compared with the control group after the addition of P60PLAD protein, and the relevant parameters of bone mass were statistically significant compared with the control group . In the mouse model of ovariectomized osteoporosis, the experimental group injected with P60PLAD protein showed a significant increase in bone mass (BMD) and other parameters related to bone mass compared with the control group. CONCLUSION: P60PLAD protein can effectively inhibit the local bone resorption induced by UHMWPE-induced mouse calvarial bone resorption and inhibit the osteoclastic resorption of local osteoclasts, and can partially reverse the osteoporosis of osteoporotic mice , Improve bone mineral density, and have good clinical application prospect in osteoclast inhibiting activity and bone protection.