猪源性骨支架材料的制备及性能研究

Preparation of pig bone carrier and its properties

目的:制备猪源性骨支架材料,并与人源性骨支架材料对比检测其理化性能和组织相容性。方法:经低温深冻、超声清洗、H2O2、酒精浸泡、冻干、辐照制备猪源性骨支架材料和人源性骨支架材料。扫描电镜观察,测定材料孔隙率、蛋白质含量、钙磷含量及弹性模量。2种材料的浸提液与脂肪源间充质干细胞复合培养,观察细胞一般形态,流式细胞仪PI法检测细胞生命周期。皮下植入2种材料,在植入4、8、12、16周取材做病理切片观察和扫描电镜观察。结果:2种材料均具有骨本身的天然网状三维支架系统。猪源性骨支架材料的孔隙率高于人源性骨支架材料,蛋白含量低于人源性骨支架材料,弹性模量分别为无显著差异。材料浸提液组及空白对照组的细胞生长状态良好。流式细胞仪PI法检测细胞周期见G1期、G2期细胞百分率接近。皮下植入试验表明,随着植入时间的延长,炎症反应逐步减轻,材料降解增加,新生软骨样结构逐渐增多。结论:猪源性异种骨支架材料在理化性能和材料毒性等方面与同种异体骨支架材料接近,具有良好的应用前景。

Objectivc：To prepare pig bone carrier and explore its properties and biocompatibility by comparing it with human bone carrier. Methods： Pig and human bone carriers were prepared by the process of ultra low temperature freezing, ultrasonic cleaning, H2O2 and alcohol soaking, freeze drying and radiation treatment. For different carriers, the following performances were concerned： scanning electron microscope, factor of porosity, Young＇s modulus, the contents of protein, Ca and P. Meanwhile, adipose tissue-derived mesenchymal stem cells were cultured with leaching solution of two kinds of bone carriers. The shape of cells was observed under inverted microscope, and cell life cycle was explored by flow cytometry. After that, bone materials were implanted into subcutaneous tissue of mouse and analyzed after 4, 8, 12 and 16 week. Results： Natural three dimensional network structures can be found in two kinds of carriers. The interval porosity of pig bone carrier was higher than that of human. The contents of protein of pig carrier were lower than that of the human. The Young＇s modulus was the same for two kinds of bone carriers. Cells survived well in leaching solution from pig and human bone carrier, as well the blank control group. The PI method proved the better proliferation ability of cells supplied to different leaching solutions. Inflammation reaction of implantation materials reduced, degradation increased, while chondroid structures increased gradually. Conclusions： Pig bone carrier is the same as human bone carrier in properties and toxicity, which can be used in the field of bone tissue engineering.