可注射性富血小板纤维蛋白混合软骨颗粒制备支撑移植物实验研究

Experimental study on preparation of supporting graft by injectable platelet-rich fibrin mixed with cartilage granule

目的探讨可注射性富血小板纤维蛋白(i-PRF)混合软骨颗粒覆盖高密度多孔聚乙烯(HPDE)制备复合软骨支撑移植物的方法。方法获取i-PRF和软骨颗粒,准备HPDE材料,分组制备复合支撑移植物,分别为A组(单纯HPDE材料组)、B组(i-PRF+HPDE材料组)、C组(软骨颗粒+HPDE材料组)和D组(i-PRF+软骨颗粒+HPDE材料组)。将移植物埋在9只新西兰白兔背部皮下(每只白兔取背部的上下左右共4个位置,分别埋入各组的1个移植物),进行动物体内实验和移植物组织学观察评估。结果肉眼观察结果显示,术后12周时,相较于C组,D组移植物从皮下取出后软骨颗粒丢失较少,表面纤维血管更丰富。苏木素-伊红(HE)染色和番红O-固绿染色结果显示,含有i-PRF的D组复合软骨支撑移植物,软骨颗粒相较C组能更早连接成片。免疫组化结果显示,含有i-PRF的D组软骨移植物中Ⅱ型胶原之间连接比C组更快。Masson染色结果显示,含有i-PRF的B组HPDE材料,其孔径内的胶原纤维长入比A组更密集。免疫荧光结果显示,含有i-PRF的B组HPDE材料,其孔径内新生血管长入比A组更密集。结论i-PRF混合软骨颗粒所形成的复合软骨支撑移植物,其软骨颗粒可更快连接成片,且软骨颗粒固定在位情况较好。相较于单纯HPDE材料,含有i-PRF的HPDE材料胶原和纤维血管在材料孔隙中的长入增多,符合临床使用要求。

Objective To investigate the preparation of composite cartilage supported grafts made by injectable platelet rich fibrin(i-PRF)mixed with cartilage particles covering on high-density porous polyethylene(HPDE).Methods Cartilage particles and i-PRF were obtained and HPDE materials were prepared.Composite supporting grafts were prepared and were divided into group A(pure HPDE material group),group B(i-PRF+HPDE material group),group C(cartilage particles+HPDE material group)and group D(i-PRF+cartilage particles+HPDE material group).The grafts were buried subcutaneously in the back of 9 New Zealand white rabbits(each white rabbit was implanted one graft on the upper,lower,left and right sides of the back),and the in vivo experiments and histological observation of the grafts were conducted.Results Visual observation results showed that group D had less cartilage particles lost,and the surface fibrous vessels were more abundant compared with the group C at 12 weeks after surgery.The results of hematoxylin and eosin(HE)staining and saffron O-solid green staining showed that the composite cartilage supported graft containing i-PRF in the group D could form cartilage particles sooner than group C.Immunohistochemical results showed the group D whose cartilage grafts contained[(CM)]i-PRF had faster typeⅡcollagen connection than group C.Masson staining results showed that collagen fibers in the pore diameter of HPDE material containing i-PRF in group B were more dense than group A.Immunofluorescence results showed that neovasculation in its aperture of HPDE material containing i-PRF in group B was more dense than group A.Conclusion The cartilage particles of composite cartilage particles formed by i-PRF mixed with cartilage particles in support graft are faster in connecting into pieces,and the cartilage particles are better fixed in place.Compared with the simple implantation of HPDE material,the collagen and fiber vascular grow more into porosity of HPDE material containing i-PRF,which is suitable for clinical requirements in application.