不同比例激活剂对富血小板凝胶的生物学影响

Effect of different proportions of activators on biological functions of platelet-rich gel

摘要:

目的比较不同比例激活剂对富血小板凝胶的生物学影响，研究其用于角膜基质再生的可行性。方法抽取兔全血，经二次离心法制备富血小板血浆（PRP），将PRP 与激活剂按不同比例激活，制备PRP 凝胶。实验分为3 个实验组，分别为9：1 组（PRP：激活剂=9：1）、11：1 组（PRP：激活剂=11：1）及5：1 组（PRP：激活剂=5：1）。将兔角膜基质细胞与PRP凝胶共同培养，进行组织形态学观察用细胞增殖- 毒性检测试剂盒法，分别检测3个实验组PRP凝胶释放的复合生长因子促角膜基质细胞的增殖作用。对不同实验组制备的PRP凝胶与双层猪角膜基质进行黏附性观察。结果9：1 组制备的PRP 凝胶促进兔角膜基质细胞的增殖和活化作用强于其他实验组。3 个实验组中，只有9：1 组制备的PRP 凝胶苏木精-伊红染色法染色结果显示，呈有序的网状支架结构，能紧密黏附并固定双层猪角膜基质。结论PRP与激活剂以9：1比例制备的PRP凝胶促角膜基质细胞增殖作用最强，同时具有有序网状结构和良好的组织黏附性及相容性，是一种可以用于角膜基质再生的良好支架。

Abstract:

Objective To compare the effect of different proportions of activators on the biological function of platelet-rich gel and to reaserch the feasibility for the reconstruction of corneal stroma. Methods Rabbit blood was extracted and platelet-rich plasma (PRP) was prepared by two-step centrifugation. Platelet-rich gels were prepared by activation of PRP with thrombin activators in different proportions and divided into three groups. There were group 9:1 (PRP : activators = 9 : 1), group 11:1 (PRP : activators = 11 : 1) and group 5:1 (PRP : activators = 5 : 1). The rabbit keratocytes were seperately co-cultured with the three platelet-rich gels and the morphological features were observed. CCK-8 assay was used to analyze the influences of composite growth factors released from the platelet-rich gel on the proliferation of rabbit keratocytes in the three groups.The adhesive properties of the three platelet-rich gels were evaluated after adhesion to the double-layer porcine corneal stroma. Results The platelet-rich gel of the group 9:1 had better effect on promoting cell proliferation than those of other two groups. HE staining displayed that only the platelet-rich gel prepared in the group 9:1 formed an orderly network scaffold that could closely attach to the double-layer porcine corneal stroma. Conclusions The platelet-rich gel, prepared by combining PRP with activators in 9 : 1 proportion, can effectively promote the proliferation of keratocytes, and has the nature of orderly network structure, effective adhesiveness and favorable biocompatibility. These properties indicate that the platelet-rich gel prepared by combining PRP with activators in 9 : 1 proportion is an excellent scaffold for the reconstruction of corneal stroma.