利福平-聚乳酸-羟基乙酸-磷酸钙骨水泥缓释复合体的实验研究

An experimental study on a slow-release complex with rifampicin-polylactic-co-glycolic acid-calcium phosphate cement

目的:制备利福平(rifampicin,RFP)-聚乳酸-羟基乙酸(polylactic-co-glycolic acid,PLGA)-磷酸钙骨水泥(calcium phosphate cement,CPC)缓释复合体(RFP-PLGA-CPC复合体),并研究其理化性质及体外释药性能。方法:采用乳化-溶剂挥发法制备RFP-PLGA缓释微球。实验分为CPC组、包埋了RFP的CPC组(RFP-CPC组)、载有RFP的PLGA缓释微球与自固化CPC复合体组(RFP-PLGA-CPC复合体组)。测定3组材料的凝固时间﹑孔隙率。通过体外药物释放实验观察释药前后的抗压强度、断面形态的变化以及体外释药情况。结果:CPC组的凝固时间最短,RFP-PLGA-CPC复合体组的凝固时间最长。CPC组的孔隙率同RFP-CPC组比较,差异有统计学意义(P<0.05);CPC组和RFP-CPC组的孔隙率与RFP-PLGA-CPC复合体组比较,差异均有统计学意义(均P<0.01)。RFP-PLGA-CPC复合体组的抗压强度与CPC组比较,差异有统计学意义(P<0.01);而RFP-CPC组和CPC组之间的抗压强度随着时间的变化逐渐表现出显著性差异(3 d:P<0.05;30和60 d:P<0.01)。CPC组在降解过程中的抗压强度的变化不大。PLGA微球的大小均一,粒径基本在100~150μm之间,微球的形态呈现出球体或是类球体,微球的表面圆润光滑,无杂质附着;CPC组的断面空隙在浸泡3 d直至60 d都没有明显变化;而RFP-CPC组的微结构变化亦不大,其断面均是小的微粒形成的;RFP-PLGA-CPC复合体组断面的孔隙明显增多,一直到60 d时PLGA微球逐渐消失,剩下空洞。RFP-PLGA-CPC复合体组无明显短时间内药物大量释放现象,60 d累计释药率达到近95%,将该复合体释药行为进行线性拟合,发现药物以恒速进行局部释放,符合零级动力学方程F=0.168×t。结论:RFP-PLGA-CPC复合体孔隙率显著高于CPC,能够持续缓慢释放有效抗结核药物,并能较长时间维持一定的力学强度。

Objective: To prepare the slow-release complex with rifampicin(RFP)-polylactic-co-glycolic acid(PLGA)-calcium phosphate cement(CPC)(RFP-PLGA-CPC complex), and to study its physical and chemical properties and drug release properties in vitro.Methods: The emulsification-solvent evaporation method was adopted to prepare rifampicin polylactic acid-glycolic acid(RFP-PLGA) slow-release microspheres, which were divided into 3 groups: a calcium phosphate bone cement group(CPC group), a CPC embedded with RFP group(RFP-CPC group), and a PLGA slow-release microspheres carrying RFP and the self-curing CPC group(RFP- PLGA-CPC complex group). The solidification time and porosity of materials were determined. The drug release experiments in vitro were carried out to observe the compressive strength, the change of section morphology before and after drug release. Results: The CPC group showed the shortest solidification time, while the RFP-PLGA-CPC complex group had the longest one. There was statistical difference in the porosity between the CPC group and the RFP-CPC group(P<0.05); Compared to the RFP-PLGA-CPC complex group, the porosity in the CPC group and the RFP-CPC group were significantly changed(both P<0.01). There was significant difference in the compressive strength between the RFP- PLGACPC complex group and the CPC group(P<0.01), while there was significant difference in the compressive strength between the RFP-CPC group and the CPC group(3 days: P<0.05; 30 and 60 days: P<0.01). The change of the compressive strength in the CPC was not significant in the whole process of degradation. The sizes of PLGA microspheres were uniform, with the particle size between 100–150 μm. The microspheres were spheres or spheroids, and their surface was smooth without the attached impurities. There was no significant change in the section gap in the CPC group after soaking for 3 to 60 days. The microstructure change in the RFP-CPC group was small, and the cross section was formed by small particles. The pores of section in the RFP-PLGA-CPC complex group increased obviously, and PLGA microspheres gradually disappeared until the 60 th day when there were only empty cavities left. The RFP-PLGA-CPC complex group had no obvious drugs sudden release, and the cumulative drug release rate was nearly 95% in the 60 days. The linear fitting was conducted for the drug release behavior of the complex, which was in accordance with zero order kinetics equation F=0.168×t.Conclusion: The porosity of RFP-PLGA-CPC complex is significantly higher than that of CPC, and it can keep slow release of the effective anti-tuberculosis drugs and maintain a certain mechanical strength for a long time.