CAJ | 학술논문

目的制备丝素蛋白(SF)/重组人骨形成蛋白2 (rhBMP-2)缓释微球并观察其异位成骨活性.方法冷冻干燥法制备SF/rhBMP-2缓释微球,扫描电镜观察.酶联免疫吸附试验(ELISA)法测定不同时间点rhBMP-2浓度,并计算累积释放量及微球载药率.将SF/rhBMP-2微球及rhBMP-2分别植入24只大鼠大腿肌袋,SF微球为对照.术后1、2、4周行组织学观察.结果 SWrhBMP-2微球呈规则圆形,当SF与交联剂比例为1∶20、5∶20、10∶20,rhBMP-2与SF质量比为0.5 μg∶1 mg时分别为:(835.00 ±97.23)、(715.30±193.80)、(398.70±99.86) nm;rhBMP-2释放符合双相动力学释药规律;其载药率分别为(4.54±0.13)％、(4.55±0.05)％、(4.53±0.08)％,各组差异无统计学意义(P＞0.05).组织形态学见实验组有软骨细胞及成骨细胞形成.结论 SF/rhBMP-2微球可保持rhBMP-2的活性,能延缓rhBMP-2的释放时间,释放具有双相规律,具有持续诱导骨形成的特点.
목적 제비사소단백(SF)/중조인골형성단백2 (rhBMP-2)완석미구병관찰기이위성골활성.방법 냉동간조법제비SF/rhBMP-2완석미구,소묘전경관찰.매련면역흡부시험(ELISA)법측정불동시간점rhBMP-2농도,병계산루적석방량급미구재약솔.장SF/rhBMP-2미구급rhBMP-2분별식입24지대서대퇴기대,SF미구위대조.술후1、2、4주행조직학관찰.결과 SWrhBMP-2미구정규칙원형,당SF여교련제비례위1∶20、5∶20、10∶20,rhBMP-2여SF질량비위0.5 μg∶1 mg시분별위:(835.00 ±97.23)、(715.30±193.80)、(398.70±99.86) nm;rhBMP-2석방부합쌍상동역학석약규률;기재약솔분별위(4.54±0.13)％、(4.55±0.05)％、(4.53±0.08)％,각조차이무통계학의의(P＞0.05).조직형태학견실험조유연골세포급성골세포형성.결론 SF/rhBMP-2미구가보지rhBMP-2적활성,능연완rhBMP-2적석방시간,석방구유쌍상규률,구유지속유도골형성적특점.
Objective To prepare silk fibroin (SF) microspheres loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2) and evaluate the ectopic bone formation performance.Methods The rhBMP-2-loaded fibroin particles were prepared with freeze-drying methods.The microspheres were observed by scanning electron microscope.They were prepared by the volume ratio of silk fibroin and cosslinking agent was 1∶20,5∶20,10∶20,and the mass ratio of rhBMP-2 and SF was 0.5 μg∶1 mg.The concentration of rhBMP-2 was measured by enzyme linked immunosorbent assay (ELISA) at different time points in the phosphate buffer (PBS),calculate the cumulative release of rhBMP-2 and microsphere drug loading rate.The SF microspheres and SF/rhBMP-2 particles were implanted in 24 rats thigh muscle tissue and the SF microspheres group was the control group.The rats were killed during 4 weeks and the thigh muscle tissue were observated after hematoxylin and eosin (HE) staining.Results The shape of SF/rhBMP-2 microspheres were regular circular and the diameter decreases with the concentration of ethanol increase.Their diameters were (835.0 ±97.23),(715.30 ± 193.80),(398.70 ±99.86) nm.The rhBMP-2 release meets biphasic kinetics of drug release pattern.Drug loading were (4.54 ±0.13)％,(4.55 ±0.05)％,(4.53 ± 0.08) ％ with no significant difference (P ＞ 0.05).Tissue morphology found that lots of cartilage cells and chondrocytes were generated in experimental group.Conclusion The regenerated silk fibroin microspheres which carried the rhBMP-2 can not only maintain the activity of rhBMP-2 but also slow its release and can continue to induce bone formation.