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丝素蛋白对聚左旋乳酸-共-ε-己内酯电纺丝支架体内降解及生物相容性的作用
程自申1,杨成伟1,刘莉2,杨洋1,邓国英1,叶晓健1*
0
(1. 第二军医大学长征医院骨科, 上海 200003;
2. 河北省保定市第一中心医院妇产科, 保定 071000
*通信作者)
摘要:
目的 探讨丝素蛋白(SF)对聚左旋乳酸-共-ε-己内酯[P(LLA-CL)]电纺丝支架的体内降解及生物相容性的作用。方法 运用静电纺丝技术分别制备[P(LLA-CL), (w/w=1:1)及混有含25% SF的P(LLA-CL)[SF/P(LLA-CL)]纳米纤维支架, 将两种支架分别植入至45只6个月龄大鼠皮下6个月, 以观察评估SF对P(LLA-CL)降解及生物相容性的影响。结果 病理切片显示, 支架植入3个月时, P(LLA-CL)支架明显肿胀, 并开始分层, 6个月时支架已支离破碎;而SF/P(LLA-CL)支架植入6个月时仍能保持相对完整的结构。免疫组化切片显示, 在支架植入1个月时 P(LLA-CL)支架表面及内部有大量的巨噬细胞, 3个月时仍有大量的巨噬细胞, 同时伴有异物巨细胞生成;而SF/P(LLA-CL)支架组巨噬细胞及异物巨细胞在各个时间点表达均不明显。炎症基因相对表达结果显示, 在支架植入1周时 P(LLA-CL)支架组TNF-α 及 IL-10相对表达高于SF/P(LLA-CL)支架组(P<0.05), 1个月时P(LLA-CL)支架组TNF-αIL-1βIL-10相对表达高于SF/P(LLA-CL)支架组(P<0.05), 2个月时 P(LLA-CL)支架组TNF-αIL-10相对表达高于SF/P(LLA-CL)支架组(P<0.05), 3个月时P(LLA-CL)支架组TGF-β相对表达高于SF/P(LLA-CL)支架组(P<0.05), 6个月时P(LLA-CL)支架组IL-1βTGF-β相对表达高于SF/P(LLA-CL)支架组(P<0.05)。结论 SF能延缓P(LLA-CL)降解, 减轻炎症反应, 改善生物相容性。
关键词:  丝素蛋白  聚左旋乳酸-共-ε-己内酯  降解  炎症反应  巨噬细胞  异物巨细胞  生物相容性
DOI:10.3724/SP.J.1008.2015.00500
投稿时间:2014-11-08修订日期:2015-01-19
基金项目:国家自然科学基金(81472071).
Effect of silk fibroin on degradation and in vivo biocompatibility of poly (L-lactic-co-ε-caprolactone) electronspun nanofibrous scaffolds
CHENG Zi-shen1,YANG Cheng-wei1,LIU Li2,YANG Yang1,DENG Guo-ying1,YE Xiao-jian1*
(1. Department of Orthopedics, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China;
2. Department of Obstetrics and Gynecology, First Central Hospital of Baoding, Baoding 071000, Hebei, China
*Corresponding author)
Abstract:
Objective To investigate the effect of silk fibroin (SF) on degradation and biocompatibility of poly (L-lactic acid-co-ε-caprolactone)(P[LLA-CL]) in vivo. Methods The scaffolds of P(LLA-CL) (w/w=1:1) blended with 25% of SF (SF/P[LLA-CL]) and P(LLA-CL) were prepared by electrospinning. Both kinds of scaffolds were subcutaneously implanted in 45 6-month-old rats for up to 6 months to evaluate their degradation and biocompatibility characteristics. Results Pathological sections showed P(LLA-CL) scaffold become swollen and began to separate into different layers after 3 months, and then become broken after 6 months; while SF/P(LLA-CL) scaffold largely maintained its structure after 6 months. Immunohistochemical staining showed a large number of macrophages on the surface and in P(LLA-CL) scaffolds 1 month after implantation, and they could still be found 3 months after implantation, accompanied by foreign body giant cells; while no obvious macrophages or foreign body giant cells were found in SF/P(LLA-CL) scaffolds at different time points. Examination of inflammatory gene expression showed that TNF-α and IL-10 expression in P(LLA-CL) scaffolds was significantly higher than that in SF/P(LLA-CL) scaffolds 1 week after implantation (P<0.05), the same was also true for TNF-α, IL-1β and IL-10 expression 1 month after implantation (P<0.05), for TNF-α and IL-10 expression 2 months after implantation (P<0.05), for TGF-β expression 3 months after implantation (P<0.05), and for IL-1β and TGF-β expression 6 months after implantation (P<0.05). Conclusion SF incorporation can delay degradation, reduce inflammation, and improve the biocompatibility of P(LLA-CL) scaffolds, which may provide reference for scaffold design in tissue engineering.
Key words:  silk fibroin  poly(L-lactic-co-ε-caprolactone)  degradation  inflammation  macrophage  foreign body giant cells  biocompatibility