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凹凸棒石/Ⅰ型胶原/聚己内酯复合材料修复兔骨缺损的实验研究

[EXPERIMENTAL STUDY ON BONE DEFECT REPAIR WITH COMPOSITE OF ATTAPULGITE/COLLAGEN TYPE I/POLY (CAPROLACTONE) IN RABBITS].

作者信息

Zhang Xiaomin, Song Xuewen, Wang Wei, Li Zhenjun, Zhao Hongbin

机构信息

Institute of Orthopedics, Lanzhou General Hospital of Lanzhou Military Command of Chinese PLA, Lanzhou Gansu, 730050, P. R. China.

Collage of Life Sciences and Technology, Gansu Agriculture University.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2016 May 8;30(5):626-633. doi: 10.7507/1002-1892.20160126.

Abstract

OBJECTIVE

To investigate the effect of repairing radial bone defect with scaffold material of attapulgite/collagen type I/poly (caprolactone) (ATP/Col I/PCL) in rabbits and the possibility as bone graft substitutes.

METHODS

ATP/Col I/PCL materials were prepared via adding ATP to hexafluoroisopropanol after dissolved Col I/PCL (3:2), and Col I/PCL materials via dissolving Col I/PCL (3:2) in hexafluoroisopropanol served as control. The structure of scaffolds was observed under scanning electron microscope (SEM). Twenty-four Japanese white rabbits (male, 2 months old) were used to establish the bilateral radius defect model of 15 mm in length, and randomly divided into group A (6 rabbits, 12 defects), group B (9 rabbits, 18 defects), and group C (9 rabbits, 18 defects); then the Col I/PCL scaffold was implanted in the bone defect area in group B, the ATP/Col I/PCL scaffold in group C, no treatment was done in group A as control. The general condition of rabbits was observed after operation, and bone defect repair was evaluated by X-ray at 4, 8, and 12 weeks. At 12 weeks, the tissue of defect area was harvested for the general, SEM, Micro-CT, histological, and immunohistochemical staining to observe defect repair and material degradation.

RESULTS

SEM observation showed that two kinds of materials were porous structure, ATP/Col I/PCL structure was more dense than Col I/PCL. All animals survived to the end of experiment, and no incision infection occurred during repair process.X-ray films showed that the bone marrow cavity was re-opened in defect area of group C with time, the repair effect was superior to that of groups A and B. At 12 weeks after operation, general observation showed that scaffold material had good fusion with the surrounding tissue in groups B and C, defect was filled with connective tissue in group A. SEM indicated that the surface and pore of the scaffold were covered with a large number of cells and tissues in groups B and C. Micro-CT demonstrated that the new bone volume, bone mineral content, tissue mineral content, and connectivity density of group C were significantly higher than those of groups A and B (<0.05). The observation of histology and immunohistochemical staining indicated that there were lots of connective tissues in defect area of group A, and ALP, Col I, and OPN were weakly expressed; there were many collagen fibers in scaffold degradation area in group B, and the expression levels of ALP, Col I, and OPN were higher than those of group A; there was few new bone in group C, the degradation rate of the scaffold was slower than that of group B, and the expression of Col I and OPN were enhanced, while ALP was weakened when compared with groups A and B.

CONCLUSIONS

ATP/Col I/PCL composite scaffold material can degrade , and has dense three-dimensional porous structure, good biocompatibility, and high potentiality of bone repair, so it can be used as bone substitute material.

摘要

目的

探讨凹凸棒石/Ⅰ型胶原/聚己内酯(ATP/Col I/PCL)支架材料修复兔桡骨缺损的效果及作为骨移植替代物的可能性。

方法

将Col I/PCL(3∶2)溶解于六氟异丙醇后加入ATP制备ATP/Col I/PCL材料,将Col I/PCL(3∶2)溶解于六氟异丙醇制备Col I/PCL材料作为对照。在扫描电子显微镜(SEM)下观察支架结构。选用24只2月龄雄性日本大耳白兔建立双侧桡骨15 mm长缺损模型,随机分为A组(6只兔,12处缺损)、B组(9只兔,18处缺损)和C组(9只兔,18处缺损);B组将Col I/PCL支架植入骨缺损区,C组植入ATP/Col I/PCL支架,A组不做处理作为对照。术后观察兔的一般情况,于4、8、12周时通过X线评估骨缺损修复情况。12周时,取缺损区组织进行大体、SEM、Micro-CT、组织学及免疫组织化学染色观察缺损修复及材料降解情况。

结果

SEM观察显示,两种材料均为多孔结构,ATP/Col I/PCL结构比Col I/PCL更致密。所有动物均存活至实验结束,修复过程中无切口感染发生。X线片显示,C组缺损区骨髓腔随时间重新开放,修复效果优于A组和B组。术后12周大体观察显示,B组和C组支架材料与周围组织融合良好,A组缺损区被结缔组织填充。SEM显示,B组和C组支架表面及孔隙覆盖大量细胞和组织。Micro-CT显示,C组新骨体积、骨矿物质含量、组织矿物质含量及连通性密度均显著高于A组和B组(P<0.05)。组织学及免疫组织化学染色观察显示,A组缺损区有大量结缔组织,碱性磷酸酶(ALP)、Ⅰ型胶原(Col I)和骨桥蛋白(OPN)表达较弱;B组支架降解区有较多胶原纤维,ALP、Col I和OPN表达水平高于A组;C组新骨较少,支架降解速度慢于B组,Col I和OPN表达增强,与A组和B组相比ALP表达减弱。

结论

ATP/Col I/PCL复合支架材料可降解,具有致密的三维多孔结构、良好的生物相容性和较高的骨修复潜力,可作为骨替代材料。

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