[1]王远庆 钱浩明 陈强 吴莎莎.自体脂肪基质细胞组织-脱细胞骨基质-壳聚糖支架修复胫骨缺损及其生物相容性研究[J].中国中医骨伤科杂志,2017,25(08):11-17.
　WANG Yuanqing QIAN Haoming CHEN Qiang WU Shasha.Repair and Its Biocompatibility Effect of SVF-Decellularized Bone Matrix-Chitosan Scaffold in Repairing Tibia Defect[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2017,25(08):11-17.
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自体脂肪基质细胞组织-脱细胞骨基质-壳聚糖支架修复胫骨缺损及其生物相容性研究()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第25卷
期数:
2017年08期
页码:
11-17
栏目:
实验研究
出版日期:
2017-08-03

文章信息/Info

Title:
Repair and Its Biocompatibility Effect of SVF-Decellularized Bone Matrix-Chitosan Scaffold in Repairing Tibia Defect
文章编号:
1005-0205(2017)08-0011-07
作者:
王远庆1 钱浩明1 陈强1 吴莎莎1
1.上海市宝山区罗店医院中医骨伤科(上海,201908)
Author(s):
WANG Yuanqing1 QIAN Haoming1 CHEN Qiang1 WU Shasha1
1.Traumatology Luodian Hospital of Baoshan District of Shanghai,Shanghai 201908 China.
关键词:
自体脂肪基质细胞组织;  脱细胞骨基质;  壳聚糖支架;  胫骨缺损;  生物相容性
Keywords:
autologous adipose stromal cells;  decellularized bone matrix;  chitosan scaffold;  tibia defect;  biocompatibility
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨自体脂肪基质细胞组织(SVF)-脱细胞骨基质-壳聚糖支架修复胫骨缺损及其生物相容性。方法:于2016年4月9日选取辽宁医学院动物实验中心4月龄大白兔40只,随机选取其中1只提取兔SVF组织10 g,体外分离培养脂肪基质细胞并行脂肪基质细胞多向分化(成脂和成骨诱导),以种植密度3×107个/cm2种植于脱细胞骨基质-壳聚糖支架上并制成SVF-脱细胞骨基质-壳聚糖支架,将P3代的兔纤维软骨细胞分别种植到SVF-脱细胞骨基质-壳聚糖支架(作为SVF组)和脱细胞骨基质-壳聚糖支架(作为对照组)并进行组织学和生物相容性检测,同时将所有大白兔的建立兔胫骨缺损模型,并用上述两种支架进行移植修复治疗,统计分析所有支架第0,4,7,14天的DNA含量、细胞存活率和第0,2,4,8周时兔胫骨骨矿物、骨密度水平及修复疗效、第8周时最大弯曲度负荷、抗弯刚度、破坏扰度水平。结果:SVF组第4,7,14天时DNA含量明显高于对照组,差异有统计学意义(P<0.05); SVF组第4,7,14天时细胞存活率明显高于对照组,差异有统计学意义(P<0.05); 术后第8周,SVF组大体观察可见仿生骨膜大部分完整、有骨痂生长,组织学观察可见软骨内骨化明显、缺损基本修复,免疫组织化学观察可见成熟骨基质部分BMP染色阳性、着色变浅,对照组大体观察可见膜降解呈碎片状、骨端骨痂较硬,组织学观察可见膜材大部分降解且出现幼稚髓腔样结构,免疫组织化学观察BMP轻度表达、软骨基质BMP染色阳性,前者第2,4,8周时骨矿物、骨密度和第8周时最大弯曲度负荷、抗弯刚度、破坏扰度水平明显高于后者,差异有统计学意义(P<0.05)。结论:SVF-脱细胞骨基质-壳聚糖支架具有良好的生物相容性,SVF可作为骨组织工程的种子细胞,可能通过协同诱导、传导效应等增强骨再生麓力,且具有良好的生物力学效果,有利于促进胫骨骨缺损的修复,值得临床进一步推广。
Abstract:
Objective:To discuss the repair and its biocompatibility effect of autologous adipose stromal cells(SVF)-decellularized bone matrix-chitosan scaffold in repairing tibia defect.Methods:White rabbit on April 9,2016 were selected in Liaoning medical school animal experiment center,Aaccording to the random distribution,one extract rabbit were selected to extract 10 g SVF organization of the rabbit,adipose stromal cell cultivation in vitro in parallel adipose stromal cells(induced)into fat and osteogenesis,multi-directional differentiation to planting density 3×107/cm2 were grown in cell-free bone matrix-chitosan scaffold and made into SVF-decellularized bone matrix-chitosan scaffold,P3 generation of rabbit fibrocartilage cells were grown respectively to SVF-decellularized bone matrix-chitosan scaffold(as SVF group),and decellularized bone matrix-chitosan scaffold(as control group),and they were given histological and biological compatibility testing,at the same time,all the big white rabbit rabbit tibia defect model was established,and the above two stents transplant repair treatment, statistical analyzed all stents 0,4,7,14 d of DNA content,cell survival and 0,2,4,8 weeks of rabbit tibia bone mineral,bone mineral density levels and repair effect,8 weeks maximum bending load,bending stiffness,destroy the immunity levels.Results:4,7,14 d DNA content of SVF group were significantly higher than those of control group,the difference were statistically significant(P<0.05); 4,7,14 d cell survival rate of SVF group were significantly higher than those of control group,the difference were statistically significant(P<0.05); Postoperative 8 weeks,gross observation visible bionic periosteum most complete,SVF group had the callus growth,histological observation endochondral ossification clearly visible,basic repair defect,immunohistochemical observations were cooked bone matrix section BMP dyeing,color becomes shallow,membranous degradation fragmented,and control group in extremities callus harder, histological observation most visible membrane material degradation and appear naive medullary cavity structure,BMP immunohistochemical observation mild cartilage matrix expression,BMP staining positive,2,4,8 weeks bone mineral density and bone minerals at 8 weeks maximum bending load, bending stiffness,destroy the immunity levels of the former were significantly higher than those of the latter,the difference were statistically significant(P<0.05).Conclusions:SVF-decellularized bone matrix-chitosan stent has good biocompatibility,SVF can be used as bone tissue engineering seed cells,it possibly through coordination induction,conduction effect enhancement foot of bone regeneration,and it has a good biomechanical effect,promote the tibia bone defect repair, it's worth for further clinical promotion.

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更新日期/Last Update: 2017-08-15