中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (3): 559-562.doi: 10.3969/j.issn.1673-8225.2010.03.044

• 材料生物相容性 material biocompatibility • 上一篇    下一篇

聚乳酸乙醇酸/RNA Ⅲ抑制肽缓释微球的组织相容性

李静东，张小斌，郝立波，邢庆昌，王继芳   

  1. 解放军总医院骨科，北京市   100853
  • 出版日期:2010-01-15 发布日期:2010-01-15
  • 通讯作者: 郝立波，博士，副主任医师，解放军总医院骨科，北京市 100853
  • 作者简介:李静东，男，1956年生，吉林医科大学毕业，主任医师，主要从事骨关节疾患及骨病的临床工作，尤其是髋关节、膝关节病的关节置换工作以及关节翻修手术。
  • 基金资助:

    国家自然科学基金资助项目(30640088)*

Histocompatibility of poly (lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide sustained release microspheres

Li Jing-dong, Zhang Xiao-bin, Hao Li-Bo, Xing Qing-chang, Wang Ji-fang   

  1. Department of Orthopedics, Chinese PLA General Hospital, Beijing  100853, China
  • Online:2010-01-15 Published:2010-01-15
  • Contact: Hao Li-bo, Doctor, Associate chief physician, Department of Orthopedics, Chinese PLA General Hospital, Beijing 100853, China libo_hao@hotmail.com
  • About author:Li Jing-dong, Chief physician, Department of Orthopedics, Chinese PLA General Hospital, Beijing 100853, China
  • Supported by:

     the National Natural Science Foundation of China, No. 30640088*

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摘要：

目的：通过实验评价聚乳酸乙醇酸/ RNA Ⅲ抑制肽缓释微球的组织相容性。
方法：采用Fmoc法由C端至N端先合成粗品肽；采用反相液相色谱法对RNA Ⅲ抑制肽粗品进行纯化分析，按紫外吸收峰收集组分，冷冻干燥，得到RNA Ⅲ抑制肽纯品。再采用液相复乳法制备直径50~70 μm的聚乳酸乙醇酸/RNA Ⅲ抑制肽微球。以急性全身毒性试验、MTT细胞毒性试验、肌肉内植入试验、过敏试验、热原试验对其组织相容性进行初步评价。
结果： ①急性全身毒性试验结果显示3组动物无中毒反应，无动物死亡，体质量无明显变化。②MTT细胞毒性试验结果显示两种浸提液的平均细胞增殖率均大于85%，细胞毒等级1级，不具有细胞毒性。③肌肉内植入试验结果显示RNA Ⅲ抑制肽粉末和聚乳酸乙醇酸/RNA Ⅲ抑制肽微球植入4 周后，组织未见明显充血、变性或坏死。RNA Ⅲ抑制肽粉末完全降解，微球部分降解，主要细胞成分为纤维母细胞，未见中性粒细胞及多核巨细胞等炎症细胞浸润。④过敏试验结果显示3组动物的平均原发刺激指数分别为0.38、0.33和0.31，3组间无显著差别。⑤热原试验结果显示每种材料测试的3只新西兰大白兔中，体温升高均在0.5 ℃以下，并且体温升高总度数在1.3 ℃以下，符合热原实验的评价标准。
结论：聚乳酸乙醇酸/RNA Ⅲ抑制肽微球具有良好的组织相容性。

关键词: 聚乳酸乙醇酸/RNA Ⅲ抑制肽微球, 组织相容性, 动物实验

Abstract:

OBJECTIVE: To evaluate the histocompatibility of poly (lactic-co-glycolic acid)/RNA III inhibiting peptide (PLGA/RIP) sustained release microspheres. 
METHODS: The crude peptide comprising N to C-terminals was synthesized using Fmoc method. The crude synthetic RNAIII peptide was purified by reverse phase high performance liquid chromatography, followed by component harvesting according to ultraviolet absorption peak, and freeze-drying. PLGA/RIP sustained release microspheres with a diameter of 50-70 μm were prepared using liquid-phase multiple emulsion method. The histocompatibility of PLGA/RIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test, MTT cytotoxicity test, intramuscular implantation test, sensitivity test, and pyrogen test.
RESULTS: Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass. MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%, with cytotoxicity grade 1, which indicates no cytotoxicity. Intramuscular implantation tests showed that at 4 weeks after implantation of RIP powder or PLGA/RIP microscopes, no obviously congested, degenerated, or necrotic tissue was observed. All RIP powder and a part PLGA/RIP microscopes were degraded. Fibroblasts accounted for a large proportion in all cells. No inflammatory cell infiltration, involving neutrophils and multinucleated giant cells, was observed. Sensitivity test results displayed that the average primary irritation index was 0.38, 0.33, and 0.31 in the eluent stock solution, 2% dinitoflruorobenzene, and physiological saline-administerd groups, respectively. Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5 ℃ and the sum of fervescence was under 1.3 ℃.This is in coincidence with evaluation criteria of pyrogen test.
CONCLUSION: PLGA/RIP sustained release microspheres exhibit good histocompatibility.

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