中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (8): 1353-1356.doi: 10.3969/j.issn.1673-8225.2010.08.006

• 组织工程血管材料 tissue-engineered vascular materials • 上一篇    下一篇

管状材料对血小板激活作用的体外评价

许建霞，王春仁，奚廷斐   

  1. 中国药品生物制品检定所医疗器械检验中心，北京市 100050
  • 出版日期:2010-02-19 发布日期:2010-02-19
  • 作者简介:许建霞★，女，1976年生，山西省稷山县人，汉族，2003年中国药品生物制品检定所毕业，硕士，助理研究员，主要从事医疗器械生物学评价。 xujianxia@nicpbp.org.cn

In vitro evaluation of platelet activation by tubular biomaterials

Xu Jian-xia, Wang Chun-ren, Xi Ting-fei   

  1. National Institute for the Control of Pharmaceutical and Biological Products, Beijing  100050, China
  • Online:2010-02-19 Published:2010-02-19
  • About author:Xu Jian-xia★, Master, Assistant researcher, National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China xujianxia@nicpbp.org.cn

PDF

426

被引次数

4

摘要：

背景：ISO10993-4及GB/T 16886.4中将血液相容性的评价分为5个方面：血栓形成、凝血、血小板、补体、血液学。目前国内较为确定和成熟的体外血液相容体外评价方法有溶血试验、凝血试验及血小板黏附试验，而对血小板激活及补体系统激活方面的研究很少。
目的：评价聚乙烯、聚氯乙烯及聚甲基乙烯基硅氧烷3种基础材料管在体外对血小板的激活作用，初步建立一种体外评价管状材料对血小板激活作用的方法。
方法：制备聚氯乙烯管、聚乙烯管、硅橡胶管的内径3.7 mm，外径5 mm，长35 cm。每管1 mL血液注入聚乙烯、聚氯乙烯、硅橡胶管，管的两端用两通连接，置于恒温培养振荡器中，30°倾斜，接口向上，37 ℃，140 r/min，振荡3.5 h。放射免疫法检测材料与血液接触后贫血小板血浆中血小板α颗粒蛋白水平，流式细胞仪检测材料与血液接触后血液中血小板α颗粒蛋白阳性血小板百分率、活化的gpⅡb/Ⅲa复合物阳性血小板百分率。
结果与结论：放射免疫法检测结果显示聚乙烯、聚氯乙烯管与血液接触后贫血小板血浆中血小板α颗粒蛋白水平大于硅橡胶管(P < 0.05)。聚乙烯和聚氯乙烯管与血液接触后贫血小板血浆中血小板α颗粒蛋白水平差异无显著性意义(P > 0.05)。流式细胞术检测结果显示聚乙烯、聚氯乙烯管与血液接触后血液中血小板α颗粒蛋白阳性血小板百分率大于硅橡胶管(P < 0.05)，聚乙烯管与血液接触后血液中血小板α颗粒蛋白阳性血小板百分率大于聚氯乙烯管(P < 0.05)。3种材料与血液接触后血液中活化的gpⅡb/Ⅲa复合物阳性血小板百分率差异无显著性意义(P > 0.05)。实验初步建立一种管材料与血液较为合理的接触方式，并可以考虑血浆血小板α颗粒蛋白是较好的反映血小板激活程度的评价指标。用流式细胞术检测血浆血小板α颗粒蛋白阳性血小板百分率更为敏感。

关键词: 管状材料, 血小板激活, 血浆血小板&alpha, 颗粒蛋白, 血液相容性, 生物材料

Abstract:

BACKGROUND: In according to ISO-10993-4 and GB/T 16886.4, the in vitro hemo-compatibility evaluation on biomaterials includes thrombosis, coagulation factors, platelets and platelet functions, hematology and complement system. However, in the case of China, the in vitro hemo-compatibility evaluations were performed only thrombosis, coagulation factors and platelet attachment, the investigation on evaluation of platelet and complement activations is less reported.
OBJECTIVE: To evaluate the effect of polyethylene, polyvinyl chloride and polymethylvinylsiloxane tubes on platelet activation, and establish a useful method to evaluate the effect of tubular materials on platelet activation.
METHODS: Tubes of polyethylene, polyvinyl chlorid and silastic were established by 3.7 mm inner diameter, 3.5 mm external diameter, and 35 cm length, respectively. 1 mL blood was injected into the tube of polyethylene, polyvinyl chlorid and silastic, respectively. The tubes were connected using a two-way tube, shaken at 140 r/min by 30° sloping for 3.5 hours at 37 ℃. Radioimmunoassay was employed to detect α-granules protein level of platelet poor plasma, while flow cytometry was used to detect the percentage of positive platelet of α-granules protein and that of activated gpⅡb/Ⅲa composite.
RESULTS AND CONCLUSION: Radioimmunoassay showed that α-granules protein level of platelet poor plasma in the polyethylene and polyvinyl chlorid tubes was significantly greater than that in the silastic tube (P < 0.05). There were no significant differences in α-granules protein between polyethylene and polyvinyl chlorid (P > 0.05). Flow cytometry indicated that percentage of positive platelet of α-granules protein in the polyethylene and polyvinyl chlorid tubes was significantly greater than that in the silastic tube (P < 0.05); the percentage in the polyethylene tube was significantly greater than that in the polyvinyl chlorid tube (P < 0.05). There was no significant differences in the percentage of positive platelet of activated gpⅡb/Ⅲa composite between the three materials (P > 0.05). A useful blood-material contact model was established, and it was considered that α-granules protein is an available parameter for evaluating platelet activation. The percentage of positive platelet of α-granules protein determined by flow cytometry was a more sensitive parameter for evaluating platelet activation.

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