中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (36): 6755-6758.doi: 10.3969/j.issn.1673-8225.2010.36.025

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

液氮冻融促进血小板源性生长因子AA与转化生长因子β1的释放

李洪涛1，段建民1，周  谋1，马玉霞1，片山直○2   

  1. 1解放军广州军区广州总医院口腔科，广东省广州市    510010；2明海大学齿学部机能保存恢复学讲座，日本琦玉县 350-0283
  • 出版日期:2010-09-03 发布日期:2010-09-03
  • 通讯作者: 段建民，硕士生导师，主任医师，解放军广州军区广州总医院口腔科， 广东省广州市 510010 duanjianmin3@ 126.com
  • 作者简介:李洪涛★，男，1976年生，河南省洛阳市人，南方医科大学在读硕士，主治医师，主要从事牙体牙髓病的基础与临床研究。 lihongtao1976@hotmail.com
  • 基金资助:

    广东省科技计划项目国际合作基金资助项目(2007B050200017)。

Freeze-thaw by liquid nitrogen promotes platelet-derived growth factor-AA and transforming growth factor-β1 release 

Li Hong-tao1, Duan Jian-min1, Zhou Mou1, Ma Yu-xia1, Katayama Tadashi2   

  1. 1 Department of Stomatology, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA, Guangzhou  510010, Guangdong Province, China; 2 School of Dentistry, Meikai University, Saitama  350-0283, Japan
  • Online:2010-09-03 Published:2010-09-03
  • Contact: Duan Jian-min, Master’s supervisor, Chief physician, Department of Stomatology, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA, Guangzhou 510010, Guangdong Province, China duanjianmin3@ 126.com
  • About author:Li Hong-tao★, Studying for master’s degree, Attending physician, Department of Stomatology, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA, Guangzhou 510010, Guangdong Province, China lihongtao1976@ hotmail.com
  • Supported by:

     the International Cooperation Foundation Program of Science and Technology Project of Guangdong Province, No. 2007B050200017

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摘要：

背景：血小板被认为是人体自身的生长因子库，目前多采用牛凝血酶激活血小板，促进释放其富含的多种生长因子，牛凝血酶的临床应用存在一定的风险。
目的：与牛凝血酶激活方式比较，观察液氮冻融促进血小板释放血小板源性生长因子AA、转化生长因子β1的作用效果。
方法：抽取5名健康志愿者静脉血，采用二次离心法制备富血小板血浆及乏血小板血浆，洗涤去除富血小板血浆中的血浆成分，制成洗涤血小板。洗涤血小板采用液氮反复冻融的方法促进其释放生长因子，而富血小板血浆采用1 000，500，250，125，62.5，31.25 U/mL的牛凝血酶-氯化钙溶液进行激活。应用酶联免疫吸附试验法定量检测洗涤血小板、富血小板血浆及乏血小板血浆中血小板源性生长因子AA、转化生长因子β1的质量浓度。
结果与结论：液氮冻溶促进血小板释放血小板源性生长因子AA和转化生长因子β1的质量浓度分别为(8.973±1.213)，(27.445±2.273) μg/L，与500~1 000 U/mL牛凝血酶激活血小板促进其释放血小板源性生长因子AA和转化生长因子β1的质量浓度差异无显著性意义(P >0.05)。提示液氮冻融作为一种安全、有效的激活洗涤血小板方式可以替代目前使用的牛凝血酶。

关键词: 液氮, 冻融, 富血小板血浆, 洗涤血小板, 血小板源性生长因子, 转化生长因子, 酶联免疫吸附试验

Abstract:

BACKGROUND: Platelet is a rich source of autologous growth factors. Bovine thrombin is the most common used activator to promote growth factors release from platelet. The clinical use of thrombin has some risks.
OBJECTIVE: To observe the effect of freeze-thaw by liquid nitrogen to promote platelet-derived growth factor-AA and transforming growth factor-β1 release from platelet in comparison with bovine thrombin.
METHODS: Platelet rich plasma and platelet poor plasma were manufactured from venous blood of five healthy volunteers by secondary centrifugation. The platelet rich plasma was then washed to get rid of the plasma and to manufacture the washed platelet. Washed platelet was activated by freeze-thaw method using liquid nitrogen to promote growth factor release and platelet rich plasma was activated by 1 000, 500, 250, 125, 62.5, 31.25 U/mL bovine thrombin/calcium chloride. Mass concentrations of platelet-derived growth factor-AA and transforming growth factor-β1 in washed platelet, platelet rich plasma and platelet poor plasma were assayed using enzyme linked immunosorbent assay. 
RESULTS AND CONCLUSION: The concentrations of platelet-derived growth factor-AA and transforming growth factor-β1 in the washed platelet activated by freeze-thaw were (8.973±1.213) and (27.445±2.273) μg/L. There was no significant difference from the concentration in the bovine thrombin of 500-1 000 U/mL activated platelet rich plasma (P > 0.05). These suggested that freeze-thaw can be used as a safe and effective technique to active washed platelet instead of bovine thrombin.

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