冻干同种异体骨的免疫原性评价及免疫实验动物模型的选择  

Immunological evaluation into the bone allograft and selection of experimental animal model on immunogenicity

作　　者：李明东[1] 奚延斐[2] 金丹[1] 裴国献[1] 

机构地区：[1]南方医科大学南方医院创伤骨科,广州510515 [2]中国药品生物制品检定所

出　　处：《中华创伤骨科杂志》2009年第7期671-674,共4页Chinese Journal of Orthopaedic Trauma

摘　　要：目的探讨冻干同种异体骨材料的免疫原性，分析何种动物更适合做免疫相关实验研究。方法人淋巴细胞来源于10名健康青年志愿者外周静脉血，Balb／c、C57小鼠（各10只）的淋巴细胞为其脾脏所分离，新西兰兔（10只）的淋巴细胞为其外周静脉血所分离。人、Balb／c小鼠、C57小鼠、新西兰兔的淋巴细胞转化实验分别分为六组：阳性对照组（植物血球凝集素／刀豆蛋白加淋巴细胞），阴性对照组（羟基磷灰石加淋巴细胞）、骨粉A组（冻干骨粉2．0g／L加淋巴细胞）、骨粉B组（冻干骨粉1．0g／L加淋巴细胞）、骨粉c组（冻干骨粉0．5g／L加淋巴细胞）及空白组（单纯培养液加淋巴细胞），采用淋巴细胞转化实验Alamarblue法进行细胞与材料共培养，72h后酶标仪570、600nm双波长下读取吸光度值，计算各组淋巴细胞转化率。并将三种动物与人淋巴细胞转化率均数做Pearson相关分析。结果人、Balb／c小鼠骨粉A、B、c组淋巴细胞转化率与阴性对照组比较，差异无统计学意义（P〉0．05）；与阳性对照组比较，差异有统计学意义（P〈0．01）；骨粉A、B、C组之间两两比较，差异无统计学意义（P〉0．05）。C57小鼠及新西兰免各组之间淋巴细胞转化率比较差异均无统计学意义（P〉0．05）。Balb／c小鼠与人淋巴细胞转化率均数的相关系数为0．959，P=0．003，二者之间存在显著正相关；C57小鼠与人的相关系数为0．527，P=0．283，新西兰兔与人的相关系数为0．866，P=0．026。结论本研究中所采用的冻干同种异体骨粉的免疫原性对淋巴细胞的影响很低，不足以引起明显的免疫反应；在以上三种动物中，Balb／c小鼠更适合做免疫原性相关的实验研究。Objective To explore the effect of immunogenicity of freeze-dried bone allograft on different in vitro experimental models. Methods The lymphocytes were obtained respectively from 10 healthy young human volunteers, 10 Balb/c and 10 C57 mice and 10 New Zealand rabbits. The experiment was carried out in 6 groups： positive control group （PHA/ConA ＋lymphocyte）, negative control group （Hydroxyapatite powder ＋ lymphocyte）, allogeneic bone group A （Freeze-dried bone powder 2.0 g/L ＋ lymphocyte）, allogeneic bone group B （Freeze-dried bone powder 1.0 g/L ＋ lymphocyte）, allogeneic bone group C （Freeze-dried bone powder 0.5 g/L ＋lymphocyte）, and negative control group （culture solution ＋ lym- phocyte）. Lymphocyte transformation test （Alamarblue） was conducted to culture the 6 kinds of experimental materials in vitro. After 72 hours, samples were scanned with ELISA muhiscan at wave lengths 570 nm and 600 nm to read the light absorption value. Pearson analyses were performed to determine the relationships a- mong the 3 animals and 1 human groups and find out which animal would be highly correlated to human. Results In the human and Balb/c mice lymphocyte transformation tests, there was no significant difference （ P 〉 0.05） between allogeneic bone groups A, B, C and negative control group （HA）; but there was sig- nificant difference （P 〈 0. 001） between allogeneic bone groups A, B, C and positive control group （PHA/ COMA）; there was no significant difference between the 3 allogeneic bone groups （P 〉 0. 05）. There was no significant difference among the 6 groups of C57 mice and New Zealand rabbits （ P 〉 0.05）. The coefficient r between Balb/c mice and human groups was 0. 959, P =0. 003, showing a highly positive correlation. The coefficient r between C57 mice and human groups was 0. 527, P = 0. 283, while the coefficient r between New Zealand rabbits and human groups was 0. 866, P =0. 026. Conclusions The immunogenicity of freeze-dried bone powder in this

关 键 词：淋巴细胞活化 细胞毒性 免疫 同种异体骨 

分 类 号：R686[医药卫生—骨科学]