重组人骨形成蛋白2骨诱导中神经生长因子及其受

目的 观察重组人骨形成蛋白2(recombined human bone morphogenetic protein 2, rhBMP-2)异位诱导成骨过程中神经生长因子(nerve growth factor, NGF)及其高亲和力受体(tyrosine kinase receptor A, TrkA)的表达,探讨NGF在BMP骨诱导中的作用。 方法 ICR小鼠36只,随机分

・组织工程・

重组人骨形成蛋白2骨诱导中神经生长因子

及其受体的表达

夏胜利　戴　戎　汤亭亭

　　【摘　要】　　目的　观察重组人骨形成蛋白2(recom b ined hum an bone mo rphogenetic p ro tein 2,rhBM P 22)异位诱导成骨过程中神经生长因子(nerve grow th facto r ,N GF )及其高亲和力受体(tyro sine k inase recep to r ,T rkA )的表达,探讨N GF 在BM P 骨诱导中的作用。　方法　I CR 小鼠36只,随机分为实验组和对照组,每组18只,均制备右侧股后部肌袋异位成骨模型。实验组植入含rhBM P 22胶原复合物,对照组仅植入相同体积的胶原海绵。分别于术后7、14和21d 取材,行大体、组织学、免疫组织化学观察及R T 2PCR 检测。　结果　大体观察实验组术后7d ,右股后部可扪及较硬肿块;术后14、21d ,肿块硬度增加。对照组各时间点未发现肿块。组织学观察实验组中rhBM P 22胶原复合物具有良好的诱导成骨能力,免疫组织化学结果显示骨诱导材料植入后7d ,N GF 于成纤维细胞、成软骨细胞、软骨细胞、肥大软骨细胞和成骨细胞中均有阳性表达;14d ,N GF 阳性表达局限于部分成骨细胞、幼稚骨细胞和成骨样细胞,同时在骨髓中单核巨噬细胞、多核巨噬细胞和破骨细胞中有明显表达;21d ,只有少量成骨样细胞和破骨细胞以及骨髓中多核巨噬细胞有N GF 表达;T rkA 免疫组织化学染色与N GF 的表达基本一致。对照组术后各时间点未见成骨现象。实验组N GF mRNA 的R T 2PCR 检测显示,术后7d N GF 的mRNA 表达最高,14d 表达下降,21d 只有微量表达。　结论　rhBM P 22复合物是一种有效的诱导成骨材料。在外源性BM P 诱导成骨过程中有明显的N GF 及其高亲和力受体T rkA 的表达,N GF 可以通过直接和间接的方式参与BM P 的诱导成骨过程。

【关键词】　　骨形成蛋白　　骨诱导　　神经生长因子　　受体　　免疫组织化学

中图分类号:R 318　Q 516　　文献标识码:A

EXPRESSI ON OF NERVE GR OW TH FACT OR AND ITS RECEPT OR D UR ING OSTEO IND UCTI ON OF RECOM B INED HU M AN B ONE MORPHOGENETI C PR OTE IN 2 X IA S heng li ,DA I K erong ,TA N G T ing ting .D ep a rt m en t of O rthop aed ics ,N in th P eop le ’s H osp ita l ,M ed ica l Colleg e of S hang ha i ,J iaotong U n iversity ,S hang ha i ,200011,P .R .Ch ina

Corresp ond ing au thor :DA I K erong ,E 2m a il :k rd a i @163.co m

【Abstract 】　　Objective 　To iden tify the exp ressi on of nerve grow th facto r (N GF )and its h igh affin ity recep to r (tyro sine k inase recep to r A ,T rkA )du ring bone inducti on by recom b ined hum an bone mo rphogenetic p ro tein 2(rhBM P 22)by i m m unoh istochem istry and reverse tran scri p tase 2po lym erase chain reacti on (R T 2PCR )and to discu ss the ro le of N GF on the bone inducti on of the BM P .M ethods 　T h irty 2six I CR m ice w ere divided in to the experi m en tal group and the con tro l group at random .rhBM P 22 co llagen sponge and co llagen sponge w ere i m p lan ted in to the righ t th igh m u scle pouches of the m ice in the experi m en tal group and the con tro l group ,respectively .T he tissues in the i m p lan ted site of the tw o group s w ere removed on the 7th ,14th and 21st day after the i m p lan tati on .H isto logical ,i m m unoh istochem ical and R T 2PCR analyses w ere perfo rm ed to detect o steo inductive effects of rhBM P 22and the exp ressi on of N GF and T rkA .Results 　Gro ss ob servati on show ed that a so lid lump w as found in the righ t th igh in the experi m en tal group on the 7th day and becam e harder on the 14th and 21st day ,w h ich w as no t found in the con tro l group .rhBM P 22 co llagen sponge disp layed a po ten t ab ility to induce bone fo rm ati on ,w h ile i m m uno stain ing fo r N GF and T rkA w as ob served in the cou rse of o steo inducti on by rhBM P 22.O n the 7th day in the experi m en tal group ,N GF po sitive i m m uno stain ing reached the peak in the stage of chondrogenesis and there w ere a large num ber of cells exp ressing N GF ,including fib rob lasts ,chondrob lasts ,chondrocytes ,and o steob lasts ;then ,there w as a decrease in the num ber of the po sitive cells and in the in ten sity of i m m uno stain ing on the 14th and 21st day .Stain ing of T rkA w as 作者单位:上海交通大学医学院附属第九人民医院骨科(上海,200011)

通讯作者:戴　戎,教授,中国工程院院士,博士导师,研究方向:骨创伤与修复、人工关节置换,E 2m ail:k rdai@ ・454・Ch inese Journal of R eparative and R econstructive Surgery ,2006,V o l 20(4)