微小颗粒骨移植骨细胞活性的实验研究  

Osteocyte bioactivity in morselized bone grafts

作　　者：王新涛[1] 闫景龙[1] 杨显声[1] 麻松[1] 周长龙[1] 奚春阳[1] 

机构地区：[1]哈尔滨医科大学附属第一医院骨科,150001

出　　处：《中华创伤骨科杂志》2008年第3期260-265,共6页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然基金资助项目（30371441）;黑龙江省青年基金资助项目（QC06C071）

摘　　要：目的观察微小颗粒骨在移植修复骨缺损过程中的骨细胞存活情况和生物活性。方法建立大鼠桡骨骨缺损模型，近交系DA大鼠88只，其中雄性大鼠28只，作为供体；雌性大鼠60只，作为受体。将受体随机分为块状骨组（n=56）、微小颗粒骨组（n=56）和空白对照组（n=4），取雄性大鼠髂骨为供体骨，分别制成直径为2mm的骨块和直径为300～500μm的微小颗粒骨，植入骨缺损，于术后1d、4d、1周、2周、4周、6周、10周取材，采用原位杂交的方法观察受体内Y染色体性别决定基因（Sry）的表达情况，应用免疫组化法观察各组骨形态发生蛋白-2（BMP-2）、转化生长因子-β1（TGF-β1）、碱性磷酸酶（ALP）和Ⅰ型胶原的表达情况。结果块状骨组在移植早期Sry的表达逐渐减少，至1周消失，4周后再次出现，并且随时间延长表达逐渐增多；微小颗粒骨组各时间段均有Sry的表达，在同一时间点，微小颗粒骨组Sry的阳性细胞数多于块状骨组（P〈0．05），两种骨移植物中参与修复骨缺损的细胞类型不同。微小颗粒骨内和周围组织中BMP-2、TGF-β1、ALP和Ⅰ型胶原的阳性细胞数在术后2周内多于块状骨组（P〈0．05）。结论微小颗粒骨与块状骨修复骨缺损时均有供体骨细胞参与，但微小颗粒骨内有更多的骨细胞存活。微小颗粒骨内存活的骨细胞具有生物学活性，合成并分泌骨生长因子和骨基质蛋白，可以加速骨缺损的修复。Objective To investigate the bioactivity of osteocytes in tiny morselized bone grafts for bone defect repair. Methods Models of radial defect were established on 60 female syngeneic inbred DA rats which were divided into 3 groups randomly： structural bone grafting group （n=56）, tiny morselized bone grafting group （n=56） and control group （without bone grafting, n=4）. The ilia of 28 male inbred DA rats were harvested as donors and made into structural bone grafts 2mm in diameter and tiny morselized bone grafts 300- 500μm in diameter to be transplanted into the radial defects of the female receptors. Samples were harvested on 1 d, 4 d, 1 w, 2 w, 4 w, 6 w and 10 w after transplantation. The presence and relative amounts of genes specific to the sex-determining region of the Y-chromosome （Sry） originating from the bone grafts were measured by in situ hybridization（ISH）. The expressions of bone morphogenetic proteins-2 （BMP-2）, trans-forming growth factor-beta1 （TGF-β1）, alkaline phosphatase （ALP） and Collagen I were semiquantified by immunohistochemistry. Results In structural bone grafting group, the expression of Sry decreased early and disappeared at 1 w. But after 4 w, Sry appeared again and its expression increased with lapse of time. In tiny morselized bone grafting group, Sry was detected all the time. At each time point, its expression was more than in Group I （P〈0.05）. Donor bone cells were different during repair in both groups. Tiny morselized bone grafts were absorbed quickly, and the expressions of BMP-2 and TGF-β1 reached the peak faster than in structural bone grafting group. The positive cells of BMP-2, TGF-β1, ALP and Collagen I in tiny morselized bone grafting group were much more than in structural bone grafting group during the first 2 w （P〈0.05）. Conclusions Both bone grafts can provide donor cells to repair bone defects. Compared with bulk bone grafts, tiny morselized bone grafts contain more living bioactive osteocytes which can secrete bone gr

关 键 词：骨生成 移植 骨细胞 

分 类 号：R687.3[医药卫生—骨科学]