Chen Xueying, Li Baoxing, Li Jing, Yin Shaofang, Song Huiping, Yuan Lin
Department of Anatomy, Southern Medical University, Guangzhou Guangdong, 510515, PR China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2009 Mar;23(3):362-5.
To develop a deantigenated porcine bone graft through enzyme treatment and to study the biomechanical properties and osteoinductivity of the xenogeneic bone graft.
Deantigenated xenogeneic bone was prepared from porcine bone by a series treatment including alpha-galactosidase (alpha-Gal) treatment, freeze drying and irradiation at a does of 25 kGy. Samples were divided into 4 groups according to the treatment methods: fresh bone (group A); deantigenated bone (group B); deantigenated and freeze dried bone (group C); deantigenated-freeze dried-irradiated bone (group D). SEM observation to group B samples was performed and the diameters of the caves of the porcine cancellous bone were measured. The a-Gal contents of samples of groups A, B, D were measured by ELISA. The effects of different treatments on porcine bone mechanic properties were evaluated through compressive test of 4 groups. The prepared porcine cortical bone were demineralized and ectopically implanted into muscle groups of hind limbs of Wistar rats as experimental group, and the demineralized cortical bone which were inactivated by high temperature and pressure were implanted as control group. Histological observation was performed and ALP activity was tested at different time postoperatively to investigate the osteoinductivity of the xenogeneic bone implants.
The porous structure of prepared porcine cancellous bone was similar to that of human cancellous bone. The diameters of the caves were between 150-600 microm. The A value of the groups A, B, D was 0.358 +/- 0.027, 0.191 +/- 0.011, 0.191 +/- 0.009, respectively, with statistically differences between groups (P < 0.05). While the biomechanical properties among 4 groups had no statistically difference (P > 0.05). Histological observation showed mesenchymal cells immigrated into cartilage and converted into chondrocytes at 3 weeks postoperatively. Cartilage was formed at 4 weeks and osteoid and more adult cartilage was formed at 6 weeks within the implants of experimental group, while there was no bone formation in control group. ALP activity of experimental group at different times were obviously higher than that of the control group (P < 0.05).
The main xenogeneic antigen of the porcine bone was removed while the mechanic properties and osteoinductivity remained, thus it may be a suitable bone substitute for clinical use.
通过酶处理制备去抗原猪骨移植物,并研究该异种骨移植物的生物力学性能和骨诱导活性。
采用一系列处理方法从猪骨制备去抗原异种骨,包括α-半乳糖苷酶(α-Gal)处理、冻干及25 kGy剂量的辐照。样本根据处理方法分为4组:新鲜骨(A组);去抗原骨(B组);去抗原冻干骨(C组);去抗原冻干辐照骨(D组)。对B组样本进行扫描电子显微镜(SEM)观察并测量猪松质骨洞穴直径。采用酶联免疫吸附测定法(ELISA)检测A、B、D组样本的α-Gal含量。通过对4组样本进行压缩试验评估不同处理对猪骨力学性能的影响。将制备的猪皮质骨脱矿后异位植入Wistar大鼠后肢肌肉群作为实验组,将经高温高压灭活的脱矿皮质骨植入作为对照组。进行组织学观察并在术后不同时间检测碱性磷酸酶(ALP)活性,以研究异种骨植入物的骨诱导活性。
制备的猪松质骨多孔结构与人类松质骨相似。洞穴直径在150 - 600微米之间。A、B、D组的A值分别为0.358±0.027、0.191±0.011、0.191±0.009,组间差异有统计学意义(P<0.05)。而4组间生物力学性能无统计学差异(P>0.05)。组织学观察显示,术后3周实验组植入物中有间充质细胞迁移至软骨并转化为软骨细胞。术后4周形成软骨,6周时植入物内形成类骨质和更多成熟软骨,而对照组无骨形成。实验组不同时间的ALP活性明显高于对照组(P<0.05)。
猪骨的主要异种抗原被去除,同时保留了力学性能和骨诱导活性,因此可能是一种适合临床应用的骨替代物。