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目的建立保留钙化层结构的猪股骨滑车全厚软骨缺损模型,为观察组织工程软骨在保留钙化层的膝关节软骨缺损模型中的修复效果提供良好的实验研究平台。方法选取6月龄清洁级贵州小香猪9只,体重40～50 kg,用标准的软骨缺损制作套件在其右后肢股骨滑车切迹旁制备直径6 mm、深0.2～0.5 mm、不伤及钙化层结构的圆柱形全厚软骨缺损模型。造模4周后行3.0T MRI观察,取材后进行大体、体视显微镜观察及固绿-番红O、阿利新蓝、天狼星红组织学染色观察缺损处软骨修复情况。结果造模后实验动物均存活,术后切口无感染,无髌骨脱位;术后即可下地行走并部分负重,1周后均能自由活动,无跛行。造模后4周,MRI检查可见滑车处有明显连续信号中断,异常信号深及软骨下骨,缺损周边深层未见明显信号异常。标本大体观察示缺损底部有少量填充物、出血点,与周围正常软骨界限清楚。体式显微镜观察示钙化层基本完整,缺损局部软骨下骨板有塌陷。普通显微镜下,固绿-番红O及阿利新蓝染色示缺损处无软骨细胞及染料着色;偏光显微镜下,天狼星红染色示缺损底部被连续、强折光性的纤维组织少量填充。结论通过该造模方法制作的不伤及钙化层结构的猪股骨滑车全厚软骨缺损模型,可用于骨关节炎早期软骨病变修复的研究及猪软骨钙化层结构作用研究的动物模型。
OBJECTIVE: To establish a full-thickness cartilage defect model of porcine trochleal block with preserved calcified layer structure and to provide a good experimental research platform for observing the effect of tissue engineering cartilage in repairing the knee joint cartilage defect model with retained calcification layer. Methods Nine clean-grade Guizhou miniature pigs aged 6 months were selected and weighed 40-50 kg. They were prepared with 6 mm in diameter and 0.2-0.5 mm in diameter on the right hindlimb femoral trochanter with a standard cartilage defect kit. Calcium layer structure of the full thickness of the cartilage defect model. After 4 weeks of modeling, 3.0T MRI was performed. Gross and stereomicroscope microscopy and fixation of green-Safronin O, Aliskiren and Sirius red staining were used to observe the cartilage repair. Results After modeling, all the experimental animals survived. No postoperative wound infection and no dislocation of the patella were found. After operation, the animals were able to walk under the ground and partially supported. After 1 week, they could move freely and without clashes. Four weeks after the model establishment, there were obvious continuous signal interruption, abnormal signal depth and subchondral bone in the block at MRI. There was no obvious signal abnormality in the deep surrounding defect. Gross observation of the specimen showed a small amount of filler at the bottom, bleeding, and the surrounding normal cartilage clearly. Body type microscope showed calcified layer is basically complete, defect subchondral bone plate collapse. Under a normal microscope, solid green-safranin O and alitake blue staining showed no chondrocytes and dye coloration in the defect; under the light microscope, Sirius red staining showed that the bottom of the defect was continuously and slightly refracted by a small amount of fibrous tissue. Conclusion The full-thickness cartilage defect model of porcine trochlea block which does not hurt the calcification layer structure can be used to study the repair of early osteochondrosis and the animal model of pig cartilage calcification structure.