交联几丁糖治疗兔膝骨关节炎的实验研究

Experimental study on crosslinked-chitosan in treatment of knee osteoarthritis in rabbits

目的探讨关节腔注射交联几丁糖治疗兔膝骨关节炎的效果。方法将32只新西兰白兔随机分为A、B、C、D 4组(n=8),其中A、B、C组采用切断左后肢前交叉韧带方法制备膝骨关节炎模型;4周后,A组关节腔内注射交联几丁糖0.6 mL 1次;B、C组分别注射几丁糖、生理盐水,0.3 mL/次,每2周1次;D组为假手术组,关节腔内注射等量生理盐水。造模术后8周,行软骨大体观察、扫描电镜观察、组织学观测(HE染色、番红-固绿染色以及Mankin评分)、Ⅱ型胶原免疫组织化学染色观测。结果大体及扫描电镜观察示D组软骨结构接近正常,A、B组软骨表面结构磨损较D组严重,C组最严重;组织学染色观察示,A、B组软骨结构相似,且优于C组,但尚未达D组正常结构。B、C组软骨大体评分及Mankin评分明显高于D组(P<0.05),A、B组比较差异无统计学意义(P>0.05)。免疫组织化学染色观察示D组Ⅱ型胶原阳性软骨细胞百分比明显高于B、C组(P<0.05),A、B组比较差异无统计学意义(P>0.05)。结论交联几丁糖能明显改善兔膝骨关节炎,延缓骨关节炎病理改变,且注射次数减少。

Objective To study the effect of intraarticular injection of crosslinked-chitosan in the treatment of knee osteoarthritis in rabbits. Methods Thirty-two New Zealand white rabbits were randomly divided into 4 groups(groups A, B, C, and D;8 rabbits in each group). The knee osteoarthritis models were prepared by anterior cruciate ligament transection in the left hind in groups A, B, and C. At 4 weeks after operation, the rabbits were received intraarticular injection of 0.6 mL crosslinked-chitosan in group A, 0.3 mL chitosan(once per 2 weeks, for twice) in group B, and0.3 mL saline(once per 2 weeks, for twice) in group C. The rabbits in group D were treated with sham operation in the left hind, and received intraarticular injection of 0.3 mL saline(once per 2 weeks, for twice). At 8 weeks, the macroscopic observation, histological examination(HE staining, Safranin-fast green double staining, and Mankin score), scanning electron microscopy(SEM) observation, and immunohistochemical staining of collagen type Ⅱ were performed.Results Macroscopic and SEM observations showed that the cartilage in group D was basically the same as normal and better than that in groups A and B, and the abrasion of cartilage in group C was the most serious. The histological observation results in groups A and B were slightly similar and better than those in group C, but not up to the structure of group D. The macroscopic score and Mankin score of groups B and C were significantly higher than those of group D(P<0.05), and there was no significant difference between group A and group B(P>0.05). Immunohistochemical staining results showed that the collagen type Ⅱ positive percentage of chondrocytes was significantly higher in group D than that in groups B and C, and no significant difference was found between group A and group B(P>0.05). Conclusion The crosslinked-chitosan can significantly improve the osteoarthritis of the rabbit knee, delay the pathological changes of o steoarthritis, and decrease the frequency of injection.