掺锶磷酸钙骨水泥材料生物学性能的动物实验

In vivo study of strontium-doped calcium phosphate cement for biological properties

目的:评价新型磷酸钙骨水泥(calcium phosphate cement,CPC)的生物相容性以及成骨效果,为其进一步的临床应用提供实验数据。方法:选择新西兰大白兔30只,以其双后腿外侧髁(60个)为实验对象,随机分为CPC组、CPC+Bio-Oss组、Bio-Oss组和空白对照组4组,在兔双侧后腿外侧髁制造直径6 mm、深7 mm的骨缺损模型,按照组别分别植入CPC、Bio-Oss、CPC+Bio-Oss混合物(CPC与Bio-Oss骨粉质量比为4∶1)。实验动物分别在手术第4周、第12周、第24周处死,取骨缺损周围组织,HE染色进行组织学评价,并计算新骨生成率(bone ingrowth fraction,BIF);利用免疫组织化学染色,计算阳性区域平均光密度(mean optical density,MOD),检测术后第4周各组样本BMP-2和COL-Ⅰ的表达情况,评价各组样本在不同时间点的骨愈合情况。结果:HE染色发现,在相同时间点,与空白对照组相比,CPC组、CPC+Bio-Oss组、Bio-Oss组的BIF值明显较高(P<0.01),其中,CPC组BIF低于Bio-Oss组和CPC+Bio-Oss组(P<0.01),CPC+Bio-Oss组与Bio-Oss组相比差异无统计学意义(P>0.05)。免疫组织化学染色结果显示,与空白对照组相比,CPC组BMP-2和COL-Ⅰ的MOD值较高,但低于Bio-Oss组和CPC+Bio-Oss组(P<0.01),CPC+Bio-Oss组BMP-2和COL-Ⅰ的MOD与Bio-Oss组相比差异无统计学意义(P>0.05)。结论:新型磷酸钙骨水泥具有良好的生物相容性,可以促进早期成骨,成骨效果稳定,长期有效。

Objective:To evaluate the biocompatibility and osteogenic effect of new calcium phosphate cement(CPC)in vivo and to provide experimental basis for its further clinical application.Methods:Thirty New Zealand white rabbits were randomly divided into four groups:CPC group,CPC+Bio-Oss group,Bio-Oss group and blank control group.Bone defect models of 6 mm in diameter and 7 mm in depth were made on the lateral condyle of bilateral hind legs of the rabbits.CPC,Bio-Oss and CPC+Bio-Oss mixture were implanted into the bone defect according to the group,and the mass ratio of CPC and Bio-Oss was 4∶1.The experimental animals were sacrificed the 4th,12th and 24th week after operation.The tissue around the bone defect was taken for histological evaluation by H&E staining.Bone ingrowth fraction(BIF)was calculated.The expression of BMP-2 and COL-Ⅰwas detected by immunohis-tochemical staining by calculating the mean optical density(MOD)of the positive area the 4th week after operation,and the bone healing of each group was evaluated at different time points.The measurement data were analyzed by one-way ANOVA and LSD test was used for multiple comparison of the differences between the means by SPSS 19.0.P<0.05 was considered to be statistically significant.Results:The results of H&E staining showed that the BIF values of CPC group,CPC+Bio-Oss group and Bio-Oss group were significantly higher than those of blank control group at the same time point(P<0.01).The BIF values of CPC group were lower than those of Bio-Oss group and CPC+Bio-Oss group(P<0.01).There was no significant difference between CPC+Bio-Oss group and Bio-Oss group.Immunohistochemical staining showed that the MOD values of BMP-2 and COL-Ⅰin CPC group were higher than those in blank control group,but lower than those in Bio-Oss group and CPC+Bio-Oss group(P<0.01).There was no significant difference between BMP-2 and COL-Ⅰin CPC+Bio-Oss group and Bio-Oss group.Conclusion:The new calcium phosphate cement has good biocompatibility and can promote early osteogenesis with stable and long-term effect.