中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (50): 9364-9369.doi: 10.3969/j.issn.2095-4344.2012.50.010

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

复合配方脱钙液处理兔股骨髁标本

张春丽,侯树勋,赵彦涛,周 颖,刘 彦,耿 凌   

  • 出版日期:2012-12-09 发布日期:2012-12-09

Application of a composite decalcification solution in rabbit femoral condyle specimens  

Zhang Chun-li, Hou Shu-xun, Zhao Yan-tao, Zhou Ying, Liu Yan, Geng Ling   

  • Online:2012-12-09 Published:2012-12-09

摘要:

背景:兔骨缺损修复模型被广泛应用于骨替代材料的研究中,在实验过程中经常需要对兔股骨髁标本进行脱钙制作病理切片,但普遍存在脱钙困难的问题。
目的:尝试应用一种复合脱钙液实现软骨覆盖兔股骨髁有效脱钙。
方法:取兔股骨髁标本12个,分为2组,分别用自制复合脱钙液和普通甲酸脱钙液进行常温条件下的脱钙处理,复合脱钙液的组成比例如下:盐酸8 mL,甲酸10 mL,醋酸5 mL,中性甲醛固定液100 mL。在经过1周的常规脱钙过程之后,进行标准化的脱水、包埋、切片和染色程序。统计切片的完整率,观察苏木精-伊红染色后切片的质量和细微结构,并对两种脱钙液的经济性进行比较。
结果与结论:复合脱钙液组的切片完整率显著高于甲酸组,差异有显著性意义(74% vs18%,P < 0.05)。复合脱钙液组的切片着色优良,背景清亮,微观结构清晰,细微结构保存完整,不同组织染色对比鲜明。该复合脱钙液的应用成本较常用的甲酸脱钙液降低65%。提示自制复合脱钙液可有效解决兔骨缺损修复模型实验中的脱钙困难问题,且复合脱钙液配置简便,成本低。

Abstract:

BACKGROUND: Rabbit bone defect model is widely used in the researches of bone substitute materials. The difficulty of decalcification is common in the experiments of rabbit bone defect models because of normal decalcification and slicing procedure before observation.
OBJECTIVE: In current study, a composite decalcification solution was attempted to solve the problem.
METHODS: Twelve rabbit femoral condyle specimens were divided into test and control groups randomly. A composite decalcification solution (8 mL hydrochloric acid, 10 mL formic acid, 5 mL acetic acid, and 100 mL neutral formalin fixation solution) and a common formic acid were used to treat the specimens under normal temperature. After the 1-week decalcification process, the specimens were dehydrated, embedded, sliced and dyed. The ratio of intact slices was compared. The quality and micro structure of the slices were observed. And the cost of two decalcification solutions was evaluated.
RESULTS AND CONCLUSION: The intact ratio of slices was significantly improved in test group (74% versus 18%, P < 0.05). The dying quality was good with a clear background, and micro structures were all preserved well and indicated clearly. The contrast between different tissues was obvious. The cost of the decalcification solution was decreased by 65%. These findings indicate that the composite decalcification solution can solve the difficulty in the decalcification of rabbit bone defect repairing models effectively and the solution is convenient and advantageous in cost, which is beneficial for spreading the applications.