5%硝酸、10%EDTA用于大鼠尾椎椎间盘组织脱钙处理的效果观察  

Effect of 5%Nitric Acid and 10%EDTA on Decalcification of the Coccygeal Disc in Rats

作　　者：齐妍 李建华[2] 王咏梅[1] QI Yan;LI Jianhua;WANG Yongmei(Department of Pathology,Tianjin Haihe Hospital,Tianjin 300052,China;Department of Orthopedics,Tianjin Haihe Hospital,Tianjin 300052,China)

机构地区：[1]天津市海河医院病理科,天津300052 [2]天津市海河医院骨科,天津300052

出　　处：《继续医学教育》2021年第11期165-168,共4页Continuing Medical Education

基　　金：天津市海河医院科技基金(HHYY-202002)。

摘　　要：目的观察并比较大鼠尾椎椎间盘经5%硝酸或10%EDTA脱钙处理后的病理表现。方法成年SD雄性大鼠10只,取鼠尾Co6/7-Co9/10椎间盘,Co6/7和Co8/9为硝酸组进行5%硝酸脱钙处理,Co7/8和Co9/10为EDTA组进行10%EDTA脱钙处理。记录脱钙时间,采用HE染色、番红O固绿染色进行组织形态学观察,采用免疫组化染色观察II型胶原蛋白表达。结果硝酸组脱钙需(18.35±4.71)h,EDTA组需(22.75±6.64)h。硝酸组整体形态保持更好,但是免疫组化染色II型胶原蛋白表达明显减少。EDTA组组织形态保持尚可,纤维环组织变硬变脆,有组织松散紊乱表现,但是II型胶原蛋白表达明显。结论大鼠尾椎椎间盘经5%硝酸、10%EDTA两种脱钙液处理后,在进行普通病理及结构观察时,5%硝酸具有脱钙时间短,组织形态更完整的优势,但需要进行免疫组化染色时,10%EDTA进行脱钙处理更有优势。可以根据不同的科研需求选择脱钙方法。Objective To observe and compare the pathological features of rat coccygeal discs after decalcification with 5%nitric acid or 10%EDTA.Methods Ten adult male Sprague Dawley rats were used.The intervertebral discs of Co6/7-Co9/10 were taken from the tail of rats.The Co6/7 and Co8/9 groups were treated with 5%nitric acid decalcification,and the Co7/8 and Co9/10 groups were treated with 10%EDTA decalcification.The decalcification time was recorded.The histomorphology was observed by HE staining and safranin o-green staining.The expression of type II collagen was observed by immunohistochemical staining.Results It took(18.35±4.71)hours to decalcify in nitric acid group and(22.75±6.64)hours in EDTA group.The overall morphology of nitric acid group was better,but the expression of type II collagen was significantly reduced by immunohistochemical staining.In EDTA group,the tissue morphology remained fair,the annulus fibrosus tissue was hard and brittle,and the tissue was loose and disordered,but the expression of type II collagen was obvious.Conclusion After 5%nitric acid and 10%EDTA decalcification solution treatment,5%nitric acid has the advantages of shorter decalcification time and more complete tissue morphology in general pathological and structural observation,but 10%EDTA decalcification treatment has more advantages when immunohistochemical staining is needed.Therefore,decalcification methods can be selected according to different research needs.

关 键 词：椎间盘 硝酸 EDTA 乙二胺四乙酸二钠 脱钙 

分 类 号：R361[医药卫生—病理学]