应用甲基丙烯酸甲酯包埋不脱钙骨组织方法探析.docVIP

应用甲基丙烯酸甲酯包埋不脱钙骨组织方法探析 　作者：谭见容，杨小红　康宁，王文，陈鸿辉，梁伟国 【摘要】 [目的]本研究通过改进既往标本制作方法，探讨应用不同浓度聚甲基丙烯酸甲酯对骨组织包埋效果的影响，寻找不脱钙骨组织标本包埋制作的理想方法。[方法]骨组织标本取材后，用10%中性福尔马林固定24 h，梯度酒精脱水、二甲苯透明，转入含有15 g/L过氧化苯甲酰的甲基丙烯酸甲酯(UMMA)中，每24h换液1次。从转入UMMA渗透液开始，每天抽滤30 min，以排除组织中的气体。 MMA聚合过程分为3个实验组，分别以30 g/L、35 g/L、40 g/L三种筛选浓度加入过氧化苯甲酰，密切观察样本聚合情况。[结果]3种浓度及凝结时间观察比较，加速剂浓度为35 g/L的标本，在室温25℃左右，约7 d时间开始凝结，并逐渐变硬，组织与包埋剂的硬度均匀，效果理想，制作的切片直接在荧光显微镜下观察，可见两种清晰的矿化标记;加速剂浓度为40 g/L的标本，约3～4 d开始凝结，但出现大量气泡影响观察及制片，效果欠佳;加速剂浓度为30 g/L的标本，观察30 d仍未见凝结。[结论]过氧化苯甲酰浓度为35 g/L的包埋剂对不脱钙骨组织包埋效果最为理想，应用本研究方法能为骨科基础研究领域相关研究提供有效的硬组织制作技术平台。 【关键词】 甲基丙烯酸甲酯; 不脱钙骨; 包埋 　　Abstract: [Objective]To investigate the effect of the application of different concentrations of PMMA in embedded bone tissue by improving the producting process, it was aimed to look for the ideal embedded method of producing non- demineralized bone tissue samples. [Methods]The specimens were fixed for 24 hours by 10% neutral formaldehyde,and dehydrated by concentration gradient alcohol,and then made transparent by xylene .They were shifted in UMMA solution with 15 g/L benzyl peroxide.The solution was changed per 24 hours.After these specimens were shifted in UMMA ,they were filtered for 30 min every day,in order to eliminate the gas in tissue.The specimens were divided into 3 groups of benzoyl peroxide’s concentrations,which were 30 g/L,35 g/L,40 g/L respectively. The polymerization situation of the specimens were closely observed.[Results]Through comparing the setting time of three different concentrations,the result was clear:when the concentration of accelerator was 35 g/L ,the specimens began to condensate after 7 days and hardened gradually,the hardness of tissue and embedding medium were well-distributed.The results were satifactory.Two kinds of mimeral mark could be seen clearly while the sections were observed by fluorescence microscope;when the accelerator’s concentration was 40 gl,the specimens began to condese after 3～4 days and bubble was found which could affect the observat