摘要
目的:探讨基于超临界二氧化碳萃取的复合技术处理猪股骨松质骨的效果,旨在分析该新技术用于同种异体骨修复材料的制备可行性。方法:足量加工后的猪股骨松质骨骨块被随机分为三组:(1)非超临界组(对照组),即猪股骨松质骨经过高压水枪彻底冲洗,再经3%双氧水脱脂灭菌,制备出猪股骨松质骨修复材料,经冷冻干燥后获得1号材料;(2)超临界复合超声波组(实验组1),即猪股骨松质骨依次由超声波清洗离心、超临界二氧化碳萃取脱脂以及化学清洗(过氧化氢、氢氧化钠)等组成的复合清洗技术处理,制备出的猪股骨松质骨修复材料,经冷冻干燥后获得2号材料;(3)超临界复合高压水枪组(实验组2),即猪股骨松质骨依次由高压水枪冲洗、超临界二氧化碳萃取脱脂以及化学清洗(过氧化氢、氢氧化钠)等组成的复合清洗技术处理,制备出的猪股骨松质骨修复材料,经冷冻干燥后获得3号材料。根据实验需要将1、2、3号材料或原材料进行Micro-CT、压缩力学、细胞毒性等检测。结果:(1)从2号材料和3号材料的外观及重量变化可认为实验组1和实验组2对猪股骨松质骨的清洗效力无明显差别;(2)1、2、3号材料经Micro-CT扫描获得CT三维重建图像提示骨小梁结构完好,提示三组方法(对照组、实验组1和实验组2)未明显破坏猪股骨松质骨修复材料的微观结构;(3)1、2、3号材料与猪股骨松质骨原材料进行压缩力学比较,提示强度极限、最大载荷和弹性模量无统计学差异(P>0.05),可认为三组方法不会引起猪股骨松质骨压缩力学改变。4.1、2、3号材料细胞毒性实验OD值结果(MTT法)与阴性组OD值结果比较无统计学意义(P>0.05)。结论:基于超临界二氧化碳萃取的复合技术处理后的猪松质骨孔隙结构较为完整,抗压性好,无毒性试剂残留,有利于细胞生长。因此,该技术是一种安全、有效、绿色的复合清洗技术,不仅可用于异种骨修复材料的制备,亦可用于同种异体骨修复材料的制备。
Objective：To investigate the effect of composite technology based on supercritical carbon dioxide extraction in the processing of porcine femoral cancellous bones,and to analyze the feasibility of this new technology for the preparation of allograft bone repair materials.Methods：After adequate processing,porcine femoral cancellous bones were randomly divided into three groups：non-supercritical group（control group）,supercritical composite ultrasound group（experimental group 1）,and supercritical composite high-pressure water gun group（experimental group 2）.（1）In the control group,the porcine femoral cancellous bones were thoroughly washed using a high-pressure water gun,then degreased and sterilized with 3%hydrogen peroxide,and finally subjected to freeze-drying to obtain porcine femoral cancellous bone repair materials（No.1 materials）.（2）In experimental group 1,the porcine femoral cancellous bones were subjected to ultrasonic washing and centrifugation,supercritical carbon dioxide extraction for degreasing,and chemical washing（with hydrogen peroxide and sodium hydroxide）,followed by freeze-drying,to obtain porcine femoral cancellous bone repair materials（No.2 materials）.（3）In experimental group2,the porcine femoral cancellous bones were subjected to washing using the high-pressure water gun,supercritical carbon dioxide extraction for degreasing,and chemical washing（with hydrogen peroxide and sodium hydroxide）,followed by freeze-drying,to obtain porcine femoral cancellous bone repair materials（No.3 materials）.According to experimental requirements,No.1,2,and 3 materials or yaw materuals.were examined by Micro-CT,compression mechanics,and cytotoxicity test,and the results were statistically analyzed.Results：（1）In terms of the appearance and weight changes of No.2 and 3 materials,there was no significant difference in the cleaning efficacy for porcine femoral cancellous bones between experimental group 1 and experimental group 2.（2）The three-dimensional reconstruction images of No.1,2,and 3 materials obtained by Micro-CT showed intact trabecular bone structure,which suggested that the three methods（used in the control group,experimental group 1,and experimental group 2）did not significantly destroy the microstructure of porcine femoral cancellous bone repair materials.（3）The compression mechanics of No.1,2,and 3 materials versus raw porcine femoral cancellous bones indicated no significant differences in strength limit,maximum load,and elastic modulus（P〈0.05）,so the three methods did not cause changes in the compression mechanics of porcine femoral cancellous bones.（4）The cytotoxicity test（MTT method）showed that there was no significant difference between the optical density of No.1,2,and 3 materials and that in negative group every day（P〈0.05）.Conclusion：The composite technology based on supercritical carbon dioxide extraction is a safe,effective,and nontoxic composite cleaning technology,as the porcine femoral cancellous bones processed with this technology have intact trabecular bone structure,good pressure resistance,no residual toxic agents,and good cellular growth.It can be used for the preparation of both xenogeneic bone repair materials and allograft bone repair materials.
作者
鲜海
刘舒云
眭翔
黄靖香
郝春香
鲁晓波
郭全义
XIAN Hai;LIU Shuyun;SUI Xiang;HUANG Jingxiang;HAO Chunxiang;LU Xiaobo;GUO Quanyi(Department of Bone and Joint Surgery,the Affiliated Hospital of Southwest Medical University,Luzhou 646000,Institute of Orthopedics,Chinses PLA General Hospital)
出处
《西南医科大学学报》
2018年第3期205-209,共5页
Journal of Southwest Medical University
基金
国家重点计划(2017YFC1104102)
关键词
超临界二氧化碳
同种异体骨
异种骨
脱脂
脱蛋白
Supercriticalcarbondioxide
Allograftbone
Xenogeneicbone
Degreasing
Deproteinization
