摘要
目的制备兔半月板脱细胞基质并研究其组织形态结构及生物力学特性。方法将切取的兔半月板用双氧水浸泡、蒸馏水洗涤后,采取Triton X-100及脱氧胆酸钠等试剂制备脱细胞的兔半月板,观察其色泽、质地,用苏木精-伊红(HE)、甲苯胺蓝、番红花O、Hoechst-33258荧光染色及Ⅰ型胶原免疫组化染色等方法进行组织学检测,用倒置相差显微镜观察其微观三维结构,用生物力学机测定其在不同压缩比率下的瞬时形变恢复率、最大形变恢复率及最大受压强度。结果兔脱细胞半月板呈乳白色,结构蓬松,HE、甲苯胺蓝和番红花O染色均显示细胞陷窝内已无细胞结构,Hoechst-33258荧光染色呈阴性,Ⅰ型胶原免疫组化染色呈阳性。倒置相差显微镜观察到脱细胞半月板基质内部结构疏松,微孔较多。生物力学测定其压缩40%时,最大瞬时形变恢复率为(76.65±4.61)%,最大形变恢复率为100%,最大强度是(4.51±0.69)N。结论成功制备的兔半月板脱细胞基质有较多微孔,形变恢复好,有利于细胞附着生长,可作为组织工程支架。
Objective To prepare a rabbit meniscus acellular matrix scaffold and explore the histomorphological and biomechanical properties of the scaffold.Methods Rabbit meniscuses were collected and acellularized using a modified eight-step detergent process with hydrogen peroxide,distilled water,Triton X-100,and sodium deoxycholate.Its color and texture were observed.Histomorphological assessment was performed using routine hematoxylin-eosin stain,toluidine blue stain,Saffron stain,Hoechst-33258 stain,and immunohistochemical staining of collagen I.The ultrastructure of the specimens was observed with inverted phase contrast microscopy.Transient recovery rate of deformation,maximal recovery rate of deformation,and maximal compressive strength were tested to determine the biomechanical properties of the scaffold.Results The processed meniscus was milk-white in color with loose structure.It histologically appeared cell-free,stained positively for collagen I,and had abundant micropores according to phase-contrast microscopy.The transient recovery rate of deformation was（76.65±4.61）%,the maximal recovery rate of deformation was 100%,and themaximal compressive strength was（4.51±0.69） N when the specimens were compressed 40%.Conclusions The rabbit meniscus acellular matrix scaffold,with numerous micropores,is easy to be recovered from deformation and suitable for the adhesiveness and growth of breeding cells.This scaffold can be used as an ideal implant for future tissue engineering of the meniscus.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2011年第1期62-65,115,116,共6页
Acta Academiae Medicinae Sinicae
基金
国家高技术研究发展计划项目(863项目)(2007AA021902)~~
关键词
兔
半月板
脱细胞基质
生物力学特性
rabbit
meniscus
acelluar matrix
biomechanical properties
