脱细胞肝支架的结构和成分对细胞相容性的影响

Effects of structure and composition of decellularized liver biological scaffolds on cytocompatibility

目的 比较不同方法制备脱细胞肝支架（DLBS）的结构和成分以及细胞相容性,探讨其内部联系.方法 取50只成年F344大鼠肝脏,使用文献报道的3种方法[以十二烷基硫酸钠（SDS）、Triton X-100和NP-40为主洗脱剂]制备大鼠DLBS.进行扫描电镜和孔径计算和氨基葡聚糖（GAG）检测.后经门静脉灌注肝卵圆细胞,进行黏附率、增殖情况、免疫荧光、扫描电镜以及白蛋白分泌检测.结果 大体观察、扫描电镜及孔径计算表明NP-40方案细胞外基质较SDS和Triton X-100方案整齐；GAG检测NP-40方案[（44.19±3.35） ng/mg]高于Triton X-100方案[（36.71±2.01） ng/mg]和SDS方案[（21.63 ±2.78） ng/mg]；细胞黏附率显示NP-40方案[（95.78±2.11）％]高于SDS方案[（84.18±3.30）％]和Triton X-100方案[（91.15±2.21）％]；细胞增殖情况Triton X-100方案和NP-40方案的细胞增殖速度显著高于SDS方案（P＜0.05）；白蛋白分泌NP-40方案[（85.77±3.30） mg/106个细胞]明显好于SDS方案[（49.37±2.43） mg/106个细胞]和Triton X-100方案[（74.66 ±4.80） mg/106个细胞].结论 DLBS的结构和成分对支架的细胞相容性具有明显的影响,同时也明显观察到NP-40方案较SDS和Triton X-100方案在制备DLBS的优势.

Objective To compare the biocompatibility of decellularized liver biological scaffolds （DLBS） made by different methods and discussing the correlation between them.Methods Decellularized liver biological scaffolds were made by transfusing livers of 50 mature F344 rats with three different ways.Glycosaminoglycan （GAG） in the scaffolds was tested,and scanning electron microscope （SCM） and cytotoxicity assays were also used on the scaffolds.The liver oval cells were transfused into scaffolds through the portal vein.Cells on the scaffolds were examined by immunofluorescence （IF）,scanning electron microscope,cells adhesion test and albumin excretion detection.Results Scaffolds made by the Method C had better orderliness than other two methods in gross observation,scanning electron microscopy and pore size calculation.The Method C [（44.19 ±3.35） ng/mg] had significantly more GAG than the Method A [（21.63 ±2.78） ng/mg] and Method B [（36.71 ±2.01） ng/mg].The scaffolds made by the Method C [（95.78 ± 2.11） ％] had higher collagen than in those made by the Method B [（91.15 ± 2.21） ％] and Method A [（84.18 ± 3.30）％].The cell adhesion rate of the Method B and Method C was higher than that of the Method A.Albumin excretion in the Method C [（85.77 ± 3.30） mg/106 cells] was greater than in the Method A [（49.37 ±2.43） mg/106 cells] and Method B [（74.66 ±4.80） mg/106 cells].Conclusion DLBS can be prepared by all three ways,but scaffolds made by our new method have better biocompatibility,and our method has significant advantages over the other methods.