细胞外基质复合温敏性水凝胶与膀胱上皮细胞的生物相容性

Biocompatibility between the compound pattern of extracellular matrix and temperature sensitivity hydrogel and urothelial cells

目的检验细胞外基质/温敏性水凝胶(ECM/TSH)复合支架材料与膀胱上皮细胞(UC)的生物相容性,探索其作为泌尿道组织工程支架的可行性。方法实验分为A、B、C三组,A组为ECM/TSH复合支架种植UC组;B组为单纯ECM种植UC组;C组为单纯培养UC组,另设无细胞培养液为空白组(D组)。应用角蛋白免疫荧光染色技术鉴定细胞,应用倒置相差显微镜和扫描电镜观察细胞的黏附和生长情况,利用MTT法检测细胞的活力。结果光镜下ECM/TSH复合支架为红染的网状结构,纤维较粗,网孔较小,未见到细胞碎片;种植UC后4 h可见细胞紧密黏附于材料表面;14 h可见细胞沿支架纤维伸展;3 d时可见黏附于材料的细胞增多、增殖明显;扫描电镜下,可见发育良好的细胞伪足沿纤维伸展,附着牢固,细胞间连接紧密,胞膜表面有ECM分泌。MTT法示各组间细胞相对增殖率差异有统计学意义(P<0.05)。结论 ECM/TSH复合材料具有便于细胞黏附和生长的微孔结构,生物相容性好,无细胞毒性,是一种理想的组织工程材料。

Objective To evaluate the biocompatibility between the compound pattern of extracellular matrix （ECM） and temperature sensitivity hydrogel （TSH） and urothelial cells （UC）. Methods There were 3 groups in this experiment： UC were cultured in ECM （group A）, in the ECM/TSH compounds materials （group B） and on the plastic surfaces as a control （group C） respectively. Cells were identified with indirect immunofluorescence staining of keratin, and were examined by phase contrast microscopy and scanning electron microscope （SEM） to assess cell growth and morphology. Cell adhesion and eel1 proliferation were detected With MTT assay. Results Histologically, the ECM/TSH compounds appeared as a net-like and red-stained construction with enough meshes and without any cellular fragments. 4 h after implantation, cells were observed adhere on the compounds and extend spurious along the fibers 10 h later. Under SEM even some ECM were observed been secreted on the surface of cells. MTT assay showed there was obvious statistic difference among 3 groups （P 〈 0.05）. Conclusion ECM/TSH compounds materials appeare great biocompatibility with UC, which confirmes that it would be an ideal tissue engineering scaffolds.