Sun Daodong, Song Bo, Sun Danning
Center of Urology, PLA 324th Hospital, Changqing, 400020, P. R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2004 May;18(3):217-9.
To evaluate the cytocompatibility of collagen membranes with transitional cells of rabbit in vitro and to discuss the possibility of the collagen membranes as urologic tissue engineering scaffolds.
Primary cultured transitional cells isolated from New Zealand rabbits were implanted on collagen membranes at 1 x 10(5) cells/cm2. The changes of cell adhering were observed by inverted microscope and scanning electron microscope 2, 12 and 24 hours later. The experiment was divided into 4 groups: non-cell group (black control) culture medium group (negative control), extract medium from Polyvinyl chloride group(positive control) and extract medium from collagen membranes group(experimental group). The cells of generations 2 to 4 were implanted in 96-hole-plank at 1 x 10(4) cells every hole. And every group had 5 holes. Then absorption coefficient were detected at the wave length of 490 nm by MTT assay. Then the cytotoxicity and cytocompatibility were evaluated by comparison of the numbers of absorption coefficient.
The bladder transitional cells began to adhere to the collagen membrane 2 hours after implanting, and the number of the adhered cells increased with time. The actual absorption coefficient of experimental groups was 0.590 +/- 0.024, 1.065 +/- 0.40 and 1.129 +/- 0.074 after 24, 72 and 120 hours. The actual absorption coefficient of negative control group was 0.639 +/- 0.068, 1.022 +/- 0.044 and 1.087 +/- 0.111. The actual absorption coefficient of positive control group was 0.302 +/- 0.029, 0.653 +/- 0.083 and 0.694 +/- 0.031. There was significant difference between the experimental group and positive control (P < 0.01), and no significant difference between the experimental group and negative control(P > 0.05).
Collagen membrane has good cytocompatibility with transitional cells and no cytotoxicity. It can be used as scaffolds of urologic tissue engineering.
评估胶原膜与兔移行细胞的体外细胞相容性,并探讨胶原膜作为泌尿外科组织工程支架的可能性。
将从新西兰兔分离的原代培养移行细胞以1×10⁵个细胞/cm²接种于胶原膜上。分别于接种后2、12和24小时,用倒置显微镜和扫描电子显微镜观察细胞黏附情况。实验分为4组:无细胞组(黑色对照)、培养基组(阴性对照)、聚氯乙烯提取物培养基组(阳性对照)和胶原膜提取物培养基组(实验组)。将第2至4代细胞以每孔1×10⁴个细胞接种于96孔板中,每组5孔。然后通过MTT法在490nm波长处检测吸光度。通过比较吸光度值评估细胞毒性和细胞相容性。
接种后2小时膀胱移行细胞开始黏附于胶原膜,且黏附细胞数量随时间增加。实验组在24、72和120小时后的实际吸光度分别为0.590±0.024、1.065±0.40和1.129±0.074。阴性对照组的实际吸光度分别为0.639±0.068、1.022±0.044和1.087±0.111。阳性对照组的实际吸光度分别为0.302±0.029、0.653±0.083和0.694±0.031。实验组与阳性对照组之间差异有统计学意义(P<0.01),实验组与阴性对照组之间无显著差异(P>0.05)。
胶原膜与移行细胞具有良好的细胞相容性且无细胞毒性。它可作为泌尿外科组织工程的支架。