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目的:探讨不同网片对盆底功能障碍性疾病患者阴道前壁成纤维细胞增殖、胶原蛋白合成的影响。方法:选取2013年3~5月在该院妇产科,行盆底重建术的患者5例为研究对象。术中切取阴道前壁组织,组织块培养法于体外行成纤维细胞原代培养。用抗波形蛋白抗体和抗角蛋白抗体进行免疫组化法鉴定成纤维细胞。将培养至第4~6代成纤维细胞分为4组,设加入Prolift网片为Prolift组、加入Avaulta网片为Avaulta组、加入TVT-O网片为TVT-O组及不加网片为正常对照组。采用四甲基偶氮唑蓝比色法(MTT法)检测4组培养2、4、7天时的细胞增殖情况,采用Western bolt法检测细胞与网片共培养4天时成纤维细胞分泌Ⅰ、Ⅲ型胶原蛋白情况。结果:1网片与细胞共培养2天时,正常对照组、Prolift组、Avaulta组和TVT-O组吸光值分别为(0.790±0.023)、(0.655±0.017)、(0.644±0.022)和(0.538±0.067);培养4天时,分别为(0.843±0.014)、(0.716±0.045)、(0.673±0.025)和(0.283±0.008);培养7天时,分别为(2.040±0.035)、(1.201±0.126)、(1.109±0.087)和(0.196±0.017)。正常对照组各时期细胞吸光度值分别与其他3组比较,差异均有统计学意义(P<0.05)。Prolift组与Avaulta组各时期之间的吸光度值分别比较,差异无统计学意义(P>0.05)。培养2天时,Prolift组和Avaulta组分别与TVT-O组比较,差异均无统计学意义(P>0.05);培养4天和7天时各组比较,差异均有统计学意义(P<0.05);2网片与细胞共培养4天时,4组Ⅰ型胶原蛋白表达量分别为(7.801±0.253)、(5.421±0.191)、(5.274±0.195)和(4.041±0.154);Ⅲ型胶原分别为(1.062±0.104)、(0.870±0.051)、(0.865±0.047)和(0.651±0.073)。正常对照组的Ⅰ、Ⅲ型胶原蛋白分别与其他3组进行比较,差异均有统计学意义(P<0.05),其中Prolift组和Avaulta组分别比较,差异无统计学意义(P>0.05),但Prolift组、Avaulta组分别与TVT-O组进行比较,差异均有统计学意义(P<0.05)。结论:网片孔径越大,对成纤维细胞增殖、胶原蛋白合成影响越小。
Objective: To investigate the effect of different mesh on the proliferation of anterior vaginal fibroblasts and collagen synthesis in patients with pelvic floor dysfunction. Methods: From March to May 2013 in our hospital obstetrics and gynecology, pelvic floor reconstruction of patients in 5 cases as the research object. Intraoperative vaginal anterior wall excision, tissue culture method in vitro fibroblast primary culture. Fibroblasts were identified by immunohistochemistry with anti-vimentin and anti-keratin antibodies. The cultured fibroblasts from the 4th to 6th generation were divided into 4 groups. Prolift mesh was used as Prolift mesh, Avaulta mesh was added as Avaulta mesh, TVT-O mesh was added as TVT-O mesh and meshless mesh was Normal control group. The proliferation of cells in 4 groups was detected by MTT method. The cell proliferation of fibroblasts Ⅰ and Ⅲ Collagen condition. Results: (1) When the cells were co-cultured with the cells for 2 days, the absorbance values in the normal control group, Prolift group, Avaulta group and TVT-O group were 0.790 ± 0.023, 0.655 ± 0.017, 0.644 ± 0.022 and 0.538 ± 0.067), respectively; (4) When cultured for 4 days, they were (0.843 ± 0.014), (0.716 ± 0.045), (0.673 ± 0.025) and (0.283 ± 0.008) ), (1.109 ± 0.087) and (0.196 ± 0.017). Compared with the other three groups, the cell absorbance values of the normal control group at each time were significantly different (P <0.05). There was no significant difference in the absorbance values between Prolift group and Avaulta group (P> 0.05). There was no significant difference between the two groups in Prolift and Avaulta groups (P> 0.05). The difference was statistically significant (P <0.05) between the two groups on day 4 and day 7, ; (2) The co-culture days of 4 mesh cells were (7.801 ± 0.253), (5.421 ± 0.191), (5.274 ± 0.195) and (4.041 ± 0.154) (1.062 ± 0.104), (0.870 ± 0.051), (0.865 ± 0.047) and (0.651 ± 0.073) respectively. Compared with the other three groups, the collagen type I and type III in the normal control group were statistically significant (P <0.05), and there was no significant difference between the Prolift group and the Avaulta group (P> 0.05) However, Prolift group and Avaulta group were compared with TVT-O group, the difference was statistically significant (P <0.05). Conclusion: The larger the mesh size, the smaller the effect on fibroblast proliferation and collagen synthesis.