The Influence Of The Transvaginal Mesh On Fibroblasts Proliferation And Collagen Synthesis

Objective:To observe the influence of different mesh on fibroblasts proliferation andcollagen synthesis in the anterior vaginal wall of women with pelvic floordysfunctionm in vitro study.Methods:Choose five cases of pelvic floor reconstruction surgery, occurs in March2013-May2013in obstetrics and gynecology, Fuzhou General Hospital for the study.Thebiopsy specimens were obtained from the anterior vaginal wall of women having apelvic floor dysfunction that was used as primary cultured fibroblasts by tissue culturein vitro. Observing morphological by inverting microscope, identifing fibroblast byimmunohistochemical staining with anti-vimentin antibodies and anti-keratinantibodies. Dividing fibroblasts into four groups to join Prolift mesh (Prolift group),Avaulta mesh (Avaulta group), TVT-O mesh (TVT-O group) and without mesh(control group). Using tetrazolium blue assay (MTT method) to detect cellproliferation in2,4,7days, and using Westen bolt to observe situation of I, III collagenegg by culturing fibroblasts together with mesh in4days.Result:1. It can be successfully obtained a large number of primary fibroblasts with theanterior vaginal fascia tissue. The method is easy, the process is simple as well, andproliferation of fibroblasts is active by culturing in vitro. Adherent cells grow larger,multi-projection, spindle-shaped or star-shaped, oval nuclei rules, and more nucleoli,observed by an inverted microscope. After the cells were passaged four generationswith anti-vimentin antibodies and anti-keratin antibodies cultures performedimmunohistochemical staining, cells were positive for anti-vimentin antibodies,negative for anti-keratin an antibody, which verifies the truth of fibroblasts. 2. Cell proliferation assay (MTT method): Cultured for2days after joining amesh,The absorbance values of fibroblasts in control group,Prolift group,Avaultagroup and TVT-O group was respectively（0.790±0.023）,(0.655±0.017),(0.644±0.022)and (0.538±0.067)；cultured for4days, the absorbance value of the correspondinggroup were (0.843±0.014),(0.716±0.045),(0.673±0.025) and (0.283±0.008);cultured for7days, respectively (2.040±0.035),(1.201±0.126),(1.109±0.087) and(0.196±0.017).In normal control group, absorbance values for each period werehigher than the corresponding period of three mesh groups, the differences werestatistically significant (P <0.05) by comparing separately, which showed that thethree mesh have an impact on proliferation of fibroblasts. Fibroblasts with threedifferent mesh were cultured for2days, Prolift group absorbance values slightlyhigher than Avaulta group and TVT-O group, but among pairwise comparison, thedifference was not statistically significant (P>0.05); cultured4and7days, Proliftgroup and Avaulta absorbance values were significantly higher than the TVT-O group,the differences were statistically significant (P<0.05), respectively; Prolift group,Avaulta group have a similar absorbance values in each period, which was notstatistically significant (P>0.05).3. Western blot method: Cultured for2days after joining a mesh, the normalcontrol group, Prolift group, Avaulta group and TVT-O group of type I collagenprotein expression were respectively (7.801±0.253),(5.421±0.191),(5.274±0.195) and (4.041±0.154); Ⅲ collagen protein expression were respectively(1.062±0.104),(0.870±0.051),(0.865±0.047) and (0.651±0.073).Comparing I andⅢ collagen of normal control group with Prolift group, Avaulta group and TVT-Ogroup respectively, the differences were statistically significant (P<0.05); Comparisonbetween Prolift group and Avaulta group, the difference was not statisticallysignificant (P>0.05), but the difference between Prolift, Avaulta and TVT-O werestatistically significant between (P <0.05).Conclusion:l. Application of tissue in vitro to culture fibroblast comes from patients withpelvic floor dysfunction disease vaginal wall, which can obtain large numbers of primary fibroblasts, good proliferation as well.2. Cultures in vitro of three polypropylene meshes have an impact on fibroblastproliferation of vaginal wall and collagen synthesis.3. Comparison between the three mesh, Prolift mesh and Avaulta mesh was equalin regard to affect the proliferation of fibroblasts, collagen synthesis and which affectthem relatively small, TVT-O mesh greatest impact.4. The greater of mesh aperture, the smaller impact of fibroblasts proliferationand the synthesis of collagen.