Toxicity of candidate sealants for rupture of membrane in vitro

Abstract:

Objective：To provide safety referrences when sealing preterm premature rupture of the membranes（PPROM） by evaluating the toxicity of 3 tissue adhesives on fetal membranes. Methods：Cyanoacrylate（CA，ＣＡ group），fibrin sealant（FS，ＦＳ group） and col－lagen（Coll group） were directly laid on pieces of fetal membrane and an untreated piece was chosen as control. After being incubated for 24 h，the patches were fixed，embedded and sectioned for HE staining to observe the structural changes and TUNEL staining was applied to detect the cell apoptosis. The quantity of proteins related with apoptosis（Bax and Bcl-2） was tested by Western blot. Re－sults：（1）HE staining showed that there was consecutive rupture in the amnion layer treated with CA，while the amnion layer was complete in the other three left groups.（2）TUNEL staining showed that there were statistical differences in apoptotic rate between CA group（51.0±1.8）% and Con group（12.1±0.8）%，FS group（16.0±0.8）%，Coll group（13.0±1.0）%，P<0.01. There were statistical dif－ferences in apoptotic rate between FS group and Con group，Coll group，P<0.05. There was no statistical difference in apoptotic rate between Coll group and Con group，P >0.05.（3）The relative grey value of Bax protein was significantly increased in CA group（1.294±0.029） than in FS group（0.421±0.023），Coll group（0.479±0.024） and Con group（0.287±0.017）（P<0.05）. The relative grey values of Bcl-2 protein in CA group，FS group，Coll group and Con group were （0.350±0.034），（0.406±0.269），（0.433±0.020），（0.493±0.027） respectively. CA group had lower expression of Bcl-2 than the left groups（P<0.05），also the expression of Bcl-2 was lower in FS group and Coll group than in Con group（P<0.05）. No statistical difference in Bcl-2 expression was observed between FS group and Coll group（P >0.05）. Conclusions：（1）According to the research，CA is harmful to the amnion layer and not suitable for fe－tal membrane repair.（2）It’s better to choose FS or collagen to repair fetal membrane for safety concern.