Preclinical, translational study about the differences in peritoneal stem cells in women with and without adhesions after gyneocological surgery

Between December 2017 and February 2019, 92 patients were interviewed and informed about possible participation in the study. 50 patients declined participation or had to be excluded due to preoperative exclusion criteria. 42 patients were recruited, 16 patients showed intraoperative exclusion criteria. 26 patients out of 40 patients needed to be included in the study. The recruitment of patients was significantly delayed. Patients showed a significantly lower willingness to participate in the study than expected of less than 50%. In addition, part of the population was not available due to a concurrent study with a partially identical population, which reduced the population by about 1/3. Patient recruitment had to be terminated due to pandemic control measures. Analysis of ß-CR and EPOR receptors, RNA isolation, cDNA synthesis, gene expression (RT-qPCR), protein extraction, quantification of proteins (anti-EPOR; RRID: AB_28063149) were performed. Samples were assigned to one of four groups according to the stage of adhesion formation to distinguish adhesiogenicity. EPOR and CSF2RB as well as EPOR and CSF2RB were expressed at low levels. We observed no significant difference between patients from the "adhesiogenicity" groups. Here, 35 samples showed a very low RNA amount below 0.05ug/ul. These samples could not be analyzed by qPCR. Thirty samples analyzed showed low relative expression in all groups, with no evidence of regulation at the RNA level. An ANOVA of the ?Ct of the different groups showed no significant differences for either gene. We concluded that although the aforementioned genes could be detected in peritoneal samples, they did not show any regulation in the "resting state," i.e., at the onset of surgery, and thus did not explain any difference in the propensity for adhesion formation. A possible explanation would be a regulation only in the course of the healing process, which we could not show in our experimental setup, but also translational or post-translational interference, which would not allow a conclusion from RNA - to protein expression. Afterwards, the protein phase was isolated and the protein concentration was measured by BCA assay. The assay showed concentrations between 0.27 and 1.29 ug/ul, allowing protein analysis by Western blot.