The interactions between various tissue factors in the congenital diaphragmatic hernia affected tissue

The aim of this study was to evaluate growth factors and their receptors,muscle and nerve quality,local defense factor,programmed cell death and separate gene expression in human pathological material with congenital diaphragm hernia(CDH)in order to find to find immunohistochemical marker differences between the control and the patient group. The diaphragm material was obtained from 5 children age from 1 to 2 days.These samples were stained with immunohistochemistry method for TGF-β,bFGF,FGFR1,IGF-1,IGF-1R,HGF,NGF,NGFR,NF,dystrophin,myosin,alpha actin,ß-defensin-2,ß-defensin-4,TUNEL,Wnt-1 to detect their expression in diaphragm tissue.A semi-quantitative counting method was used for the evaluation of the tissues and structures in the stained slides. Obtained results were analyzed with IBM SPSS Statistics 23 using Mann-Whitney U test(U)and Spearmann’s rank correlation, where correlation coefficient(r)results were interpreted:0.80-1.00 very high,0.60-0.79 high,0.40-0.59 moderate,0.20-0.39 low,<0.2 very low correlation.For all test statistical significance was defined as p<0.05. Statistically significant changes between patient and control groups were found in muscle fibers:bFGF(U=10;p=0.013),FGFR1(U=9;p=0.01),in blood vessels:TGF-β(U=13.5;p=0.032), FGFR1(U=7;p=0.005),IGF-1R(U=9;p=0.01),ß-defensin-2(U=14.5;p=0.041),TUNEL(U=14.5;p=0.041),NGF(U=10.50;p=0.013), in connective tissue:HGF(U=0;p<0.001),but no significant data in mesothelium.Spearman’s rank correlation coefficient revealed multiple very strong(r 0.8–1.0),strong(r 0.6–0.79)and moderate(r 0.4–0.59)correlations between immunoreactives in different diaphragm cells and structures. Various correlations and significant changes between control and patient groups indicates that angiogenesis in diaphragm can be severely affected by CDH pathogenetic processes.CDH could also affect wnt-1 expression whereas the correlations suggest wnt-1 interactions with other growth factors could lead to dysregulation in angiogenetic processes.Notable changes between patient and control groups in bFGF and FGFR expression shows their importance in CDH pathogenesis. Immunoreactive expression showed equal distribution suggesting CDH affects all diaphragm sites equally. Correlations between IGF-1R,HGF,TGF-β and β-defensin-2/apoptosis indicate the leading role of tissue growth,anti-microbial defence and programmed cell death in the morphopathogenesis of CDH.Decreased NGF,NGFR and NF expression indicates the possible decrease of neuronal structure quality in the pathogenesis of this anomaly.CDH has little to no effect on mesothelium as well as myosin formation.