Effects Of Sperm Morphology And DNA Integrity After Semen Optimization Technique On Outcomes Of IVF-ET

Object:To explore the effects of morphology and DNA integrity after semen optimization technique on outcomes of IVF-ET.Methods:605 cycles of in-vitro fertilization and embryo transfer(IVF-ET)for oviductal infertility were enrolled in this study in Department of Reproduction and Endocrinology in Ningbo Women and Children's Hospital from January 1st 2013 to December 31st 2014.On the day of the retrieval,we examined sperm morphology,acrosome morphology and DNA integrity of protoplasmic sperm and optimized sperm using Differential Quik(Diff-Quik),Pisum sativum agglutinin-Fluorescein isothiocyanate(PSA-FITC)and sperm chromatin dispersion(SCD)method.The different levels of sperm deformity index(SDI),acrosome abnormity rate(AAR)and DNA fragmentation index(DFI)between optimized sperm group and protoplasmic sperm group,and the effects of those parameters on the fertilization failure and clinical pregnancy failure in IVF were analyzed.The correlations of outcomes of IVF-ET with SDI and DFI of optimized sperm were also analyzed.Results:The levels of SDI,AAR and DFI were 1.39±0.38,(11.95±5.79)%and(11.66±6.73)%in protoplasmic sperm group,and 1.25±0.26,(8.36±5.43)%and(9.65±5.60)%in optimized sperm group respectively.The difference between protoplasmic sperm group and optimized sperm group was statistically significant(P<0.05).SDI,AAR and DFI were negatively correlated with sperm progressive movement rate(PR)(r=-0.25,-0.22 and-0.27,P all<0.05),and optimized sperm deformity index(OSDI),optimized sperm acrosome abnormity rate(OAAR)and optimized sperm DNA fragmentation index(ODFI)were negatively correlated with PR of optimized sperm as well(r=-0.36,-0.38 and-0.45,P all<0.05).The levels of SDI,AAR,DFI,OSDI,OAAR and ODFI were 1.55±0.28,(16.71±3.64)%,(17.03±4.53)%,1.37±0.15,(12.05±3.06)%and(14.422±4.03)%in the fertilization failure group,and 1.35±0.23,(11.14±3.34)%,(10.78±3.99)%,1.21±0.20,(7.75±2.28)%and(8.67±3.17)%in the control group respectively.The difference between fertilization failure group and control group was statistically significant(P<0.05).SDI,AAR,DFI,OSDI,OAAR and ODFI were risk factors for fertilization failure(OR= 1.48,1.83,1.55,2.28,11.29 and 3.27,P all<0.01).The joint predictor(JP)and optimized sperm joint predictor(OJP)were established by Logistic equation.The area under ROC curve(AUC)of SDI,AAR,DFI,JP,OSDI,OAAR,ODFI and OJP were 0.629±0.036,0.774±0.034,0.703±0.032,0.816±0.030,0.699±0.034,0.893±0.020,0.806±0.024 and 0.925±0.018.According to the Youden index,OSDI,OAAR and ODFI were used to obtain the optimal cut-off values of fertilization failure,approximately 1.30,10%and 15%.The levels of numbers of high quality transplanted embryos,SDI,DFI,OSDI and ODFI were 1.75±0.50,1.58±0.10,(15.91 ±3.69)%,1.32±0.17 and(12.25±4.14)%in the clinical pregnancy failure group,and 1.92±0.28,1.36±0.11,(9.30±4.22)%,1.24±0.18 and(7.44±3.35)%in the control group respectively.The difference between clinical pregnancy failure group and control group was statistically significant(P<0.05).Numbers of high quality transplanted embryos,SDI,DFI,OSDI and ODFI were influence factors for clinical pregnancy failure(OR= 0.54,1.28,1.41,1.66 and 2.98,P all<0.01),DFI,OSDI and ODFI were negatively correlated with numbers of high quality transplanted embryos(r=-0.18,-0.31 and-0.66,P all<0.01),but there was no statistically significant correlation between SDI and the parameter of transplanted embryos(r=-0.03,P>0.05).The AUC of SDI,DFI,OSDI and ODFI were 0.626±0.034,0.709±0.029,0.691±0.032 and 0.835±0.022.According to the Youden index,OSDI and ODFI were used to obtain the optimal cut-off values of clinical pregnancy failure,approximately 1.30 and 10%.According to the cut-offs of ODFI,605 cycles were divided into four groups,<5%,(5-10)%,(10-15)%,?15%.The rates of fertilization,cleavage,high quality embryo,implantation,clinical pregnancy,early spontaneous miscarriage and live birth were statistical significant in the difference of comparisons among four groups(P<0.05).No statistically significant differences were observed in the rates of multiple pregnancy,premature birth and low birth weight(P>0.05).According to the cut-off of OSDI,605 cycles were divided into two groups,<1.30,?1.30.The rates of fertilization,high quality embryo,implantation,clinical pregnancy were statistical significance in the difference of comparisons between two groups(P<0.05).No statistically significant differences were observed in the other rates(P>0.05).Negative correlations were found between OSDI,OAAR,ODFI and the fertilization rate(r=-0.24,-0.50,-0.32,P all<0.01).Negative correlations were found between ODFI and the cleavage rate,high quality embryo rate,clinical pregnancy rate and live birth rate(r=-0.19,-0.40,-0.20 and-0.09,P<0.01,<0.01,<0.01 and 0.04).Positive correlation was found between ODFI and the early spontaneous miscarriage rate(r= 0.23,P<0.01),but there were no statistically significant correlations between ODFI and the multiple pregnancy rate,premature birth rate and low birth weight rate(r=-0.01,0.04 and 0.03,P= 0.83,0.54 and 0.62).OSDI was negatively correlated with the high quality embryo rate(r=-0.17,P<0.01),and was positively correlated with ODFI(r= 0.32,P<0.01).Conclusions:Our study suggests that the levels of morphology and DNA integrity of the optimized sperm are significantly improved and are closely associated with the capacity of fertilization in IVF,and acrosome morphology is more valuable than other single predictors,and OJP is the most effective.Morphology of optimized sperm is likely to be lower associated with the embryonic development and clinical pregnancy,however,DNA integrity of optimized sperm effects on embryonic development and clinical pregnancy outcomes,and the relationship between premature birth,low birth weight and those two parameters should be investigated by further research.