A Study Of Sperm Acrosome Integrity And Morphology From Males With Oligozoospermia Or Obstructive Azoospermia As Well As Their Clinical Assisted Reproductive Treatment Outcomes

Objectives1. In order to examine the predictive value of sperm acrosome integrity on the outcomes of assisted reproductive techniques (ARTs), sperm acrosome integrity (AI) was evaluated in oligozoospermic and obstructive azoospermic men. Correlation analyses were assessed between AI and results of fertilization, embryo development, pregnancy.2. Assessments of sperm morphology were performed in males with oligozoospermia or obstructive azoospermia to evaluate the effect of sperm morphology on their ART outcomes.Materials and Methods1.(1) Thirty-six sperm samples from oligozoospermic males were collected as oligozoospermia group (OG). According to the5th edition WHO semen analysis manual (2010), these males were diagnosed as oligozoospermia via semen analysis for more than twice that their sperm count was less than39×106spermatozoa per ejaculate. The husbands, whose wives were aged from27to37years, were aged from28to40years old in this group.(2) Thirty-seven obstructive azoospermia (OA) males were divided into OA group (OAG). Husbands in this group were aged from27to37and the wives were aged from25to35years. (3) Males of control group (n=34) were normozoospermia, who were treated by ART for female factors. In this group, husbands were aged from29to37and wives were aged from27to35years.2.(1) According to the5th edition WHO semen analysis manual (2010), fluorescein isothiocyanate-conjugated pisum sativum agglutinin (FITC-PSA) at a final concentration of25μg/mL was used to assess the sperm acrosome integrity. Acrosome-intact rate (AIR) was observed under fluorescence microscopy using a450nm-490nm excitation laser after a2-h incubation in wet box at4℃.(2) Sperm morphology was carried out according to papanicolaou staining procedure in WHO semen analysis manual (2010). Normal form rate (NFR) was estimated under light microscopy.3. Oligozoospermic males and obstructive azoospermic males were treated by in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) technique according to rules in Center for Reproductive Medicine of the Third Affiliated Hospital of Guangzhou Medical College.4. Data were analyzed by SPSS statistical software.Results1. Many samples were detected with small acrosome, tapered acrosome, or amorphous head on sperm smear under fluorescence microscopy in OG. Lots of small heads, tapered heads, pyniform heads, amorphous heads and tail defects were observed after papanicolaou staining. In this group, AIR was significantly lower than that of control group (27±15%vs57±15%, P<0.01). OG showed significantly lower NFR than control group (9±5%vs18±9%, P<0.01).2. Small heads, tapered heads, pyniform heads, vacuolated heads and coiled tails became much more in OAG. Observed under fluorescence microscopy, these samples contained a lot of small acrosomes, amorphous acrosomes, and acrosomes with heterogeneous fluorescence stain. OAG gave significantly lower AIR (20±7%vs57±15%) and NFR (8±2%vs18±9%) than control group (P<0.01), respectively. Furthermore, AIR of OAG was significantly lower than that of OG.3. FR (52.8%vs84.2%) and OUR (28.5%vs47.5%) in OG were significantly lower compared with control group (P<0.01), respectively. Similar results were found between OAG (FR61.5%, OUR32.4%) and control group. There was no significant difference about EAR, OER, IR and PR between groups (P>0.05).4. Strong positive correlations were found between AIR and fertilization rate (FR)(P<0.01), AIR and oocyte utilization rate (OUR)(P<0.05), NFR and FR (P<0.05), NFR and OUR (P<0.05) in OG, OAG and control group, separately.Conclusions1. Oligozoospermic male’s spermatozoa contain impaired acrosome integrity and bad sperm morphology. Acrosome integrity will affect the fertilization after IVF and ICSI. Little influence is showed by acrosome integrity on embryo development and pregnancy outcomes. It indicates that sperms from oligozoospermic males with high AIR can result in high FR while going through IVF/ICSI.2. The bad acrosome integrity and sperm morphology from men with obstructive azoospermia lead to a bad fertilization results. It is pointed out that the acrosome integrity possibly may be regarded as an additional tool in the prediction of IVF/ICSI outcome and especially may be of value in men with oligozoospermia and obstructive azoospermia.3. Because of the poor sperm quality, the FR and OUR of IVF/ICSI from oligozoospermic men and obstructive azoospermic men are low. The embryo development and pregnancy outcomes are not affected.