Nghiên cứu đặc điểm lâm sàng, cận lâm sàng và một số yếu tố liên quan đến cặp vợ chồng vô sinh nguyên phát

Mục tiêu: Nghiên cứu nhắm đánh giá đặc điểm lâm sàng, cận lâm sàng và nguyên nhân vô sinh của các cặp vợ chồng vô sinh nguyên phát. Đồng thời, tìm hiểu một số yếu tố liên quan đến tình trạng vô sinh nguyên phát. Đối tượng phương pháp nghiên cứu: Nghiên cứu mô tả cắt ngang 91 cặp vợ chồng trong độ tuổi sinh sản được chẩn đoán vô sinh nguyên phát, đến khám và điều trị tại Trung tâm Nội tiết sinh sản và Vô sinh, Bệnh viện Trường Đại học Y dược Huế từ tháng 5/2020 đến tháng 4/2021. Kết quả: Nguyên nhân vô sinh do cả 2 vợ chồng là 54,9%, trong đó vô sinh nam chiếm 21,3% và vô sinh nữ chiếm 18,7%. Nguyên nhân vô sinh nữ chủ yếu là rối loạn phóng noãn (62,6%) mà phần lớn là nguyên nhân PCOS (Polycystic Ovary Syndrome-hội chứng buồng trứng đa nang) (40,6%). Tỷ lệ vô sinh nguyên phát ở nhóm có chu kỳ kinh nguyệt không đều cao hơn so với nhóm có chu kỳ kinh nguyệt đều, tỷ lệ tương ứng là 82% so với 63,4% (OR= 2,6; 95% CI: 1,02 – 6,81; p=0,04). Nguyên nhân vô sinh nam chủ yếu là bất thường tinh trùng (78%). Ngoài ra, có 9,9% trường hợp có giãn tĩnh mạch thừng tinh. Tỷ lệ bất thường tinh dịch đồ ở nhóm đứt gãy DNA tinh trùng cao và trung bình cao hơn nhiều so với nhóm DFI (DNA fragmentation index - chỉ số phân mảnh DNA tinh trùng) thấp (tỷ lệ tương ứng là 82,1% và 81,5% so với 54,5%) với p=0,048. Không có mối liên quan giữa độ tuổi, BMI (Body mass index- chỉ số khối cơ thể) với tỷ lệ vô sinh nguyên phát ở 2 giới. Kết luận: Vô sinh nguyên phát có nguyên nhân đồng thời cả nam giới và nữ giới chiếm tỷ lệ cao nhất, trong đó nguyên nhân phổ biến nhất bao gồm rối loạn phóng noãn và tinh dịch đồ bất thường. Đặc điểm chu kỳ kinh nguyệt và kết quả phân mảnh DNA tinh trùng là những yếu tố ảnh hưởng đến chức năng sinh sản ở nữ giới và nam giới.

Effect of Sperm Dna Fragmentation Index on Clinical Outcomes of Intra-Uterine Insemination Patients

Objective: To study the effect of sperm chromatin condensation (DNA fragmentation index (DFI)) using aniline blue-eosin (AB-E) staining on pregnancy outcomes in patients facing unexplained infertility undergoing intra- uterine insemination (IUI). Our initial hypothesis states that if DNA fragmentation is high then chances of pregnancy are low/NIL hence these patients should be recommended with advanced ART procedures like IVF and ICSI. Design: Prospective study Setting: Tertiary care infertility centre Method: A total of 185 patients with age less than 40 years, non-smokers and without history of any pathogenic infection in the past 2months facing unexplained infertility i.e., males with normal semen analysis reports and females with normal ovulation and hysterosalpingography (HSG) reports were selected for the study. Patients were undergoing their first or second IUI treatment cycle between the period of June 2016 to December 2016. DNA fragmentation index (DFI) using aniline blue- eosin staining method was studied in semen samples provided on the day of IUI procedure. The patients were separated into 3 groups: low DFI (DFI<= 10%), medium DFI (DFI=11 % - 20%), and high DFI (DFI >= 21%) and clinical pregnancy outcomes of IUI were recorded. Statistical analysis was performed using Pearson correlation co-efficient, ANOVA and Shapiro Wilk Test on the above groups. Main Outcome Measures: DNA fragmentation index (DFI) (%), Clinical pregnancy rate (%) Result: The overall clinical pregnancy rate for the selected patient pool was 21.08% with an average DFI of 8.84% in the pregnant female group and 14.65% in the non-pregnant female group. Sperm DFI % and clinical outcomes in IUI treated patients were statistically significant and negatively correlated with correlation coefficient (r) of -0.1, -0.3 and -0.3 in low DFI%, medium DFI and high DFI% groups respectively. Conclusion: Our study demonstrated that DFI (%) and clinical pregnancy rate (%) are significantly and negatively correlated in patients with normal semen parameters undergoing IUI. The higher the DFI% the chances of clinical pregnancy become very low, therefore, these patients should not be recommended IUI but with advanced ART procedures like IVF and ICSI.

Correlation between Sperm Dna Fragmentation and Conventional Semen Parameters among Different Age Groups

Objective: To study the relationship between conventional semen parameters and sperm chromatin condensation (DNA fragmentation index) using aniline blue-eosin staining method among patients of different age groups visiting the In-vitro fertilization (IVF) clinic.Design: Retrospective study Setting: Tertiary care infertility centre Method: A total of 240 patient semen samples were studied between the period of May 2015 to May 2016 for conventional semen parameters (WHO criteria) and DNA fragmentation index (DFI) using aniline blue- eosin staining method. Patients were separated into three groups: <=30 years, 31-35 years and 36 years & above. Statistical analysis was performed using Pearson correlation co-efficient and regression tests on the groups. Main Outcome Measures: Sperm concentration (Millions /ml), motility(%), normal morphology(%), DFI (%). Result: In each age group, i.e., <=30years, 31-35 years and 36 years & above, there was a significant and negative correlation between DFI and sperm concentration (r= -0.50, r= -0.34, r= -0.49 respectively; P<0.05), motility(r= -0.69,r= -0.66, r= -0.54 respectively; P<0.05) and normal morphology (r= -0.86,r= -0.80, r= -0.75 respectively; P<0.05). Sperm DNA fragmentation index among the age groups was not statistically significantly (P>0.05). Conclusion: Our study demonstrated that age is not a predictor of DFI. Whereas, sperm concentration, sperm motility and normal sperm morphology showed a significant association with DFI in all the age groups i.e., better the conventional semen parameters, lower the DFI.

Evaluation of the Relationship Between Body Mass Index (BMI) and DNA Fragmentation Index Changes in Primary Infertile Patients Following Microscopic Sub Inguinal Varicocelectomy

Objectives: To investigate changes in DNA fragmentation index in primary infertile patients with varicocele, which is followed by microscopic subingual varicocelectomy in different groups based on body mass index (BMI). Methods: This study was performed in 100 patients with primary infertility with varicocele. Patients were divided into three groups (normal (N), overweight (OW), and obese (OB)) based on BMI index. DNA fragmentation index (DFI) parameters were evaluated before and 6 months after varicocelectomy. For DFI analysis, the SCD (sperm chromatin dispersion test) method was used. Data were analyzed using t-test, Chi-square, and ANOVA. Results: In this study, the mean age of participants was 33.6 and their mean BMI was 28.6, that 51 patients underwent bilateral varicocelectomy and 49 patients underwent left varicocelectomy surgery. In this study, a comparison of DFI before and 6 months after surgery showed a decrease in DFI in all three groups. The difference was 23 in the normal weight group, 11.2 in the overweight group and 9.58 in the obese group, which is statistically significant (PV < 0.05). Also, in comparison with the rate of DFI reduction between groups, the normal weight group showed a greater decrease than the overweight and obese group. This difference was statistically significant (PV < 0.05), while comparing the rate of DFI reduction between the two groups of overweight and obese, was observed no significant difference (PV = 0.635). Conclusions: Although DFI level decreased significantly 6 months after surgery in all groups with different body mass index. However, the rate of reduction was not the same in different groups and was higher in normal-weight patients than in overweight and obese individuals. But there was no significant difference in the rate of reduction between the overweight and obese groups.

Associations of Sperm mtDNA copy number, DNA fragmentation index, and reactive oxygen species with clinical outcomes in ART treatments

Background Except for sperm parameters (sperm concentration, motility, and morphology), sperm mitochondria DNA copy number (mtDNA-CN), DNA fragmentation index (DFI), and reactive oxygen species (ROS) content are essential characteristics for sperm function. However, the roles of these measurements in embryo development and pregnancy outcomes are still unclear. Methods Semen samples of 402 participants were collected. Sperm parameters, including sperm morphology, concentration, and motility, were analyzed by a computer-assisted sperm analysis system. MtDNA-CN, DFI, and ROS levels were measured using remained sperms through quantitative polymerase chain reaction method, sperm chromatin structure assay and Reactive Oxygen Species Assay Kit, respectively. The assisted reproductive technology (ART) treatments, including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), were performed in 126 couples of these participants. Results In 402 semen samples, elevated mtDNA-CN and DFI were associated with poor seminal quality. In 126 couples conducted ART, only mtDNA-CN was negatively correlated with fertilization rate in ART cases. However, it was not significant after adjusting male age, female age, seminal quality, and ART strategy. With regard to pregnancy outcomes, none of sperm mtDNA-CN, ROS and DFI was associated with clinical pregnancy rate in 79 cases transferred embryos. Conclusions Increased mtDNA-CN and DFI in sperms jointly contributed to poor seminal quality. However, none of sperm mtDNA-CN, ROS/MS and DFI were associated with clinical outcomes in ART.

Impact of Paternal Genome with a High DNA Fragmentation Index (>60%) on Early Embryonic Development

: Objectives: The objective of this study is to propose thresholds of the sperm DNA fragmentation rate (IFA≤30% IFA31%-60% IFA>60%), in order to assess the clinical effects of the paternal genome on intra cytoplasmic sperm injection parameters, in particular the effect of the latter on early embryonic development. Materials and Methods: The procedure is a retrospective study, which involved 101 patients enrolled in an ICSI program with their partners. The index of spermatic DNA fragmentation rate was measured using the Sperm Chromatin Dispersion assay. Results: There is a negative correlation between high levels of the spermatic DNA fragmentation index and spermiological characteristics: Concentration P=0.002 and mobility P=0.0001. For ICSI results, there are different observations on the existence of a correlation between the spermatic DNA fragmentation index and fertility rate. On the other hand, the rate of sperm DNA fragmentation does not seem to influence early embryonic development, and even couples whose partners have a high fragmentation index manage to obtain the best quality embryos (P=0.002). We observe a decrease in the rate of implantation with an increase in the rate of alteration of the sperm genome, but this remains insignificant P > 0.05. Conclusion: ICSI remains the only alternative for men with a high rate of sperm DNA fragmentation. Moreover, the operator seems to influence the results more than is suggested. This does not exclude the paternal effect which may influence the quality of the concepltus later on. Keywords: DNA Fragmentation Index, ICSI, Fertilization Rate, Embryos Quality.