Muharam Natadisastra

Expressions of growth differentiation factor-9 on granulosa cells of infertile women with endometriosis undergoing in vitro fertilization

Endometriosis may cause ovarian physiology disturbances and one of them, by affecting folliculogenesis that leads to a decreased oocyte quality. The oocyte plays an important role in regulating and promoting follicle growth by oocyte growth factors production. Several growth factors have been identified in human oocytes, including growth differentiation factor-9 (GDF-9). However, studies on GDF-9 expression in the granulosa cells of infertile women with endometriosis are sparse. To investigate the expression of GDF-9 mRNA in the granulosa cells of endometriosis patients undergoing in vitro fertilization (IVF) and identify the correlation between GDF-9 expression and oocyte quality. This cross-sectional study was conducted at the Yasmin IVF Clinic and Dr. Sander B Clinic, Jakarta from July 2014 to July 2017. In total, 50 granulosa cell samples were collected from 25 women with endometriosis and 25 controls who were healthy. The granulosa cell samples were collected at the time of oocyte retrieval. GDF-9 mRNA expression was investigated by real-time PCR. The numbers of retrieved oocytes, mature oocytes, and the oocyte morphology score were lower in patients with endometriosis than in controls; the difference was statistically significant. GDF-9 mRNA expression levels were quantitatively lower in the endometriosis groups than in the control group (5.05 [0.00002–3523.0] ng/mL vs. 81.93 [1.47–32450] ng/μL; p = 0.01). However, no correlation was obtained between GDF-9 expression levels and oocyte quality (oocyte morphology score and fertilization rate). The GDF9 mRNA level was lower in the endometriosis group than in the control group. However, no direct relationship was noted between individual GDF-9 level and oocyte quality. Hence, large-scale studies are needed to confirm whether GDF-9 expression is correlated with the oocyte quality and to ascertain whether GDF-9 has the potential for use as a new molecular marker to predict oocyte developmental competency.

Genetic and clinical predictors of ovarian response in assisted reproductive technology

Several factors are known to influence ovarian response to rFSH stimulation such as age, antral follicle count (AFC), and basal FSH level, Mutation of allele Ser680Asn in FSHR gene was responsible to ovarian resistance toward exogenous FSH. The aim of this study is to develop a prediction model of ovarian response to COS in IVF. This study was a prospective cohort study. One hundred and thirteen women undergoing their first cycle of IVF in Yasmin IVF Clinic Jakarta were recruited to this study. Clinical datas included were age, BMI, and AFC. Basal FSH and E2 as well as serum AMH was measured from peripheral blood taken at second day of cycle. Bsr-1 enzyme is used to identify the polymorphism in exon 10 position 680 with RFLP technique. Three genotype polymorphism, Asn/Asn (255 bp ribbon), Asn/Ser (97 bp and 158 bp), and Ser/Ser (97 bp, 158 bp, and 255 bp). AFC has the highest predictor for ovarian response with AUC 0.922 (CI 95% 0.833-1.000). AMH also showed high predicting value (AUC 0.843 CI 95% 0.663-1.000). The multivariate analysis revealed combination of AFC, AMH, age, and basal FSH is a good model for ovarian response prediction (AUC=0.97). No significant relation between Asn/Asn, Asn/Ser, or Ser/Ser genotype FSHR polymorphism with ovarian response (p = 0.866) and total dose of rRSH (p = 0.08). This study showed that model combination of AFC, AMH, patients age and basal FSH are very good to predict number of mature oocytes.

Correlation between Follicular Fluid’s Androgen Level and Fertilization Rate in Poor Responder Patients Undergone IVF: A Prospective Cohort Done in Yasmin IVF Clinic, Jakarta, Indonesia

Introduction Androstenedione (A4), testosterone (T), and dehidroepiandrosterone (DHEA) are known to be involved in folliculogenesis and follicular maturity. Lower follicular androgen levels in poor responder due to malfunctioned granulosa and theca cells and decreased inhibin B production will decrease FSH and LH. However, androgenic follicular fluid might also induce follicular atresia, decreased oocytes viability, thus affecting fertilization. The aim of current study is to find the correlation between intra-follicular androgen levels and fertilization rate, specifically in poor responder whom contributed in 84% cancelled cycle. Material and Methods This prospective cohort study was done at Yasmin IVF Clinic, Jakarta, Indonesia, in January 2014-March 2015. Infertile women undergone IVF were asked to participate, grouped into poor responder and other, and the androgen levels in the follicular fluid of each consenting patients were measured. Results From total 62 patients, aged 23-47 years old (37.6±5,068), there were 21 poor responders, whereas the other 41 patients with other indication. In poor responder group, levels of androstenedione, testosterone, and DHEA were 50.8 – 272.3 (103.5 ± 59.9), 383.2 – 1747.9 (1114.4 ± 373.2), 11.3 – 454.8 (151.3 ± 96.8), whereas in other group the androgen levels were 44.3 – 326.8 (95.1 ± 61.2), 414.1 – 1463.7 (976.9 ± 258.4), 44.6 – 265.8 (132.7 ± 61.3) with the correlation with fertilization rate 0.609, 0.095, and 0.361 respectively. Conclusion Correlation between follicular androgen levels and fertilization rate found to be not significantly different. The low number of subjects might cause this result, as well as the presence of bias, e.g.male factor and endometriosis might also affect fertilization. A multi-center study with larger sample size added with thorough analysis is needed to reconfirm current data.

Pentoxifylline as a Therapy for Thin Endometrial Lining in Infertility

Success rate of in vitro fertilization depends on two main things, namely the quality of the embryo and the atmosphere of endometrium during implantation. Endometrium has two phases during each menstrual cycle, which are proliferative and secretion phase.1 Physiologic changes in endometrial lining consists of increased functional layer thickness and increased spiral artery to prepare for implantation. If those changes fail to achieve pregnancy, menses occur.2,3 Therefore, to facilitate a successful pregnancy, an adequate growth of endometrium is necessary.3,4

Anti-Müllerian hormone as a predictor of polycystic ovary syndrome treated with clomiphene citrate

Objective This study aimed to determine the threshold of anti-Müllerian hormone (AMH) as predictor of follicular growth failure in polycystic ovary syndrome (PCOS) patients treated with clomiphene citrate (CC). Methods Fifty female subjects with PCOS were recruited and divided into two groups based on successful and unsuccessful follicular growth. Related variables such as age, infertility duration, cigarette smoking, use of Moslem hijab, sunlight exposure, fiber intake, body mass index, waist circumference, AMH level, 25-hydroxy vitamin D level, and growth of dominant follicles were obtained, assessed, and statistically analyzed. Results The AMH levels of patients with successful follicular growth were significantly lower (p=0.001) than those with unsuccessful follicular growth (6.10±3.52 vs. 10.43±4.78 ng/mL). A higher volume of fiber intake was also observed in the successful follicular growth group compared to unsuccessful follicular growth group (p=0.001). Our study found the probability of successful follicle growth was a function of AMH level and the amount of fiber intake, expressed as Y=–2.35+(–0.312×AMH level)+(0.464×fiber intake) (area under the curve, 0.88; 95% confidence interval, 0.79–0.98; p<0.001). Conclusion The optimal threshold of AMH level in predicting the failure of follicle growth in patients with PCOS treated with CC was 8.58 ng/mL.