Ten years of in vitro fertilization in Indonesia: Access to infertility care in a developing country.

OBJECTIVE: This research was conducted to assess access to assisted reproductive technologies (ART) and the current status of the in vitro fertilization (IVF) program that have been implemented in Indonesia over the last 10 years. METHODS: We established a retrospective cohort study and descriptive analysis of the current state of access to infertility care in Indonesia. The data were collected from all IVF centers, clinics, and hospitals in Indonesia from 2011 to 2020, including the number of IVF clinics, total ART cycles, retrieved fresh and frozen embryos, average age of IVF patients, IVF pregnancy rate, and causes of infertility. RESULTS: The number of reported fertility clinics in Indonesia has increased from 14 clinics in 2011 to 41 clinics by 2020. As many as 69 569 ART cycles were conducted over the past 10 years, of which 51 892 cycles used fresh embryos and 17 677 cycles used frozen embryos. The leading cause of consecutive infertility diagnosis was male infertility. Nearly half of the women who underwent IVF procedures (48.9%) were under 35 years old. The pregnancy rate outcome of women who underwent IVF ranged from 24.6% to 37.3%. CONCLUSION: Developments in ART in Indonesia have led to improvements in the ART cycles performed throughout the 10 year period. The identification of key areas that require improvement can provide an opportunity to enhance access to infertility care.

Down-Regulation of miR-93 Negatively Correlates with Overexpression of VEGFA and MMP3 in Endometriosis: A Cross-Sectional Study.

BACKGROUND: Endometriosis is identified as presence of the endometrium outside the uterine cavity. Retrograde menstruation contributes to the endometrial tissue implantation and the establishment of endometriotic lesions at ectopic sites. It has been suggested that the endometriotic lesions are rich in angiogenic growth factors, while they have an essential role in survival and invasion of these cells. We investigated regulation of microRNA-93 (miR-93) and its involvement with vascular endothelial growth factor A (VEGFA) and matrix metalloproteinase (MMP) 3 expression in women with endometriosis. MATERIALS AND METHODS: This was a cross-sectional study at Central Surgical Installation, Dr. Cipto Mangunkusumo General Hospital, Jakarta, Indonesia, between October 2020 and November 2021. Eutopic and ectopic endometrial tissues were collected from 30 subjects with laparoscopically-confirmed endometriotic women. Normal endometrial cells of non-endometriosis women served as controls. Total RNA was isolated from all samples and a quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to analyze the expression of miR-93, VEGFA and MMP. RESULTS: There was no significant difference in the expression levels of VEGFA (2.14 ± 0.50, P=0.719) and MMP3 (2.99 ± 0.42, P=0.583) between endometriotic lesions of endometriosis women and the healthy endometrium. Expression of miR-93 was significantly lower in the eutopic endometrium (16.7 fold) and ectopic endometriotic lesion (20 fold) compared to the normal endometrium (P<0.001). Furthermore, we also observed a significant correlation between miR-93, VEGFA expression in eutopic endometrium obtained from women with endometriosis (r=-0.544, P=0.029). Expression of the miR-93 was also negatively correlated with MMP3 expression in both eutopic (r=-0.412, P=0.01) and ectopic (r=-0.539, P=0.03) endometrial cells of women with endometriosis. CONCLUSION: VEGFA and MMP3 expression levels trended to be increased in both eutopic and ectopic endometrial tissues of endometriosis women, while down-regulation of miR-93 might be involved in the alteration of VEGFA and MMP3 in endometriosis.

Cytotoxic activity of peripheral blood mononuclear cells in patients with endometriosis: A cross-sectional study.

Background: Endometriosis is believed to be associated with dysfunction of the lymphocyte population and cytotoxicity of natural killer (NK) cells, induced by the production of interleukin-2 (IL-2). Objective: This study aimed to investigate T lymphocytes and NK cell activity in the peripheral blood mononuclear cells (PBMCs) of women with endometriosis. Materials and Methods: PBMCs were obtained from the peripheral venous blood samples of 14 women with and without endometriosis (n = 7 for each group). Then, the PBMCs were co-cultured for 4 days and were treated with recombinant IL-2 for cytotoxic activity toward target cells (Daudi and K562 cells). The cytotoxicity activity was determined using the 51 chromium release assay before and after stimulation. Flow cytometry measurement was used to examine the expression of T lymphocytes and NK cells before and after being treated with IL-2. Results: The concentration of CD3+CD28+ (co-stimulatory) was significantly lower in the endometriosis group (65.62 ± 5.38) compared to in its counterpart (50.24 ± 4.22) (p = 0.04) before stimulation. However, no significant differences were observed in any other T lymphocytes and NK cells. It was also found that there was a significant increase of CD3-CD28+ after treatment with IL-2 only in the healthy control but not in women with endometriosis. Conclusion: Increased expression of CD160 and decreased CD28 play a role in inhibiting NK cell activation and T cell response in women with endometriosis.

Vitamin B3 (niacin), B6, C, and iron intake are associated with the free androgen index, especially in normoandrogenic polycystic ovary syndrome

Objective: Nutritional intake is one of the most common environmental risk factors for polycystic ovary syndrome (PCOS) because it is associated with obesity and insulin resistance. The aim of this study was to determine the relationship between micronutrient intake and androgen levels associated with PCOS. Material and Methods: This cross-sectional study was performed in patients with PCOS divided into two groups, normoandrogenic (NA) and hyperandrogenic (HA), and healthy controls. Dietary intake assessment was performed using a modified 38-item semi-quantitative food frequency questionnaire. Bivariate, correlation, and multivariate analyses were performed to determine the association between study variables. Results: There were 79 patients with PCOS, of whom 50 were NA and 29 were HA. There were 66 subjects in the healthy control group. The baseline characteristics in all groups were similar, except for body mass index and hormonal profile which were elevated in the HA group compared to the other groups. There was a significant negative correlation between the free androgen index (FAI) and intake of vitamin B1, vitamin B2, niacin, vitamin B6, calcium, and iron in the NA group, while this association was absent in the HA group. Multivariate linear regression analysis showed that the intake of vitamin B6, vitamin C, niacin, and iron had a significant effect on the FAI. Conclusion: There is an effect of micronutrient intake on androgen levels in women with PCOS. The association was more significant in NA PCOS than in the HA PCOS groups. These findings suggest an association between micronutrients, androgens and PCOS at a systemic level.

Combination of electroacupuncture and pharmacological treatment improves insulin resistance in women with polycystic ovary syndrome: Double-blind randomized clinical trial.

Background: Acupuncture is a nonpharmacological treatment which has been known to improve ovulatory function in polycystic ovary syndrome (PCOS) women. Acupuncture modulates the somatic and autonomic nervous systems, which regulate endocrine and metabolic functions to impact ovulatory functions. Objective: To investigate the effectiveness of electroacupuncture (EA) and pharmacological combination therapy on improving insulin resistance in women with PCOS. Materials and Methods: This double-blind, randomized clinical trial was performed on 44 participants from March to September 2018 at Cipto Mangunkusumo National hospital, Jakarta, Indonesia. Participants were randomly allocated to treatment (true EA + medication) and control group (sham EA + medication) in a 1:1 ratio using a web-based computer random-number generator. Randomization was carried out by an independent project manager. Both groups received 12 sessions of acupuncture therapy and metformin as pharmacological therapy. Results: This study showed a significant decrease in the Homeostatic Model Assessment for Insulin Resistance index in the true EA + medication group before and after therapy (p = 0.014). Conclusion: The combination of EA and pharmacological therapy effectively improves insulin sensitivity in women with PCOS.

Knowledge, attitudes, and behaviors of practitioners supporting cancer patients in fertility preservation in DKI Jakarta: A cross-sectional study.

Background: More than 135,000 people aged under 45 yr are diagnosed with cancer annually in Indonesia. Good detection and management of cancer increase the quality of life. Objective: To determine the knowledge, attitudes, and behaviors of practitioners supporting cancer patients in fertility preservation. Materials and Methods: This cross-sectional study was conducted in 18 type D government hospitals and Dr. Cipto Mangunkusumo Hospital, in Jakarta, Indonesia, between January 2018 and August 2019. This study involved practitioners providing care to cancer patients. Data were described descriptively. Results: Most of the general practitioners, specialists, and subspecialists who participated in this study were aged 26-30 yr (65.4%), 31-35 yr (70.4%), and 31-40 yr (53.0%), respectively. The fertility treatment most known by general practitioners was in vitro fertilization with embryo cryopreservation (12.1%); for specialists it was sperm cryopreservation (24.5%). Meanwhile, subspecialists knew most about in vitro fertilization with embryo cryopreservation and sperm cryopreservation using a GnRH agonist (such as leuprolide injection) pre-cancer treatment (13%). A positive attitude towards fertility preservation as an important priority for cancer patients was shown in 72.0% of general practitioners, 73.3% of specialists, and 100% of subspecialists. General practitioners mostly referred patients to fertility specialists (44.4%). Many specialists (54.9%) and subspecialists (67%) discussed the possible impact of the patient's condition and / or treatment on fertility. Conclusion: The knowledge of and practices related to fertility preservation differed among general practitioners, specialists, and subspecialists. However, positive attitudes among them were similar.

Correlation of calprotectin serum levels with degrees of endometriosis: A cross-sectional study.

BACKGROUND: Endometriosis is closely associated with delayed diagnosis due to the lack of a definitive and sensitive noninvasive approach. The use of calprotectin in inflammatory process has been demonstrated in various inflammatory diseases. Calprotectin has a significant correlation with high-sensitivity C-reactive protein (hs-CRP) and could be used as an inflammatory marker. No study thus far has evaluated the correlation between calprotectin and endometriosis. OBJECTIVE: To determine the correlation of calprotectin with the degree of endometriosis in order to help clinicians in establishing better early detection and management. MATERIALS AND METHODS: In this cross-sectional study, 46 women referred to the Cipto Mangunkusumo, Fatmawati, and Persahabatan Hospitals in Jakarta, Indonesia between July 2017 and April 2018 were enrolled, and their blood serum were taken a day before surgery. Calprotectin serum level was treated using the PhicalⓇ ELISA method. After the diagnosis of endometriosis was confirmed through pathological examination, the final diagnosis of endometriosis could be established. The degree of endometriosis was classified according to the revised American Society for Reproductive Medicine (ASRM) classification. RESULTS: The prevalence of minimal, mild, moderate, and severe degrees of endometriosis were 15.2, 39.1, 34.8, and 10.9%, respectively. The median serum calprotectin levels for minimal, mild, moderate, and severe endometriosis were 138.98, 121.49, 124.16, and 122.82 mg/mL, respectively. No correlation was observed between calprotectin and the degrees of endometriosis (r = -0.16, p = 0.278). CONCLUSION: There is no correlation between calprotectin serum levels and the degrees of endometriosis.

Suppressing HOXA-10 Gene Expression by MicroRNA 135b During the Window of Implantation in Infertile Women.

BACKGROUND: Implantation failure has long been identified as a common problem underlying low success rate of IVF. Currently, endometrial receptivity has gained expert attention as it is demonstrated to contribute to successful embryo implantation. MicroRNAs (miRNAs) is known to affect endometrial receptivity through post-transcriptional gene expression regulation. This study aimed to evaluate the expression of miRNA 135b and HOXA-10 during the implantation window in endometrial tissue of infertile women. METHODS: A total of 14 patients diagnosed with infertility in the gynaecology clinic of Cipto Mangunkusumo and Daya Medika hospitals Jakart, Indonesia were selected as the observed group, and 9 fertile patients were enrolled in the control group. Total RNA was isolated from endometrial tissues collected at the secretory phase of the menstrual cycle. The miRNA 135b and HOXA-10 mRNA expression were measured using quantitative real-time PCR (qPCR). The correlation between these variables was then determined using Pearson's correlation coefficient. RESULTS: The expression of miRNA 135b in the infertile group was significantly higher by 1.81-fold compared to the control group (p<0.01), whereas, expression of HOXA-10 mRNA was significantly lower in the infertile group compared to the controls (p=0.047). Significant negative correlation was observed between the expression of miRNA 135b and HOXA-10 mRNA in infertile women (p=0.021; r=-0.607). CONCLUSION: Taken together, this study provides that alteration of miRNA expression is involved in regulating the implantation process partly via modulation of the expression of gene required for implantation.

Correlation of anti-Müllerian hormone levels with metabolic syndrome events in polycystic ovary syndrome: A cross-sectional study.

BACKGROUND: Various endocrine disorders have been reported in women of reproductive age, 10% of which is affected by polycystic ovary syndrome (PCOS). OBJECTIVE: This study aimed to evaluate the correlation of anti-Müllerian hormone (AMH) levels with the metabolic syndrome in patients with PCOS. MATERIALS AND METHODS: This cross-sectional study employed a consecutive sampling method using medical records from January 2013 to December 2017 at Dr. Cipto Mangunkusumo General Hospital polyclinic and Yasmin in vitro fertilization Clinic (Kencana), Jakarta, Indonesia. The primary outcome of the study was the AMH levels as independent variable correlated with metabolic syndrome. The secondary outcome was also the AMH levels correlated with each PCOS phenotype. The tertiary outcome was each PCOS phenotype as independent variable correlated with metabolic syndrome. RESULTS: Women with phenotype 1 of PCOS had a median AMH level of 13.92 (range: 3.88-34.06) ng/ml. 21% patients had metabolic syndrome, with a median AMH level 7.65 (3.77-20.20) ng/ml, higher than the women without metabolic syndrome (p = 0.38). The most frequent phenotype in women with PCOS was phenotype 4, oligo- or anovulation and polycystic ovary morphology (OA/PCOM) in 41.3%. The most frequent phenotype in women with metabolic syndrome was phenotype 1, OA + PCOM + hyperandrogenism in 56.5%. CONCLUSION: All PCOS phenotypes exhibited significant correlations with the AMH level. Phenotype 1 (OA + PCOM + hyperandrogenism) was associated with the highest AMH level and was significantly associated with metabolic syndrome.

Analysis of factors associated with ovarian reserve in a group of poor responders to in vitro fertilization: A cross-sectional study.

BACKGROUND: Poor ovarian reserve and a high rate of pregnancy failure associated with low quality and quantity of oocytes are observed in poor responders to in vitro fertilization. OBJECTIVE: To assess the effect of age, body mass index (BMI), endometriosis, and history of ovarian surgery on ovarian reserve in a group of poor responders. MATERIALS AND METHODS: In this cross-sectional study 749 women who referred to Yasmin Clinic of Dr. Cipto Mangunkusumo National General Hospital from January 2013 to June 2017 were enrolled. Two definitions of poor responders and Poseidon criteria and consecutive sampling techniques were used. Participants were divided into good and poor responder groups based on the ovarian reserve test; participant with oocyte ≤ 3 was classified as a poor responder. Based on this, 188 participants with nine (4-47) oocytes were included in the poor responder group. While, good responder comprised of two (0-3) oocytes. RESULTS: Age and anti-Mullerian hormone level (AMH) were significantly associated with ovarian reserve in the poor-responder group (p < 0.001). However, in multivariate analyses, age was the only significant predictor of ovarian response in the poor-responder group (p = 0.004). While endometriosis was the significant predictor of Poseidon groups 1 and 4, surgical history was the significant predictor of Poseidon groups 2 and 3. Meanwhile, an increase in BMI decreased the risk of classification under Poseidon group 3. CONCLUSION: Age, AMH, BMI, endometriosis, and history of ovarian surgery affected the risk of classification of the Poseidon group.

Dysregulation of Kisspeptin and Leptin, as Anorexigenic Agents, Plays Role in the Development of Obesity in Postmenopausal Women.

During the menopausal period, women have a higher tendency to develop obesity and any other metabolic syndromes. Dysregulation of leptin and kisspeptin signaling as anorexigenic agents is believed to be the connection between metabolic disorders and altered reproductive function. Therefore, this study aimed at investigating the association between leptin, soluble leptin receptor (sOBR), free leptin index, kisspeptin concentrations, and body mass index (BMI) in postmenopausal women. A cross-sectional study was carried out among 171 postmenopausal women aged 40-75 years from 2017 to 2018. Subjects were assigned into 2 groups according to their BMIs: obese group (84 subjects) and nonobese group (87 subjects). In addition to anthropometric measurement, blood sample was collected from each subject for leptin, sOBR, free leptin index (FLI), and kisspeptin evaluation. Bivariate and correlation analysis discovered that leptin and FLI were positively correlated with BMI, while sOBR and kisspeptin were negatively correlated with BMI. Among those variables, multivariate analysis found that leptin, sOBR, and kisspeptin were independently associated with obesity. Therefore, it can be concluded that higher serum leptin concentration and FLI, as well as lower serum sOBR and kisspeptin concentrations, are significantly associated with obesity in postmenopausal women.

Body fat percentage is a better marker than body mass index for determining inflammation status in polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is an endocrinopathic disorder most commonly experienced by women of reproductive age, and it is characterized by a low-grade chronic inflammatory condition. Excessive fat deposit has been long considered as an etiological factor in the pathogenesis of this inflammatory condition. Currently, body mass index (BMI) or percentage of body fat is used as a marker to assess the body fat composition of a person. OBJECTIVE: To determine whether BMI or body fat percentage (BFP) can be used as a better marker for measuring inflammation related to body fat accumulation in polycystic ovary syndrome patients. MATERIALS AND METHODS: This study took place at the Center for Reproductive Medicine, Yasmin Clinic, Cipto Mangunkusumo Hospital from January to December 2015. In this cross-sectional study, 32 reproductive age women with PCOS according to the Rotterdam criteria (2003) participated. Women with hyperandrogenism caused by non-classic congenital adrenal hyperplasia, pregnant and lactating women, etc., were excluded. Some variables such as BMI, clinical hyperandrogenism sign, BFP, and inflammatory markers were assessed and statistically analyzed. RESULTS: From a total of 32 subjects of the study, BFP had a significant positive correlation with procalcitonin levels (r=0.35; p=0.048), while BMI did not (r=0.27; p=0.131). CONCLUSION: BFP can be used as a better marker for measuring inflammation related to body fat accumulation in PCOS subjects.