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Density Gradient Centrifugation Pra-freezing Mengoptimalkan Persentase Morfologi Normal Spermatozoa Pasca-thawing Suyono, Seso Sulijaya; Hinting, Aucky; Lunardhi, Hamdani; I'tishom, Reny
Majalah Kedokteran Bandung Vol 50, No 3 (2018)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (129.265 KB) | DOI: 10.15395/mkb.v50n3.1348

Abstract

Kriopreservasi akan mengganggu struktur dan fungsi spermatozoa. Preparasi semen mampu menghasilkan spermatozoa dengan kualitas baik. Penelitian ini bertujuan menganalisis pengaruh preparasi semen dengan density gradient centrifugation (DGC) pra-freezing terhadap kualitas spermatozoa pasca-thawing. Penelitian dilakukan di boratorium Biologi Kedokteran, Fakultas Kedokteran Universitas Airlangga periode November 2017–Januari 2018. Penelitian eksperimental laboratoris dilakukan menggunakan cairan ejakulat volunter pria infertil. Semua sampel dibagi menjadi dua bagian, kelompok kontrol serta kelompok perlakuan berupa preparasi mini DGC. Setelah penambahan krioprotektan, selanjutnya dilakukan freezing. Pemeriksaan kualitas spermatozoa, meliputi motilitas, viabilitas serta persentase morfologi normal menggunakan metode WHO 2010, baik pra-freezing maupun pasca-thawing. Persentase perubahan kualitas spermatozoa pasca-thawing kedua kelompok dibandingkan menggunakan uji t dan bermakna jika nilai p<0,05. Total 20 sampel cairan ejakulat digunakan dalam penelitian ini. Persentase penurunan motilitas progresif, motilitas total, serta viabilitas pasca-thawing antara kedua kelompok tidak berbeda bermakna dengan nilai p masing-masing 0,422; 0,873 serta 0,432. Namun, penurunan persentase morfologi normal pasca-thawing pada kelompok kontrol jauh lebih besar daripada kelompok perlakuan dengan nilai p<0,001. Penelitian ini menyimpulkan bahwa preparasi semen berupa mini DGC pra-freezing mampu menghasilkan spermatozoa pasca-thawing dengan persentase morfologi normal yang lebih baik daripada protokol direct freezing.Pre-freezing Density Gradient Centrifugation Optimizes the Percentage of Post-thawed Sperms with Normal Morphology Cryopreservation impairs sperm structure and functions. Sperm preparation is a selection technique to obtain a population of high quality sperms. This study aimed to analyze the effect of pre-freezing Density Gradient Centrifugation (DGC) sperm preparation on the quality of post-thawed sperms. An experimental laboratory study was conducted using the ejaculates collected from volunteers visiting to our department. Samples were split into two fractions: control group and treatment group. In the treated group, mini-DGC sperm preparations were developed. After some cryoprotectants were added, samples were then cryopreserved using rapid freezing protocol. The evaluation of sperm quality that included evaluation on motility, viability and percentage of sperm morphology was performed by referring to 2010 WHO standardization on semen analysis. Evaluation was performed under pre-freezing and post-thawed condition. The percentage of sperm parameter changes between the two groups were compared using t-test with p-value <0.05 considered statistically significant. A total of 20 samples were included in this study. Post-thawed progressive motility, total motility, and viability considerably declined between two groups with p-values of 0.422, 0.873, and 0.432 respectively. In post-thawed observation, the percentage of sperms with normal morphology was significantly lower in the control group when compared to the treatment group  (p<0.001). In conclusion, the pre-freezing mini-DGC can optimize the post-thawed percentage of sperms with normal morphology compared to direct freezing protocol
Hyaluronan Expression on Vitrified Oocytes Before and After In Vitro Maturation (EKSPRESI HYALURONAN PADA OOSIT YANG DIVITRIFIKASI SEBELUM DAN SESUDAH MATURASI IN VITRO) Zakiyatul Faizah; R. Haryanto Aswin; Hamdani Lunardhi; Widjiati Widjiati
Jurnal Veteriner Vol 19 No 1 (2018)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (102.094 KB) | DOI: 10.19087/jveteriner.2018.19.1.71

Abstract

Oocyte vitrification is a major challenge in assisted reproductive technology. Oocyte vitrification with cumulus cells provide benefits in the process of maturation and fertilization. Vitrification leads to rapid temperature changes, therefore the decreasing in temperature could damage the cells even when the morphology was normal. Vitrification of mature oocytes is common because of its low sensitiveness towards low temperatures than immature oocytes. The aim of the research was to compare the effect of vitrification before and after in vitro maturation to the expression of hyaluronan. Maturation was operated in medium TC 50 ?L in CO2 incubators for 24 hours. Vitrification started with washing oocyte in PBS basic medium supplemented with 20% serum for 1-2 minutes, then in equilibration medium PBS + 20% serum + 10% ethylene glycol for 10-14 minutes, then transferred to 20% serum + PBS + 0.5 M sucrose + 15% ethylene glycol + PROH 15% for 25-30 seconds. Thawing was processed by submerging the oocytes in the media: 1). PBS + 20% serum + 0.5 M sucrose (K1); 2) PBS + 20% serum + 0.25 M sucrose (K2); and 3).PBS + 20% serum + 0.1 M sucrose (K3). Immunocytochemical stain was performed to evaluate the hyaluronan expression. Remmele scale index (Immunoreactive score, IRS) was used to read the result. There was no differences of hyaluronan expression in oocyte and cumulus group of K1, K2 and K3 at p< 0.05, statistically. We concluded that there was no difference of hyaluronan expression on oocyte and cumulus between vitrified oocyte of pre and post in vitro maturation which indicated that oocyte could be vitrified in the immature and mature state.
Effect of oocyte vitrification before and after in vitro maturation towards Bcl-2, Bax and Bcl-2/Bax ratio expression Zakiyatul Faizah; R. Haryanto Aswin; Hamdani Lunardhi
Majalah Obstetri dan Ginekologi Vol. 24 No. 2 (2016): May - August
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (117.057 KB) | DOI: 10.20473/mog.V24I22016.56-60

Abstract

Objectives: to compare the expression of Bcl-2, Bax and Bcl-2/Bax ratio in cumulus cell and oocyte between vitrified oocyte pre and post in vitro maturation.Materials and Methods: Maturation was operated in medium TC 100 µl for 24 hours. Vitrification begins with washing oocyte in PBS basic medium supplemented of 20% serum for 1-2 minutes, followed by equilibration medium PBS + 20% serum + 10% ethylene glycol for 10-14 minutes, then transferred to 20% serum + PBS + 0.5 M sucrose + 15% ethylene glycol + PROH 15% for 25-30 seconds. Thawing is processed by submerging the oocytes in the media: 1). PBS + 20% serum + 0.5 M sucrose, 2). PBS + 20% serum + 0.25 M sucrose, and 3). PBS + 20% serum + 0.1 M sucrose. Imunocytochemistry observed the expression of Bcl-2, bax and Bcl-2/bax ratio.Results: Bcl-2 expression on oocyte in control group differed significantly with treatment group, Bcl-2 expression on cumulus in control group differed significantly with treatment 1 group. Bax expression on oocyte in control group differed significantly with treatment group. Bax expression on cumulus in control group differed significantly with treatment group. Bcl-2/Bax expression ratio on oocyte and cumulus did not differ significantly in all groupConclusion: No difference Bcl-2/Bax expression ratio on oocyte and cumulus between vitrified oocyte pre and post in vitro maturation.
Comparison of pregnancy rates on day 3 and day 5 embryo transfer in In Vitro Fertilization (IVF) Audia Mumtaz Rifasky; Puspa Wardhani; Ashon Sa’adi; Ninik Darsini; Hamdani Lunardhi; Zakiyatul Faizah
Majalah Obstetri dan Ginekologi Vol. 29 No. 1 (2021): April
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/mog.V29I12021.14-17

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Objectives: To identify the success rates of pregnancy on the third and fifth day embryo transfer at Graha Amerta Hospital, Surabaya, Indonesia.Materials and Methods: This study used comparative cross sectional design. Data were taken from medical record of IVF participants who met the inclusion and exclusion criteria at Graha Amerta Hospital for the period of January 2016 - December 2016.Results: Successful pregnancy rates were found in this research. The embryo transfer on the third day and the fifth day were 35% and 49.3% respectively. In other words, the rates of pregnancy success were not affected by embryo transfer on the third day and the fifth day in the medical record sample as it had p value of 0.090.Conclusion: Embryo transfer on the third and fifth days had the same rates of pregnancy success in IVF participants at Graha Amerta Hospital, Surabaya, Indonesia. 
PENGARUH SPESIES BAKTERI DAN RATIO SPERMATOZOA/BAKTERI TERHADAP VITALITAS SPERMATOZOA MANUSIA SECARA IN VITRO Sukarjati Sukarjati; Hamdani Lunardhi
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 7 No 1 (2001): December 2001
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/508

Abstract

The effect of some pecies of bacteria and sperm/bacteria ratio on sperm vitality has been studied. Four species of bacteria were used in this study: Stapylococcus epidermidis, Streptococcus faecalis, Enterobacter aerogenes were obtained from semen culture of infertile men and E. coli was obtained from prostaltic fluid culture from men with prostat and urinary system disturbances. Five semen samples fulfilling the WHO criteri (1992) were used in this study. After preparation by Percoll gradient-column method, sperm were inoculated in a microplate with Stapylococcus epidermidis, Streptococcus faecalis, Enterobacter aerogenes and E. coli under the sperm/bacteria ratio 1:10 and 1:1000. Sperm vitality was observed immediately, 3 and 6 hours after inoculation. At the second experiment, the detrimental influence of bacteria on sperm was prevented by adding penicillin. Result of this study indicated that Stapylococcus epidermidis, Streptococcus faecalis, and Enterobacter aerogenes were not affected on the sperm vitality. The effect of E. coli on sperm vitality occurred at the ratio of sperm/bacteria 1:10 after 3 and 6 hours incubation and at the ratio of sperm/bacteria 1:1000 occurred after 6 hours incubation. It might be concluded that the negative influence of bacteria on sperm vitality in vitro, is dependent in species of bacteria, bacteria concentration, and time of incubation. The most detrimental effect on sperm vitality was shown by E. coli at the ratio of sperm/bacteria 1:10 after 6 hours incubation. The detrimental effecr was not prevented bt the addition of penicillin.
Factors Affecting The Outcome of in Vitro Fertilization (IVF) Fayka Putri Poempida; Jimmy Yanuar; Hamdani Lunardhi; Samsulhadi Samsulhadi; Relly Y. Primariawan
Syntax Literate Jurnal Ilmiah Indonesia
Publisher : Syntax Corporation

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (206.598 KB) | DOI: 10.36418/syntax-literate.v7i1.5780

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The high prevalence of infertility motivated researchers to find a solution, henceforth In Vitro Fertilization was invented. Factors that affect the outcome of IVF may include sperm analysis, maternal Body Mass Index (BMI), maternal smoking habits, endometriosis, and maternal age. However, there are ongoing debates about the role of said factors regarding the outcome of IVF. The objective of this research is to analyze those factors. This research is a Case-Control study with an analytical observational design. Data were retrieved from patients’ medical records undergoing IVF in Graha Amerta Fertility Clinic from January 2019-October 2020. First, the Chi-Square Test revealed sperm abnormality (p=0.212), Maternal BMI (p=0.427), endometriosis (p=0.067), meaning there was no connection with the outcome of IVF. Simultaneously, maternal age (p=0.037) showed a connection with the outcome of IVF. From the Binary Logistic Regression Test, maternal age 36-40 years old (p=0.044) affects the outcome of IVF significantly. Concurrently maternal BMI, endometriosis, and sperm abnormality have p value>0.05 meaning it is insignificant to the outcome of IVF. This research concluded that sperm abnormality, maternal BMI, and endometriosis do not affect the outcome of IVF. There was no data about maternal smoking habits. Whilst maternal age affects the outcome of IVF. Conclusion: This research concluded that sperm abnormality, maternal BMI, and endometriosis do not affect the outcome of IVF. There was no data about maternal smoking habits. Whilst maternal age affects the outcome of IVF.
Hubungan Usia Ibu, Jumlah AFC, dan Total Pemberian Gonadotropin terhadap Kualitas Embrio pada Program Fertilisasi In Vitro di Klinik Fertilitas Graha Amerta RSUD Dr. Soetomo Surabaya Periode 2018 – 2019 Chitra Devi Paramita Prasetyawati; Budi Santoso; Hamdani Lunardhi; Hendy Herdarto; Ashon Sa’ad
Syntax Literate Jurnal Ilmiah Indonesia
Publisher : Syntax Corporation

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (206.598 KB) | DOI: 10.36418/syntax-literate.v7i1.5742

Abstract

Fertilisasi In Vitro merupakan metode untuk mendapatkan kehamilan pada pasangan infertil. Meskipun dengan FIV, angka keberhasilan kehamilan masih cukup rendah yang tentu tidak terlepas dari pengaruh kualitas embrio yang didapatkan. Banyak faktor yang dapat mempengaruhinya, beberapa contohnya adalah usia ibu, jumlah AFC, dan total pemberian gonadotropin. Namun, penelitian mengenai ini masih terbatas terkait literatur dan informasinya. Mengetahui hubungan usia ibu, jumlah AFC, dan total gonadotropin terhadap kualitas embrio pada program FIV di Klinik Fertilitas Graha Amerta Periode 2018- 2019. Penelitian ini menggunakan rancangan cross sectional dengan metode total sampling. Bahan penelitian dari rekam medis pasien yang menjalami program FIV di Klinik Fertilitas Graha Amerta periode 2018-2019. Dari 52 pasangan yang menjalani FIV didapatkan hasil p-value antara usia dengan kualitas embrio baik hari ke-3 0,020, degenerasi 0,003, kualitas embrio baik hari ke-5 0,007, buruk 0,048. AFC dengan kualitas embrio baik hari ke-3 0,000, degenerasi 0,001, kualitas embrio baik hari ke-5 0,001. Total gonadotropin dengan kualitas embrio hari ke-3 dan ke-5 <0,05. Biochemical pregnancy dengan kualitas embrio baik hari ke-3 0,013. Penelitian ini membuktikan bahwa usia ibu dan jumlah AFC berhubungan dengan kualitas embrio. Total pemberian gonadotropin tidak berhubungan dengan kualitas embrio. Namun, apabila diteliti secara bersamaan terdapat hasil pengaruh signifikan secara simultan antara usia ibu, jumlah AFC dan total pemberian gonadotropin terhadap persentase kualitas embrio baik pada hari ke-3 dan hari ke-5.
Vitamin C and ubiquinone have the same ability in reducing the spermatozoa DNA fragmentation index in infertile men at Doctor Soetomo General Hospital, Surabaya, Indonesia Ayang Halim; Supardi Supardi; Hamdani Lunardhi
Intisari Sains Medis Vol. 11 No. 1 (2020): (Available online: 1 April 2020)
Publisher : DiscoverSys Inc.

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (449.449 KB) | DOI: 10.15562/ism.v11i1.546

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Background: Infertility is one of the most common health problems in the world. Malefactors contribute to 50% of cases and about 15-30% of infertile men are categorized as unexplained infertility. There have been numerous studies investigating the role of spermatozoa DNA fragmentation in male infertility. Spermatozoa nuclear DNA integrity has been suggested as a better predictor of male infertility and oxidative stress has been known related to it. This study was conducted to determine the oral supplementation of Vitamin C and Ubiquinone in reducing the spermatozoa DNA fragmentation index (DFI) among infertile men. Methods: This was an experimental clinical trial with a pre-test and post-test group design. Thirty infertile men were randomized double-blindingly into 3 groups: Vitamin C and Ubiquinone; Vitamin C and Placebo; Ubiquinone and Placebo groups, each treatment was given for 35 days. The DFI was evaluated by Sperm Chromatin Dispersion (SCD) test, before and after treatment. Data were analyzed using SPSS version 21 for Windows.Results: The average age of respondents was 32.6 years old, followed by 4.6 years of infertility duration in 5.8 years of marriage. Most of the respondents had a normal Body Mass Index (BMI) (46.7%), no smoking history (75.0%), no history of heat (90.0%) or chemical (73.3%) exposure, and diagnosed with primary male infertility (90.0%). Vitamin C group showed a significant reduction in the spermatozoa DFI (95% CI, p<0.05), while the other two groups showed no differences in the spermatozoa DFIs (95% CI, p>0.05). Comparison analysis showed no differences in the reduction of the spermatozoa DFIs (95% CI, p>0.05) among these three groups. Conclusion: The recent findings suggest that oral supplementation of vitamin C can reduce the spermatozoa DFI, while ubiquinone and the combination of vitamin C and ubiquinone could not reduce the spermatozoa DFI.
Glutathione administration reduces the DNA fragmentation index in sperm preparation with the Swim-Up (SU) and mini-Density Gradient Centrifugation (mini-DGC) methods at Doctor Soetomo General Hospital, Surabaya, Indonesia Lutfi Hardiyanto; Amarudin Amarudin; Indra Gusti Mansur; Supardi Supardi; Wahyuning Ramelan; Hamdani Lunardhi
Intisari Sains Medis Vol. 11 No. 3 (2020): (Available online: 1 December 2020)
Publisher : DiscoverSys Inc.

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (295.245 KB) | DOI: 10.15562/ism.v11i3.739

Abstract

Background: Spermatozoa requires Reactive Oxygen Species (ROS) in physiological quantities for capacitation for fertilization purposes. However, if the balance between ROS production and antioxidant capacity is disrupted, it can result in oxidative stress, negatively affecting chromatin spermatozoa, leading to male infertility. This study evaluates the glutathione administration reduces the DNA fragmentation index in sperm preparation with the Swim-Up (SU) and mini-Density Gradient Centrifugation (mini-DGC) methods at Doctor Soetomo General Hospital, Surabaya, Indonesia.Methods: A true experimental study by post-test only control group was conducted using ejaculate fluid of 9 infertile patients with normozoospermic at the Department of Medical Biology, Faculty of Medicine, Universitas Indonesia, Jakarta in January - February 2019 period. The spermatozoa DNA fragmentation was examined by following the instructions for using Spermfunc® DNAf Kit for Determination of the DNA Fragmentation Level in Spermatozoa by Sperm Chromatin Dispersion (SCD) for both preparation methods. Data were analyzed using SPSS version 16.0 for Windows.Results: SU vs. SU+glutathione method gave a lower but insignificant DNA fragmentation effect (p>0.05). The mini-DGC vs. mini-DGC+glutathione method showed significant results (p<0.05). SU+glutathione vs mini-DGC+glutathione showed no significant results (p>0.05). Meanwhile, the SU vs. mini-DGC method showed significant results (p <0.05). The recovery rate (RR) of the mini-DGC method has a higher value and is even better if glutathione is given compared to the SU method.Conclusion: Spermatozoa preparation in the SU method gives a lower DNA fragmentation effect than the mini-DGC method. The administration of glutathione can be a particular consideration in carrying out spermatozoa preparation to protect spermatozoa from damage during preparation.
IN VITRO FERTILITY TEST OF HUMAN SPERMATOZOA MEMBRANE PROTEIN FERTILIN BETA ANTIBODY IN MICE (Mus musculus Balb/c) AS IMMUNOCONTRACEPTIVE CANDIDATE I'tishom, Reny; Soebadi, Doddy M; Hinting, Aucky; Lunardhi, Hamdani; Yudiwati, Rina
Folia Medica Indonesiana Vol. 52 No. 3 (2016): JULY - SEPTEMBER 2016
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (104.598 KB) | DOI: 10.20473/fmi.v52i3.5453

Abstract

One of the materials as potential candidates immunocontraception material is spermatozoa. Fertilin beta is spermatozoa membrane protein and is found only in mature spermatozoa and ejaculate, which serves as an adhesion molecule. Spermatozoa membrane protein that is used as an ingredient immunocontraception candidate, must have specific criteria that the specificity of spermatozoa, the role of antigen in the fertilization process, which includes the formation of immunogenicity sufficient antibody response has the potential to block fertilization. Antibodies against spermatozoa affect the stages before fertilization of the reproductive process and can hinder the development of the embryo after fertilization. Until now very little research data spermatozoa membrane protein as an ingredient immunocontraception are up to the test of experimental animals. The research objective is to prove the role of the resulting antibody induction of antibodies fertilin beta protein in the membrane of human spermatozoa induce agglutination and reduce motility thus reducing the number of in vitro fertilization. Research conducted at the IVF Laboratory, Department of Biology of Medicine, Faculty of Medicine, University of Airlangga. This research includes: Test the potential of antibody protein beta fertilin membrane of human spermatozoa and inhibit the role of antibodies in vitro fertilization in mice (Mus musculus Balb/c). In vitro studies have resulted in fertilization figure of 25% is smaller than the number that is equal to control fertilization of 58.7%, whereas previously the spermatozoa were incubated first with a beta membrane protein antibody fertilin human spermatozoa. While the percentage of inhibition of sperm to fertilize an oocyte by 33.75%. Potential imunokontraseptif considered effective if it decreased significantly (P <0.05) than the numbers fertilization in the treatment group compared with the control group. This shows fertilin beta membrane protein antibody has the ability to inhibit human spermatozoa to fertilize oocytes that reduce the number of fertilization.