Restu Syamsul Hadi
Fakultas Kedokteran Universitas YARSI

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Apoptosis Pada Sperma Sebagai Petanda Adanya Gangguan Kesuburan Pria Hadi, Restu Syamsul
Majalah Kesehatan Pharmamedika Vol 3, No 2 (2011): VOL 3 No 2 tahun 2011
Publisher : Majalah Kesehatan Pharmamedika

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Semen analysis includes assessment of volume, color, viscosity, pH, concentration, motility, and morphology as standard procedures to semen analysis. It has known that correlation between apoptosis and spermatogenesis. Apoptosis is a mechanism regulating spermatogenesis in humans, and there are clear differences in molecular markers of apoptosis between males with normal sperm parameters and those with abnormal sperm parameters. Sperm apoptosis has been shown to increase after testicular injury, such as exposure to toxics, varicocle, testicular torsion, hormonal deprivation and genetic abnormalities. It has been found that the number of sperm with Fas expression was low in subjects with normal sperm parameters but high in men with abnormal sperm parameters. It has been reported that the number sperm with Fas expression was low in subjects with normal sperm parameters but high in men with abnormal sperm parameters. Male infertility appears to be positively correlated with increased levels of apoptotic sperm. Sperm DNA damage and sperm apoptosis have been considered as useful marker of male fertility disorder.Keywords : infertility, spermatogenesis, apoptosis, DNA damage
Mekanisme Apoptosis Pada Regresi Sel Luteal Hadi, Restu Syamsul
Majalah Kesehatan Pharmamedika Vol 3 N0 1 2011
Publisher : Majalah Kesehatan Pharmamedika

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Corpus luteum is a transient endocrine gland and its presence plays pivotal role in ovulation process, implantation, and luteinization. The regression of corpus luteum or luteolysis is actually needed for next cycle survival and playing a role in providing new follicles.  Luteal regression represents a broad definition of the process of demise of the corpus luteum that is capable of accommodating all new knowledge evolved on the molecular mechanisms activated or inhibited during the process of regression of the corpus luteum. Apoptosis is controlled by a number of regulator genes such as bcl-2 family. The ratio of Bax expression to Bcl-2 is a pivotal factor for a cell to survive or apoptosis. The increasing Bcl-2 expression will lengthen cell life, whereas increasing of Bax expression promotes cell death.
Curcumin Analogue (Pentagamavunone-0) Induces Luteal Cell Apoptosis by Increased Bax/Bcl-2 Protein Ratio Hadi, Restu Syamsul; Soejono, Sri Kadarsih
Majalah Kesehatan Pharmamedika Vol.2 No.1 2010
Publisher : Majalah Kesehatan Pharmamedika

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The present study was conducted to investigate the role of curcumin analoguePentagamavunone-0 (PGV-0) of luteal cell function in apoptosis by examined Bcl-2 and Baxprotein expressions. Corpora lutea of rat Sprague-Dawley at 28 days of age after induced by10 IU pregnant mare’s serum gonadotropin (PMSG) were dispersed enzimatically bycollagenase. Isolated luteal cells were seeded in 24 well culture plates containing minimalessential medium (MEM) and fetal bovine serum (FBS) until cell had grown to confluence.PGV-0 (400 μM) was immediately treated after they were stimulated by luteinizing hormone(LH), prostaglandin-F2 (PGF2) and LH+PGF2. All these treatments were incubated for 24hours. Apoptotic luteal cells were analyzed for DNA break by terminal deoxy-nucleotidyltransferase mediated dUTP-biotin nick end labeling (TUNEL) assay. To examine of Bcl-2 andBax protein expression, sample of plated luteal cells on coverslip were stained byimmunocytochemical method. The administration of PGV-0 and/or PGF2α induced lutealcells apoptosis. LH (50 ng/ml) decreases Bax protein expression and inhibits PGV-0-inducedluteal cell apoptosis by decresed Bax/Bcl-2 protein ratio. Treatment of PGV-0 (400 μM)decreased Bcl-2 protein expression, increased Bax protein expression, increased Bax/Bcl-2protein ratio and induced apoptotic luteal cells. It was concluded that PGV-0 induces lutealcells apoptosis by increased Bax/Bcl-2 protein ratio and decreased apoptotic inhibition by LH.Keywords : curcumin, PGV-0, Bcl-2, Bax, apoptosis, luteal cell
Geraniin supplementation increases human keratinocyte proliferation in serum-free culture Kusuma, Indra; Hadi, Restu Syamsul
Universa Medicina Vol 32, No 1 (2013)
Publisher : Faculty of Medicine, Trisakti University

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BACKGROUND Various products used in cellular therapy utilize tissue culture techniques requiring keratinocyte culture. An efficient and clinically acceptable keratinocyte culture system requires supplements with mitogenic activity. Geraniin is a phytochemical with the potential as a supplement for expansion culture of keratinocytes. The objective of the present study was to verify the mitogenic activity of geraniin on human keratinocytes. METHODS This was an experimental study using two samples of human foreskin obtained by circumcision of a male child. Epidermal keratinocytes were isolated from the foreskin samples and were divided into paired groups, comprising intervention and control groups. The intervention groups were cultured with geraniin supplementation, whereas the control groups with standard supplements, without the addition of geraniin. Mitochondrial activity of the cells was evaluated by means of the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium-bromide (MTT) proliferation assay. Absorbance values in each of the groups was measured at 450 nm. Data analysis was performed with the paired t-test. RESULTS Geraniin supplementation significantly increased the keratinocyte proliferation rates at dosages of 0.8 to 3.1 μM. An increase of 57% in the proliferation rate was obtained at a dosage of 1.6 μM, while at a dosage of 12.5 μM toxic effects were starting to appear. Geraniin presumably causes increased cellular energy status, resulting in increased proliferation rates. CONCLUSION The findings in this study provide evidence in support of the utilization of geraniin as a supplement for expansion culture of keratinocytes. Further studies may presumably identify the molecules acting as geraniin receptors and the intracellular mechanisms underlying the increase in proliferation rates.
Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture Hadi, Restu Syamsul; Kusuma, Indra; Sandra, Yurika
Universa Medicina Vol 33, No 2 (2014)
Publisher : Faculty of Medicine, Trisakti University

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BACKGROUNDTransplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC).METHODSIn this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4) expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1) proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test.RESULTSDermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05). Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacityCONCLUSIONSAllogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.
Isolasi Sel Punca Pluripoten dengan Penanda CD105+ dan SSEA3+ dari Sel Fibroblas Kulit asal Jaringan Preputium CHURIYAH, CHURIYAH; KUSUMA, INDRA; KUSUMASTUTI, SISKA A; HADI, RESTU SYAMSUL; WIBOWO, AGUNG ERU; FABIOLA, FAIZA KARA
JURNAL ILMU KEFARMASIAN INDONESIA Vol 14 No 2 (2016): JIFI
Publisher : Fakultas Farmasi Universitas Indonesia

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The development and knowledge of stem cell research are growing quickly. Stem cells are now likely to become a modern treatment as a regenerative approach that has potential treatment for various diseases that are difficult to cure, such as genetic diseases, degenerative, trauma, and malignancy. The aimed of this study was to isolate pluripotent stem cells using human dermal fibroblast cell obtained from preputium tissue by enzymatic and tissue explants methods, followed by double positive characterization for specific markers of cluster differentiation (CD) 105 and surface-stage embryonic antigen (SSEA)-3 using magnetic-based purification system (MACS). The fibroblast cells resulted from both method were cultured and expanded by variations medium and passage cells, then were characterized for double positive CD 105+ and SSEA3+. Fibroblast cell culturing in conditioned medium, lower passage and higher density has the highest percentage of CD105+ subpopulation cells compared than fibroblast cell in standard medium, higher passage and lower density. Subpopulation of both of CD 105+ and SSEA3+ cells revealed a positive response to alkaline phosphatase dye which proving a population of stem cells.
Gangguan fungsi sitoskeleton pada proses vitrifikasi keratinosit primer manusia Kusuma, Indra; Hadi, Restu Syamsul; Sandra, Yurika
Jurnal Kedokteran YARSI Vol 25, No 2 (2017): MEI - AGUSTUS 2017
Publisher : Lembaga Penelitian Universitas YARSI

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Abstract

Keratinosit basal memiliki sifat multipotent, dibutuhkan kultur bebas-serum agar terhindar dari diferensiasi spontan. Kultur keratinosit memberikan peluang untuk berbagai jenis aplikasi riset dan terapi seperti bioengineered skin. Penyimpanan sel dengan metode vitrifikasi terbukti dapat melindungi fungsi embrio pada layanan bayi tabung. Penggunaan vitrifikasi pada penyimpanan keratinosit diharapkan dapat menjadi melindungi fungsi sel.            Sampel kulit diperoleh dari preputium anak usia 4-9 tahun sebanyak 7 orang yang diperoleh dengan informed consent dari orang tua atau wali. Isolasi keratinosit menggunakan metode enzimatik dengan dispase dan trypsin/EDTA. Viabilitas dan proliferasi sel di ukur secara kalorimetrik dengan reagen WST-1 pada panjang gelombang 450 nm dan tehnik tryphan blue exclusion test. Data yang diperoleh diolah secara statistic dengan uji student t-test.            Kriopreservasi dengan tehnik vitrifikasi dapat mempertahankan viabilitas pasca thawing sebesar 80% tidak ada perbedaan bermakna dengan tehnik slow-freezing (p>0,05). Meski demikian hanya 30% dari sel tersebut dapat melakukan perlekatan. Hal ini jauh lebih rendah daripada tehnik slow-freezing yang dapat melakukan perlekatan hingga 70% (p<0,05). Fotomikrograph yang diambil pasca thawing menunjukkan keratinosit yang mengalami blebbing. Disfungsi sitoskeleton akibat syok hiperosmotik dapat menyebabkan cell blebbing.            Pembekuan sel dengan metode vitrifikasi mempengaruhi viabilitas, perlekatan dan kemampuan proliferasi sel dalam kultur. Syok hiperosmotik diperkirakan menyebabkan disfungsi sitoskeleton sehingga menjadi penyebab rendahnya kemampuan perlekatan dan hilangnya daya proliferasi pasca thawing yang dialami sel dengan perlakuan vitrifikasi. Penelitian selanjutnya dapat dilakukan dengan modifikasi komponen kriomedium yang dapat melindungi fungsi keratinosit.