Articles
<![CDATA[Gangguan fungsi sitoskeleton pada proses vitrifikasi keratinosit primer manusia ]]>
<![CDATA[Kusuma, Indra]]>;
<![CDATA[Hadi, Restu Syamsul]]>;
<![CDATA[Sandra, Yurika]]>
<![CDATA[ Jurnal Kedokteran YARSI Vol 25, No 2 (2017): MEI - AGUSTUS 2017 ]]>
Publisher : <![CDATA[Lembaga Penelitian Universitas YARSI ]]>
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DOI: 10.33476/jky.v25i2.118
<![CDATA[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.]]>
<![CDATA[Pengaruh Madu terhadap Migrasi dan Diferensiasi Sel Human Dermal Fibroblast (HDF ) sebagai Model Uji Luka In Vitro ]]>
<![CDATA[Rahmah Aprilia, Yoan]]>;
<![CDATA[Nadira, Nadira]]>;
<![CDATA[Syamsul Hadi, Restu]]>
<![CDATA[ Majalah Kesehatan Pharmamedika Vol 10, No 2 (2018): DESEMBER 2018 ]]>
Publisher : <![CDATA[Lembaga Penelitian Universitas YARSI ]]>
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DOI: 10.33476/mkp.v10i2.725
<![CDATA[Penelitian ini bertujuan mengetahui pengaruh madu terhadap kemampuan migrasi dandiferensiasi sel HDF sebagai model luka in vitro. Sel Human Dermal Fibroblast(HDF)ditanam dalam cawan kultur 6 sumuran untuk uji migrasi dengan scratch assay dan 24 sumuran untuk uji diferensiasi. Sel HDF untuk uji migrasi diberi madu dengan konsentrasi bervariasi selanjutnya diinkubasi selama 18, 44, dan 90 jam. Kecepatan penutupan model luka dihitung dengan prosentase. Untuk uji diferensiasi, sel HDF yang ditanam pada plate 24 dibagi menjadi 6 perlakuan yaitu kontrol tanpa serum dan medium adipogenesis (K1), kontrol serum dan medium adipogenesis (K2),. Setelah konfluens + 70%, diberikan perlakuan yang berisi medium tanpa serum dan madu dosis 0,5% (K3), 1% (K4), 2% (K5), dan 4% (K6) diinkubasi hingga hari ke-7. Kemudian sumuran yang berisi medium madu pada hari ke-8 diganti dengan medium adipogenesis. Pengamatan dilakukan pada hari ke-14, ke-21 dan dilanjutkan dengan pewarnaan Oil Red-O. Hasil penelitian menunjukkan pemberian madu dosis 1% mempercepat migrasi fibroblast sehingga mempengaruhi keberhasilan penyembuhan luka. Pemberian madu juga menyebabkan peningkatan jumlah diferensiasi sel HDF menjadi sel adiposit secara signifikan (p0,05) pada dosis madu 1%. Pemberian suplementasi madu dosis 1% dapat meningkatkan kemampuan migrasi dan diferensiasi sel HDF pada model luka in vitro.]]>
<![CDATA[PENGARUH PLATELET-RICH PLASMA (PRP) TERHADAP PROLIFERASI DAN VIABILITAS HUMAN DERMAL FIBROBLAST (HDF) DALAM KONSENTRASI GLUKOSA TINGGI ]]>
<![CDATA[Hadi, Restu Syamsul]]>;
<![CDATA[Kusumah, Indra]]>;
<![CDATA[Sandra, Yurika]]>
<![CDATA[ JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA ]]>
Publisher : <![CDATA[Perhimpunan Biologi Indonesia ]]>
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DOI: 10.14203/jbi.v15i2.3815
<![CDATA[ABSTRACTThe administration of Platelet Rich Plasma (PRP) is expected to be a supplement for treatment of diabetic wounds and hyperglycemia by increasing growth factors. The purpose of this study was to examine the effect of Platelet Rich Plasma (PRP) on the proliferation and viability of human dermal fibroblast (HDF) in high glucose conditions, as a model for healing diabetic wounds in vitro. HDF cells are grown in DMEM medium containing high glucose which are then with PRP. To measure the effect of PRP on the HDF cell proliferation, CCK-8 kit was used and evaluated by using a microplate reader. To evaluate the viability of HDF, an automated cell counter. The results of the research showed that PRP stimulate the HDF cells proliferation. The optimal dose of PRP to increase HDF cell proliferation is at dose of PRP 10%. Supplementation of PRP is significantly increased cell viability and HDF cell counts within 48 hours. The results showed PDGF growth factor secreted by PRP is increased significantly. The conclusion is PRP stimulated HDF cell proliferation and viability in a high glucose condition. This finding support the used of PRP as a therapy for diabetic wounds. Keywords: proliferation, viability, human dermal fibroblast, platelet-rich plasma, diabetic]]>
<![CDATA[PENGARUH PLATELET-RICH PLASMA (PRP) TERHADAP PROLIFERASI DAN VIABILITAS HUMAN DERMAL FIBROBLAST (HDF) DALAM KONSENTRASI GLUKOSA TINGGI ]]>
<![CDATA[Hadi, Restu Syamsul]]>;
<![CDATA[Kusumah, Indra]]>;
<![CDATA[Sandra, Yurika]]>
<![CDATA[ JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA ]]>
Publisher : <![CDATA[Perhimpunan Biologi Indonesia ]]>
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DOI: 10.14203/jbi.v15i2.3815
<![CDATA[ABSTRACTThe administration of Platelet Rich Plasma (PRP) is expected to be a supplement for treatment of diabetic wounds and hyperglycemia by increasing growth factors. The purpose of this study was to examine the effect of Platelet Rich Plasma (PRP) on the proliferation and viability of human dermal fibroblast (HDF) in high glucose conditions, as a model for healing diabetic wounds in vitro. HDF cells are grown in DMEM medium containing high glucose which are then with PRP. To measure the effect of PRP on the HDF cell proliferation, CCK-8 kit was used and evaluated by using a microplate reader. To evaluate the viability of HDF, an automated cell counter. The results of the research showed that PRP stimulate the HDF cells proliferation. The optimal dose of PRP to increase HDF cell proliferation is at dose of PRP 10%. Supplementation of PRP is significantly increased cell viability and HDF cell counts within 48 hours. The results showed PDGF growth factor secreted by PRP is increased significantly. The conclusion is PRP stimulated HDF cell proliferation and viability in a high glucose condition. This finding support the used of PRP as a therapy for diabetic wounds. Keywords: proliferation, viability, human dermal fibroblast, platelet-rich plasma, diabetic]]>
<![CDATA[Evaluasi Kemampuan Proliferasi, Diferensiasi dan Viabilitas Sel Punca Asal Pulpa Gigi Manusia Pasca Vitrifikasi ]]>
<![CDATA[Hadi, Restu Syamsul]]>;
<![CDATA[Kusuma, Indra]]>
<![CDATA[ Majalah Kesehatan Pharmamedika Vol 11, No 1 (2019): JUNI 2019 ]]>
Publisher : <![CDATA[Lembaga Penelitian Universitas YARSI ]]>
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DOI: 10.33476/mkp.v11i1.951
<![CDATA[Sejumlah keterbatasan sumber sel pulpa gigi manusia diantaranya harus dari pulpa gigi sehat dengan jumlah sangat terbatas. Teknologi penyimpanan sel dan pengembangan metode vitrifikasi pada DPSC dan MSC sangat bermanfaat untuk keperluan terapi. Penelitian ini bertujuan mengkaji pertumbuhan, proliferasi dan diferensiasi serta pengaruh simpan beku metode vitrifikasi terhadap sel punca asal pulpa gigi. Sampel yang digunakan adalah pulpa gigi yang diisolasi. Untuk mengukur proliferasinya digunakan Cell Proliferation Reagent WST-1 Absorbansi diukur menggunakan mikroplate ELISA Zenyth pada panjang gelombang 450 nm. Untuk proses simpan beku dengan metode vitrifikasi sel MSC dipanen dan di-cryopreservasi dengan dua langkah yaitu diberikan dengan Equilibration (EQ) dan vitrifikasi solutions (VS). Untuk diferensiasi sel adipogenik digunakan StemPro Adipogenesis dan diuji dengan pewarnaan Oil Red O. DPSC berhasil diisolasi dan tumbuh dengan baik pada medium alfa MEM dengan serum 10%. Kemampuan proliferasi sel DPSC pada inkubasi 24 jam lebih tinggi secara signifikan dibandingkan inkubasi 18 jam. DPSC dapat terdiferensiasi menjadi neural like cells dan pada sel MSC dengan pemberian differentiation Kit StemPro Adipogenesis, diferensiasi MSC ke arah sel adipogenik setelah pewarnaan menggunakan Oil Red O. Teknik vitrifikasi sebagai metode simpan beku pada MSC menunjukan viabilitas yang tinggi ( 80%) dan sel mampu berproliferasi dan berdiferensiasi dengan baik.]]>
<![CDATA[Pengaruh Glukosa Tinggi terhadap Proliferasi, Migrasi dan Ekspresi Gen OCT-4 pada Kultur Sel Dermal Fibroblast Manusia ]]>
<![CDATA[Hadi, Restu Syamsul]]>;
<![CDATA[Sandra, Yurika]]>
<![CDATA[ Majalah Kesehatan Pharmamedika Vol 12, No 1 (2020): JUNI 2020 ]]>
Publisher : <![CDATA[Lembaga Penelitian Universitas YARSI ]]>
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DOI: 10.33476/mkp.v12i1.1604
<![CDATA[Human dermal fibroblasts (HDF) termasuk sel mesenchymal yang diisolasi dari lapisan dermis kulit. HDF berpotensi digunakan untuk pengobatan penyembuhan luka berdasarkan pengobatan regeneratif. Peningkatan konsentrasi glukosa dapat merusak fungsi sel dan menghambat terapi penyembuhan luka. Penelitian ini dilakukan untuk mengkaji pengaruh glukosa tinggi pada proliferasi, migrasi dan ekspresi gen OCT-4 sel HDF sebagai model penyembuhan luka diabetes in vitro. Pada penelitian eksperimental ini, fibroblast diisolasi dari kulit setelah sirkumsisi, kemudian ditumbuhkan dalam medium Minimal Essential Medium (DMEM) Dulbecco lengkap dengan serum 10%. Untuk menguji efek glukosa tinggi pada proliferasi sel HDF dilakukan dengan uji CCK-8. Migrasi sel HDF dievaluasi menggunakan uji scratch-assay. RT-PCR digunakan untuk menentukan ekspresi gen OCT-4. Hasilnya menunjukkan bahwa glukosa tinggi (25 mM-50 mM) meningkatkan kemampuan proliferasi dan migrasi sel HDF. Efek glukosa tinggi tergantung pada dosis. Perlakuan glukosa pada dosis 75 mM akan menghambat kemampuan pertumbuhan HDF. Ekspresi gen OCT-4 meningkat secara bermakna pada pemberian glukosa dosis 25-50 mM. Hasil penelitian ini memberikan dasar untuk pengembangan terapi dalam kondisi diabetes bahwa terapi sel HDF dapat meningkatkan penyembuhan luka meskipun dalam kondisi glukosa tinggi.]]>
<![CDATA[Mekanisme Apoptosis Pada Regresi Sel Luteal ]]>
<![CDATA[Syamsul Hadi, Restu]]>
<![CDATA[ Majalah Kesehatan Pharmamedika Vol 3, No 1 (2011): JANUARI - JUNI 2011 ]]>
Publisher : <![CDATA[Lembaga Penelitian Universitas YARSI ]]>
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DOI: 10.33476/mkp.v3i1.442
<![CDATA[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.]]>
<![CDATA[Apoptosis Pada Sperma Sebagai Petanda Adanya Gangguan Kesuburan Pria ]]>
<![CDATA[Syamsul Hadi, Restu]]>
<![CDATA[ Majalah Kesehatan Pharmamedika Vol 3, No 2 (2011): JULI - DESEMBER 2011 ]]>
Publisher : <![CDATA[Lembaga Penelitian Universitas YARSI ]]>
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DOI: 10.33476/mkp.v3i2.449
<![CDATA[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 apoptotic 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.]]>
<![CDATA[Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture ]]>
<![CDATA[Restu Syamsul Hadi]]>;
<![CDATA[Indra Kusuma]]>;
<![CDATA[Yurika Sandra]]>
<![CDATA[ Universa Medicina Vol. 33 No. 2 (2014) ]]>
Publisher : <![CDATA[Faculty of Medicine, Universitas Trisakti ]]>
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DOI: 10.18051/UnivMed.2014.v33.91-99
<![CDATA[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.]]>
<![CDATA[Geraniin supplementation increases human keratinocyte proliferation in serum-free culture ]]>
<![CDATA[Indra Kusuma]]>;
<![CDATA[Restu Syamsul Hadi]]>
<![CDATA[ Universa Medicina Vol. 32 No. 1 (2013) ]]>
Publisher : <![CDATA[Faculty of Medicine, Universitas Trisakti ]]>
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DOI: 10.18051/UnivMed.2013.v32.3 - 10
<![CDATA[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.]]>
