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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

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2013.v32.3 - 10

Abstract

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2014.v33.91-99

Abstract

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2117.621 KB)

Abstract

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (363.168 KB) | DOI: 10.33476/jky.v25i2.118

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.
PENGARUH PLATELET-RICH PLASMA (PRP) TERHADAP PROLIFERASI DAN VIABILITAS HUMAN DERMAL FIBROBLAST (HDF) DALAM KONSENTRASI GLUKOSA TINGGI Hadi, Restu Syamsul; Kusumah, Indra; Sandra, Yurika
JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i2.3815

Abstract

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
Perilaku Kader dalam Penemuan Suspek Tuberkulosis Fadhilah, Nur; Nuryati, Elmi; Duarsa, Artha; Djannatun, Titiek; Hadi, Restu Syamsul
Jurnal Kesehatan Masyarakat Nasional Vol. 8 No. 6 Januari 2014
Publisher : Faculty of Public Health Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (253.363 KB) | DOI: 10.21109/kesmas.v0i0.381

Abstract

Dukungan berbagai pihak meliputi perubahan perilaku masyarakat dan pemberdayakan masyarakat sangat diharapkan untuk penanggulangan tuberkulosis (TB). Pimpinan Pusat Muhammadiyah dan ?Aisyiyah Provinsi Lampung terpanggil untuk bergerak bersama dalam program penanggulangan penyakit TB agar keberhasilan penanggulangan TB dapat tercapai. Penelitian ini bertujuan mengetahui berbagai faktor yang berhubungan dengan perilaku kader dalam menemuan suspek TB di Kabupaten Lampung Tengah. Penelitian dengan metode kuantitatif dan kualitatif ini menggunakan desain potong lintang, data primer dikumpulkan dari sampel 72 kader TB ?Aisyiyah Kabupaten Lampung Tengah. Analisis dilakukan secara univariat, bivariat dengan menggunakan metode kai kuadrat, dan multivariat dengan regresi logistik. Penelitian ini menemukan lima variabel yang meliputi pengetahuan, sikap, pelatihan, dukungan pemegang program dan motivasi yang mendukung perilaku penemuan suspek. Tiga variabel yang meliputi pendidikan, pendapatan dan pekerjaan tidak mendukung perilaku penemuan suspek. Untuk meningkatkan penemuan suspek TB disarankan untuk lebih meningkat dukungan pengelola program yang berkelanjutan.The support of various parties, peoples behavior and empower communities in the implementation of TB countermeasures highly expected by Muhammadiyah Central Executive and Aisyiyah Lampung Province omit to move together in a tuberculosis prevention program for successful TB control can be achieved. This study aimed to determine the related factors of behavior cadres to detect suspected tuberculosis in Lampung districy middle. The quantitative and qualitative with study design a cross sectional was conducted using primary data on samples 72 Aisyiyah tuberculosis cadres Lampung district middle. The statistical analyses were performed by chisquare and logistic regression. The study results showed a significant five variable (support program managers, knowledge of cadre, motivation of cadre, attitude of cadre, training cadre) with the discovery suspected tuberculosis cases in Lampung Province. Logistic regression analysis found a good support program holders associated with the case of suspected tuberculosis. Program holders support is the most dominant factor of the discovery of suspected tuberculosis cases. Therefore the need for tangible support over again that the findings by the cadre suspected tuberculosis increased.
PENGARUH PLATELET-RICH PLASMA (PRP) TERHADAP PROLIFERASI DAN VIABILITAS HUMAN DERMAL FIBROBLAST (HDF) DALAM KONSENTRASI GLUKOSA TINGGI Hadi, Restu Syamsul; Kusumah, Indra; Sandra, Yurika
JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i2.3815

Abstract

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
EVALUASI KEMAMPUAN PROLIFERASI, DIFERENSIASI DAN VIABILITAS SEL PUNCA ASAL PULPA GIGI MANUSIA PASCA VITRIFIKASI Hadi, Restu Syamsul; Kusuma, Indra
Majalah Kesehatan Pharmamedika Vol 11, No 1 (2019): JUNI 2019
Publisher : Lembaga Penelitian Universitas YARSI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33476/mkp.v11i1.951

Abstract

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.