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All Journal Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Berkala Ilmu Kesehatan Kulit dan Kelamin Indonesian Journal of Biotechnology Mutiara Medika: Jurnal Kedokteran dan Kesehatan The Indonesian Biomedical Journal Cermin Dunia Kedokteran JKKI : Jurnal Kedokteran dan Kesehatan Indonesia Indonesian Journal of Cancer Journal of General Procedural Dermatology and Venereology Indonesia Cermin Dunia Kedokteran
Yohanes Widodo Wirohadidjojo
Departemen Dermatologi Dan Venereologi, Fakultas Kedokteran, Keperawatan Dan Kesehatan Masyarakat, Universitas Gadjah Mada, Rumah Sakit Umum Pusat Dr. Sardjito, Yogyakarta, Indonesia
Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Public Health

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The effect of metformin on proliferation and glucose uptake in keloid fibroblast culture Nur Dwita Larasati, Sunardi Radiono Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 41, No 04 (2009)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Background: Metformin as an antihyperglycemic agent has a potential effect in increasing type I collagen synthesis and decreasing MMP, so that it has a potential to be an antiaging agent. One of aging failure processes is the development of keloids. Keloids are formed due to hyperproliferation of fibroblasts, an increase of collagen synthesis, particularly type I and III, and a decrease in MMP-1 and MMP-2. Fibroblast proliferation process and collagen synthesis need glucose uptake. The study on metformin ability to aggravate or stimulate the formation of keloid has never been conducted before. Objective: The aim of this study was to know the difference of proliferation and glucose uptake between keloid fibroblasts given metformin and without metformin. Method: A simple experiment was conducted using 3rdpassage keloid fibroblasts culture. Keloid fibroblasts were divided into 2 groups, the first group was treated with metformin in the dose of 100 pg/mL, 200 pg/mL, 300 pg/ mL, 400 pg/mL, and control. Keloid fibroblasts proliferation in the first group was measured using spectrophotometer with MTT assay, and glucose uptake of keloid fibroblast in the other group was measured using glucometer. The difference in proliferation and glucose uptake of keloid fibroblast was analyzed using one-way anova. Result: The result of this study showed that the average keloid fibroblast proliferation in the metformin treatment groups was not increased compared to that in control group. Meanwhile, the average keloid fibroblast glucose consumption in metformin treatment group significantlyincreased, at the dose of 300 ig/mL (p =0.044) and 400 I!g/mL (p = 0.0081. Conclusion: Metformin could not increase keloid fibroblasts proliferation, but it could increase glucose uptake of keloid fibroblasts.
The effect of vitamin C on fibroblast proliferation and VEGFexpression in fibroblast culture Munira, Sunardi Radiono Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 41, No 03 (2009)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Background: One of the factors that determine the success of efficient woul')d healing is wound healing rate, that can be achieved by increasingcell proliferation, angiogenesisor neovascularisation,and extracellular matrix production. Dermalfibroblast is a cell that plays an important role in wound healing. Fibroblast proliferation and neovascularisation are critical elements in granular tissue formation. Local hypoxia causes fibroblasts to express HIF-1a that will induce fibroblast VEGFexpression. The nature of vitamin C makes it easily oxidized. The addition of vitamin Con fibroblast culture medium is expected to produce local hypoxia condition that will induce fibroblast expression of HIF-1a, so that the expression of fibroblast VEGFwill be increased. Vitamin C may modulate the growth of various types of cells. The effect of vitamin Con normal fibroblast proliferation and fibroblast VEGFexpression is still unknown. Objective: This study was aimed to know whether vitamin Ccan increase normal human fibroblast proliferation and expression of VEGF. Method: A simple experimental study was conducted by using preputial skin fibroblast culture from 10-year-old donor, subculture passage 3. Fibroblast culture was divided into 6 groups, each group received vitamin C treatment with the dose of 50pg/mL, 100pg/mL, 150pg/mL, 200pg/mL, and 300pg/mL, and one group without treatment acting as control. Measurement of fibroblast proliferation was conducted by spectrophotometer using MTT, and fibroblast expression of VEGFwas measured using ELISA. The average of difference in fibroblast proliferation and VEGFexpression was analyzed with one-way analysis of variance. Result: There was a significant increase in fibroblast proliferation rate in the groups receiving vitamin C with the dose of 200 mg/mL (p = 0.016) and 300 mg/mL (p = 0.005), whereas in the group with the dose of 50 mg/mL, 100 mg/mL and 150 mg/mL there was no significant difference compared to the control (p = 0.933, p = 0.961, P = 0.301, respectively). Average fibroblast VEGFexpression between various concentrations of vitamin C compared to the control showed no significant difference (p > 0.05). Conclusion: Vitamin Ccould be considered to be used as an agent to accelerate wounds healing. Keywords: vitamin C, skin fibroblast culture, fibroblast proliferation, fibroblast VEGFexpression
The effect of topically extravirgin olive oil on the UVB-induced immunosuppression Vohanes Widodo Wirohadidjojo AriefBudiyanto, Irianiwati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 41, No 03 (2009)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Background: UVBradiation may act as an immunosuppressive agent through Langernans cells (LCs)depletion correlated with cyclobuthane pyrimidine dimer (CPD), as the most mutagenic photoproducts. Other studies showed that olive oil can prevent various human cancers, which are defect of immune-surveillance. The effect of olive oil in the UVBinduced LCs depletion is still unclear. Objective: To discover the topical effect of extravirgin-olive-oil in the LCs depletion. Methods: A simple experimental study was performed on foreskins collected from 8 circumcised boys. Each of them was cut into 3 pieces of 0.5 cm2, grouped randomly to: (Olive oil+ UVB)group, (placebo + UVB)group, and unirradiated group. They were cultured in DMEM for 24 hours, irradiated with 100mJs/cm2 UVB, and olive oil or placebo were then applied immediately after irradiation. All specimens were fixed in buffered-formalin, paraffin blocked, and cut to be 2 slides, and each of them was stained with anti-CD 1a or anti-CPD antibody. The pixels of AEC signats were digitally counted based on Photoshop's selected color histogram.The pixel means of various groups were analyzed by Kruskal-Wallis test and post hoc analysis by Mann Whitney test. Results: Olive oil treated groups showed a very significant (P
The combination effect of triamcinolone acetonide and tamoxifen citrate on fibroblast populated collagen lattice contractions Agung Pranoto; Satiti Retno Pudjiati; Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 02 (2008)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Background: Keloid is caused by fibroblast hyperproliferation stimulated by transforming growth factor-IH ITGF-131 I, and it is usually treated with triamcinolone acetonide (TAl, which has the ability to inhibit TGF131 synthesis. However, the clinical results is still unsatisfied. Another drug that may inhibit keloid fibroblast TGF-131 synthesis is tamoxifen citrate (TCI, but the effect of the combination on keloid fibroblast activities has never been published.Objective: To find out the effect of combined triamcinolone acetonide and tamoxifen citrate on fibroblast keloid activities in vitro.Methods: It was a parallel post-test only study. The third passage keloid fibroblasts were isolated from a patient with keloid, cultivated in collagen lattice, and treated with several combinations of 5, 10, and 20 pM TA and 10, and 20 pM TC. Lattice contractions were measured based on digital image using scion image.Results: Among TA groups, the best inhibition of lattice contraction was found among 20 pM treated group and among TC groups. The best inhibition of lattice contraction was found among 20 pM TC. The best combination was found in the combination of 20 pM TA plus 20 pM TC.Conclusion: The result indicated that a combination of triamcinolone acetonide and tamoxifen citrate had a significant role in suppressing fibroblast activity, better than triamcinolone acetonid or tamoxifen citrate alone.Key words: tamoxifen - triamcinolone - collagen lattice - keloid fibroblast.
The Combination of suprakeloidal flap and pulsed light heat energy in keloid management: a Case report Yohanes Widodo Wirohadidjojo; Kristiana Etnawati; Dwi Retno Adiwinarni
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 01 (2008)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

The role of chronic tissue hypoxia in the keloid patho-mechanism has been widely accepted. Whereas, the pulsed-light heat energy (LHE) has been developed which has the capacity to generate reactive oxygen species on exposed skin. Although the supra keloidal flap technique has a high recurrence rate, it was used because of its capacity to prevent suturing hypoxia, thereby the formation of lager recurrent keloid after surgery.The combination of supra keloidal flap and pulsed light heat energy was done I the treatment of postvaricella keloid on the right ear lobe of a 9 year old girl. The keloid was excised two times a year ago, but observation one month after the surgery showed a recurrent larger keloid. The performance of supra keloidal technique followed by pulsed-light heat energy treatment in dose 2.5 J/cm2, was administered on day 3r
The Effect of Topical Extravirgin Olive Oil on Ultraviolet B-Induced 8-Hydroxy-2'-Deoxyguanosine Formation in Human Skin Catharina Sagita Yohanes Widodo Wirohadidjojo Arief Budiyanto, Irianiwati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 39, No 02 (2007)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Background: Ultraviolet B (UVBI may cause DNA damage due to photooxydative reaction and form 8hydroxy-2'-deoxyguanosine (8-0HdGI as a genotoxic product. Topically extra virgin olive oil (EVOOI can decrease 8-0HdG formation among UVB irradiated-hairless mice epidermal cells. Such effect in human skin has never been discovered.Objective: To know the effect of topically EVOO in UVB induced-8-0HdG formation in human epidermal cells.Method: A simple experiment was performed on cultured human foreskins. The foreskins were divided into 3 groups: group was treated with 100mJ/cm2 UVB + topical EVOO, group B with 100mJ/cm2 UVB, and group C as control. All skin was cultured in completed DMEM and fixation was performed at one and 24 hours. 8-0HdG expression was determined immunohistochemically with antibody anti 8-0HdG and measured based on histogram of selected colour of Adobe photoshop.Results: The mean of 8-0HdG expression in experimental groups: (8.02:t 13.81, (26.95:t 23.231. and 14.96:t 7.141, respectively (p =0.0001 at an hour; (14.91 :t30.63I, (37.58:t 25.631. and (8.76:t 10.941 respectively (p = 0.0001 at 24 hours.Conclusion: Extra virgin olive oil was proved to reduce the 8-0HdG formation in exposed human skin-UVB. and may be developed to be anti-photocarcinogenesis active ingredient topically.Kata kunci:Extra virgin olive oil, UVB, 8-OHdG, human Skin
The Effect of Narrow and Broad Band Ultraviolet B Onto Keloid Fibroblast-VEGF Expressions Yohanes Widodo Wirohadidjojo Ishandono Dahlan
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 39, No 02 (2007)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Background: Collagenase inactivation of keloid lesions is due to plasminogen activator inhibitors which are synthesized under stimulation of vascular endothelial growth factor (VEGF), a released protein under hypoxia conditions. On the other hand, ultraviolet B (UVB) may generate various oxidative molecules of irradiated chromophores. The effect of UVB in VEGF synthesis is unclear.ObJective. To know the effect of narrow and broad band ultraviolet-B on keloid fibroblast-VEGF expression. Materials and methods: Materials in this study were keloid materials collected from dermatosurgery and plastic surgery keloid revision. A parallel simple experiment study was performed to compare the effect of 0, 50, 75, and 100mJ/cm2 of broad band (BBUVB) as well as narrow band UVB (NBUVB) in VEGF synthesis of passage-3 keloid fibroblasts isolated from 4 patients. Samples were stained with monoclonal antibody anti VEGF. The selected DAB-brown colors of cytoplasm were computed based on Photoshop software histogram.Results: Compared to untreated group, all of various NBUVB showed a very significant (P0.05) between NBUVB and BBUVB groups.Conclusion: NBUVB as well as BBUVB can suppress VEGF synthesis among irradiated keloid fibroblasts. The 50 mJ/cm2 of NB UVB as well as 75 mJ/cm2 of BBUVB may be developed as the modality in keloid prevention or treatment.Keywords: keloid-fibroblasts, broad-band UVB, narrow-band UVB, VEGF
Multiple mini punch grafts for extensive ulcer: a case report Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 44, No 02 (2012)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Multiple mini punch grafts is the placing of mini size of full thickness skins on to ulcer bed. Theyconsist of epidermal and dermal component composed with hair follicles and other skin appendiceswhere epidermal stem cells are located. The epidermal stem cells are the best source of epidermalcells in reconstruction of skin equivalent that is usually used for replacing classic split thicknessskin graft in recovering extensive ulcer. In this article, the application of multiple mini punchgrafts onto extensive ulcer is reported. A case of extensive ulcer was suffered by a 6-year-oldboy whose left foot is injured in a traffic accident. His toes had already been amputated bysurgeon but a classic skin graft failed to recover the ulcer. Multiple mini punch grafts had beenharvested from his inguinal and buttock skin and they were placed onto his ulcer. Pre and postmini punch grafting photographs were reviewed. After eight weeks, placed multiple mini punchtissues onto large ulcer reveals lateral extensions and more than 90% of epithelialization. Multiplemini punch grafts can be used as a method to cover large ulcer.Key words: mini punch grafts-large ulcer-epithelialization-epidermal-stem cells
Collagen synthesis on ultraviolet A irradiated human skin fibroblast treated with insulin Febrina Rismauli Panggabean Satiti Retno Pudjiati Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 43, No 01 (2011)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Ultraviolet (UV) irradiation from the sun can stimulate premature skin aging because UV irradiation inhibits collagensynthesis, promotes collagen degradation and inhibits fibroblast proliferation. Insulin is capable to stimulate fibroblastgenes collagen expression, DNA synthesis, and collagen synthesis. The effect of insulin in reducing collagen synthesisamong repeated-UVA irradiation on human skin fibroblast has never been studied. This study aims to investigate theeffect of insulin in collagen synthesis among repeated-UVA irradiation on normal human skin fibroblast. To asses thecollagen synthesis collagen degradation, collagen deposition and fibroblast proliferationweremeasured. Experimentalstudy was performed among passage 3 of fibroblast which was isolated from a circumcised skin of a 6-year-old boy.Fibroblasts were irradiated with 3 repeated exposurewith total cumulative dose 9000 mJ/cm2 and treated withinsulin 0.5; 1; 2 μg/mL and placebo. Cellswere then incubated for 48 hours, collagen degradation, collagen depositionand fibroblast proliferation were read colorimetric by using Spectroscopy 550 nm. The effect of insulin 0.5; 1 and2 ìg/mL in collagen synthesis among repeated-UVA irradiation on normal human skin fibroblast with cumulative dose9000 mJ/cm2 was not capable to reduce collagen degradation, nor capable to increase collagen and fibroblastproliferation. Insulin dose 0,5 μg/ml-2 μg/ml among repeated-UVA irradiation on normal human skin fibroblast wasnot capable to increase collagen synthesis.Key words: photoaging-DNA synthesis-proliferasion-aging process-gene expression
α-Lipoic acid can not prevent effet on cell viability, collagen synthesis inhibition, collagen degradation induction in ultraviolet A irradiated human fibroblast cell Putu Dyah Ayu Saraswati Soedirman Sastrodiprodjo Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 43, No 02 (2011)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Ultraviolet A (UVA) irradiation on human skin can generate free radical and stimulate matrix metalloproteinaseproduction resulting in collagen degradation and transforming growth factor beta (TGF-β) inhibition. It causes synthesiscollagen inhibition and induces cell death. α-Lipoic acid (ALA) is an universal antioxidant and a powerful scavengerof free radicals. In this study we investigated the effect of ALA on viability, collagen synthesis and degradation inUVA irradiated human fibroblasts. Normal human skin fibroblasts cell culture were irradiated with UVA for threetimes with each dose of 3000 mJ/cm2 UVA. α-Lipoic acid in various concentration was added to the culturefollowing UVA irradiation and incubated for 48 hours. The cell viability was determined by MTT-assay while collagensynthesis and degradation were determined by Sirius red binding assay. The difference of cell viability and collagensynthesis and degradation between fibroblasts cell after and without UVA irradiation were analyzed using paired-ttest with 95% confidence interval (p<0.05). The results showed that UVA irradiation decreased cell viability,inhibited collagen synthesis and induced collagen degradation in fibroblasts cell. However, ALA was not sufficient toincrease viability, to increase collagen synthesis and to inhibit collagen degradation in fibroblasts cell due to UVAirradiation. In conclusion, ALA can not prevent UVA irradiation effect on human skin fibroblasts cell.Key words: UVA – irradiation - human skin fibroblasts – antioxidants – α-lipoic acid
Co-Authors . Suswardana Agnes Sri Siswati AGUNG PRANOTO Andayani, Raden Roro Rini Anggatama, Marcella Anggreana, Farisa Arief Budiyanto Budiyanto, Arief Budiyanto Christiana Tri Nuryana Dwi Retno Adiwinarni Endra Yustin E. S Faber, William R Fajar Waskito Fitriani, Katia Flandiana Yogianti Graciella Regina Hady Anshory Hanggoro Tri Rinonce Hardyanto Soebono Ishandono Dachlan Jesslyn Amelia Kartika Kemala Kristiana Etnawati Listya Paramita Mae Sri Hartati Wahyuningsih Maria Vianny Sansan Muhamad Eko Irawanto Niken Indrastuti Nita Damayanti Sulistianingrum Nur Arfian Oktarina, Dyah Ayu Mira Paramita, Listya Prasta Bayu Putra Radiono, Sunard Regina, Graciella Rosmelia Rosmelia Satiti Retno Pudjiati Satiti Retno Pudjiati Shelly, Wan Gifanni Efmadian Sofia Mubarika Haryana Suryaningsih, Betty Ekawati Teguh Aryandono Tiara Puspita Agustin Tuntas Rayinda Wening Setyani William Faber Winarni, Dwi Retno Adi Yefta