Yohanes Widodo Wirohadidjojo
Departemen Dermatologi Dan Venereologi, Fakultas Kedokteran, Keperawatan Dan Kesehatan Masyarakat, Universitas Gadjah Mada, Rumah Sakit Umum Pusat Dr. Sardjito, Yogyakarta, Indonesia

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The effect of metformin on proliferation and glucose uptake in keloid fibroblast culture Yohanes Widodo Wirohadidjojo, Nur Dwita Larasati, Sunardi Radiono
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.
Multiple mini punch grafts for extensive ulcer: a case report Wirohadidjojo, Yohanes Widodo
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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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
The combination effect of triamcinolone acetonide and tamoxifen citrate on fibroblast populated collagen lattice contractions Yohanes Widodo Wirohadidjojo, Agung Pranoto Satiti RetnoPudjiati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 02 (2008)
Publisher : Universitas Gadjah Mada

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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.
Collagen synthesis on ultraviolet A irradiated human skin fibroblast treated with insulin Yohanes Widodo Wirohadidjojo, Febrina Rismauli Panggabean Satiti Retno Pudjiati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 43, No 01 (2011)
Publisher : Universitas Gadjah Mada

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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
The effect of vitamin C on fibroblast proliferation and VEGFexpression in fibroblast culture Yohanes Widodo Wirohadidjojo, Munira, Sunardi Radiono
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 41, No 03 (2009)
Publisher : Universitas Gadjah Mada

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Abstract

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 Combination of suprakeloidal flap and pulsed light heat energy in keloid management: a Case report Dwi retno Adiwinarni, Yohanes Widodo Wirohadidjojo Kristiana Etnawati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 01 (2008)
Publisher : Universitas Gadjah Mada

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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 Narrow and Broad Band Ultraviolet B Onto Keloid Fibroblast-VEGF Expressions Ishandono Dahlan, Yohanes Widodo Wirohadidjojo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 39, No 02 (2007)
Publisher : Universitas Gadjah Mada

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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
α-Lipoic acid can not prevent effet on cell viability, collagen synthesis inhibition, collagen degradation induction in ultraviolet A irradiated human fibroblast cell Yohanes Widodo Wirohadidjojo, Putu Dyah Ayu Saraswati Soedirman Sastrodiprodjo
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 43, No 02 (2011)
Publisher : Universitas Gadjah Mada

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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
α-Lipoic acid inhibit the decrease of collagen deposition in ultravioled B-irradiated cultured normal human skin fibroblasts cell culture Yohanes Widodo Wirohadidjojo, Arum Krismi Satiti Retno Pudjiati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 43, No 02 (2011)
Publisher : Universitas Gadjah Mada

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Repeated ultraviolet B (UVB) irradiation on human skin has been considered to be responsible in premature agingprocess because UVB has been proved to inhibit collagen deposition and accelerates collagen degradation. Clinicalstudies showed that topical usage of 5% α-lipoic acid (ALA) improved the clinical appearance of photoaged skin.However, the effect of ALA on collagen deposition and degradation in UVB-irradiated normal human skin fibroblastsculture has not been reported. The aim of the study was to investigate the effect of ALA on collagen deposition anddegradation in UVB-irradiated cultured normal human skin fibroblasts. Culture of normal human skin fibroblasts weretreated with 0, 125, 250, 500 μM ALA diluted in complete Dulbecco’s Modified Eagle’s Medium (DMEM) andirradiated with 300 mJ/cm2 UVB. The mean collagen deposition and degradation’s level were measured by Siriusred assay and read with spectrophotometer at λ 550 nm. Mean difference of collagen deposition as expressed byoptical density (OD) between normal human skin fibroblasts cell after UVB irradiation and without UVB irradiationwas analyzed by Wilcoxon signed-ranks test and Friedman test, while mean difference collagen degradation wasanalyzed by one way analysis of variance (ANOVA) and paired t test with 95% confidence level (p<0.05). Theresults showed that ALA 125 μM inhibited the decrease of collagen deposition significantly (p<0.05), though higherconcentrations did not. However, ALA did not inhibit collagen degradation increment (p>0.05). In conclusion, ALAinhibited the decrease of collagen deposition, but did not inhibit collagen degradation in UVB-irradiated normalhuman skin fibroblasts culture.Key words: α-lipoic acid - collagen - human skin - fibroblasts – UVB - irradiation
Papuloerythroderma of Ofuji: A first case report from Indonesia Graciella Regina; Listya Paramita; Sunardi Radiono; Yohannes Widodo Wirohadidjojo; William Faber
Journal of General - Procedural Dermatology and Venereology Indonesia Vol 1, No 3 (2016): December
Publisher : Universitas Indonesia

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Papuloerythroderma (PE) is a rare skin disease which was first described by Ofuji et al. in 1984, with a typical sign that the lesions spare the large cutaneous folds, known as the deck chair sign. Due to its recent identification, this disease is still underrecognized and may lead to misdiagnosis. We reported the first case report of PE of Ofuji from Indonesia in which the diagnosis was delayed for two years. Besides the deck chair sign in the large cutaneous fold, we also found that the area between and above his eyebrows that was relatively spared in contrast to the sparing of the cutaneous folds, and it may be considered as pseudo-deck chair sign. The patient showed good response with combination therapy of phototherapy with Narrow-Band Ultraviolet B (NBUVB), oral methotrexate, and corticosteroids. The deck chair sign disappeared after six months therapy, but the patient’s skin was still xerotic. Keywords: Papuloerythroderma of Ofuji, deck chair sign, pseudo-deck chair sign