Abstract
Keloid is a skin lesion caused by an abnormal proliferation of fibroblasts and the deposition of collagen in the wound healing process. Therapy for keloids is relatively limited and mostly has side effects. Recent studies show that 5α-oleandrin from N. indicum Mill. (N. indicum) has anti-keloid effect by inhibiting (proliferation, migration activity and TGF-β1 expression) keloid fibroblast. Further studies with N. indicum extract standardized 5α-oleandrin should be conducted to develop it as an anti-keloid agent by performing at proliferative effects, collagen accumulation, and keloid fibroblast migration. The antifibrotic activity of N. indicum extract on the proliferation of keloid fibroblasts was measured by MTT assay, whereas collagen deposits were observed by Sirius Red method. Observation of keloid fibroblast migration by scratch assay is according to Liang et al. (2009). Ethanol extract of N. indicum standardized 5α-oleandrin can inhibit the proliferation of keloid fibroblas with IC50 0.458 μg/mL, also inhibited collagen deposits with IC50 0.055 μg/mL at incubation for 72 hours. In incubation 48 hours after treatment with ethanol extract of 1.0 μg/mL occurred inhibition of migration significantly compared with control.
Highlights
a skin lesion caused by an abnormal proliferation of fibroblasts
Recent studies show that 5α-oleandrin from N. indicum Mill
Further studies with N. indicum extract standardized 5α-oleandrin should be conducted to develop it as an anti-keloid agent
Summary
Pembuatan kadar uji ekstrak etanol N. indicum Ekstrak etanol N. indicum Mill sebanyak 5 mg dilarutkan dengan 100 μL DMSO sehingga didapatkan konsentrasi larutan stok 50.000 μg/mL. Kemudian dibuat 6 serial kadar (0.3125; 0.625; 1.25; 2.5; 5 dan 10) μg/mL. Perlakuan uji ekstrak etanol N. indicum Mill Sejumlah 2x105sel fibroblas keloid/. Sumuran dimasukkan kedalam 96 well plate diinkubasi dalam inkubator CO2 5%, suhu 37oC selama 24 jam. Setelah inkubasi 24 jam, semua media pada 96 well plate dibuang dan diganti dengan ekstrak N. indicum dengan serial kadar (0.3125; 0.625; 1.25; 2.5; 5 dan 10) μg/mL, yang masing-masing seri kadar ekstrak diberikan rangkap tiga (triplikasi). Sel Fibroblas keloid didalam 96 well plate diinkubasi selama tiga hari pada inkubator CO2 dengan suhu 37oC. Dilakukan pengukuran hambatan proliferasi fibroblas keloid dengan MTT Assay
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