| dc.description.abstract | Partial hydrolysis of Virgin Coconut Oil (VCO) produced free fatty acids and 2-monoglycerides especially lauric acid and 2-monolaurin which are active as antibacterial, antiviral and wound healing agents. Chitosan could accelerate fibroblast formation thereby increasing the wound healing process. This study aimed to determine the activity of HVCO, chitosan and the combination of HVCO and chitosan toward wound healing parameters in vitro on NIH 3T3 cells.
NIH 3T3 cells were incubated for 24 hours and treated with various concentration series of test materials and positive control (Aloclair®) starting concentrations of 15.625 μg/ml to 500 μg/ml. Each series of concentrations was incubated for 24 hours and cell proliferation activity was performed using the MTT method [3- (4,5-dimethylthiazol-2-il) -2,5-diphenyl tetrazolium bromide]. Proliferation activity of 3T3 NIH cells was measured by calculating cell viability after 24 hours incubation. The concentration with the highest cell viability of the proliferation test results was then referred to as the best concentration. The best concentration was combined to determine the percent wound closure by cell migration assay using the scratch wound healing method. Combination ratio of HVCO and chitosan in the cell migration assay were 1:1; 1:3; and 3:1. Combination of HVCO and chitosan which results in the highest percentage of wound closure followed by COX-2 and VEGF protein expression testing with immunocytochemical methods.
The test materials increased the proliferation activity of NIH 3T3 cells in incubation for 24 hours. The highest cell viability was obtained at concentrations of 31.25 μg/ml namely 151.16% for HVCO, 124.06% for chitosan, which did not differ significantly from Aloclair® 31,25 μg/ml namely 153.35%. The combination of HVCO 31.25 μg/ml and chitosan 31.25 μg/ml with a ratio of 1:1 produced the most optimal percent of wound closure, which was 86.30% after 24 hours incubation and 100% after 48 hours incubation, which was significantly different from Aloclair® 31,25 μg/ml. The immunocytochemical examination of the combination of HVCO 31.25 μg/ml and chitosan 31.25 μg/ml (1:1) resulted in COX-2 protein expression was 45.65% and VEGF protein expression was 35.77%, which was significantly different from Aloclair® 31,25 μg/ml.
Based on the results of in vitro testing of wound healing parameters, the combination of HVCO 31.25 μg/ml and chitosan 31.25 μg/ml (1:1) was the best combination to accelerate the wound healing process. | en_US |
| dc.description.abstract | Hidrolisis parsial Virgin Coconut Oil (VCO) menghasilkan asam lemak bebas dan 2-monogliserida terutama asam laurat dan 2-monolaurin yang aktif sebagai agen antibakteri, antivirus dan penyembuhan luka. Kitosan dapat mempercepat pembentukan fibroblas sehingga meningkatkan proses penyembuhan luka. Penelitian ini bertujuan untuk mengetahui aktivitas HVCO, kitosan serta kombinasi HVCO dan kitosan terhadap parameter penyembuhan luka pada sel NIH 3T3 secara in vitro.
Sel NIH 3T3 diinkubasi selama 24 jam kemudian diterapi dengan bahan uji dan kontrol positif (Aloclair®) berbagai seri konsentrasi dimulai konsentrasi 15,625 μg/ml sampai 500 μg/ml. Setiap seri konsentrasi diinkubasi selama 24 jam dan dilakukan uji aktivitas proliferasi sel menggunakan metode MTT [3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazolium bromida]. Aktivitas proliferasi sel NIH 3T3 diukur dengan menghitung viabilitas sel setelah inkubasi 24 jam. Konsentrasi dengan viabilitas sel paling tinggi hasil uji proliferasi selanjutnya disebut sebagai konsentrasi terbaik. Konsentrasi terbaik selanjutnya dikombinasi untuk penentuan persen penutupan luka dengan uji migrasi sel menggunakan metode scratch wound healing. Perbandingan HVCO dan kitosan pada uji migrasi sel adalah 1:1; 1:3; dan 3:1. Perbandingan HVCO dan kitosan yang menghasilkan persen penutupan luka paling tinggi dilanjutkan uji ekspresi protein COX-2 dan VEGF dengan metode imunositokimia.
Bahan uji meningkatkan aktivitas proliferasi sel NIH 3T3 pada inkubasi selama 24 jam. Viabilitas sel paling tinggi diperoleh pada konsentrasi 31,25 μg/ml yaitu 151,16% untuk HVCO, 124,06% untuk kitosan, yang tidak berbeda secara signifikan dengan Aloclair® 31,25 μg/ml (153,35%). Kombinasi HVCO 31,25 μg/ml dan kitosan 31,25 μg/ml dengan perbandingan 1:1 menghasilkan persen penutupan luka paling optimal yakni 86,30 % setelah inkubasi 24 jam dan 100% setelah inkubasi 48 jam, yang berbeda secara signifikan dengan Aloclair® 31,25 μg/ml. Uji imunositokimia kombinasi HVCO 31,25 μg/ml dan kitosan 31,25 μg/ml (1:1) menghasilkan ekspresi protein COX-2 45,65% dan ekspresi protein VEGF 35,77%, yang berbeda secara signifikan dengan Aloclair® 31,25 μg/ml.
Berdasarkan hasil pengujian in vitro terhadap parameter penyembuhan luka maka kombinasi HVCO 31,25 μg/ml dan kitosan 31,25 μg/ml (1:1) merupakan kombinasi terbaik untuk mempercepat proses penyembuhan luka. | en_US |
