Uji Sitotoksisitas Metronidazol Berbasis Hidrogel Kitosan Terhadap Viabilitas Sel Fibroblas 3T3 Secara In Vitro

Abstract

Background: Metronidazole is a local antibiotic that is often used in periodontal disease. Chitosan hydrogels-based metronidazole shows considerable antibacterial effects. However, the cytotoxic effects on fibroblast have yet to be studied. Objective: To determine the in vitro cytotoxicity effect of chitosan hydrogel-based metronidazole on fibroblast. Material and Method: Chitosan hydrogel-based metronidazole (0.12%, 0.15%, 0.25%) and chitosan without metronidazole are made by dissolving lactic acid to reach a gel consistency, and then adding powdered metronidazole according to the concentration. 3T3 fibroblast cells were cultured in well mikroplate containing RPMI-1640 media inside an incubator. The cell viability tests were carried out using the MTT assay method and repeated 5 times. The colour degradation was read with a microplate reader. Result: The mean viability of fibroblasts mixed with chitosan hydrogel-based metronidazole at concentration of 0.12%, 0.15%, and 0.25%, and chitosan without metronidazole were 96%, 93%, 91% and 98.8% respectively. ANOVA statistical test results for the 0.12% concentration group, where p < 0.05 represents the cell’s viability, significantly differs from the results for the 0.15% and 0.25% concentration groups. Conclusion:Chitosan hydrogel-based metronidazole shows non toxic effects on fibroblast cells.

Latar Belakang: Metronidazol merupakan antibiotik lokal yang sering digunakan pada penyakit periodontal. Metronidazol berbasis hidrogel kitosan menunjukkan efek antibakteri yang cukup besar. Namun belum ada penelitian terhadap efek sitotoksisitas metronidazol berbasis hidrogel kitosan terhadap sel fibroblas 3T3. Tujuan : untuk mengetahui sitotoksisitas metronidazol berbasis hidrogel kitosan terhadap viabilitas sel fibroblas 3T3 secara invitro. Metode : Pembuatan metronidazol 0.12%; 0.15%; 0.25% berbasis hidrogel kitosan dan hidrogel kitosan tanpa metronidazol diawali dengan melarutkan asam laktat sampai berbentuk gel lalu penambahan bubuk metronidazol sesuai dengan konsentrasi. Sel fibroblas 3T3 dibiakkan pada media RPMI-1640 dalam inkubator. Pengujian viabilitas sel fibroblas 3T3 dilakukan dengan metode MTT assay dan diulangi sebanyak 5 kali. Perubahan warna dibaca dengan microplate reader. Hasil : Rerata viabilitas sel fibroblas 3T3 yang diaplikasikan metronidazol berbasis hidrogel kitosan dengan konsentrasi 0.12%; 0.15%; 0.25% dan hidrogel kitosan 3% tanpa metronidazol berturut-turut sebesar 96%; 93%; 91% dan 98.8%. Hasil uji statistik ANOVA dengan p<0.05. Metronidazol berbasis hidrogel kitosan 0.12% menunjukkan rerata viabilitas sel yang paling tinggi. Kesimpulan : Metronidazol berbasis hidrogel kitosan menunjukkan tidak toksik terhadap sel fibroblas 3T3.