POTENSI SENYAWA SERI BENZOIL KALIKS[4]RESORSINARENA SEBAGAI TABIR SURYA: KAJIAN EFEK PROTEKSI, SITOTOKSISITAS DAN PEMBENTUKAN KOLAGEN PADA BIAKAN FIBROBLAS AKIBAT PAJANAN SINAR ULTRAVIOLET B

ANDI AIDAWATY, Prof. Dr. Mustofa, M.Kes., Apt.

2015 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Latar Belakang. Pajanan UVB berlebihan pada kulit manusia merupakan etiologi terjadinya eritema, edema, dan supresi imun yang pada akhirnya sebagai pemicu kanker kulit. Penggunaan tabir surya dibutuhkan untuk mencegah efek buruk radiasi UVB. Senyawa seri benzoil kaliks[4]sorsinarena yang diuji mempunyai efek proteksi terhadap UVB adalah benzoil C-fenil kaliks[4]resorsinarena, benzoil C-metil kaliks[4]resorsnarena dan C-fenil kaliks(4)resorsinaril okta benzoat. Tujuan. Mengkaji aktifitas efek proteksi dan toksisitas senyawa uji seri benzoil kaliks[4]resorsinarena, serta mengkaji senyawa uji ini terhadap aktifitas terbaik terhadap deposisi kolagen dan peningkatan penghambatan degradasi kolagen yang mempengaruhi sintesis kolagen. Metode. Uji efek proteksi, disiapkan senyawa uji dengan konsentrasi 0,01%, 0,02%, 0,04%, 0,08%, 0,16%, dan 0,32% dalam bentuk sediaan krim. Selanjutnya dioles diatas pada tutup 96-well plate di atas sel fibroblas, well plate dipajan sinar UVB 300 mJ/cm2. Viabilitas sel dinilai dengan MTT-assay. Berdasarkan rerata DO dihitung EC50. Uji sitotoksisitas kultur fibroblas dalam 96 well plate diberi senyawa uji 0,04%, 0,08%, 0,16%, 0,32% dan 0,64%. Setelah inkubasi 24 jam, viabilitas sel dinilai dengan MTT assay, IC50 dihitung menggunakan analisis probit. Untuk uji deposisi dan degradasi kolagen, disiapkan kultur fibroblas dalam 96-well plate dan dioles krim di atas tutup well plate dengan konsentrasi 1/3EC50, EC50, dan 3EC50 senyawa uji, lalu well plate dipajan dengan sinar UVB 300 mJ/cm2. Pengukuran deposisi dan degradasi kolagen dilakukan dengan metode sirius red assay. Uji efek proteksi dan sitotoksisitas dianalisa secara probit dan uji kolagen menggunakan uji repeated ANOVA, Uji Friedman untuk melihat rerata perbedaan densitas optis. Hasil. Uji efek proteksi, sitotoksisita serta indeks sitoksitas menghasilkan EC50,IC50 dan IS masing-masing senyawa uji 1= 0,125%, 0,324%, 2,592, senyawa uji 2= 0,250%, 0,947%, 3,788 dan senyawa uji 3= 0,339%, 1,219%, 3,596.. Sedangkan pada uji deposisi dan degradasi kolagen peningkatan deposisi dan peningkatan penghambatan degradasi kolagen masing-masing pada konsentrasi untuk senyawa uji 1= 0,042% dan 0,125%, senyawa uji 2= 0,750% dan 0,250%, dan senyawa uji 3= 0,339% dan 0,113%. Hasil uji deposisi dan degradasi tidak ada perbedaan bermakna. Kesimpulan. Ketiga senyawa uji memberikan efek proteksi terhadap fibroblas yang diberi pajanan UVB 300 mJ/cm2 dan memiliki sifat tidak toksik terhadap fibroblas, serta dapat meningkatkan deposisi dan meningkatkan penghambatan degradasi kolagen walaupun secara statistik tidak bermakna. Kata kunci: Benzoil C-fenil kaliks[4]resorsinarena, benzoil C-metil kaliks[4]resorsinarena, C-fenil kaliks[4]resorsinarena, Fibroblas, Sitotoksisitas, efek proteksi, viabilitas, sinar UVB, deposisi kolagen, degradasi kolagen

Background. Excessive UVB exposure is the etiology of human skin erythema, edema, and immune suppression, that will ultimately become as a skin cancer trigger. The use of sunscreen is needed to prevent the adverse effects of UVB radiation. The benzoyl calix[4]resorcinarene series compounds tested in this study for their protective effects against UVB are benzoyl C-phenylcalix[4]resorcinarene, benzoyl C-methylcalix[4]resorcinarene and C-phenylcalix[4]resorsinaryl octa benzoate. Objective. Reviewing the protective effect and toxicity of test compounds (benzoyl calix[4]resorcinarene series compounds), and elucidating which compound has the best activity against collagen deposition and increased inhibition of collagen degradation that affect the synthesis of collagen. Methods. Protective effect assessment was initiated with the preparation of test compounds with concentrations of 0.01%, 0.02%, 0.04%, 0.08%, 0.16%, and 0.32% in cream dosage form. Test compounds were smeared on the lids of 96-wells plate,just above the cultured fibroblasts. The fibroblasts were then exposed to UVB rays with the dose of 300 mJ/cm2. Cell viability was assessed using MTT assay. Based on the average optical density (OD), EC50 values were then calculated. The cytotoxicity assay in 96 well plate was done by treating the fibroblasts with the test compounds (with the concentrations of 0.04%, 0.08%, 0.16%, 0.32% and 0.64%). Cell viability was then assessed using MTT assay after 24 hours of incubation.The IC50 values were calculated using probit analysis. The collagen deposition and degradation assays were begun with the preparation of fibroblast cultures in 96-wells plate. Tested compounds was smeared on top of the lids of 96-wells plate at the concentrations of 1/3EC50, EC50, and 3EC50. The plate wast han exposed to UVB rays (300 mJ/cm2). Measurements of collagen deposition and degradation were conducted using sirius red assay. Protective effect and cytotoxicity test were analyzed using probit analysis and collagen assays were analyzed using repeated ANOVA test, mean while Friedman test was used to elucidate the mean optical density difference. Result. The values of EC50, IC50 and IS for each of the test compounds areas follows: test compound 1= 0.125%, 0.324%, 2.592; test compound 2= 0.250%, 0.947%, 3.788; and test compound 3= 0.339%, 1.219%, 3.596, respectively. There are no significant differences in deposition and degradation assays results. Conclusion. All of the test compounds provide protective effect to fibroblasts against UVB exposure with the dose of 300 mJ/cm2, without toxic properties to the fibroblasts. The compounds can improve the deposition and increase the inhibition of collagen degradation although the differences in the assays results are not statistically significant. Keywords: Benzoyl C-phenylcaliks[4]resorcinarene, Benzoyl C-methylcalix[4]resorcinarene, C-Phenylcalix[4]resorcinaryl octa benzoate, fibroblast, cytotoxicity, index cytoxity, protective effect, viability, UVB, collagen deposisition, degradation collagen

Kata Kunci : Benzoil C-fenil kaliks[4]resorsinarena, benzoil C-metil kaliks[4]resorsinarena, C-fenil kaliks[4]resorsinarena, Fibroblas, Sitotoksisitas, efek proteksi, viabilitas, sinar UVB, deposisi kolagen, degradasi kolagen; Benzoyl C-phenylcaliks[4]resorcinarene, B