Achatina fulica mucous improves cell viability and increases collagen deposition in UVB-irradiated human fibroblast culture

Ch Tri Nuryana; Sofia Mubarika Haryana; Yohanes Widodo Wirohadidjojo; Nur Arfian

doi:10.46582/jsrm.1601005

Achatina fulica mucous improves cell viability and increases collagen deposition in UVB-irradiated human fibroblast culture

J Stem Cells Regen Med. 2020 May 27;16(1):26-31. doi: 10.46582/jsrm.1601005. eCollection 2020.

Authors

Ch Tri Nuryana¹, Sofia Mubarika Haryana², Yohanes Widodo Wirohadidjojo³, Nur Arfian⁴

Affiliations

¹ Doctoral program of Medicine and Health, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
² Department of Histology and Cell Biology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
³ Department of Dermatology and Venereology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
⁴ Department of Anatomy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.

Abstract

Introduction: Ultraviolet radiation induces skin photoaging by increasing matrix metalloproteinase-1 (MMP-1). MMP-1 degrades type I and III collagen that comprise the dermal connective tissue. Achatina fulica mucous (AFM) is a natural remedy that has protective effects on fibroblasts and collagen. Objective: To investigate the effects of AFM on cell viability and collagen deposition in UVB-irradiated human fibroblast culture. Methods: The mucous was extracted from 50 Achatina fulica snails that were stimulated by a 5-10 Volt electricity shock for 30-60 seconds and converted into powder by the freeze-drying process. The human dermal fibroblast culture was divided into six groups: group 1 were normal fibroblasts without UVB irradiation as normal control, groups 2-5 consisted of 100 mJ/cm² UVB-irradiated fibroblasts. Group 2 had no treatment as negative control, group 3 was treated by PRP 10% as positive control group and groups 4-6 were treated by various concentrations of AFM (3.9; 15.625 and 62.5 μg/mL). At the end of the experiment, the proliferation was assessed with MTT assay, furthermore collagen deposition was measured by Sirius red assay. Real Time-PCR (RT-PCR) was performed to quantify Coll I, Coll III and MMP-1 mRNA expression, then to measured COL 1/COL III ratio. Results: UVB induced significant lower viability, upregulated MMP-1 and downregulated COL I and COL III mRNA expressions. Meanwhile AFM treated groups demonstrated higher cell viability with downregulation of MMP-1 and upregulation of COL I and COL III mRNA expressions. The ratio of COL I/ III expression was significantly (p<0.05) lower in the AFM treated groups compared to the UVB group. Among AFM treated groups, administration of 62.5 μg/mL AFM represented the best result. Conclusion: AFM may ameliorate viability of UVB-irradiated human fibroblast culture which associates with downregulating MMP-1, upregulating COL I and Col III, and reducing COL I/III ratio.

Keywords: Achatina fulica; Collagen; MMP-1; UVB-irradiated human fibroblast culture; Viability.