QUALITATIVE METHYLENE BLUE MICROSCOPIC COMPARISON OF TRITON X 100 AND SDS DECELLULARIZATION PROTOCOLS FOR <i>CHANNA STRIATA</i> SKIN

Terry Previo Avianto; Noraini Ahmad; Refan Maulana Putra; Afrisya Idzni Nurashiddiqy

Terry Previo Avianto Department of Marine, Faculty of Fisheries and Marine, Universitas Airlangga, 60115, Surabaya, Indonesia.
Noraini Ahmad Department of Chemistry, Faculty of Science, Universiti Malaya, 50603, Kuala Lumpur Malaysia.
Refan Maulana Putra Fisheries Product Technology Study Program, Faculty of Fisheries and Marine, Universitas Airlangga, 60115, Surabaya, Indonesia.
Afrisya Idzni Nurashiddiqy Fisheries Product Technology Study Program, Faculty of Fisheries and Marine, Universitas Airlangga, 60115, Surabaya, Indonesia.

Keywords: Decellularization, channa striata skin, methylene blue microscopy, responsible consumption and production

Abstract

Fish skin-derived extracellular matrix (ECM) has emerged as a promising biomaterial for chronic wound management and commercial grafts. Compared to mammalian xenografts, acellular fish skin demonstrates favourable clinical outcomes alongside a reduced risk of disease transmission and fewer cultural limitations. Channa striata (snakehead fish) skin, naturally rich in type I collagen, is an excellent candidate for developing these biopolymer dressings. Effective decellularization is critical to remove cellular material while preserving the underlying ECM architecture; however, common detergents like sodium dodecyl sulphate (SDS) and Triton X-100 impact ECM composition and mechanics differently. This exploratory study qualitatively compared three detergent-based decellularization protocols for C. striata skin: 1 % Triton X-100, 1 % SDS, and a combined 0.5 % Triton X-100/0.5 % SDS solution. Fresh skin was divided into matched pairs of treated and untreated groups, processed accordingly, and evaluated via methylene blue (MB) staining and bright-field microscopy. Untreated controls exhibited abundant MB-positive nuclei and dense background staining. Treatment with 1 % Triton X-100 reduced nuclear staining while largely preserving the fibrillar collagen network. Conversely, 1 % SDS produced the greatest reduction in visible nuclei but caused apparent swelling and structural disruption of the collagen architecture. The combined 0.5 % Triton/0.5 % SDS protocol achieved intermediate nuclear clearance while better maintaining collagen organization than SDS alone. These findings support MB microscopy as a rapid, qualitative screening tool for decellularization strategies. They also suggest that mixed, low-concentration Triton/SDS protocols best balance cell removal with ECM preservation in C. striata skin. Future quantitative DNA, glycosaminoglycan, and mechanical testing will be required to optimize this protocol for scaffold development in diabetic ulcer applications.