Which detergent is most suitable for the generation of an acellular pancreas bioscaffold?

• Published in: Brazilian journal of medical and biological research Vol. 57; p. e13107
• Main Authors: Mantovani, M C, Damaceno-Rodrigues, N R, Ronatty, G T S, Segovia, R S, Pantanali, C A, Rocha-Santos, V, Caldini, E G, Sogayar, M C
• Format: Journal Article
• Language: English
• Published: Brazil Associacao Brasileira de Divulgacao Cientifica (ABDC) 01.08.2024; Associação Brasileira de Divulgação Científica
• Subjects: Analysis; Animals; Bioengineering; BIOLOGY; Care and treatment; Collagen; Comparative analysis; Decellularized Extracellular Matrix - chemistry; Deoxycholic acid; Deoxycholic Acid - chemistry; Deoxycholic Acid - pharmacology; Detergents - chemistry; Detergents - pharmacology; Detergents, Synthetic; Diabetes Mellitus, Type 1; Extracellular Matrix; Glycosaminoglycans; Health aspects; Human acellular pancreatic matrix; Humans; MEDICINE, RESEARCH & EXPERIMENTAL; Methods; Methylene blue; Mice; Microscopy, Electron, Scanning; Octoxynol - chemistry; Pancreas; Pancreas - cytology; Pancreatic matrix recellularization; Physiological aspects; Rat acellular pancreatic matrix; Sodium deoxycholate (SDC); Sodium dodecyl sulfate (SDS); Sodium Dodecyl Sulfate - pharmacology; Sulfates; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Triton X-100; Type 1 diabetes; Brazil; Sodium deoxycholate (SDC); Sodium dodecyl sulfate (SDS); Rat acellular pancreatic matrix; Human acellular pancreatic matrix; Pancreatic matrix recellularization; Triton X-100
• ISSN: 0100-879X; 1414-431X; 1414-431X
• DOI: 10.1590/1414-431X2024e13107

Pancreatic bioengineering is a potential therapeutic alternative for type 1 diabetes (T1D) in which the pancreas is decellularized, generating an acellular extracellular matrix (ECM) scaffold, which may be reconstituted by recellularization with several cell types to generate a bioartificial pancreas. No consensus for an ideal pancreatic decellularization protocol exists. Therefore, we aimed to determine the best-suited detergent by comparing sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), and Triton X-100 at different concentrations. Murine (n=12) and human pancreatic tissue from adult brain-dead donors (n=06) was harvested in accordance with Institutional Ethical Committee of the University of São Paulo Medical School (CEP-FMUSP) and decellularized under different detergent conditions. DNA content, histological analysis, and transmission and scanning electron microscopy were assessed. The most adequate condition for pancreatic decellularization was found to be 4% SDC, displaying: a) effective cell removal; b) maintenance of extracellular matrix architecture; c) proteoglycans, glycosaminoglycans (GAGs), and collagen fibers preservation. This protocol was extrapolated and successfully applied to human pancreas decellularization. The acellular ECM scaffold generated was recelullarized using human pancreatic islets primary clusters. 3D clusters were generated using 0.5×104 cells and then placed on top of acellular pancreatic slices (25 and 50 μm thickness). These clusters tended to connect to the acellular matrix, with visible cells located in the periphery of the clusters interacting with the ECM network of the bioscaffold slices and continued to produce insulin. This study provided evidence on how to improve and accelerate the pancreas decellularization process, while maintaining its architecture and extracellular structure, aiming at pancreatic bioengineering.