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

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 pancrea...

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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) 01-01-2024 Revista Brasileira de Pesquisas Medicas Associação Brasileira de Divulgação Científica
Subjects:	Analysis Animals Beta cells Bioengineering BIOLOGY Care and treatment Collagen Comparative analysis Decellularized Extracellular Matrix - chemistry Deoxycholic acid Deoxycholic Acid - chemistry Deoxycholic Acid - pharmacology Detergents Detergents - chemistry Detergents - pharmacology Detergents, Synthetic Diabetes mellitus (insulin dependent) 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 Proteoglycans Rat acellular pancreatic matrix Scanning electron microscopy Sodium deoxycholate Sodium deoxycholate (SDC) Sodium dodecyl sulfate (SDS) Sodium Dodecyl Sulfate - pharmacology Sodium lauryl sulfate 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
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0100-879X 1414-431X 1414-431X 1678-4510
DOI:	10.1590/1414-431X2024e13107