Tissue-Specific Decellularization Protocols: A Comparative Histological Analysis

Abstract

Background: Obtaining tissue-engineered constructs by decellularization can become an alternative method for restoring and regenerating tissues and organs with impaired functions. The quality of the extracellular matrix depends on the protocol used.
Objective: The objective of this study is to identify a quick and cost-effective protocol for decellularization of organs containing muscular and epithelial tissues.
Methods: This paper describes two methods of decellularization: the classical method of immersing the tissue in a solution and stirring it until the cells are completely removed from the tissue. This method is only suitable for flat tissues up to 5 mm thick, so we used it to clean the diaphragms. To decellularize larger tissues with the preservation of their structures, we used the native vascular system for the perfusion of solutions inside by cannulating the artery. For decellularization, a modified detergent method was used using sodium deoxycholate, sodium dodecyl sulfate, EDTA, and Triton X-100. For morphological studies, sections were stained using three methods: hematoxylin-eosin, Van Gieson, and Masson’s trichrome staining. Histological sections were studied under a Leica DM 1000 microscope with microphotography using a Leica ICC50 HD digital camera.
Results: Using this method, we successfully decellularized kidneys while preserving the vasculature. Such organs can be called cell-free vascularized bioscaffolds. Morphological studies confirmed the preservation of the tissue architecture, the absence of cells, and nuclear material. The main result of this work is the development of two simple methods for creating biodegradable, biocompatible, vascularized tissue (organ) scaffolds with the same degree of
complexity as in nature. The main result of this work is the development of two simple methods for creating biodegradable, biocompatible, vascularized tissue (organ) scaffolds with the same degree of complexity as in nature. Using a peristaltic pump, the kidney was flushed with SDS. Using this method, we successfully decellularized kidneys while preserving the vasculature. Such organs can be called cell-free vascularized bioscaffolds—the rat diaphragm decellularization protocol, perfusion with SD. Morphological studies confirmed the preservation of the tissue architecture, the absence of cells, and nuclear material.
Conclusion: The obtained results proved that the method of decellularization depends on the origin of the tissue. To achieve the best results on different tissue structures, it is necessary to use different protocols with specific detergents.