The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffolds

Abstract

Biologic materials from various species and tissues are commonly used as surgical meshes or scaffolds for tissue reconstruction. Extracellular matrix (ECM) represents the secreted product of the cells comprising each tissue and organ, and therefore provides a unique biologic material for selected regenerative medicine applications. Minimal disruption of ECM ultrastructure and content during tissue processing is typically desirable. The objective of this study was to systematically evaluate effects of commonly used tissue processing steps upon porcine dermal ECM scaffold composition, mechanical properties, and cytocompatibility. Processing steps evaluated included liming and hot water sanitation, trypsin/SDS/TritonX-100 decellularization, and trypsin/TritonX-100 decellularization. Liming decreased the growth factor and glycosaminoglycan content, the mechanical strength, and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for all). Hot water sanitation treatment decreased only the growth factor content of the ECM (p ≤ 0.05). Trypsin/ SDS/TritonX-100 decellularization decreased the growth factor content and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). Trypsin/TritonX-100 decellularization also decreased the growth factor content of the ECM but increased the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). We conclude that processing steps evaluated in the present study affect content, mechanical strength, and/or cytocompatibility of the resultant porcine dermal ECM, and therefore care must be taken in choosing appropriate processing steps to maintain the beneficial effects of ECM in biologic scaffolds.

Cellular Localization, Invasion, and Turnover Are Differently Influenced by Healthy and Tumor-Derived Extracellular Matrix

“…The ability to maintain the presence of glycosaminoglycans is mandatory for the proper function of ECM, although many protocols of decellularization lack this important component due to of the use of harsh detergents and aspecific enzymes. 18,38,39 Indeed, all the above reported features of tissue-derived ECMs are relevant for assessing the role of microenvironment on tissue homeostasis, as (i) intrinsic domains within stromal proteins might act as ligands for canonical growth factor receptors, (ii) many growth factors bind to glycosaminoglycan chains, 3 and (iii) the proper 3D structure allow for the specific and organized localization of ligands 40 and associated proteins, such as mucins, complement, metallo-proteases, metallo-protease inhibitors, latent TGF-b, growth factors, and chemokines. 41,42 To establish the availability of tissue-derived ECMs to be employed as scaffolds for the study of tumorigenesis we used monocytes and tumor epithelial cell lines.…”

Creation of an injectable in situ gelling native extracellular matrix for nucleus pulposus tissue engineering

“…Removal of cells and antigens is essential to preventing an immune response; however, decellularization must also maintain the composition and structure of the native tissue [33][34][35][36]. Decellularization techniques have been optimized for the heart [37], lung [38], nerve [39], skin [40], brain [41], and spinal cord [42], among other tissues, and can involve physical, chemical, and biological methods to lyse cells, and subsequently remove cellular debris. Our laboratory has previously developed gentle decellularization techniques for nerve tissue to preserve essential proteins, proteoglycans, and microstructure [39,43], which would be ideal for the delicate nucleus pulposus.…”

Chemical decellularization: a promising approach for preparation of extracellular matrix

“…In addition, agitation can also be used for rinsing residual detergents, by means of PBS washing cycles. It may be used for heart valves 28 , muscles 46 , peripheral nerves 47 , trachea 48 and dermis 49 .…”