Biological evaluations of decellularized extracellular matrix collagen microparticles prepared based on plant enzymes and aqueous two-phase method

Abstract

For patients with extensive full-thickness burns who do not have sufficient autologous split-thickness skin for skin grafts, the application of biological skin substitutes may be considered. The aim of this study was to find an optimal new type method for the production of a biovital skin substitute based on acellular dermal matrix (ADM) and preclinical evaluations. In this work, 25 methods of ADM production were assessed. The proposed methods are based on the use of the following enzymes: papain, Carica papaya lipase (CPL), and purification using a polymer/salt aqueous two-phase system. The obtained ADM samples were characterized via scanning electron microscopy (SEM), porosity measurement and water vapor transmission test. Results showed that the collagen bundles of ADM microparticles were intact and orderly. Through differential scanning calorimetry (DSC), thermo gravimetric analysis (TGA) and biocompatibility tests, the results indicated that the proportion of papain and CPL was the same and 5 h processing time are the optimum conditions for ADM preparation and the material showed good biocompatibility. Our results suggested that the potential of developing this kind of decellularization process to manufacture ADM scaffolds for clinical application.

Characteristics and Preparation of Designed Alginate-Based Composite Scaffold Membranes with Decellularized Fibrous Micro-Scaffold Structures from Porcine Skin

“…Numerous synthetic materials and natural materials have been proposed, modified, and employed for medical applications, such as 3D bioprinting scaffolds for skin and bone tissue reconstruction [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. Three-dimensional (3D) bioprinting shows potential in tissue engineering and regenerative applications due to its overwhelming advantages over other approaches.…”

Design And Preparation of New Decellularized Extracellular Matrix/Alginate Composite Membranes For Biomimetic Bioinks Via Supercritical Carbon Dioxide Fluid Treatments

“…Treatments with supercritical carbon dioxide before preparation of a decellularized extracellular matrix Supercritical uids of carbon dioxide (ScCO 2 ) was empolyed for preparation of new designed decellularized extracellular matrix scaffolds in this study. The ScCO 2 was employed before enzyme treatments for removing most fatty acids and tissues [4].…”

“…Numerous materials have been proposed, modi ed and empolyed for medical bioprinting applications such as scaffolds for skin and bone tissue reconstruction such as synthetic materials and natural materials [4][5][6][7][8][9][10][11][12][13][14][15][16]. Sodium alginate is a naturally occurring biopolymer extracted from different species of marine brown algae [12].…”

Stem cell culture media enriched with plant‐derived compounds: Cell differentiation enhancement

“…Fortunately, simpler and cheaper techniques such as aqueous two‐phase systems (ATPSs) have been successfully implemented for the separation of multiple compounds from different plants, 86‐88 representing cost‐effective strategies with high biological compatibility. In this sense, ATPSs have been used for the purification of enzymes derived from papaya that are able to remove biological material from a tissue resulting in a compact and complete structure with extracellular matrix components, useful as a scaffold for skin replacement 89 . Most importantly, this is another example that highlights the need for replacement of animal enzymes such as trypsin and pepsin with plant derivatives such as papain and Carica papaya lipase as novel decellularization methods.…”

“…In this sense, ATPSs have been used for the purification of enzymes derived from papaya that are able to remove biological material from a tissue resulting in a compact and complete structure with extracellular matrix components, useful as a scaffold for skin replacement. 89 Most importantly, this is another example that highlights the need for replacement of animal enzymes such as trypsin and pepsin with plant derivatives such as papain and Carica papaya lipase as novel decellularization methods.…”