Design of epidermal growth factor immobilization on 3D biocompatible scaffolds to promote tissue repair and regeneration

Abstract

Exogenous application of human epidermal growth factor (hEGF) stimulates epidermal wound healing. The aim of this study was to develop bioconjugates based on hEGF mimicking the protein in its native state and thus suitable for tissue engineering applications, in particular for treating skin-related disorders as burns. Ribonuclease A (RNase A) was used to investigate a number of different activated-agarose carriers: cyanogen bromide (CNBr)-activated-agarose and glyoxyl-agarose showed to preserve the appropriate orientation of the protein for receptor binding. EGF was immobilized on these carriers and immobilization yield was evaluated (100% and 12%, respectively). A peptide mapping of unbound protein regions was carried out by LC–MS to take evidence of the residues involved in the immobilization and, consequently, the flexibility and surface accessibility of immobilized EGF. To assess cell proliferative activities, 10, 25, 50, and 100 ng/mL of each immobilized EGF sample were seeded on fibroblast cells and incubated for 24, 48 and 72 h. The immobilized growth factor showed significantly high cell proliferative activity at 50 and 100 ng/mL compared to control and soluble EGF. Although both of the immobilized samples show dose-dependency when seeded with high number of fibroblast cells, CNBr-agarose-EGF showed a significantly high activity at 100 ng/mL and 72 h incubation, compared to glyoxyl-agarose-EGF.

Changes in the Transcriptome Profiles of Human Amnion-Derived Mesenchymal Stromal/Stem Cells Induced by Three-Dimensional Culture: A Potential Priming Strategy to Improve Their Properties

“…Gothelf and collaborators demonstrated that the CM derived from neurotrophic factor-secreting MSCs was enriched with BDNF, VEGF-A, and HGF and has been used effectively to have protective effects in several animal models of neurodegenerative diseases [68]. EGF stimulates the growth of numerous epidermal and epithelial tissues [69], and activation of TGF-β is also involved in the recruitment of stem/progenitor cells in tissue regeneration/remodeling processes [70]. Furthermore, it has been proven that LIF, which plays a crucial role in blastocyst implantation, is able to regulate some regenerative processes after injury in several tissues [71].…”

Polymeric Biomaterials for the Treatment of Cardiac Post-Infarction Injuries

“…Thus, scaffold stiffness and geometry are important features to be considered, with square pores being more efficient in promoting cell adhesion rather than hexagonal pores [ 32 ]. Moreover, biochemical cues, such as growth factors [ 33 ], cytokines, and adhesion molecules, should be inserted to induce effective maturation of cardiac cells [ 21 ]. Finally, in order to produce the optimal material for tissue interfaces, the design of the cardiac biomaterial should fit with the complex electrical pathways of the myocardium [ 34 ], obtained by a fine coordination between membrane potential depolarization, pacemaker conduction system, and specific intracellular communication networks [ 35 ].…”

Potent Particle-Based Vehicles for Growth Factor Delivery from Electrospun Meshes: Fabrication and Functionalization Strategies for Effective Tissue Regeneration

“…10,11 Covalent conjugation also typically results in negligible GF release and is not ideally suited for applications where GFs internalization is required for cells to exert their intended biological effects. 60,61 In summary, extant modes of GF incorporation and delivery from electrospun meshes suffer from several limitations such as the use of additional processing steps, promotion of sharp burst release profiles for surface-adsorbed proteins, direct coupling of GF release kinetics with intrinsic fiber properties for blend electrospun meshes, potential loss of GF conformation-activity, significantly delayed release from coaxially prepared core−shell meshes, and possible negative impact on bulk mesh properties.…”

“…While covalent conjugation avoids exposure of GFs to electrospinning conditions, it can potentially interfere with GF activity should access to the binding epitope be reduced during the conjugation step. Moreover, most synthetic polymers used for electrospinning often lack active sites for direct covalent conjugation; therefore, additional surface treatments or the inclusion of natural polymers is required prior to protein immobilization. , Covalent conjugation also typically results in negligible GF release and is not ideally suited for applications where GFs internalization is required for cells to exert their intended biological effects. , …”