Coating Electrospun Collagen and Gelatin Fibers with Perlecan Domain I for Increased Growth Factor Binding

Abstract

Electrospun natural polymer membranes were fabricated from collagen or gelatin coated with a bioactive recombinant fragment of perlecan, a natural heparan sulfate proteoglycan. The electrospinning process allowed the facile processing of a three-dimensional, porous fibril (2-6 microm in diameter) matrix suitable for tissue engineering. Laser scanning confocal microscopy revealed that osteoblast-like MG63 cells infiltrated the depth of the electrospun membrane evenly without visible apoptosis. Tissue engineering scaffolds ideally mimic the extracellular matrix; therefore, the electrospun membrane must contain both structural and functional matrix features. Fibers were coated, after processing, with perlecan domain I (PlnDI) to improve binding of basic fibroblast growth factor (FGF-2), which binds to native heparan sulfate chains on PlnDI. PlnDI-coated electrospun collagen fibers were ten times more effective than heparin-BSA collagen fibers at binding FGF-2. Because FGF-2 modulates cell growth, differentiation, migration and survival, the ability to effectively bind FGF-2 to an electrospun matrix is a key improvement in creating a successful tissue engineering scaffold.

Engineering on the Straight and Narrow: The Mechanics of Nanofibrous Assemblies for Fiber-Reinforced Tissue Regeneration

“…63 Additional studies have modified fiber surfaces to enhance cell binding and=or growth factor retention. [64][65][66] Further, methacrylate-based copolymers have been electrospun to form nanofibrous coatings that can be crosslinked after formation. 67,68 We have recently reported on the electrospinning of several elements of a library of 120 poly(b-aminoester)s that were photo polymerized after formation 69 as well as novel photocrosslinkable and hydrolytically degradable elastomers.…”

Porous Polymeric Bioresorbable Scaffolds for Tissue Engineering

“…Mainly PLA and PLAGA scaffolds were produced with this approach. Salts are the most commonly used porogen [64,92], but also sugar [93], paraffin [94] and gelatine spheres [94] have been employed. This method enables to tune independently pore size (up to 500 µm in diameter) and porosity (up to 90%) by controlling particle dimensions and porogen/polymer ratio respectively.…”

“…In the literature other types of GFs have been employed in combination with ES scaffolds. However, most of these studies concern the immobilization of such biomolecules at the fibre surface [94][95][96]. Leong and coworkers incorporated GFs that were intended to promote nerve regeneration into ES fibres of caprolactone-ethylethylene phosphate copolymer [97,98].…”

Recent advances in annular pathobiology provide insights into rim-lesion mediated intervertebral disc degeneration and potential new approaches to annular repair strategies

“…Perlecan's chondrogenic, matrix organisational and stabilising roles and ability to sequester a range of anabolic growth factors in the pericellular matrix indicates that it has important attributes relevant to annular remodelling and repair processes, reinforcing its potential as an effector molecule worthy of further investigation in the context of repair biology. It will be very interesting in this regard to ascertain whether the therapeutic potential of 3D scaffolds of electrospun fibres of collagen and gelatin functionalised with recombinant domain-I of perlecan are fully realised [13].…”

“…Collagen sponge, collagen gel, agarose, alginate or fibrin gel cell carriers, alginate/chitosan hybrid fibre scaffolds [129], scaffolds assembled from electrospun collagen and gelatin functionalised with perlecan domain-I [13], chitosan salts cross-linked to genipen [95], a chitosan glycerophosphate hydrogel [114], a porous calcium polyphosphate carrier [128], elastic poly (1, 8-octanediol malate) 3D scaffolds [151], a composite demineralised bone matrix gelatin-(polycaprolactone triol malate)-poly(caprolactone triol malate) biphasic scaffold [152] and collagen/hyaluronan hybrid scaffolds [5] have all been examined in IVD and/or annular repair strategies. Exciting possibilities also exist for the use of replacement total IVDs using biocompatible and biomechanically competent PLGA and PLA scaffolds seeded with AF and NP cells [88,89].…”