Electrospun, Biofunctionalized Fibers as Tailored in vitro Substrates for Keratinocyte Cell Culture

Abstract

Cell adhesion preventing fiber surfaces were tailored differently with bioactive peptides (a fibronectin fragment (GRGDS), a collagen IV fragment (GEFYFDLRLKGDK) and a combination of both) to provide an artificial extracellular matrix as a substrate for HaCaT keratinocyte cell culture. Therefore, a polymer blend containing a six-arm star-shaped statistical copolymer of ethylene oxide and propylene oxide in the ratio 80:20 (NCO-sP[EO-co-PO]) and poly-[D,L-(lactide-co-glycolide)] (PLGA) was electrospun. The resulting fibers were biofunctionalized and investigated as in vitro substrates using the HaCaT kerationcyte cell line. Appropriate surface chemistry on these electrospun fibers proved to prevent adhesion of keratinocytes, while additional immobilization of certain peptide sequences induced cell adhesion. These specific fibers enable investigation of immobilized active molecules and the subsequent cellular response to the scaffold. HaCaT keratinocytes were found to selectively adhere to those fibers modified with either collagen IV segment GEFYFDLRLKGDK or a mixture of the two peptide sequences GEFYFDLRLKGDK and GRGDS (1:1). However, the synergistic effects of both (the fibronectin fragment and the collagen IV fragment) seem to significantly increase the numbers of adherent keratinocytes.

In situ guided tissue regeneration in musculoskeletal diseases and aging

“…Exciting developments in material science are capable of perfectly matching these scenarios using intelligent and functional materials, which can also be designed for perfect timely release of factors involved in regeneration (Astachov et al 2011; Chen et al 2010; Di Maggio et al 2011; Dvir et al 2011; Gilbert and Blau 2011; Grafahrend et al 2010; Klinkhammer et al 2010; Lutolf et al 2009; Meinel et al 2009; Votteler et al 2010). Cellular and organismal aging phenomena in an elderly and diseased target population for regenerative strategies may be serious obstacles for successful treatment regimens.…”

“…Endogenous stem cell homing and retransplantation of ex vivo amplified precursors have been addressed as a means of in situ tissue engineering as well as the engineering of new, partially functionalized scaffolds especially for bone tissue engineering, among them also injectable scaffolds for regeneration induction (Chen et al 2011; Grafahrend et al 2010, 2011; Pennesi et al 2011; Shekaran and Garcia 2011; Uskokovic and Uskokovic 2011). …”

Current Treatment Limitations in Age-Related Macular Degeneration and Future Approaches Based on Cell Therapy and Tissue Engineering

“…However, cell adhesion peptides can be immobilized on the surface of these fibres, so that specific adhesion of cells onto the fibrous sheets results. With this method, scaffolds could be produced that influence the reaction of immune cells [169] and that mimic the basal membrane in skin [170]. Hence, such strategies bear great potential for TE approaches in AMD, where materials that mimic the BM are one key factor for success.…”

Enzymatic Activity at the Surface of Biomaterials via Supramolecular Anchoring of Peptides: The Effect of Material Processing

“…[15] The electrospinning of meshes containing bioactive molecules either by mixing or by covalent attachment is reported as well as the generation of electrospun meshes applicable for covalent functionalization after processing. [16,17] Although this allows for the incorporation of bioactive moieties into the meshes, due to their covalent attachment the meshes do not possess the dynamic nature of the ECM. However, the supramolecular UPy-modified biomaterials developed in our group are based on noncovalent interactions, generating meshes that are potentially dynamic.…”