Structural and functional characterization of proteins adsorbed on hydrophilized polylactide-co-glycolide microfibers

Abstract

Background: Hydrophobic biopolymers such as polylactide- co -glycolide (PLGA, 85:15) have been extensively explored as scaffolding materials for tissue engineering applications. More recently, electrospun microfiber-based and nanofiber-based scaffolds of PLGA have received increased attention because they act as physical mimics of the fibrillar extracellular matrix. However, the hydrophobicity of the PLGA microfiber surface can limit its use in biomedical applications. Therefore, in a previous study, we fabricated Pluronic ® F-108 (PF-108)-blended PLGA microfibrous scaffolds that alleviated the hydrophobicity associated with PLGA by enriching the surface of microfibers with the ethylene oxide units present in PF-108. Methods: In this study, we report the influence of the extent of surface enrichment of PLGA microfibers on their interaction with two model proteins, ie, bovine serum albumin (BSA) and lysozyme. BSA and lysozyme were adsorbed onto PLGA microfiber meshes (unmodified and modified) and studied for the amount, secondary structure conformation, and bioactivity of released protein. Results: Irrespective of the type of protein, PF-108-blended PLGA microfibers showed significantly greater protein adsorption and release than the unblended PLGA samples. However, in comparison with BSA, lysozyme showed a 7–9-fold increase in release. The Fourier transform infrared spectroscopy studies for secondary structure determination demonstrated that irrespective of type of microfiber surface (unblended or blended), adsorbed BSA and lysozyme did not show any significant change in secondary structure (α-helical content) as compared with BSA and/or lysozyme in the free powder state. Further, the bioactivity assay of lysozyme released from blended PLGA microfiber meshes demonstrated 80%–85% bioactivity, indicating that the process of adsorption did not significantly affect biological activity. Therefore, this study demonstrated that the decreased hydrophobicity of blended PLGA microfibrous meshes not only improved the amount of protein adsorbed (lysozyme and BSA) but also maintained the secondary structure and bioactivity of the adsorbed proteins. Conclusion: Modulating the hydrophobicity of PLGA via blending with PF-108 could be a viable strategy to improve its interaction with proteins and subsequent cell interaction in tissue engineering applications.

Heat-Treated Lysozyme Hydrochloride: A Study on Its Structural Modifications and Anti-SARS-CoV-2 Activity

“…Fourier transform infrared (FTIR) spectroscopy is widely used in the structural characterization of proteins or peptides. This study, applied to the amide I region between 1700 and 1600 cm −1 of the spectrum, is utilized in order to evaluate the secondary structure of proteins, as it enables the resolution of the amide peak into smaller component peaks, each representative of a different secondary structure [ 15 , 16 ]. In this region falls the larger contribution of C=O stretching vibrations of the amide group on the protein backbone.…”

Improved Neural Regeneration with Olfactory Ensheathing Cell Inoculated PLGA Scaffolds in Spinal Cord Injury Adult Rats

“…They tend to provide a better cell growth milieu in the conveyance of exogenous cells into the lesion site, promoting and supporting axonal growth towards the damaged neural cells or tissues [21]. However, the poor hydrophilicity of the PLGA complex coupled with the lack of cell surface recognition motifs [16, 22] can negatively influence its protein interaction and subsequent cellular behavior [22]. …”