Scanning electron microscopy-based quantification of keratin and hyaluronic acid microstructure in electrospun scaffolds

Abstract Background The extracellular matrix (ECM) structural deficiencies in chronic wounds prevent the wounds from healing through natural physiological processes. Electrospun biocompatible polymers offer a platform to produce microstructure wound dressing materials that mimic the ECM containing various bioactives to address the deficiencies in the chronic wound healing process. Quantitative characterization of the electrospun fiber microstructure could provide valuable information on using fiber constructs to facilitate wound healing. This work employed a validated image analysis tool to quantitatively explain various parameters for the microstructure of six electrospun fiber constructs, D1{Polycaprolactone (PCL), Polyvinyl alcohol (PVA), Keratin}, D2{PCL, PVA, keratin, Co-enzyme Q10 (CoQ10)}, D3 (PCL, PVA, keratin, mupirocin), D4 (PCL, PVA, keratin, CoQ10, mupirocin, valsartan), D5 {PVA, Hyaluronic acid (HA)}, and D6 (PVA), using scanning electron microscopy imaging modality. Results The fiber intersection density (FID) parameter was quantified in the formulations, e.g., 0.272% for D5 and 0.416% for D4. Orientation histograms for D1 and D6 are characteristic of isotropic materials, while orientations for D2 and D3 indicate anisotropy with 2 preferred orientations in each formulation. D4 and D5 present orientations characteristic of transversely isotropic materials. The tortuosity for D2 and D4 indicates almost straight fiber segments, in contrast with undulated fiber segments in all other formulations. Furthermore, the mean fiber diameter was quantified, e.g., 1.414 and 1.630 mm for D3 and D4, respectively. Conclusion Co-electrospun PVA/PCL microfibers offer great potential for controlled delivery of bioactives needed to accelerate the healing of chronic wounds. This image-based analysis technology quantitatively characterized different formulations of electrospun fiber scaffolds. This analysis sets the stage for future study that utilizes microstructural information in finite element biomechanical modeling, to investigate possible influence of structure-based mechanical factors on the ECM restorative potential of wound dressings. Adjustment of electrospinning conditions could produce fabricated constructs like the native ECM structural components with a functional role in wound healing.