Cross-linked electrospun gelatin nanofibers for cell-based assays

Abstract

The present study evaluates the crosslinking of electrospun gelatin nanofibers by physical and chemical methods to further elucidate the importance of the application of gelatin scaffold platforms for cell-based assays. The dehydrothermally cross-linked electrospun gelatin scaffolds were unable to retained their structure morphology and integrity upon exposure to 1X PBS or cell-culture media. The DHT and EDC/Sulfo-NHS cross-linked gelatin scaffolds exhibited fiber diameter on average in the nanometer range. Subsequently, we utilized 1X PBS and cell culture media to evaluate the stability of the nanofibers in solution. The immersion evaluation indicated that the chemically cross-linked gelatin nanofibers maintained their random nanofiber distribution and morphology. However, a high degree of swelling was observed in the presence of cell culture media. Overall, the gelatin scaffold demonstrated good performance in PBS and cell culture media. Hence, EDC/Sulfo-NHS cross-linked electrospun gelatin nanofibrous scaffolds have good biocompatibility and are promising bio-scaffolds for cell-based assays.

Fabrication, Characterization, and Biological Evaluation of Airbrushed Gelatin Nanofibers

“…In recent years, several papers related to the preparation of gelatin nanofibers using electrospinning method have been reported. ,,, There is no report of successful preparation of gelatin nanofibers from commercially available animal origin (porcine) gelatin using the airbrush/solution-blow spinning method. Very recently, Liu et al.…”

“…Cells on the TCPS and nanofiber mats were fixed (4% paraformaldehyde at RT for 15 min), permeabilized (with Triton X-100 solution for 10 min), and stained using phalladoin and DAPI and finally washed with PBS (pH 7.4). Stained samples were imaged using a fluorescence microscope (Carl Zeiss Axio Observer Z1). , …”

Electrospun hydrogels for dynamic culture systems: advantages, progress, and opportunities

“…100 Another effective method to stabilize collagen/gelatin-based fibers leverages carbodiimide chemistry, such as EDC/NHS crosslinking, to introduce 'zero-length' crosslinks. [101][102][103] Chemical crosslinking has also been used to stabilize nanofibers formed from synthetic hydrophilic materials, 104 for example using glutaraldehyde to crosslink polyacrylamide (PA) 105 and poly(vinyl alcohol) (PVA). [106][107][108] Glutaraldehyde as a crosslinker readily reacts with pendant groups on PA and PVA to form linkages, and offers the potential to provide user-defined control over the stiffness and swelling of resultant electrospun fibers.…”

“…• Degree (extent) of chemical crosslinking 97 Gelatin Chemical: Biochemical: • Glutaraldehyde 98 and diisocyanate crosslinkers 100 • Gelatin provides natural bioactive sites for cell adhesion and interaction 96 • Carbodiimide crosslinking (EDC/NHS) 101,103 • Pendant methacrylates (chain-growth polymerization) [115][116][117] Physical:…”

Degradation and Mechanical Behavior of Fish Gelatin/Polycaprolactone AC Electrospun Nanofibrous Meshes

“…[10] Additives, such as rather expensive elastin and fibronectin, may be required to improve the scaffold elasticity, promote cell proliferation, and control the degradation rate. [11,12] Gelatin, a derivative of collagen, has more attractive properties [13,14] for a nanofibrous ECM mimic. Nanofibrous gelatin is naturally elastic, [14] stimulates cell proliferation, [15,16] and its biodegradation rate can be controlled by the degree of crosslinking.…”