New Three-Dimensional Poly(decanediol-co-tricarballylate) Elastomeric Fibrous Mesh Fabricated by Photoreactive Electrospinning for Cardiac Tissue Engineering Applications

Abstract

Reactive electrospinning is capable of efficiently producing in situ crosslinked scaffolds resembling the natural extracellular matrix with tunable characteristics. In this study, we aimed to synthesize, characterize, and investigate the in vitro cytocompatibility of electrospun fibers of acrylated poly(1,10-decanediol-co-tricarballylate) copolymer prepared utilizing the photoreactive electrospinning process with ultraviolet radiation for crosslinking, to be used for cardiac tissue engineering applications. Chemical, thermal, and morphological characterization confirmed the successful synthesis of the polymer used for production of the electrospun fibrous scaffolds with more than 70% porosity. Mechanical testing confirmed the elastomeric nature of the fibers required to withstand cardiac contraction and relaxation. The cell viability assay showed no significant cytotoxicity of the fibers on cultured cardiomyoblasts and the cell-scaffolds interaction study showed a significant increase in cell attachment and growth on the electrospun fibers compared to the reference. This data suggests that the newly synthesized fibrous scaffold constitutes a promising candidate for cardiac tissue engineering applications.

Keywords: biocompatibility; cardiac tissue engineering; particulate leaching; photo-crosslinking; poly(diol-tricarballylate); reactive electrospinning.

Figure 1

Schematic illustration of the PDET synthesis, acrylation, and further crosslinking to prepare (A) electrospun fibers (ESF) using UV-based photoreactive electrospinning (PRES) or (B) UV photocrosslinked scaffolds using the NaCl particulate leaching technique (NA).

Figure 2

FT-IR analysis of PDET-based polymer (5190 KDa). (A) Before acrylation (PDET), (B) after acrylation (APDET), and (C) after photoreactive electrospinning (crosslinked APDET based electrospun fibers). The two arrows correspond to the CH₂=CH₂ bands formed.

Figure 3

H-NMR Spectra of (A) PDET and (B) APDET. Black line represents the peaks with their corresponding chemical shifts. Red vertical lines represent the integrals of those peaks.

Figure 4

SEM micrographs of APDET scaffolds produced by particulate leaching using NaCl as a porogen. (A,B) Blank films, (C,D) Top view of porous APDET scaffolds, and (E,F) Sectional view of porous APDET scaffolds.

Figure 5

Morphological structure and analysis of ADPT/PVP electrospun fibers with a 2× acrylation degree at different PVP molecular weights. (Upper) SEM micrographs (A,B) with PVP 90 KDa, (C,D) with PVP 1300 kDa. (Lower) plot of (E) Fiber diameter and (F) Average pore size (E). Data were represented as mean ± SEOM (n = 5), * p < 0.05.

Figure 6

The effect of different synthesized scaffolds on H9C2 cell viability. Cells were incubated with both ESF and NA for 24 and 48 h and cell number was assessed by automated quantitation of DAPI positive nuclei using ArrayScan XTI (Target activation module). (Upper) the number of nuclei of viable cells represented as percentages relative to untreated control. Data presented as Mean ± SEOM, n = 6. Statistical significance: * p < 0.05 compared to the control. (Lower) Representative Images of the DAPI stained nuclei of viable cells after incubation with the scaffolds.

Figure 7

Qualitative assessment of cell/scaffold interaction in non-tissue culture treated plates. H9C2 cells were directly seeded on the scaffolds and incubated for 14 days in non-tissue culture treated plates. At day 14, cells were stained with Calcein-AM and representative images of H9C2 cells on ESF (top) and NA (bottom) were captured using a fluorescent microscope. Live cells appear as a fluorescent green color.