Abstract

Cardiac patch implantation helps maximize the paracrine function of grafted cells and serves as a reservoir of soluble proangiogenic factors required for the neovascularization of infarcted hearts. We have previously fabricated a cardiac patch, EF-HAM, composed of a human amniotic membrane (HAM) coated with aligned PLGA electrospun fibers (EF). In this study, we aimed to evaluate the biocompatibility and angiogenic effects of EF-HAM scaffolds with varying fiber thicknesses on the paracrine behavior of skeletal muscle cells (SkM). Conditioned media (CM) obtained from SkM-seeded HAM and EF-HAM scaffolds were subjected to multiplex analysis of angiogenic factors and tested on HUVECs for endothelial cell viability, migration, and tube formation analyses. All three different groups of EF-HAM scaffolds demonstrated excellent biocompatibility with SkM. CM derived from SkM-seeded EF-HAM 7 min scaffolds contained significantly elevated levels of proangiogenic factors, including angiopoietin-1, IL-8, and VEGF-C compared to plain CM, which was obtained from SkM cultured on the plain surface. CM obtained from all SkM-seeded EF-HAM scaffolds significantly increased the viability of HUVECs compared to plain CM after five days of culture. However, only EF-HAM 7 min CM induced a higher migration capacity in HUVECs and formed a longer and more elaborate capillary-like network on Matrigel compared with plain CM. Surface roughness and wettability of EF-HAM 7 min scaffolds might have influenced the proportion of skeletal myoblasts and fibroblasts growing on the scaffolds and subsequently potentiated the angiogenic paracrine function of SkM. This study demonstrated the angioinductive properties of EF-HAM composite scaffold and its potential applications in the repair and regeneration of ischemic tissues.

Highlights

  • Skeletal myoblasts are an attractive cell source for cellular cardiomyoplasty because of their autologous availability, high proliferative properties, and high tolerance to ischemia [1]

  • We aimed to investigate the proangiogenic potential of skeletal muscle cells (SkM)-seeded electrospun fibers (EF)-human amniotic membrane (HAM) scaffolds for the future treatment of ischemic tissue by studying their angiogenic paracrine effects in vitro

  • Cells growing on Electrospun fiber-coated human amniotic membrane (EF-HAM) scaffolds were attached to PLGA fibers and elongated along the alignment of the fibers, whereas cells growing on HAM were attached to the exposed extracellular matrix (ECM) and repopulated the membrane in a random orientation

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Summary

Introduction

Skeletal myoblasts are an attractive cell source for cellular cardiomyoplasty because of their autologous availability, high proliferative properties, and high tolerance to ischemia [1]. It has been suggested that the improvement in cardiac function observed following myoblast transplantation might be exerted through paracrine properties, rather than the replacement of damaged cardiac muscle with new contractile tissue [1]. Several strategies have been used to enhance the survival and engraftment efficiency of skeletal myoblasts in infarcted hearts. These strategies include transplantation of myoblast cell sheets or tissue-engineered scaffolds, genetic modification of skeletal myoblasts, and preconditioning of skeletal myoblasts prior to transplantation [7,8,9,10]. The use of myoblast cell sheets or tissue-engineered scaffolds may alleviate the risk of arrhythmia while facilitating cardiac functional recovery through consistent delivery of angiogenic, anti-apoptotic, and antifibrotic cytokines for a longer period of time

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