Electrospun Fiber-Coated Human Amniotic Membrane: A Potential Angioinductive Scaffold for Ischemic Tissue Repair

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.

The Effect of Hydroxytyrosol in Type II Epithelial-Mesenchymal Transition in Human Skin Wound Healing

“…Then, the cell culture inserts were incubated in the dark with 1ml 4′,6-diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific, Carlsbad, CA, USA) at 1:15,000 dilution for each well for 40 min at room temperature, and then were washed thrice with PBS. Pictures of DAPI-stained nuclei at the bottom of the inserts were taken at five different fields using an immunofluorescence microscope (Nikon, Tokyo, Japan) before the numbers of nuclei were averaged [ 44 ].…”

Emerging Strategies for the Biofabrication of Multilayer Composite Amniotic Membranes for Biomedical Applications

“…Both synthetic and natural polymers were used as secondary materials to create electrospun multilayered composite AM (Table 1). The synthetic electrospun polymers were polycaprolactone (PCL) [74,84,86,88,89,[97][98][99], poly(lactic-co-glycolic acid) (PLGA) [90,91,97], poly(lactic acid) (PLA) [97], and poly-(Llactide-co-E-caprolactone) (PCLC) [85]. They were all biodegradable and biocompatible synthetic polymers.…”

“…They also reported that skeletal muscle cells seeded on AM-PLGA scaffolds were oriented along the alignment of the electrospun PLGA fibers, whereas cells seeded on AM displayed random orientation, thereby guiding the orientation and migration of skeletal muscle cells owing to their aligned topography. In a further study, they investigated the proangiogenic potential of the same skeletal muscle cell-seeded composite scaffolds according to three different thicknesses of PLGA fibers [91]. Thus, they compared in vitro the angiogenic paracrine effects of (i) these three skeletal muscle cell-seeded composite scaffolds, (ii) skeletal muscle cellseeded AM, and (iii) non-seeded skeletal muscle cells on human umbilical vein endothelial cells (HUVECs).…”