Fabrication of Cell-Laden AME-Loaded Collagen-Based Hydrogel Promotes Fibroblast Proliferation and Wound Healing In Vitro

Document Type : Original Article


1 Department of Tissue Engineering, Faculty of Basic Sciences and Advanced Technologies in Medicine, Royan Institute, ACECR, Tehran, Iran

2 Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran

3 Department of Mathematics, Statistics, and Computer Science, Faculty of Science, University of Tehran, Tehran, Iran

4 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

5 Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

6 Department of Cell Engineering, Stem Cells and Developmental Biology, Cell Science Research Center, ACECR, Royan Institute, Tehran, Iran


Objective: The biological factors secreted from cells and cell-based products stimulate growth, proliferation, and
migration of the cells in their microenvironment, and play vital roles in promoting wound healing. The amniotic membrane
extract (AME), which is rich in growth factors (GFs), can be loaded into a cell-laden hydrogel and released to a wound
site to promote the healing of the wound. The present study was conducted to optimize the concentration of the
loaded AME that induces secretion of GFs and structural collagen protein from cell-laden AME-loaded collagen-based
hydrogels, to promote wound healing in vitro.
Materials and Methods: In this experimental study, fibroblast-laden collagen-based hydrogel loaded with different
concentrations of AME (0.1, 0.5, 1, and 1.5 mg/mL, as test groups) and without AME (as control group), were incubated
for 7 days. The total proteins secreted by the cells from the cell-laden hydrogel loaded with different concentrations of
AME were collected and the levels of GFs and type I collagen were assessed using ELISA method. Cell proliferation
and scratch assay were done to evaluate the function of the construct.
Results: The results of ELISA showed that the concentrations of GFs in the conditioned medium (CM) secreted from the
cell-laden AME-loaded hydrogel were significantly higher than those secreted by only the fibroblast group. Interestingly,
the metabolic activity of fibroblasts and the ability of the cells to migrate in scratch assay significantly increased in the
CM3-treated fibroblast culture compared to other groups. The concentrations of the cells and the AME for preparation
of CM3 group were 106 cell/mL and 1 mg/mL, respectively.
Conclusion: We showed that 1 mg/ml of AME loaded in fibroblast-laden collagen hydrogel significantly enhanced the
secretion of EGF, KGF, VEGF, HGF, and type I collagen. The CM3 secreted from the cell-laden AME-loaded hydrogel
promoted proliferation and scratch area reduction in vitro.


Main Subjects

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