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Cell–matrix and cell–cell interactions of human gingival fibroblasts on three‐dimensional nanofibrous gelatin scaffolds
Author(s) -
Sachar Ashneet,
Strom T. Amanda,
San Miguel Symone,
Serrano Maria J.,
Svoboda Kathy K. H.,
Liu Xiaohua
Publication year - 2014
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.1588
Subject(s) - focal adhesion , extracellular matrix , gelatin , confocal microscopy , confocal , chemistry , microbiology and biotechnology , 3d cell culture , vinculin , cell , biomedical engineering , matrix (chemical analysis) , micropatterning , cell adhesion , regeneration (biology) , cell culture , tissue engineering , materials science , nanotechnology , biology , biochemistry , medicine , geometry , mathematics , genetics , chromatography
Abstract An in‐depth understanding of the interactions between cells and three‐dimensional (3D) matrices (scaffolds) is pivotal to the development of novel biomaterials for tissue regeneration. However, it remains a challenge to find suitable biomimetic substrates and tools to observe cell–material and cell–cell interactions on 3D matrices. In the present study, we developed biomimetic nanofibrous 3D gelatin scaffolds (3D‐NF‐GS) and utilized confocal microscopy combined with a quantitative analysis approach to explore cell–matrix and cell–cell interactions on the 3D‐NF‐GS. Human gingival fibroblasts (HGFs) migrated throughout the 3D‐NF‐GS by 5 days and formed stable focal adhesions by 14 days. The focal adhesions were detected using integrin‐ β 1, phospho‐paxillin and vinculin expression, which were quantified from specific wavelength photon data generated using a spectral separation confocal microscope. As the cells became more confluent after 14 days of culture, cell–cell communication via gap junctions increased significantly. Collagen I matrix production by HGFs on 3D‐NF‐GS was visualized and quantified using a novel approach incorporating TRITC label in the scaffolds. Based on confocal microscopy, this study has developed qualitative and quantitative methods to study cell–matrix and cell–cell interactions on biomimetic 3D matrices, which provides valuable insights for the development of appropriate scaffolds for tissue regeneration. Copyright © 2012 John Wiley & Sons, Ltd.