Cell-matrix and cell-cell interactions of human gingival fibroblasts on three-dimensional nanofibrous gelatin scaffolds.
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-β1, 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.
Department of Biomedical Sciences, Baylor College of Dentistry, Texas A&M University Health Sciences Center, Dallas, TX, USA.
Ashneet
A
Sachar
T Amanda
TA
Strom
Symone
S
San Miguel
Maria J
MJ
Serrano
Kathy K H
KK
Svoboda
Xiaohua
X
Liu
D002450:Cell Communication / Q000187:drug effects*; D002452:Cell Count; D048430:Cell Shape / Q000187:drug effects; D021982:Cell-Matrix Junctions / Q000187:drug effects*; D024042:Collagen Type I / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D017629:Gap Junctions / Q000187:drug effects / Q000378:metabolism; D005780:Gelatin / Q000494:pharmacology*; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D057139:Nanofibers / Q000737:chemistry / Q000648:ultrastructure; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
D024042:Collagen Type I; D005780:Gelatin