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Primary osteoblasts colonize and mineralize 3D nanofibrous gelatin scaffolds in 14 days
Author(s) -
Sachar Ashneet,
Strom Amanda,
Serrano Maria,
Miguel Symone San,
Liu Xiaohua,
Svoboda Kathy K.H.
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.917.2
Subject(s) - vinculin , osteoblast , chemistry , focal adhesion , paxillin , integrin , microbiology and biotechnology , extracellular matrix , gelatin , tyrosine phosphorylation , phosphorylation , cell , biology , biochemistry , in vitro
To enhance the outcome of bone tissue engineering, it is important to understand the interactions between osteoblasts and scaffolds. In this study, we analyzed and quantified cell‐matrix interactions and tissue formation by primary cultured osteoblasts on 3D nanofibrous gelatin scaffolds (3D‐NF‐GS) using confocal microscopy. 3D‐NF‐GS were cultured with osteoblasts for 5 or 14 days and immunolabeled with specific antibodies: Integrin b1, vinculin, tyrosine phosphorylated paxillin and collagen type I. A Leica Confocal SP5 microscope was used to analyze the protein distribution. Alizarin red staining was used for mineral detection. Osteoblasts on 3D‐NF‐GS exhibited uniform migration by 5 days and the cells proliferated to an increased confluence by the 14th day. They adhered to 3D‐NF‐GS via focal adhesions by 5 days demonstrated by integrin b1, vinculin and phosphorylated paxillin. Integrin b1 and phosphorylated paxillin adhesions increased in number by 14 days and vinculin adhesions turned more fibrillar. The osteoblasts started arranging into nodular aggregations by 14 days and abundant mineralization of 3D‐NF‐GS was documented. In conclusion, osteoblasts’ adhesion, migration and proliferation on 3D‐NF‐GS lead to secretion of the primary osteoid protein, type I collagen and matrix mineralization. This demonstrates favorable osteoblast behavior on this 3D matrix material. Grant Funding Source : NIH/NIDCR P30 DE020742