Differentiation of Rat Mesenchymal Stem Cells toward Osteogenic Lineage on Extracellular Matrix Protein Gradients

Soraya Rasi Ghaemi, Bahman Delalat, Alex Cavallaro, Agnieszka Mierczynska-Vasilev, Krasimir Vasilev, Nicolas H. Voelcker

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10 Citations (Scopus)


This report addresses the issue of optimizing extracellular matrix protein density required to support osteogenic lineage differentiation of mesenchymal stem cells (MSCs) by culturing MSCs on surface-bound density gradients of immobilized collagen type I (COL1) and osteopontin (OPN). A chemical surface gradient is prepared by tailoring the surface chemical composition from high hydroxyl groups to aldehyde groups using a diffusion-controlled plasma polymerization technique. Osteogenesis on the gradient surface is determined by immunofluorescence staining against Runx2 as an early marker and by staining of calcium phosphate deposits as a late stage differentiation marker. The Runx2 intensity and calcified area increase with increasing COL1 density up to a critical value corresponding to 124.2 ng cm−2, above which cell attachment and differentiation do not rise further, while this critical value for OPN is 19.0 ng cm−2. This gradient approach may facilitate the screening of an optimal biomolecule surface density on tissue-engineered scaffolds, implants, or tissue culture ware to obtain the desired cell response, and may generate opportunities for more cost-effective regenerative medicine.

Original languageEnglish
Article number1900595
Number of pages11
JournalAdvanced Healthcare Materials
Issue number17
Publication statusPublished - 1 Sept 2019


  • high throughput screening
  • mesenchymal stem cells
  • osteogenesis
  • protein gradients

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