Abstract
The influence of polymer blend coatings on the differentiation of mouse mesenchymal stem cells was investigated. Polymer blending is a common means of producing new coating materials with variable properties. Stem cell differentiation is known to be influenced by both chemical and mechanical properties of the underlying scaffold. We therefore selected to probe the response of stem cells cultured separately on two very different polymers, and then cultured on a 1:1 blend. The response to mechanical properties was probed by culturing the cells on polybutadiene (PB) films, where the film moduli was varied by adjusting film thickness. Cells adjusted their internal structure such that their moduli scaled with the PB films. These cells expressed chondrocyte markers (osterix (OSX), alkaline phosphatase (ALP), collagen X (COL-X), and aggrecan (ACAN)) without mineralizing. In contrast, cells on partially sulfonated polystyrene (PSS28) deposited large amounts of hydroxyapatite and expressed differentiation markers consistent with chondrocyte hypertrophy (OSX, ALP, COL-X, but not ACAN). Cells on phase-segregated PB and PSS28 films differentiated identically to those on PSS28, underscoring the challenges of using polymer templates for cell patterning in tissue engineering.
Highlights
Controlled delivery of growth factors and cytokines has been proposed as adjunctive therapy for bone regeneration
Using the genetically engineered mouse mesenchymal stem cells (MSCs) line, C9 [15], where BMP-2 can be chemically regulated, we studied the response of the cells, first on Si wafers coated with films of PB or PSS28 separately, where we were able to determine that the different scaffolds had a profound influence on the cellular response to BMP-2 expression
To expand C9, cells were plated, directly onto the polymer coated Si substrates at a density of 2000/cm2 and cultured in “growth medium” composed of Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco) supplemented with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA), 2 mM l-glutamine, 100 U mL−1 penicillin, 100 μg/mL streptomycin (Invitrogen, Carlsbad, CA, USA), and 1 μg/mL doxycycline
Summary
Controlled delivery of growth factors and cytokines has been proposed as adjunctive therapy for bone regeneration. Additional treatment options for BMP-2-driven functions have been considered. One such option is the application of mesenchymal stem cells (MSCs). These cells can undergo chondrogenesis or osteogenesis dependent upon the environmental niche. Work carried out with a mouse embryonic MSC cell line (C3H10T1/2) demonstrated that when grown in micromass cultures chondrogenesis could be induced with BMP-2 [13]. In experiments using C9, a a cell line derived from C3H10T1/2 by genetically engineering to express rhBMP-2 in the absence of tetracyclines or its analogues (e.g., doxycycline), MSCs will undergo osteochondrogenesis in vitro and will support bone healing in vivo [14,15,16,17].
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