chapter  Chapter 3
20 Pages

Growth of Mesenchymal Stem Cells on Surface-Treated 2d Poly(Glycerol-Sebacate) Bio-Elastomers of Varying Stiffness

WithRaju B. Maliger, Peter J. Halley, Justin J. Cooper-White, Donna Dinnes

Poly(glycerol-sebacate) bioelastomers are regarded as potential materials for tissue engineering applications. This paper deals with the effect of poly(glycerol-sebacate) substrate stiffness and surface treatment methods on the morphology and lineage of mesenchymal stem cells (MSC). Three types of PGS films of varying stiffness (glycerol: sebacic acid = 0.6, 0.8, 1.0) were subjected to aminolysis, hydrolysis, and multilayering (HA-Chitosan). Cell culture studies were performed on treated PGS substrates at four different time points (4 h, 24 h, 7 d, 14 d). The cell-seeded untreated, aminolyzed, hydrolyzed, and multilayered films were analyzed using fluorescence microscopy to investigate cell attachment, proliferation, and morphology. After removing unattached cells, cell lysates were centrifuged and the protein and DNA contents of the supernatants were quantified using Bradford and DNA assays, respectively. At different time-points, the adherent cells of treated PGS substrates (of varying stiffness) exhibited varying morphology. At the highest time-point (14 d) for this 44study, the PGS surfaces became more confluent with cells (~60–70%) on untreated and hydrolyzed surfaces and less confluent on aminolyzed and multilayered surfaces. Statistical analysis of DNA assay was performed using the Software SAS, and the significant variables were determined from it. Bradford protein assay at different time-points indicated that PGS surfaces did not appear to be cytotoxic towards the cells. For better attachment and proliferation of MSCs on the PGS substrates, the optimal treatment methods and time points required are assessed.