RESUMO
Pectin polysaccharides have significant potential as all-natural, non-toxic "green" coatings that exhibit thermally-cued swelling behavior. Herein, ultra-thin coatings of highly-esterified pectin polysaccharides were cross-linked with calcium chloride (CaCl2) and their swelling in water was investigated with ellipsometry. At low temperatures, the coatings swell to 2-3 times their dry layer thickness. As the temperature is increased, the coatings show a pronounced decrease in swollen thickness, reminiscent of the hydrophilic-hydrophobic transition observed in lower critical solution temperature (LCST) polymers. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy establishes that this transition is driven by dehydration of the esterified galacturonic acid residues along the pectin backbone. By adjusting both the CaCl2 concentration used to crosslink the pectin coatings as well as pH of swelling medium, the pectin coatings could be judiciously tuned for a desired swelling response as a function of temperature. Due to their non-toxic and responsive nature, it was further demonstrated that such coatings could be used in applications to control cell adhesion.
Assuntos
Materiais Revestidos Biocompatíveis/química , Pectinas/química , Polissacarídeos/química , Animais , Reagentes de Ligações Cruzadas/química , Fibroblastos/citologia , Camundongos , Células NIH 3T3 , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Rational design of bioactive tissue engineered scaffolds for directing bone regeneration in vivo requires a comprehensive understanding of cell interactions with the immobilized bioactive molecules. In the current study, substrates possessing gradient concentrations of immobilized peptides were used to measure the concentration-dependent proliferation and osteogenic differentiation of human bone marrow stromal cells. Two bioactive peptides, one derived from extracellular matrix protein (ECM), GRGDS, and one from bone morphogenic protein-2 (BMP-2), KIPKASSVPTELSAISTLYL, were found to synergistically enhance cell proliferation, up-regulate osteogenic mRNA markers bone sialoprotein (BSP) and Runt-related transcription factor 2, and produce mineralization at densities greater than 130 pmol cm(-2) (65 pmol cm(-2) for each peptide). In addition, COOH-terminated self-assembled monolayers alone led to up-regulated BSP mRNA levels at densities above 200 pmol cm(-2) and increased cell proliferation from day 3 to day 14. Taking further advantage of both the synergistic potentials and the concentration-dependent activities of ECM and growth-factor-derived peptides on proliferative activity and osteogenic differentiation, without the need for additional osteogenic supplements, will enable the successful incorporation of the bioactive species into biorelevant tissue engineering scaffolds.