Maximizing phenotype constraint and extracellular matrix production in primary human chondrocytes using arginine–glycine–aspartate concentration gradient hydrogels

New systematic approaches are necessary to determine and optimize the chemical and mechanical scaffold properties for hyaline cartilage generation using the limited cell numbers obtained from primary human sources. Peptide functionalized hydrogels possessing continuous variations in physico-chemical...

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Bibliographic Details
Published in:Acta biomaterialia 2013-07, Vol.9 (7), p.7420-7428
Main Authors: Smith Callahan, Laura A., Childers, Erin P., Bernard, Sharon L., Weiner, Scott D., Becker, Matthew L.
Format: Article
Language:English
Subjects:
actin
Cartilage
Cells, Cultured
chondrocytes
Chondrocytes - classification
Chondrocytes - drug effects
Chondrocytes - metabolism
Combinatorial methods
Dose-Response Relationship, Drug
extracellular matrix
Extracellular Matrix - metabolism
Gradient
Humans
hydrocolloids
Hydrogel
Hydrogels - chemistry
Materials Testing
Oligopeptides - chemistry
Oligopeptides - pharmacology
phenotype
physicochemical properties
polyethylene glycol
primary productivity
Tissue engineering
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Summary:New systematic approaches are necessary to determine and optimize the chemical and mechanical scaffold properties for hyaline cartilage generation using the limited cell numbers obtained from primary human sources. Peptide functionalized hydrogels possessing continuous variations in physico-chemical properties are an efficient three-dimensional platform for studying several properties simultaneously. Herein, we describe a polyethylene glycol dimethacrylate (PEGDM) hydrogel system possessing a gradient of arginine–glycine–aspartic acid peptide (RGD) concentrations from 0mM to 10mM. The system is used to correlate primary human osteoarthritic chondrocyte proliferation, phenotype maintenance and extracellular matrix (ECM) production to the gradient hydrogel properties. Cell number and chondrogenic phenotype (CD14:CD90 ratios) were found to decline in regions with higher RGD concentrations, while regions with lower RGD concentrations maintained cell number and phenotype. Over three weeks of culture, hydrogel regions containing lower RGD concentrations experience an increase in ECM content compared to regions with higher RGD concentrations. Variations in actin amounts and vinculin organization were observed within the RGD concentration gradients that contribute to the differences in chondrogenic phenotype maintenance and ECM expression.
ISSN:1742-7061
1878-7568
DOI:10.1016/j.actbio.2013.04.005