Mesenchymal Stromal Cell Chondrogenic Differentiation Induced by Continuous Stiffness Gradient in Photocrosslinkable Hydrogels
Sabrina C. Mierswa
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Department of Biomedical Engineering, University of California, Davis, California, USA
Search for more papers by this authorErika E. Wheeler
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Department of Biomedical Engineering, University of California, Davis, California, USA
Search for more papers by this authorAyla N. Apsey
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Search for more papers by this authorOju Jeon
Department of Biomedical Engineering, University of Illinois at Chicago, Chicago, Illinois, USA
Search for more papers by this authorEben Alsberg
Department of Biomedical Engineering, University of Illinois at Chicago, Chicago, Illinois, USA
Department of Orthopaedic Surgery, University of Illinois at Chicago, Chicago, Illinois, USA
Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, Illinois, USA
Department of Mechanical and Industrial Engineering, University of Illinois at Chicago, Chicago, Illinois, USA
Jesse Brown Veterans Affairs Medical Center (JBVAMC), Chicago, Illinois, USA
Search for more papers by this authorCorresponding Author
J. Kent Leach
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Department of Biomedical Engineering, University of California, Davis, California, USA
Correspondence:
J. Kent Leach ([email protected])
Search for more papers by this authorSabrina C. Mierswa
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Department of Biomedical Engineering, University of California, Davis, California, USA
Search for more papers by this authorErika E. Wheeler
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Department of Biomedical Engineering, University of California, Davis, California, USA
Search for more papers by this authorAyla N. Apsey
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Search for more papers by this authorOju Jeon
Department of Biomedical Engineering, University of Illinois at Chicago, Chicago, Illinois, USA
Search for more papers by this authorEben Alsberg
Department of Biomedical Engineering, University of Illinois at Chicago, Chicago, Illinois, USA
Department of Orthopaedic Surgery, University of Illinois at Chicago, Chicago, Illinois, USA
Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, Illinois, USA
Department of Mechanical and Industrial Engineering, University of Illinois at Chicago, Chicago, Illinois, USA
Jesse Brown Veterans Affairs Medical Center (JBVAMC), Chicago, Illinois, USA
Search for more papers by this authorCorresponding Author
J. Kent Leach
Department of Orthopaedic Surgery, UC Davis Health, Sacramento, California, USA
Department of Biomedical Engineering, University of California, Davis, California, USA
Correspondence:
J. Kent Leach ([email protected])
Search for more papers by this authorFunding: This work was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health under award numbers R01 DE025899 (J.K.L., E.A.) and R01 AR079211 (J.K.L.). S.C.M. was supported by a National Institute of Arthritis and Musculoskeletal and Skin Diseases funded training program in Musculoskeletal Health Research (T32 AR079099) and the Floyd and Mary Schwall Fellowship in Medical Research. E.E.W. received support from the California Institute for Regenerative Medicine grant number EDUC4-12792 (CIRM EDUC4 Research Training Program).
Sabrina C. Mierswa and Erika E. Wheeler have shared first authorship.
ABSTRACT
Chondrogenic differentiation of stem and progenitor cells is dependent on the biophysical properties of the surrounding matrix. Current biomaterials-based approaches for chondrogenesis are limited to discrete platforms, slowing our ability to interrogate the role of mechanical cues such as substrate stiffness and other signals. Thus, novel platforms must incorporate a range of biophysical properties within a single construct to effectively assess changes in cell response. We encapsulated human mesenchymal stromal cells (MSCs) within biodegradable, photocurable oxidized, and methacrylated alginate (OMA). Cell-laden hydrogels were crosslinked when exposed to light through a grayscale photomask to form substrates with a continuous stiffness gradient. We also tested the influence of the adhesive ligand Arg-Gly-Asp (RGD) on chondrogenic differentiation. Compared to unmodified gels possessing uniform biophysical properties, RGD-modified OMA hydrogels with the same modulus promoted chondrogenic differentiation of MSCs as evidenced by gene expression, matrix deposition, and histological analysis. MSCs entrapped in OMA hydrogels exhibiting a biologically relevant stiffness gradient (2–13 kPa over 8 mm) demonstrated increased chondrogenic differentiation with increases in stiffness. MSC chondrogenic differentiation was dependent upon the ability to mechanosense the modulus of the surrounding matrix, confirmed by the addition of Latrunculin A (LatA), a soluble inhibitor of actin polymerization. These findings validate a methodology for customizing hydrogel platforms for chondrogenic differentiation and identifying the interplay of key variables to instruct cell function.
Conflicts of Interest
The authors declare no conflicts of interest.
Open Research
Data Availability Statement
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Supporting Information
Filename | Description |
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jbma37928-sup-0001-supinfo.docxWord 2007 document , 91.1 KB |
Figure S1. DNA content and metabolic activity in bulk and stiffness gradient hydrogels with and without LatA treatment. (A) DNA content in OMA versus RGD OMA round bulk hydrogels. (B) Metabolic activity measured via alamarBlue assay normalized to DNA in OMA versus RGD OMA bulk hydrogels. DNA content (C) and alamarBlue fluorescence normalized to DNA (D) in square bulk and stiffness gradient hydrogels without LatA treatment. DNA content (E) and alamarBlue fluorescence normalized to DNA (F) in square bulk and stiffness gradient hydrogels after LatA treatment. Data are mean ± SD (n > 3). Significance is determined by Student’s t-test with significance of p < 0.05 (A, B) and one-way ANOVA (C–F). Groups that are significantly different are denoted with different letters; ns = not significant. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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