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Title of Journal: Cell Tissue Res

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Abbravation: Cell and Tissue Research

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Springer Berlin Heidelberg

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DOI

10.1002/clc.4960060313

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ISSN

1432-0878

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Functional nucleus pulposuslike matrix assembly b

Authors: Michelle S Gupta Steven B Nicoll
Publish Date: 2014/08/05
Volume: 358, Issue: 2, Pages: 527-539
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Abstract

Intervertebral disc IVD degeneration is associated with several pathophysiologic changes of the IVD including dehydration of the nucleus pulposus NP Tissue engineering strategies may be used to restore both biological and mechanical function of the IVD following removal of NP tissue during surgical intervention Recently photocrosslinked carboxymethylcellulose CMC hydrogels were shown to support chondrogenic NPlike extracellular matrix ECM elaboration by human mesenchymal stromal cells hMSCs when supplemented with TGFβ3 however mechanical properties of these constructs did not reach native values Fabrication parameters ie composition crosslinking density can influence the bulk mechanical properties of hydrogel scaffolds as well as cellular behavior and differentiation patterns The objective of this study was to evaluate the influence of CMC macromer concentration 15 25 and 35  weight/volume on bulk hydrogel properties and NPlike matrix elaboration by hMSCs The lowest macromer concentration of 15  exhibited the highest gene expression levels of aggrecan and collagen II at day 7 corresponding with the largest accumulation of glycosaminoglycans and collagen II by day 42 The ECM elaboration in the 15  constructs was more homogeneously distributed compared to primarily pericellular localization in 35  gels The 15  gels also displayed significant improvements in mechanical functionality by day 42 compared to earlier time points which was not seen in the other groups The effects of macromer concentration on matrix accumulation and organization are likely attributed to quantifiable differences in polymer crosslinking density and diffusive properties between the various hydrogel formulations Taken together these results demonstrate that macromer concentration of CMC hydrogels can direct hMSC matrix elaboration such that a lower polymer concentration allows for greater NPlike ECM assembly and improvement of mechanical properties over timeThis work has been supported by the National Science Foundation CAREER Award CBET 0747968 and DMR 1207480 to SBN The IIII6B3 monoclonal antibody developed by T Linsenmayer 5C6 monoclonal antibody developed by E Engvall and XAC9 monoclonal antibody developed by TF Linsenmayer were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa Department of Biological Sciences Iowa City IA The authors would also like to thank Dr James Iatridis and Dr Devina Purmessur at the Mount Sinai School of Medicine and Dr Gerard Ateshian at Columbia University for helpful discussions

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