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Tissue transglutaminase regulates chondrogenesis in mesenchymal stem cells on collagen type XI matrices

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Abstract

Tissue transglutaminase (tTG) is a multifunctional enzyme with a plethora of potential applications in regenerative medicine and tissue bioengineering. In this study, we examined the role of tTG as a regulator of chondrogenesis in human mesenchymal stem cells (MSC) using nanofibrous scaffolds coated with collagen type XI. Transient treatment of collagen type XI films and 3D scaffolds with tTG results in enhanced attachment of MSC and supports rounded cell morphology compared to the untreated matrices or those incubated in the continuous presence of tTG. Accordingly, enhanced cell aggregation and augmented chondrogenic differentiation have been observed on the collagen type XI-coated poly- (L-lactide) nanofibrous scaffolds treated with tTG prior to cell seeding. These changes implicate that MSC chondrogenesis is enhanced by the tTG-mediated modifications of the collagen matrix. For example, exogenous tTG increases resistance to collagenolysis in collagen type XI matrices by catalyzing intermolecular cross-linking, detected by a shift in the denaturation temperature. In addition, tTG auto-crosslinks to collagen type XI as detected by western blot and immunofluorescent analysis. This study identifies tTG as a novel regulator of MSC chondrogenesis further contributing to the expanding use of these cells in cartilage bioengineering.

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Abbreviations

BSA:

Bovine serum albumin

Ca2+ :

Calcium

CO2 :

Carbon dioxide

cont-tTG:

Collagen type XI films continuously treated with tTG

pre-TG:

Collagen type XI films pre-treated with tTG

DSC:

Differential scanning calorimetry

DMEM:

Dulbecco’s modified eagle medium

EDTA:

Ethylenediaminetetraacetic acid

MSC:

Human mesenchymal stem cells

HCl:

Hydrochloric acid

PFA:

Paraformaldehyde

PBS:

Phosphate buffered saline

PLLA-coll XI:

PLLA scaffold coated with collagen type XI

coll XI-pre tTG:

PLLA-coll XI pre-treated with tTG

coll XI-cont tTG:

PLLA-coll XI in the continuous presence of tTG

PLLA:

Poly (L-lactide)

GAG:

Sulphated glycosaminoglycan

3D:

Three- dimensional

tTG:

Tissue transglutaminase

2D:

Two-dimensional

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Acknowledgments

We would like to thank Dr. Kimberly Griswold, of Picatinny Arsenal, NJ for DSC measurements. Also, Dr. Michael Jaffe and Dr. George Collins of New Jersey Institute of Technology, NJ for helpful discussions on DSC data. This work was supported by NIH grants R56DK071920 and R03AR057126 and a grant from Maryland Stem Cell Research Fund to M. Nurminskaya.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    Shobana Shanmugasundaram & Maria Nurminskaya

  2. Department of Anatomy and Cell Biology, Tufts University School of Medicine, Boston, MA, 02111, USA

    Sheila Logan-Mauney

  3. Department of Biology, Towson University, Towson, MD, 21252, USA

    Kaitlin Burgos

Authors
  1. Shobana Shanmugasundaram
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  2. Sheila Logan-Mauney
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  3. Kaitlin Burgos
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  4. Maria Nurminskaya
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Corresponding author

Correspondence to Maria Nurminskaya.

Additional information

S. Shanmugasundaram, S. Logan-Mauney contributed equally.

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Shanmugasundaram, S., Logan-Mauney, S., Burgos, K. et al. Tissue transglutaminase regulates chondrogenesis in mesenchymal stem cells on collagen type XI matrices. Amino Acids 42, 1045–1053 (2012). https://doi.org/10.1007/s00726-011-1019-7

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