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Mesenchymal Stromal/Stem Cell and Minocycline-Loaded Hydrogels Inhibit the Growth of Staphylococcus aureus that Evades Immunomodulation of Blood-Derived Leukocytes

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Abstract

Mesenchymal stromal/stem cells (MSCs) have demonstrated favorable wound healing properties in addition to their differentiation capacity. MSCs encapsulated in biomaterials such as gelatin and polyethylene glycol (PEG) composite hydrogels have displayed an immunophenotype change that leads to the release of cytokines and growth factors to accelerate wound healing. However, therapeutic potential of implanted MSC-loaded hydrogels may be limited by non-specific protein adsorption that facilitates adhesion of bacterial pathogens such as planktonic Staphylococcus aureus (SA) to the surface with subsequent biofilm formation resistant to immune cell recognition and antibiotic activity. In this study, we demonstrate that blood-derived primary leukocytes and bone marrow-derived MSCs cannot inhibit colony-forming abilities of planktonic or biofilm-associated SA. However, we show that hydrogels loaded with MSCs and minocycline significantly inhibit colony-forming abilities of planktonic SA while maintaining MSC viability and multipotency. Our results suggest that minocycline and MSC-loaded hydrogels may decrease the bioburden of SA at implant sites in wounds, and may improve the wound healing capabilities of MSC-loaded hydrogels.

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Acknowledgments

The authors would like to thank the University of Wisconsin Carbone Cancer Center Experimental Pathology Laboratory for their assistance in MSC histology. This research was supported in part by the University of Wisconsin-Madison School of Pharmacy, the National Institutes of Health HL115482 grant, the National Institutes of Health Biotechnology Training Program HL115482 grant, and the University of Wisconsin Science and Medicine Graduate Research Scholars Fellowship.

Conflict of Interest

The authors indicate no potential conflicts of interest.

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Authors and Affiliations

  1. School of Pharmacy, Division of Pharmaceutical Sciences, Pharmacy Practice Division, University of Wisconsin-Madison, 777 Highland Avenue, 7123 Rennebohm Hall, Madison, WI, 53705, USA

    Alberto Daniel Guerra, David Antonio Cantu, Warren E. Rose & Weiyuan John Kao

  2. Department of Biomedical Engineering, College of Engineering, University of Wisconsin-Madison, Madison, WI, 53705, USA

    David Antonio Cantu, Joseph T. Vecchi & Weiyuan John Kao

  3. Department of Medicine, School of Medicine and Public Health, University of Wisconsin-Madison, 1685 Highland Avenue, Madison, WI, 53705, USA

    Peiman Hematti

  4. Department of Surgery, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI, 53705, USA

    Weiyuan John Kao

  5. University of Wisconsin Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI, 53705, USA

    Peiman Hematti & Weiyuan John Kao

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  1. Alberto Daniel Guerra
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  2. David Antonio Cantu
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  3. Joseph T. Vecchi
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  4. Warren E. Rose
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  5. Peiman Hematti
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  6. Weiyuan John Kao
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Corresponding author

Correspondence to Weiyuan John Kao.

Additional information

David Antonio Cantu contributed significantly to this work

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Supplementary Figure 1

Monocyte and MSC viability in the presence of 24 h established biofilm-associated SA after 12 h of co-culture at 200x magnification. Adherent monocyte viability in co-culture with (a): Gel-PEG-Cys encapsulated MSCs and biofilm-associated SA; (b): with biofilm-associated SA; (c): with collagen encapsulated MSCs and biofilm-associated SA. Gel-PEG-Cys encapsulated MSC viability in co-culture with (d): biofilm-associated SA; (e): polycarbonate-adherent monocytes and biofilm-associated SA. Collagen encapsulated MSC viability in co-culture with (f): biofilm-associated SA; (g): adherent monocytes and biofilm-associated SA (GIF 98 kb)

High Resolution image (TIFF 699 kb)

Supplementary Figure 2

Mature biofilm-associated SA Log CFU/mL after 6 h (black rectangle) and 12 h (gray rectangle) MSC-monocyte/macrophage co-culture. P < 0.05 versus SA-only condition for 6 h co-culture (Asterisk) and for 12 h co-culture (Dagger). (GIF 105 kb)

High Resolution image (TIFF 493 kb)

Supplementary Figure 3

Cytokine concentrations for planktonic SA (OD600 = 0.5) in co-culture with encapsulated MSCs and adherent PMNs after 4 h of co-culture. * indicates below the detection range of the bioplex protein quantification assay (GIF 265 kb)

High Resolution image (TIFF 1265 kb)

Supplementary Figure 4

MSC viability imaged with live/dead stain after different doses of minocycline over 7 days. MSCs maintained viability up to 0.2 mg/ml minocycline after 7 days exposure (GIF 514 kb)

High Resolution image (TIFF 968 kb)

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Guerra, A.D., Cantu, D.A., Vecchi, J.T. et al. Mesenchymal Stromal/Stem Cell and Minocycline-Loaded Hydrogels Inhibit the Growth of Staphylococcus aureus that Evades Immunomodulation of Blood-Derived Leukocytes. AAPS J 17, 620–630 (2015). https://doi.org/10.1208/s12248-015-9728-6

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  • DOI: https://doi.org/10.1208/s12248-015-9728-6

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