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Fluorinated Methacrylamide Chitosan Hydrogels Enhance Cellular Wound Healing Processes

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Abstract

Low availability of oxygen can lead to stalled wound healing processes and chronic wounds. To address local hypoxia and to better understand direct cellular benefits, a perfluorocarbon conjugated chitosan (MACF) hydrogel that delivers oxygen was created and applied for the first time to in vitro cultures of human dermal fibroblasts and human epidermal keratinocytes under both normoxic (21% O2) and hypoxic (1% O2) environments. Results revealed that local application of MACF provided 233.8 ± 9.9 mmHg oxygen partial pressure at 2 h and maintained equilibrium oxygen levels that were approximately 17 mmHg partial pressure greater than untreated controls. Cell culture experiments showed that MACF oxygenating gels improved cellular functions involved in wound healing such as cell metabolism, total DNA synthesis and cell migration under hypoxia in both fibroblasts and keratinocytes. Adenosine triphosphate (ATP) quantification also revealed that MACF treatments improved cellular ATP levels significantly over controls under both normoxia and hypoxia (p < 0.005). In total, these studies provide new data to indicate that supplying local oxygen via MACF hydrogels under hypoxic environments improves key wound healing cellular functions.

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Abbreviations

ATP:

Adenosine triphosphate

BSA:

Bovine serum albumin

DMEM:

Dulbecco’s modified eagle medium

dsDNA:

Double-stranded DNA

DTT:

Dithiothreitol

ECM:

Extracellular matrix

MAC:

Methacrylamide chitosan

MACF:

Fluorinated methacrylamide chitosan

nHDF:

Neonatal human dermal fibroblast

nHEK:

Neonatal human epidermal keratinocytes

NMR:

Nuclear magnetic resonance

PO2 :

Oxygen partial pressure

PBS:

Phosphate buffered saline

PFCs:

Perfluorocarbons

ROS:

Reactive oxygen species

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Acknowledgments

We would like to acknowledge funding from the National Institute of General Medical Sciences of the National Institutes of Health under award number R15GM104851. We would like to thank Steve Roberts for design and construction of the tri-gas incubator, Judy Fulton from Akron General Hospital/Serena Group for tissue donation and valuable feedback, as well as thank group members Ashley Wilkinson, Andrew McClain, Mahmoud Farrag, Pritam Patil and Trevor Ham for assistance with various techniques, biomolecular assays and interpretation.

Conflict of interest

Sridhar Akula, Ivy Brosch and Nic D. Leipzig declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, University of Akron, 200 East Buchtel Common, Akron, OH, 44325-3906, USA

    Sridhar Akula, Ivy K. Brosch & Nic D. Leipzig

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  1. Sridhar Akula
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  2. Ivy K. Brosch
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  3. Nic D. Leipzig
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Contributions

S.A. and N.D.L. designed the study, analyzed data, and wrote the manuscript. S.A. performed all experiments, while I.K.B. established techniques and performed initial experiments for PrestoBlue, PicoGreen, and microBCA experiments.

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Correspondence to Nic D. Leipzig.

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Associate Editor Michael S. Detamore oversaw the review of this article.

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Akula, S., Brosch, I.K. & Leipzig, N.D. Fluorinated Methacrylamide Chitosan Hydrogels Enhance Cellular Wound Healing Processes. Ann Biomed Eng 45, 2693–2702 (2017). https://doi.org/10.1007/s10439-017-1893-6

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  • DOI: https://doi.org/10.1007/s10439-017-1893-6

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