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Glutathione-Conjugated Hydrogels: Flexible Vehicles for Personalized Treatment of Bacterial Infections

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Abstract

Purpose

Skin and soft tissue infections are increasingly prevalent and often complicated by potentially fatal therapeutic hurdles, such as poor drug perfusion and antibiotic resistance. Delivery vehicles capable of versatile loading may improve local bioavailability and minimize systemic toxicities yet such vehicles are not clinically available. Therefore, we aimed to expand upon the use of glutathione-conjugated poly(ethylene glycol) GSH-PEG hydrogels beyond protein delivery and evaluate the ability to deliver traditional therapeutic molecules.

Methods

PEG and GSH-PEG hydrogels were prepared using ultraviolet light (UV)-polymerization. Hydrogel loading and release of selected drug candidates was examined using UV-visible spectrometry. Therapeutic molecules and GST-fusion protein loading was examined using UV-visible and fluorescent spectrometry. Efficacy of released meropenem was assessed against meropenem-sensitive and -resistant P. aeruginosa in an agar diffusion bioassay.

Results

For all tested agents, GSH-PEG hydrogels demonstrated time-dependent loading whereas PEG hydrogels did not. GSH-PEG hydrogels released meropenem over 24 h. Co-loading of biologic and traditional therapeutics into a single vehicle was successfully demonstrated. Meropenem-loaded GSH-PEG hydrogels inhibited the growth of meropenem-sensitive and resistant P. aeruginosa isolates.

Conclusion

GSH ligands within GSH-PEG hydrogels allow loading and effective delivery of charged therapeutic agents, in addition to biologic therapeutics.

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Notes

  1. No correction for the GSH consumption of PEGDA end groups was used similar to previous publications (23).

Abbreviations

AR:

Antibiotic resistances

CAMHB:

Cation-adjusted Mueller-Hinton cation-adjusted Mueller-Hinton broth

C d :

Donor cell drug concentration

CDC:

Center for Disease Control and Prevention

CLSI:

Clinical and Laboratory Standards Institute

C n :

Characteristic ratio

D i :

Diffusion coefficient

GSH:

Glutathione

GSH-PEG:

Glutathione-conjugated polyethylene glycol

GST:

Glutathione-S-transferase

I :

Ionic strength

IMF:

Intermolecular force

j ss :

Flux of drug through membrane at steady state

K i :

Partition coefficient

l :

Bond length along the polymer backbone

L :

Membrane thickness

LMS:

Low molecular weight salt

m 0 :

Mass of the dried hydrogel

M c :

Molecular weight between crosslinks

m d,a :

Mass of dry hydrogel in air

m i :

Initial relaxed hydrogel mass

MIC:

Minimum inhibitory concertation

M n :

Number average molecular weight

M r :

Molecular weight of the repeating unit

m r,b :

Mass of relaxed hydrogel in butanol

MRSA:

methicillin-resistant Staphylococcus aureus

m s,b :

Mass of swollen hydrogel in butanol

m t :

Swollen mass at a given time

p :

Statistical p value

PBS:

Phosphate buffered saline

PEG:

Poly(ethylene glycol)

PEGDA:

Poly(ethylene glycol) diacrylate

P i :

Permeability coefficient

q :

Hydrogel mass swelling ratio

SSTI:

Skin and soft tissue infection

t lag :

Lag time to steady state conditions

UV:

Ultraviolet

v :

Specific volume of the polymer

V1 :

Molar volume of the swelling agent, water

Δm:

Change in mass (%)

ν2,r :

Polymer volume fraction in the relaxed state

ν2,s :

Polymer volume fraction in the swollen state

ξ:

Mesh size of the hydrogel

ρb :

Density of 1-butanol

ρp :

Density of polymer

χ:

Flory polymer-solvent interaction factor

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Acknowledgements and Disclosures

The authors would like to acknowledge and thank Alec C. Thompson for his help in preparing hydrogels, Shitalben R. Patel for her help conducting the microbiology experiments, and Dr. Zackery P. Bulman for generous sharing of instrumentation and feedback. The authors also thank Catherine F. Dial and Timothy D. Langridge for their support and feedback during the development of this work. Karol Sokolowski: Methodology; Conceptualization; Investigation; Formal Analysis; Writing-Original Draft. Hai M. Pham: Investigation; Formal Analysis; Writing-Review & Editing. Eric Wenzler: Methodology; Formal Analysis; Supervision; Writing-Review & Editing. Richard Gemeinhart: Methodology; Conceptualization; Formal Analysis; Supervision; Writing-Review & Editing; Funding Acquisition. This work was supported in part by the W.C. and May Preble Deiss Fund for Biomedical Research (to K.S.). This investigation was conducted in a facility constructed with support from Research Facilities Improvement Program Grant Number C06 RR15482 from the National Center for Research Resources, NIH. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The datasets generated during and/or analyzed during the current study are available from the corresponding authors on reasonable request. E.W. serves on the speaker’s bureau for Melinta Therapeutics, Astellas Pharma, and Allergan Plc., and on the advisory board for GenMark Diagnostics and Shionogi. All other authors certify no potential conflicts of interest.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Illinois at Chicago, Chicago, Illinois, USA

    Karol Sokolowski, Hai M. Pham & Richard A. Gemeinhart

  2. Department of Pharmacy Practice, University of Illinois at Chicago, Chicago, Illinois, USA

    Eric Wenzler

  3. Department of Bioengineering, University of Illinois at Chicago, Chicago, Illinois, USA

    Richard A. Gemeinhart

  4. Department of Chemical Engineering, University of Illinois at Chicago, Chicago, Illinois, USA

    Richard A. Gemeinhart

  5. Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, USA

    Richard A. Gemeinhart

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  1. Karol Sokolowski
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  2. Hai M. Pham
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  4. Richard A. Gemeinhart
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Correspondence to Eric Wenzler or Richard A. Gemeinhart.

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Supplementary Information

ESM 1

Supplemental chemical schema and results include hydrogel swell behavior in isotonic PBS, agar-well diffusion data against carbapenem-susceptible P. aeruginosa, representative images of hydrogels, and physical characteristics of slab-cut hydrogels. Supplementary data to this article can be found online at TBD. (PDF 1128 kb)

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Sokolowski, K., Pham, H.M., Wenzler, E. et al. Glutathione-Conjugated Hydrogels: Flexible Vehicles for Personalized Treatment of Bacterial Infections. Pharm Res 38, 1247–1261 (2021). https://doi.org/10.1007/s11095-021-03057-1

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