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The Effect of Glutathione as Chain Transfer Agent in PNIPAAm-Based Thermo-responsive Hydrogels for Controlled Release of Proteins

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ABSTRACT

Purpose

To control degradation and protein release using thermo-responsive hydrogels for localized delivery of anti-angiogenic proteins.

Methods

Thermo-responsive hydrogels derived from N-isopropylacrylamide (NIPAAm) and crosslinked with poly(ethylene glycol)-co-(L-lactic acid) diacrylate (Acry-PLLA-b-PEG-b-PLLA-Acry) were synthesized via free radical polymerization in the presence of glutathione, a chain transfer agent (CTA) added to modulate their degradation and release properties. Immunoglobulin G (IgG) and the recombinant proteins Avastin® and Lucentis® were encapsulated in these hydrogels and their release was studied.

Results

The encapsulation efficiency of IgG was high (75–87%) and decreased with CTA concentration. The transition temperature of these hydrogels was below physiological temperature, which is important for minimally invasive therapies involving these materials. The toxicity from unreacted monomers and free radical initiators was eliminated with a minimum of three buffer extractions. Addition of CTA accelerated degradation and resulted in complete protein release. Glutathione caused the degradation products to become solubilized even at 37°C. Hydrogels prepared without glutathione did not disintegrate nor released protein completely after 3 weeks at 37°C. PEGylation of IgG postponed the burst release effect. Avastin® and Lucentis® released from degraded hydrogels retained their biological activity.

Conclusions

These systems offer a promising platform for the localized delivery of proteins.

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Abbreviations

Acry-PEG-SVA:

Acryloyl-(ethylene glycol)-succinimidyl valerate

Acry-PLLA-b-PEG-b-PLLA-Acry:

Poly(ethylene glycol)-co-(L-lactic acid) diacrylate

AMD:

Age related macular degeneration

APS:

Ammonium persulfate

BME:

β-mercaptoethanol

CTA:

Chain transfer agent

DCM:

Dichloromethane

EDTA:

Ethylenediaminetetraacetic acid

IgG:

Immunoglobulin G

LCST:

Lower critical solution temperature

NIPAAm:

N-isopropylacrylamide

PEG:

Poly(ethylene glycol)

PLLA:

Poly(L-lactic acid)

PNIPAAm:

Poly(N-isopropylacrylamide)

TEA:

Triethylamine

TEMED:

N,N,N′,N′-tetramethylethylenediamine

VPTT:

Volume phase transition temperature

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ACKNOWLEDGMENTS AND DISCLOSURES

We would like to thank Chen Zhang and Michael Turturro for help with PEGylation and radiolabeling of IgG. This research was made possible through the funding provided by The Lincy Foundation, The Macula Foundation and the Veterans Administration.

Author information

Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Illinois Institute of Technology, 10 West 33rd Street, Chicago, Illinois, 60616-3793, USA

    Pawel W. Drapala & Victor H. Pérez-Luna

  2. Department of Biomedical Engineering, Illinois Institute of Technology, Chicago, Illinois, USA

    Bin Jiang, Yu-Chieh Chiu, Eric M. Brey & Jennifer J. Kang-Mieler

  3. Department of Research, Hines V.A. Hospital, Hines, Illinois, USA

    Eric M. Brey

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

    William F. Mieler

Authors
  1. Pawel W. Drapala
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  2. Bin Jiang
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  3. Yu-Chieh Chiu
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  4. William F. Mieler
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  5. Eric M. Brey
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  6. Jennifer J. Kang-Mieler
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  7. Victor H. Pérez-Luna
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Correspondence to Victor H. Pérez-Luna.

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Drapala, P.W., Jiang, B., Chiu, YC. et al. The Effect of Glutathione as Chain Transfer Agent in PNIPAAm-Based Thermo-responsive Hydrogels for Controlled Release of Proteins. Pharm Res 31, 742–753 (2014). https://doi.org/10.1007/s11095-013-1195-0

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  • DOI: https://doi.org/10.1007/s11095-013-1195-0

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