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Characterization of a Microsphere Formulation Containing Glucose Oxidase and its In Vivo Efficacy in a Murine Solid Tumor Model

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Abstract

Purpose

This work focused on the characterization and in vitro/in vivo evaluation of an alginate/chitosan microsphere (ACMS) formulation of glucose oxidase (GOX) for the locoregional delivery of reactive oxygen species for the treatment of solid tumors.

Methods

The GOX distribution and ACMS composition were determined by confocal laser scanning microscopy and X-ray photoelectron spectroscopy. The mechanism of GOX loading and GOX-polymer interactions were examined with Fourier transform infrared spectroscopy and differential scanning calorimetry. In vitro cytotoxicity and in vivo efficacy of GOX-encapsulated ACMS (ACMS-GOX) were evaluated in EMT6 breast cancer cells and solid tumors.

Results

GOX was loaded into calcium alginate (CaAlg) gel beads via electrostatic interaction and the CaAlg-GOX-chitosan complexation likely stabilized GOX. Higher concentrations of GOX near the surface of ACMS were detected. GOX retained its integrity upon adsorption to CaAlg gel beads during the coating and after release from ACMS. ACMS-GOX exhibited cytotoxicity to the breast cancer cells in vitro and their efficacy increased with increasing incubation time. Intratumorally delivered ACMS-GOX significantly delayed tumor growth with much lower general toxicity than free GOX.

Conclusion

The results suggest that the ACMS-GOX formulation has the potential for the intratumoral delivery of therapeutic proteins to treat solid tumors.

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Abbreviations

ACMS:

Alginate/chitosan microspheres

ACMS-GOX:

GOX encapsulated in alginate/chitosan microspheres

CaAlg:

Calcium alginate

CLSM:

Confocal laser scanning microscope

DD:

Degree of deacetylation

DDI:

Distilled and deionized

DSC:

Differential scanning calorimetry

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

FR-GOX:

Free glucose oxidase

FT-IR:

Fourier transform infrared

GOX:

Glucose oxidase

i.t.:

Intratumoral

MDR:

Multidrug resistance

NaAlg:

Sodium alginate

ROS:

Reactive oxygen species

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

TGD:

Tumor growth delay

TPLD:

Tumor plus leg diameter

XPS:

X-ray photoelectron spectroscopy

α-MEM:

Alpha-minimal essential medium

η:

Intrinsic viscosity

ηsp :

Specific viscosity

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ACKNOWLEDGEMENT

This work was supported in part by the Canadian Institutes of Health Research. University of Toronto Open Fellowships and Ben Cohen Fund offered to Q. Liu are also acknowledged. The author would like to thank Dr. T. Chalikian for use of the lyophilizer, Dr. J. Uetrecht for use of the plate reader, Dr. R. Sodhi for XPS experiment and consultation, Mr. H. Huang for DSC analysis, and Mr. B. Calvieri for CLSM imaging analysis. The author would also like to thank Dr. R.Y. Cheung, Dr. H.L. Wong and Mr. A. Shuhendler for their generous help in cell culture work. The authors also thank Mr. Bob Kuba for assistance in animal work.

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Authors and Affiliations

  1. Leslie Dan Faculty of Pharmacy, University of Toronto, 144 College Street, Toronto, Ontario, M5S 3M2, Canada

    Qun Liu, Karlo Babakhanian, Xinyue Wang, Reina Bendayan & Xiao Yu Wu

  2. Division of Experimental Therapeutics, Ontario Cancer Institute, Toronto, Ontario, M5G 2M9, Canada

    Andrew Michael Rauth & Jiang Liu

Authors
  1. Qun Liu
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  2. Andrew Michael Rauth
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  3. Jiang Liu
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  7. Xiao Yu Wu
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Correspondence to Xiao Yu Wu.

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Liu, Q., Rauth, A.M., Liu, J. et al. Characterization of a Microsphere Formulation Containing Glucose Oxidase and its In Vivo Efficacy in a Murine Solid Tumor Model. Pharm Res 26, 2343–2357 (2009). https://doi.org/10.1007/s11095-009-9951-x

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  • DOI: https://doi.org/10.1007/s11095-009-9951-x

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