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Formulation and In Vitro-In Vivo Evaluation of Black Raspberry Extract-Loaded PLGA/PLA Injectable Millicylindrical Implants for Sustained Delivery of Chemopreventive Anthocyanins

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Abstract

Purpose

The objective of this study was to formulate and evaluate freeze-dried black raspberry (FBR) ethanol extract (RE) loaded poly(DL-lactic-co-glycolic acid) (PLGA) and poly(DL-lactic acid) (PLA) injectable millicylindrical implants for sustained delivery of chemopreventive FBR anthocyanins (cyanidin-3-sambubioside (CS), cyanidin-3-glucoside (CG) and cyanidin-3-rutinoside (CR)).

Methods

Identification and quantitation of CS, CG, and CR in RE was performed by mass spectroscopy and HPLC. RE:triacetyl-β-cyclodextrin (TA-β-CD) inclusion complex (IC) was prepared by a kneading method and characterized by X-ray diffraction (XRD), nuclear magnetic resonance spectroscopy (NMR) and UV-visible spectroscopy. RE or RE:TA-β-CD IC-loaded PLGA or PLA implants were prepared by a solvent extrusion method. In vitro and in vivo controlled release studies were conducted in phosphate-buffered saline Tween-80 (pH 7.4, 37°C) and after subcutaneous administration in male Sprague-Dawley rats, respectively. Anthocyanins were quantified by HPLC at 520 nm.

Results

The content of CS, CG, and CR in RE was 0.2, 1.5, and 3.5 wt%, respectively. The chemical stability of anthocyanins in solution was determined to be pH-dependent, and their degradation rate increased with an increase in pH from 2.4 to 7.4. PLGA/PLA millicylindrical implants loaded with 5 or 10 wt% RE exhibited a high initial burst and short release duration of anthocyanins (35–52 and 80–100% CG + CR release after 1 and 14 days, respectively). The cause for rapid anthocyanins release was linked to higher polymer water uptake and porosity associated with the high osmolytic components of large non-anthocyanin fraction of RE. XRD, 1H NMR and UV-visible spectroscopy indicated that the non-anthocyanin fraction molecules of RE formed an IC with TA-β-CD, decreasing the hydrophilicity of RE. Formation of an IC with hydrophobic carrier, TA-β-CD, provided better in vitro/in vivo sustained release of FBR anthocyanins (16–24 and 97–99% CG + CR release, respectively, after 1 and 28 days from 20 wt% RE:TA-β-CD IC/PLA implants) over 1 month, owing to reduced polymer water uptake and porosity.

Conclusion

PLA injectable millicylindrical implants loaded with RE:TA-β-CD IC are optimal dosage forms for 1-month slow and continuous delivery of chemopreventive FBR anthocyanins.

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ACKNOWLEDGEMENT

This study was supported by NIH R01 CA95901 and NIH R01 CA129609. We thank Dr. Scott Woehler, College of Pharmacy, University of Michigan, for the technical assistance with the NMR analysis.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Michigan, 428 Church St., Ann Arbor, USA

    Kashappa Goud H. Desai, Karl F. Olsen & Steven P. Schwendeman

  2. Department of Oral Maxillofacial Surgery and Pathology, College of Dentistry and the Comprehensive Cancer Center and Solove Research Institute, The Ohio State University, Columbus, Ohio, USA

    Susan R. Mallery

  3. Department of Internal Medicine, College of Medicine and the Comprehensive Cancer Center and Solove Research Institute, The Ohio State University, Columbus, Ohio, USA

    Gary D. Stoner

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  1. Kashappa Goud H. Desai
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Correspondence to Steven P. Schwendeman.

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Desai, K.G.H., Olsen, K.F., Mallery, S.R. et al. Formulation and In Vitro-In Vivo Evaluation of Black Raspberry Extract-Loaded PLGA/PLA Injectable Millicylindrical Implants for Sustained Delivery of Chemopreventive Anthocyanins. Pharm Res 27, 628–643 (2010). https://doi.org/10.1007/s11095-009-0038-5

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