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Intestinal Uptake and Transport of Vitamin B12-loaded Soy Protein Nanoparticles

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Abstract

Background

Intestinal absorption of vitamin B12 (VB12) is a major challenge in combating pernicious anemia due to intrinsic factor (IF) deficiency.

Purpose

The aim of this study was to explore the feasibility of using soy protein isolates (SPI) nanoparticles to improve the intestinal transport and absorption of VB12.

Methods

Three different sized VB12-loaded SPI nanoparticles were produced by modulating preparation conditions using a cold-gelation method. The intestinal uptake and transport mechanisms of SPI nanoparticles for VB12 delivery were investigated and related to particle size.

Results

SPI nanoparticles were not cytotoxic to Caco-2 cells and were effectively internalized into the cytoplasm via multiple endocytosis pathways including clathrin- and/or caveolae-mediated endocytosis and macropinocytosis routes. VB12 transport across the Caco-2 cell monolayers was increased to 2–3 times after nanoencapsulation, which was dependent on particle size, in the increasing order of 30 > 100 > 180 nm. Using inhibitor block method, the transport of 30 and 100 nm SPI nanoparticles appeared to be clathrin-mediated transcytosis and macropinocytosis routes. The intestinal transport of VB12, assessed using rodent jejunum in Ussing chambers, was improved up to 4-fold after being encapsulated into 30 nm SPI nanoparticles.

Conclusions

The findings suggest that SPI nanoparticles could be a promising carrier to facilitate the oral delivery of VB12.

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Abbreviations

COL :

Colchicine

COUM-6 :

Coumarin 6

CPZ :

Chlorpromazine hydrochloride

CyD :

Cytochalasin D

DAPI :

4′, 6-Diamidino-2-phenylindole

DLS :

Dynamic light scattering

DMEM :

Dulbecco’s modified eagle medium

DMSO :

Dimethyl sulfoxide

FBS :

Fetal bovine serum

FI :

Fluorescence intensity

FLI :

Flilipin III

HBSS :

Hank’s balanced salt solution buffer

HEPES :

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

IF :

Intrinsic factor

LC :

Loading capacity

LE :

Loading efficiency

PBS :

Phosphate buffered saline

PLGA :

Poly(lactic-co-glycolic) acid

PMS :

Phenazine methosulfate

SPI :

Soy protein isolates

TEER :

Transepithelial electrical resistance

VB 12 :

Vitamin B12

VD 3 :

Vitamin D3

WGA :

Wheat germ agglutinin

XTT :

2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide

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ACKNOWLEDGMENTS AND DISCLOSURES

We are grateful to the Natural Sciences and Engineering Research Council of Canada (NSERC) for financial support and Canada Foundation for Innovation (CFI) for equipment support. Jing Zhang thanks the China Scholarship Council for providing a scholarship for her PhD program. The authors thank Mr. Jingzhou Huang for helping with Flow cytometry. The authors also thank Dr. Xuejun Sun and Ms. Geralidine Barron for their assistance with Confocal laser scan microscopy and Ms. Arlene Oatway for help with TEM.

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Authors and Affiliations

  1. Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, T6G 2P5, Canada

    Jing Zhang, Catherine J. Field, Donna Vine & Lingyun Chen

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  1. Jing Zhang
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  2. Catherine J. Field
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  3. Donna Vine
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  4. Lingyun Chen
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Correspondence to Lingyun Chen.

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Zhang, J., Field, C.J., Vine, D. et al. Intestinal Uptake and Transport of Vitamin B12-loaded Soy Protein Nanoparticles. Pharm Res 32, 1288–1303 (2015). https://doi.org/10.1007/s11095-014-1533-x

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  • DOI: https://doi.org/10.1007/s11095-014-1533-x

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