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Local Sustained Delivery of 25-Hydroxyvitamin D3 for Production of Antimicrobial Peptides

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Abstract

Purpose

This study seeks to develop fiber membranes for local sustained delivery of 25-hydroxyvitamin D3 to induce the expression and secretion of LL-37 at or near the surgical site, which provides a novel therapeutic approach to minimize the risk of infections.

Methods

25-hydroxyvitamin D3 loaded poly(L-lactide) (PLA) and poly(ε-caprolactone) (PCL) fibers were produced by electrospinning. The morphology of obtained fibers was characterized using atomic force microscope (AFM) and scanning electron microscope (SEM). 25-hydroxyvitamin D3 releasing kinetics were quantified by enzyme-linked immunosorbent assay (ELISA) kit. The expression of cathelicidin (hCAP 18) and LL-37 was analyzed by immunofluorescence staining and ELISA kit. The antibacterial activity test was conducted by incubating pseudomonas aeruginosa in a monocytes’ lysis solution.

Results

AFM images suggest that the surface of PCL fibers is smooth, however, the surface of PLA fibers is relatively rough, in particular, after encapsulation of 25-hydroxyvitamin D3. The duration of 25-hydroxyvitamin D3 release can last more than 4 weeks for all the tested samples. Plasma treatment can promote the release rate of 25-hydroxyvitamin D3. Human keratinocytes and monocytes express significantly higher levels of hCAP18/LL-37 after incubation with plasma treated and 25-hydroxyvitamin D3 loaded PCL fibers than the cells incubated with around ten times amount of free drug. After incubation with this fiber formulation for 5 days LL-37 in the lysis solutions of U937 cells can effectively kill the bacteria.

Conclusions

Plasma treated and 25-hydroxyvitamin D3 loaded PCL fibers induce significantly higher levels of antimicrobial peptide production in human keratinocytes and monocytes without producing cytotoxicity.

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Abbreviations

AFM:

Atomic force microscope

CFUs:

Colony-forming units

DAPI:

4′,6-diamidino-2-phenylindole

DCM:

Dichloromethane

D-MEM:

Dulbecco’s modified eagle medium

DMF:

N, N-dimethylformamide

DMSO:

Dimethylsulfoxide

ELISA:

Enzyme-linked immunosorbent assay

FBS:

Fetal bovine serum

FDA:

U.S. Food and Drug Administration

FITC:

Fluorescein isothiocyanate

LB:

Luria-Bertani

PBS:

Phosphate buffer solution

PCL:

Poly(ε-caprolactone)

PLA:

Poly(L-lactide)

RPMI:

Roswell Park Memorial Institute

SEM:

Scanning electron microscope

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ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported partially from startup funds from University of Nebraska Medical Center and National Institute of General Medical Science (NIGMS) grant 2P20 GM103480-06.

Author information

Authors and Affiliations

  1. Department of Surgery and Mary & Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, Omaha, Nebraska, 68198, USA

    Jiang Jiang & Jingwei Xie

  2. Bruker Nano Surface Division, Santa Barbara, California, 93117, USA

    Guojun Chen

  3. Department of Orthopaedic Surgery, Joan C. Edwards School of Medicine, Marshall University, Huntington, West Virginia, 25755, USA

    Franklin D. Shuler

  4. Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, 117585, Singapore

    Chi-Hwa Wang

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  1. Jiang Jiang
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  2. Guojun Chen
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  3. Franklin D. Shuler
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Correspondence to Jingwei Xie.

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Jiang, J., Chen, G., Shuler, F.D. et al. Local Sustained Delivery of 25-Hydroxyvitamin D3 for Production of Antimicrobial Peptides. Pharm Res 32, 2851–2862 (2015). https://doi.org/10.1007/s11095-015-1667-5

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  • DOI: https://doi.org/10.1007/s11095-015-1667-5

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