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Tacrolimus Loaded PEG-Cholecalciferol Based Micelles for Treatment of Ocular Inflammation

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Abstract

Purpose

Poor corneal permeability, nasolacrimal drainage and requirement of chronic administration are major drawbacks of existing therapies for ocular inflammation. Hence, we designed topical micelles of PEG2000 conjugated with cholecalciferol (PEGCCF).

Methods

Integrin targeted tacrolimus loaded PEGCCF micelles (TTM) were prepared by solvent diffusion evaporation method and characterized for particle size, osmolality, encapsulation efficiency and drug loading. Therapeutic potential of TTM was evaluated in benzalkonium chloride induced ocular inflammation model in BALB/c mice. Corneal flourescein staining and histopathological analysis of corneal sections was performed.

Results

TTM had a particle size of 45.3 ± 5.3 nm, encapsulation efficiency (88.7 ± 0.9%w/w) and osmolality of 292–296 mOsmol/Kg. TTM significantly reduced the corneal fluorescence as compared to tacrolimus suspension (TACS). H&E staining showed that TTM could restore corneal epithelial thickness, reduce stromal edema (p < 0.05) and decrease number of inflammatory cells (p < 0.01) compared with TACS. Immunohistochemistry analysis demonstrated lower expression of Ki67 + ve cells (p < 0.05) and IL-6 throughout the cornea against TACS (p < 0.01) and the control (p < 0.001).

Conclusions

TTM is an innovative delivery system for improving ocular inflammation due to a) integrin targeting b) PEGCCF in the form of carrier and c) anti-inflammatory and synergistic effect (due to Pgp inhibition) with TAC.

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Fig. 1: Characterization of TTM.
Fig. 2: Representative slit-lamp captured fluorescein staining and quantitative analysis
Fig. 3: Assessment of corneal integrity through H&E staining of mouse cornea after different treatments and quantitative analysis
Fig. 4
Fig. 5: Immunohistochemistry of mouse eye for Ki67 after different treatments and quantitative analysis
Fig. 6: Immunohistochemistry of mouse eye for IL-6 after different treatments and quantitative analysis

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Abbreviations

APC:

Antigen presenting cells

BKC:

Benzalkonium chloride

CRP:

C-reactive protein

DCC:

Dicyclohexylcarbodiimide

DMAP:

4-dimethylaminopyridine

DSPE-PEG2000:

1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]

HCEC:

Human corneal epithelial cells

IFN- γ:

Interferon-γ

IHC:

Immunohistochemistry

iOP:

Intraocular pressure

LC:

Langerhans cells

mPEG2000:

Polyethylene glycol methyl ether (average Mn~2000)

NFAT:

Nuclear factor of activated T cells

PDI:

Poly-dispersity index

PEG:

Polyethylene glycol

PEGCCF:

PEG2000 conjugated with cholecalciferol

Pgp:

P-glycoprotein

RGD:

Arg-Gly-Asp

SF:

Sodium fluorescein

TAC:

Tacrolimus

TACS:

Tacrolimus suspension

TEA:

Triethylamine

TNF-α:

Tumor necrosis factor

TTM:

Targeted tacrolimus loaded micelles

VD3:

Cholecalciferol

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Authors and Affiliations

  1. College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, Florida, 32307, USA

    Shallu Kutlehria, Imran Vhora, Arvind Bagde, Nusrat Chowdhury, Gautam Behl, Ketan Patel & Mandip Singh

  2. College of Pharmacy and Health Sciences, St. John’s University, Queens, New York, 11439, USA

    Ketan Patel

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  1. Shallu Kutlehria
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  2. Imran Vhora
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  3. Arvind Bagde
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Correspondence to Mandip Singh.

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Kutlehria, S., Vhora, I., Bagde, A. et al. Tacrolimus Loaded PEG-Cholecalciferol Based Micelles for Treatment of Ocular Inflammation. Pharm Res 35, 117 (2018). https://doi.org/10.1007/s11095-018-2376-7

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  • DOI: https://doi.org/10.1007/s11095-018-2376-7

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