research article

Topical Application of Yolk Lecithin Liposomes Reinforces Skin Barrier Function Against Chemical Agents Such as Psoriasis-inducing IMQ and Alleviates Disease Phenotype

R.C. Lai1#, R.W.Y. Yeo2#, B. Zhang1#, M. Koh3, S.S. Tan4, W. K. Sim1, S. B. Ang5, Y.W.A Foong5, M. J. Ng6, K. H. Tan7 and S. K. Lim1,8*

1Institute of Medical Biology, A*STAR, 8A Biomedical Grove, 138648, Singapore

2Esco Regenerative Medicine Pte Ltd, Block 71 Ayer Rajah Crescent #07-26 s139951, Singapore

3Dermatology Service, KK Women’s and Children’s Hospital, 100 Bukit Timah Road, 229899, Singapore

4Vesiderm Pte Ltd, 8A Biomedical Grove, 138648, Singapore

5Family Medicine Service, KK Women’s and Children’s Hospital, 100 Bukit Timah Road, 229899, Singapore

6Division of Obstetrics & Gynaecology, KK Women’s and Children’s Hospital, 100 Bukit Timah Road, 229899, Singapore

7Department of Maternal Fetal Medicine, KK Women’s and Children’s Hospital, 100 Bukit Timah Road, 229899, Singapore

8Department of Surgery, YLL School of Medicine, NUS, 5 Lower Kent Ridge Road, 119074, Singapore

#Contributed equally

*Corresponding author: Sai Kiang Lim, Institute of Medical Biology, 8A Biomedical Grove, #05-505 Immunos,

138648, Singapore. Tel: +6564070150; +6564070161; Fax: +6564642048; Email: saikiang.lim@imb.a-star.edu.sg

Received Date: 23 May, 2018; Accepted Date: 31 May, 2018; Published Date: 08 June, 2018

Citation: Lai RC, Yeo RWY, Zhang B, Koh M, Tan SS, et al. (2018) Topical Application of Yolk Lecithin Liposomes Reinforces Skin Barrier Function Against Chemical Agents Such as Psoriasis-inducing IMQ and Alleviates Disease Phenotype. Clin Exp Dermatol Ther: CEDT-146. DOI: 10.29011/2575-8268/100046

1.       Abstract

1.1.  Background: Compromised skin barrier is a major driver of skin diseases such as psoriasis. We hypothesized that Yolk Lecithin Liposomes (YLLs) being water soluble bi-lipid membrane vesicles rich in skin barrier lipid and their precursors would be efficient in enhancing skin barrier. We tested this hypothesis by applying YLLs topically to Imiquimod (IMQ)-induced mouse model of psoriasis for evidence of skin barrier protection against IMQ and alleviation of the disease phenotype.

1.2.  Materials and Methods: 100 nm YLLs were first tested for effects on human MSC proliferation, collagen production in human dermal fibroblasts and penetration into human skin. They were then tested for efficacy in enhancing skin barrier against IMQ infiltration in a psoriasis mouse model. After 6 daily topical applications of IMQ and then a cream with YLLs or cream alone, barrier efficacy was assessed by the severity of psoriatic symptoms and cytokine induction in the skin lesion.

1.3.  Results: YLLs are non-toxic, and can enhance cell proliferation and collagen production. They readily permeated the stratum corneum. They alleviated psoriatic skin pathology and reduced IL-23 and TNF-a in the skin lesion of an IMQ-induced psoriasis mouse model.

1.4.  Conclusion: YLLs are effective in enhancing the skin barrier as evidenced by the alleviation of IMQ-induced psoriasis.

2.       Keywords: Liposome; Psoriasis; Skin barrier; Yolk lecithin



Figures 1 (A-B): YLLs are nano-sized lipid vesicles(A) Size distribution of YLLs as measured by Nanoparticle Tracking Analysis on a Zetaview over a size range of 5 to 1000 nm at 25°C. (B) YLLs as observed under TEM.



Figures 2(A-B): Cellular growth promotion and uptake of YLLs (A) (Left panel) Immortalized human mesenchymal stem cells (E1-MYC) were cultured in a Chemically Defined Medium (CDM) with increasing YLL concentration (mM PC) over 5 days. Cell proliferation was assessed by MTS assay. Data are presented as mean ± SD, * P< 0.05 as compared to control (0 mM PC) by Student’s t-test. (Right panel) Phase contrast images of cells treated with 0.025 mM PC and without YLLs (vehicle). Scale bar = 100 µm. (B) E1-MYC cells were incubated with Alexa Fluor 488-labeled YLLs for 0.5, 1, 2, and 4 hours. The control was cells that were not incubated with the labeled YLLs. The cells were analysed by flow cytometry. Histogram represents a plot of fluorescence intensity against number of events (cells).



Figures 3(A-C): YLLs increased collagen production in dermal fibroblasts(A) Phase contrast images of primary human dermal fibroblasts cultured in the presence of vehicle, 0.25 mM PC YLLs, 0.1% or 1% Matrixyl, fixed and stained with picro-sirius red solution to visualize collagen. Scale bar = 100 µm. (B) Collagen content per cell in each culture was normalized to that in untreated control. Collagen content per cell was determined by normalizing the amount of stain in each culture against cell numbers by MTS assay. Data are presented as mean ± SD(C) Cell numbers relative to untreated control, as estimated by MTS assay. Data are presented as mean ± SD



Figures 4(A-B): Topical application of YLLs on skin organ culture. Human skin organ culture was topically treated with PBS (Control) or Alexa Fluor 488-labeled YLLs, washed, frozen in OCT medium and sectioned. (A) H&E staining with arrowheads indicating position of stratum corneum. Scale bar = 100 µm. (B) Fluorescence imaging of sections counterstained with DAPI (blue). White broken lines denote epidermal-dermal junction. Scale bar = 50 µm.



Figures 5 (A-D): Topical application of YLLs on an IMQ-induced mouse model of psoriasis. IMQ was applied to the shaved backs and right ears of mice from day 0 to day 5. This was followed by the liposome cream, base cream (vehicle control), or 3 mg/Kg dexamethasone, i.p. (positive control). (A) Weight of mice over six days of study;(B) The spleen from each mouse was removed, weighed and the ratio of spleen to body weight for each mouse was calculated; (C) Erythema, scaling, and thickness in each mouse were determined daily on a scale from 0 to 4, and combined to generate the cumulative score; (D)The back skin of each mouse was harvested on day 6, homogenized and the homogenized solution was assayed for TNF-α, IL-17and IL-23 by ELISA. Data are presented as mean ± SE, * P< 0.05.

Ingredients

Liposome

Base

(% v/v)

Liposome solution in 10% (v/v) Ethanol (2.6 x 10E13 particles/mL)

50.00

0

10% (v/v) Ethanol

0

50

Distilled Water

33.25

33.25

Grape Seed Oil (Making Cosmetics, OIL-GRAP-01)

10.00

10.00

GelMaker EMU (Making Cosmetics, EMF-GEMK-01)

3.00

3.00

CreamMaker Wax (Making Cosmetics, EMF-EMWX-01)

1.00

1.00

Glycerin (Making Cosmetics, HUM-GLYC-01)

1.00

1.00

Dimethicone Satin (Making Cosmetics, ELL-DIMSAT-01)

1.00

1.00

Benzylalcohol-DHA (Making Cosmetics, PRV-BADHA-01)

0.50

0.50

Vitamin E (dl-alpha tocopherol) (Making Cosmetics, VIT-VITE-01, Activity 750 IU per 1 ml)

0.250

0.250

 

Supplementary Table 1: Composition of the liposome and base cream.

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Clinical & Experimental Dermatology and Therapies

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