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.
8.
Budai L, Kaszás N, Gróf P, Lenti K, Maghami K, et al. (2013) Liposomes for Topical
Use: A Physico-Chemical Comparison of Vesicles Prepared from Egg or Soy
Lecithin. Scientia Pharmaceutica 81: 1151-1166.
12.
Asserhøj LL, Mosbech H, Krøigaard M, Garvey LH (2016) No evidence for contraindications
to the use of propofol in adults allergic to egg, soy or peanut †. British
Journal of Anaesthesia 116: 77-82.
14.
Sze SK, de Kleijn DP, Lai RC, Khia Way Tan E, et al. (2007) Elucidating
the secretion proteome of human embryonic stem cell-derived mesenchymal stem
cells. Molecular & Cellular Proteomics 6: 1680-1689.
16.
Maquart FX, Bellon G, Pasco S, Monboisse JC (2005) Matrikines
in the regulation of extracellular matrix degradation. Biochimie 87: 353-360.
17.
Ricard-Blum S, Salza R (2014) Matricryptins and matrikines:
biologically active fragments of the extracellular matrix. Experimental
Dermatology 23: 457-463.
18.
DeLouise LA (2012)
Applications of Nanotechnology in Dermatology. The Journal of investigative
dermatology132: 964-975.
19.
Brewer J, Bloksgaard M, Kubiak J, et al. (2013) Spatially
Resolved Two-Color Diffusion Measurements in Human Skin Applied to Transdermal
Liposome Penetration. Journal of Investigative Dermatology133: 1260-1268.
20.
Elmore SA (2006)
Enhanced Histopathology of the Spleen. Toxicologic Pathology 34: 648-655.
21.
Baliwag J, Barnes DH,
Johnston A (2015) Cytokines in psoriasis. Cytokine 73: 342-350.
22.
Cameron DJ, Tong Z, Yang Z, Kaminoh J, Kamiyah S, et al.
(2007) Essential role of Elovl4 in very long chain fatty acid synthesis, skin
permeability barrier function, and neonatal survival. International Journal of
Biological Sciences 3: 111-119.
23.
Ansari MNA, Nicolaides N,
Fu HC (1970) Fatty acid composition of the living layer and stratum corneum
lipids of human sole skin epidermis. Lipids 5: 838-845.
24.
Hara M, Ma T,
Verkman AS (2002) Selectively Reduced Glycerol in Skin of Aquaporin-3-deficient
Mice May Account for Impaired Skin Hydration, Elasticity, and Barrier Recovery.
Journal of Biological Chemistry 277: 46616-46621.
25.
Flutter B, Nestle FO (2013) TLRs to cytokines: Mechanistic
insights from the imiquimod mouse model of psoriasis. European Journal of
Immunology 43: 3138-3146.
26.
Sano S (2015) Psoriasis as
a barrier disease. Dermatologica Sinica 33: 64-69.
27.
Boguniewicz M, Leung DY (2011) Atopic Dermatitis: A Disease
of Altered Skin Barrier and Immune Dysregulation. Immunological Reviews 242: 233-246.
28.
Lavrijsen APM, Bouwstra JA, Gooris GS, Weerheim A, Boddé HE, et al. (1995) Reduced
Skin Barrier Function Parallels Abnormal Stratum Corneum Lipid Organization in
Patients with Lamellar Ichthyosis. Journal of Investigative Dermatology105: 619-624.
© by the Authors & Gavin Publishers. This is an Open Access Journal Article Published Under Attribution-Share Alike CC BY-SA: Creative Commons Attribution-Share Alike 4.0 International License. Read More About Open Access Policy.