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Supplementary Materials
Table S1. Constituents of the Vehicles used for NP penetration study.
Eucerin Smoothing Essentials (Fast Absorbing Lotion for Dry Skin)
Beiersdorf Inc. (Based in Germany)
Ingredients: Gluco-Glycerol Enhanced Formula
Water, Glycerin, Caprylic/Capric Triglyceride, Cetearyl Alcohol, Urea, Hydrogenated Cocoglycerides,
Isopropyl Stearate, Octyldodecanol, Sodium Lactate, Dimethicone, Arginine HCL, Glyceryl Stearate SE,
Myristyl Myristate, Carnitine, Chondrus Crispus (Carrageenan), Sodium Cetearyl Sulfate, Lactic Acid,
Sodium Citrate, Citric Acid, Acrylates/C10–30 Alkyl Acrylate Crosspolymer, Phenoxyethanol, Benzyl
Alcohol, Propylparaben, Potassium Sorbate.
Eucerin Calming (Itch Relief Treatment)
Beiersdorf Inc. (Based in Germany)
Ingredients: Active Ingredient: Menthol
Inactive Ingredients: Water, Glycerin, Octyldodecanol, Caprylic/Capric Triglyceride, Isopropyl Stearate,
Myristyl Myristate, Cetyl Alcohol, Colloidal Oatmeal, Cetearyl Alcohol, Dimethicone, Glyceryl Stearate SE,
Oenothera Biennis (Evening Primrose) Oil, PEG-40 Castor Oil, Caprylyl Glycol, , Sodium Cetearyl Sulfate,
Sodium Citrate, Citric Acid, Polyglyceryl-3 Methyl Glucose Distearate, Benzyl Alcohol, Phenoxyethanol,
DMDM Hydantoin, Potassium Sorbate.
Eucerin Daily Protection SPF 15 Sunscreen Lotion
Beiersdorf Inc. (Based in Germany)
Ingredients: Active Ingredients (sunscreens): Homosalate 5.0%, Octinoxate 5.0%, Octisalat 4.0%, Titanium
Dioxide 1.0%.
Other Ingredients: Water, Glycerin, Butylene Glycol, Cetearyl Alcohol, Urea, C18-36 Acid Triglyceride,
Caprylic/Capric Triglyceride, Dimethicone, Octyldodecanol, Sodium Lactate, Arginine HCL, Tocopheryl
Acetate, Glyceryl Stearate SE, Chondrus Crispus (Carrageenan), PEG-40 Castor Oil, Sodium Cetearyl
Sulfate, Disodium EDTA, Sodium Citrate, Lactic Acid, Carbomer, Citric Acid, Trimethoxycaprylysilane,
Benzyl Alcohol, Phenoxyethanol, Methylparaben, Propylparaben, Potassium Sorbate.
Vaseline Intensive Rescue Clinical Therapy Lotion
Unilever
Ingredients: Water, Glycerin, Petrolatum, Stearic acid, Glycol stearate, Dimethicone, Isopropyl isostearate,
Dihydroxypropyltrimonium chloride, Hydroxyethyl urea, Tapioca starch, Cetyl alcohol, Glyceryl stearate,
Magnesium aluminum silicate, Stearamide amp, Carbomer, Isopropyl myristate, Cedrol, triethanolamine,
Disodium edta, Phenoxyethanol, Ethylparaben, Propylparaben.
Dermovan
Healthpoint Ltd
Ingredients: Water, Glyceryl stearate (and) stearanidoethyl diethylamine, glycerin, mineral oil, cetyl esters,
cetyl alcohol, butylparaben, methylparaben, propylparaben.
a
b
c
Figure S1. Comparison between the exposure models show that the mouse skin swells up after a 24
h duration in the Franz diffusion cell. (a) Mouse skin placed in the Franz Chamber for swelled up
after a 24 h exposure. (b) Mouse skin removed from the Franz chamber after a 24 h exposure shows
that the portion of the skin exposed to the bottom chamber with the circulating media swells up.
(c) No gross physical changes were observed in the mouse skin treated using the petri dish protocol.
Cosmetics 2016, 3, 6; doi:10.3390/cosmetics3010006
www.mdpi.com/journal/cosmetics
Cosmetics 2016, 3, 6
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Vaseline
0 Hours after
application
Eucerin Smoothing Lotion
24 Hours after
application
0 Hours after
application
Glycerol
Dermovan
24 Hours after
application
0 Hours after
application
24 Hours after
application
24 Hours after
application
0 Hours after
application
Figure S2. Samples under a hand-held UV Lamp. Mouse skin samples were photographed under a
hand-held UV lamp immediately after and 24 h after the application of the vehicle with QDs in the
Petri dish protocol. The vehicles do not affect the QD intensity in the 24 h duration. Lowest QD
penetration was observed in the Vaseline group after confocal imaging, however it can be observed
that QD intensity is intact in this vehicle. Pictures are not to scale.
Human Skin-Control
Mouse Skin-Control
Total Intensity (QD 605)
1.40
1.20
1.00
0.80
0.60
0.40
0.20
0.00
0
5
10
15
20
25
30
35
40
Depth from Stratum Corneum (µm)
Figure S3. No autofluorescence was measured in untreated ex vivo mouse and human skin using
CLSM. No autofluorescence was measured in the two skin types using the QD605 filter.
Cosmetics 2016, 3, 6
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a
Total Intensity (QD 605) between 0-40 µm
9000
*
8000
7000
6000
5000
4000
3000
2000
*
1000
*
*
*
*
0
Eucerin
Dermovan
Smoothing
Lotion
Glycerol
Eucerin
Calming
Itch Relief
Eucerin
SPF15
Lotion
Vaseline
Intensive
Care
Water
Only LotionNo QD
b
Total Intensity (QD 605) between 0-40 µm
40000
35000
*
30000
25000
20000
15000
10000
5000
*
0
Eucerin
Smoothing
Lotion
Dermovan
Glycerol
Water
Only Lotion-No
QD
Figure S4. Eucerin Smoothing lotion enhances QD penetration in ex vivo mouse skin using the petridish protocol. (a) Integrated QD fluorescence signal in the Eucerin smoothing lotion group was 2 fold
higher than water. QD penetration in the Dermovan group was comparable to water. QD presence
between 0 and 40 µ m depth in all other vehicle groups was significantly lower compared to water
alone; (b) Integrated QD fluorescence signal in the Eucerin smoothing lotion group was 2.2 fold higher
than water. QD penetration in the Dermovan and glycerol test groups was comparable to water.
Means ± SEM (N = 4, n = 3). * p < 0.05 vs. Water. Lotion with no QD is the Control.
Figure S5
Cosmetics 2016, 3, 6
a
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Side View-Section
Top View-Stack
630 um
Epidermis
Epidermis
Epidermis
40 um
Dermis
Dermis
Dermis
b
3
1
2
iv
i
630
µm
ii
iii
v
40 µm
40 µm
c
Injection QD 605
Diffusion QD 605
600000
Total Intensity (QD605)
500000
400000
300000
200000
100000
0
0
5
10
15
20
25
30
35
40
Depth from the Stratum Corneum (µm)
45
50
Figure S5. Injection and diffusion of QDs in ex vivo human skin demonstrate that epidermal scattering
and absorption generate a decay in the intensity profile with an increase in depth from the stratum
corneum. (a) QDs were either injected intradermally into ex vivo human skin or the skin was immersed
in a QD solution for 24 h prior to CLSM imaging. The top view was obtained using the stacks from 0
to 50 µ m. Skin was cryosectioned and imaged to obtain the side profile views.
(b) 1 & 2. Cryosection of human skin showing high QD presence at the periphery of the tissue. The
QD diffusion front produced over the 24 h immersion was sufficient to yield homogenous QD
presence to depth exceeding 40 µ m. (b) 3. Cryosection of human skin showing high QD presence in
the dermis after a sub-dermal injection. Scale Bars (i) 50 µ m (ii) 120 µ m (iii) 170 µ m (iv) 40 µ m (v) 150
µ m. (c) Stacks obtained using CLSM were quantified using ImageJ. Similar trends were observed for
both exposure conditions with intensity values peaking around 10 µ m followed by a subsequent
decay in the intensity profile; Means ± SEM (N = 4, n = 3).
Cosmetics 2016, 3, 6
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3 Hours
6 Hours
24 Hours
3500
Total Intensity (QD 605)
3000
2500
2000
1500
1000
*
500
0
0
5
10
15
20
25
30
35
40
Depth from Stratum Corneum (µm)
Figure S6. Less QD presence is observed in the ex vivo mouse skin treated using the petri dish protocol
at 3 h compared to a 6 and 24 h exposure. Significantly lower QD presence was observed in mouse
skin treated for 3 h compared to 24 h. There were no significant differences between the 6 and 24 h
exposure group; Means ± SEM (N = 4, n = 3). * p < 0.05 vs. 24 h time point.
3 Hours
6 Hours
24 Hours
1400
*
Total Intensity (QD 605)
1200
1000
*
800
600
400
*
200
0
0
5
10
15
20
25
30
35
40
Depth from Stratum Corneum (µm)
Figure S7. Less QD presence is observed in the ex vivo mouse skin treated using the Franz diffusion
cell at 24 hours compared to a 3 and 6 h exposure. Significantly higher QD presence was observed in
mouse skin treated for 3 and 6 h compared to 24 h. There were no significant differences between the
3 and 6 hour exposure group; Means ± SEM (N = 4, n = 3). * p < 0.05 vs. 24 h time point.
Cosmetics 2016, 3, 6
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a
Franz Chamber Protocol 24 Hours
Petri Dish Protocol 24 Hours
b
Epidermal Thickness (µm)
30.00
*
*
25.00
20.00
15.00
10.00
5.00
0.00
Control Skin
Petri-Dish 24 Franz Chamber
(No Treatment) Hour Exposure
24 Hour
Exposure
Figure S8. Quantification of epidermal thickness of mouse skin using ImageJ. (a) Mouse skin was
exposed to the vehicle Eucerin Smoothing lotion and GSH QDs using the Petri dish protocol and
Franz diffusion chamber. 24 h after exposure the vehicle was wiped off and the skin was
cryosectioned. Brightfield images obtained at 20× magnification were used to quantify the epidermal
thickness in the two exposure protocols. (b) The epidermal thickness of skin exposed to the vehicle
using the petri dish protocol was significantly lower than the skin exposed to the lotion using the
Franz diffusion chamber and was of similar thickness to control skin—measured immediately posteuthanasia—no treatment. * p < 0.05, 2-talied t-Test, unpaired with unequal variances. Means ± SEM
(N = 3, n = 3).
Cosmetics 2016, 3, 6
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Human Skin (Rest)
Franz Diffusion Cell
Total Intensity (QD 605) between 0-40 µm
30000
Human Skin (Rest)
Petri Dish Protocol
*
25000
20000
15000
10000
5000
0
Eucerin Smoothing
Lotion
Glycerol
Water
Only Lotion-no QD
Figure S9. Comparable QD presence was observed in ex vivo human skin (rest t = 24 h) in the Eucerin
smoothing lotion and water test groups treated using the Franz diffusion cell and petri dish protocol.
No significant differences were observed in the two treatment protocols in the Eucerin smoothing
lotion and water test groups. A significantly higher QD presence (1.7 fold) was observed in skin
treated using the petri dish protocol compared to the Franz diffusion cell in the glycerol treatment
group; Means ± SEM (N = 4, n = 3). * p < 0.05.