56 HAIR CARE
Nasturtium officinale Leaf/Stem Extract significantly increases the RSPO1 dose- dependently even with the presence of Dihydrotestosterone (DHT) which is considered a hair loss trigger in scalp hair. The addition of 1% Nasturtium officinale Leaf/Stem Extract to the cells increased RSPO1 secretion by 42% compared to the control supplemented with DHT alone, indicating that this extract, which enhances RSPO1 secretion, is useful as a treatment for androgenetic alopecia (AGA), one of the most common type of hair loss, since it is not affected by DHT.
Suppression of DKK1 secretion It has been documented that DHT induces DKK1 secretion from DP of hair follicles and that DKK1 is upregulated in the balding scalp compared with the haired scalp of patients with AGA.10 To examine the capability of Nasturtium officinale Leaf/Stem Extract of inhibiting DKK1 secretion, 0.25 and 0.5% extract were added to cultured human DP cells in the presence of DHT, after 3 days the supernatant was collected and the DKK1 protein amount was measured by using ELISA. The result shown in Figure 4 proves that DHT increases the DKK1 and that Nasturtium officinale Leaf/Stem Extract significantly suppresses the secretion of DKK1 which is related to the balding scalp of patients with AGA.
Hair follicle elongation ex vivo To substantiate the efficacy of Nasturtium officinale Leaf/Stem Extract an ex vivo test was conducted. HFs were collected from a female subject and cultured. On the next day, HFs were cultured in a control medium or medium containing 1% Nasturtium
1.5 * 1.0 1.0 0.5 0.5 0
Nasturtium officinale Leaf/Stem Extract
DHT
– + 0.5 1.0% – + + +
**p< 0.01, *p<0.05 vs. -/+ (Dunnett’s test). Values indicate the mean±SE.
Figure 3:Nasturtium officinale Leaf/Stem Extract promoting RSPO1 secretion. Cultured human dermal papilla cells were seeded on 96-well plate at a density of 10000 cells/well. After starving for one day, the culture medium was replaced with fresh medium containing Nasturtium officinale Leaf/Stem Extract and/or DHT. After 2 days, cell culture supernatant was collected and analysed by ELISA
PERSONAL CARE EUROPE 1.5 ** ** **
1.0
0.5
0
Wnt -3a DKK1 RSP01
– – –
+ – –
+ + –
+ + +
**p< 0.01 VS. -/+ (Dunnett’s test). Values indicate the mean±SE.
Figure 2: RSPO1 enhancing Wnt activity. Normal Human Epidermal Keratinocytes were seeded on 96-well plate at a density of 5000 cells/well. After starving for one day, the culture medium was replaced with fresh medium containing Wnt-3a, DKK1, and RSPO1. After 3 days, cell viability was measured using the CCK-8 (DOJINDO) assay.
Officinale Leaf/Stem Extract for 5 days, and the length of the HFs was measured. The picture of the HFs are shown in Figure 5. The elongation of HFs in the 1% Nasturtium Officinale Leaf/Stem Extract medium was measured at 758 μm up by 40% compared to the control HF elongation of 539 μm. Using immunofluorescence staining (data not shown) the RSPO1 secretion in treated hair was promoted and bonding with its receptor LGR5 which promotes outer root sheath cells self-renewal therefore 1% Nasturtium Officinale Leaf/Stem Extract enhanced the elongation of the HFs significantly compared to the control.
** 1.5 ** ** **
Human clinical trial Finally, a clinical trial to test the efficacy of the special processed Nasturtium officinale Leaf/Stem Extract on actual people with hair loss problems, a double blind and randomised controlled trial was conducted by a third party at a dermatological clinic in Bangalore, India. A total of 40 male volunteers aged 20 to 55 years with grade 3 to 6 hair pattern loss (MSCR Photonumeric linear scale,10 point scale) were divided into two groups (20 people per group); placebo group using a control hair lotion and the active group using a hair lotion with 2% Nasturtium officinale Leaf/Stem Extract. Both applied the hair lotion twice a day for
0
Nasturtium officinale Leaf/Stem Extract
DHT – – 0.25 0.50% – + + + **p< 0.01 vs. -/+ (Dunnett’s test). Values indicate the mean±SE.
Figure 4: Nasturtium officinale Leaf/Stem Extract suppression of DKK1 secretion .Cultured human dermal papilla cells were seeded on 96-well plate at a density of 10000 cells/well. After starving for one day, the culture medium was replaced with fresh medium containing Nasturtium officinale Leaf/Stem Extract and/or DHT. After 3 days, cell culture supernatant was collected and analysed by ELISA.
June 2020
RSP01 secretion
Cell proliferation
DKK1 secretion
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