34 HAIR CARE
RPMI 1640 or DMEM medium completed with 10% FBS and 1% PS (penicillin streptomycin solution; Invitrogen, CA) at 37°C in a humidified CO2
Anti-inflammatory experiments on cells M. furfur stains ATCC 44344 (Beijing zhongkezhijian Biotechnology, China) were maintained in LAN medium.19
The cultures
were heat-killed by subjecting to temperatures of 85°C for 30 mins as an inducer of cellular inflammation on THP-1 and NHEK.20
Cells were
seeded in a 96-well plate at a density of 5 or 1×104 cells per well overnight. And then were treated with VGP (1% to 0.016% by two-fold dilution), with or without M. furfur stimulation for 24h.20-23
and MTT (Sigma, USA) assay.23,24 B
Cell viability was detected by CCK-8 MCP-1, IL-1β,
PGE-2 and IL-6 were determined with the corresponding ELISA kit (Neobioscience, China) using the culture medium.
Anti-inflammatory experiments on 3D reconstructed skin The M. furfur and H2
O2 (100 mM) was applied in
the epidermis of 3D reconstructed skin (Episkin Biotechnology, China) for 30 min, and then VGP with different concentration from 0.25% to 2.0% was added to the surface, cocultured with Episkin for another 42 h.25
After treatment, the skin biopsy
was measured by HE staining, and ELISA was used to detect the expression of cytokines (IL-8, LTB4 and PGE-2) in the supernatant. Cell viability was tested by MTT assay according to manual.26
Scalp care test in vivo A clinical study of 16 subjects was performed to evaluate the effects of VGP on scalp.27
In
recuperation period, vehicle shampoo was used to balance and maintain the condition of scalp in two weeks. During 4 week’s treatment, participants applied test shampoo (contained 0.5% VGP) thrice a week. Photos were taken and data were collected by Vapometer and Submeter at the following time points: baseline (T0w), recuperation (T2w), after 2 and 4 weeks of use (T4w and T6w). The redness and dandruff was valued by a dermatologist according to ASFS criterion, while the itching was evaluated by consumer through questionnaire survey.
Statistical analysis All experiments were repeated at least three times with different batches of cells. Data were evaluated statistically using Student’s t-test, and the statistical significance was set at P < 0.05.
Results Anti-inflammatory efficacy of VGP against M. furfur on cells The toxicity of VGP on THP-1 and NHEK cells were detected firstly. As shown in Table 1, it has toxicity
A (5%) chamber (Thermo Fisher Scientific, US).
500 400 300 200 100 0
80 60 40 20 0
C
*
** ** Control M.f 0.016 0.031 (VGP)% 0.063 0.125 Ref
** ** ** Control M.f 0.016 0.031 (VGP)%
80 60 40 20 0
0.063 ** 0.125 Ref
**
**
** ** Control M.f 0.016 0.031 (VGP)%
Figure 2: The effect of VGP on cytokines induced by M.furfur in THP1 cells. (* Significant differences compared to M.f group, P<0.05; ** P<0.01)
on both THP-1 and NHEK when above 0.25%, while the cell viability was reached to 80% under the concentration of 0.125%. Focusing on the inflammation stimulated by
M. furfuron NHEK cells, the expression of cytokines, such as IL-6 and PGE-2 were significantly up-regulated (Fig 1, M.f). When co- cultured with VGP from 0.016% to 0.125% for 24 h, the secretion of IL-6 was reduced from 36.8 to 18.9 pg/ml, PGE-2 from 1228.3 to 542.0 pg/ml as well. And the inhibitory effect was stronger when compared to dipotassium glycyrrhizinate (DG) at 0.125% (Fig 1, Ref). On THP-1 cells, the secretion of MCP-1, IL-1β
and PGE-2 were also promoted by M. furfur. As seen in Fig 2, the expression of MCP-1 was from
TABLE 1: EFFECT OF VGP ON THE VIABILITY OF TWO CELLS. Cell Viability
%
NHEK THP1
0
100.00±1.75 100.00±1.87
PERSONAL CARE June 2021 0.0625
95.19±3.10 96.07±0.78
w(VGP),% 0.125
88.31±3.75 84.23±0.50
92.9 to 368.4 pg/ml, IL-1β from 14.0 to 64.3 pg/ml and PGE-2 from 22.2 to 65.3 pg/ml after M. furfur stimulation (M.f). When treated with VGP from 0.016% to 0.125% for 24 h, the secretion of MCP-1, IL-1β and PGE-2 were down to 138.9, 9.7 and 26.9 pg/ml respectively, while the inhibitory effect of DG under 0.125% concentration was weaken than VGP, especially for MCP-1 and PGE-2. Results strongly supported VGP’s anti-inflammatory effect on epidermal keratinocytes and dermal monocytes.
VGP inhibited the inflammation and cell damage on reconstructed skin M. furfur cannot induce inflammation on Episkin model even in high concentration treated for 24 to 42 h (data not shown), while H2
O2 , a peroxide
often used in hair dyeing and perming, promoted the cell damage and secretion of cytokines, such as LTB4, PGE-2 and IL-8.28-30
In addition, these 0.25
30.58±1.32 29.21±1.10
0.5
5.68±0.27 20.66±1.47
inflammatory factors can act on various receptors on the c-type nerve fibres of the skin, transmitting itching and pain31-33
to cause various scalp
problems. Firstly, MTT and HE stain was used to verify the cell damage and structure. Cell viability
www.personalcaremagazine.com 0.063 0.125 Ref
PGE-2, ng,L-1
IL-1ẞ, ng,L-1
MCP-1, ng,L-1
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68 |
Page 69 |
Page 70 |
Page 71 |
Page 72 |
Page 73 |
Page 74 |
Page 75 |
Page 76 |
Page 77 |
Page 78 |
Page 79 |
Page 80 |
Page 81 |
Page 82 |
Page 83 |
Page 84 |
Page 85 |
Page 86 |
Page 87 |
Page 88 |
Page 89 |
Page 90 |
Page 91 |
Page 92
