26 MEN’S GROOMING
result of these initial inflammatory events. After invading the area, the immune cells start to play an important role in ‘amplifying’ the inflammatory cell–cell communication. Redness, edema, pain and itch occur as a consequence, and papules are developed. Additionally, many immune cells invading the area play an active role in the formation of what is known as pus. This is a key feature in the formation of pustules (Fig 4). LPM Complex was tested for its activity
on reducing sebum production and its ability to reduce the growth and biofilm forming potential of virulent strains of Cutibacterium acnes.
In vitro and microbiological assays Sebum production – gene expression of 5-reductase Elevation of sebum production is a result of the increased production of testosterone. Testosterone is converted to dihydrotestosterone (DHT) by 5- reductase. DHT then interacts with androgen receptors, eventually leading to the elevation of sebum production. The critical step in this process is the conversion of testosterone to DHT. Reduction of the presence of 5-reductase will reduce the likelihood that this conversion will take place. The reduction of 5-reductase production will therefore be beneficial in reducing sebum production. Human follicle dermal papilla cells (HFDPC) were treated with 0.01% LPM Complex for 24 hours. After that, total RNA was purified, quantified, and used to synthesize complementary DNA (cDNA). This cDNA from treated or untreated cells (control) served to determine the relative gene expression of the genes SRD5A1 (codes for 5-reductase type 1), SRD5A2 (type 2) and SRD5A3 (type 3) through RT-qPCR.
Results As shown in Figure 5, the expression of the genes of all three types of 5-reductase was reduced considerably. In the context of acne, these results suggest that LPM Complex can reduce the probability of the growth of virulent phylotypes of C. acnes, which can only thrive when sufficient sebum is produced.
Growth and biofilm forming activity Three virulent phylotypes of C. acnes, HM- 552 (HL110PA1), HM-492 (HL005PA1), HM- 523 (HL053PA2) and one “neutral” phylotype (ATCC 6919™) were analyzed. Studies were performed as follows.
Growth Tubes with test agent were inoculated with bacterial suspensions at a final concentration of 1000 CFU/ml and incubated at 37° C under anaerobic
PERSONAL CARE NORTH AMERICA Growth (Log increase at t = 24hrs vs t=0 HM-552 (HL110PA1)
Biofilm-forming potential at t=24hrs (Optical Density, OD590
)
HM-523 (HL053PA2)
HM-492 (HL005PA1)
ATCC 6919TM
-1
0
1
2
3
4 0.0
0.1
0.2
0.3
0.4
0.5
0.6
Figure 6: Reduction of growth and biofilm-forming ability of C. acnes phylotypes by LPM Complex. n Carrier control n 1% LPM Complex n 3% LPM Complex
conditions. Samples were taken at t = 0 and t = 24 hrs, processed and plated to obtain total viable counts. A carrier control included bacterial suspensions with 3% Tween 80 (used in other tubes to solubilize LPM Complex).
Biofilm formation Bacterial inocula were prepared to a cell density of 1 x 108 CFU/ml. A 96-well plate was prepared with individual wells containing 100 μl of test compound and carrier control. A total of 100 μl of bacterial inoculum was added to each test well. The well plates were then incubated at 37° C for 24 hrs. Subsequently, wells were processed to remove any non-adherent bacteria (bacteria in biofilm adhere). Finally, 2 % crystal violet (CV) dye solution was added and wells were rinsed and dried to remove unbound dye. Bound dye was solubilized and biofilm biomass was quantified with a microplate reader at an optical density of 595 nm (OD595). Background interference was negated by subtracting CV-stained blank wells from test absorbance values.
Results: Figure 6 shows that LPM Complex was able to halt the growth of the neutral phylotype and 2 out of 3 virulent phylotypes. Only the growth of HM-552 phylotype appeared to be more difficult to stop. Interestingly, the growth kinetics of the 4 phylotypes were relatively similar, indicating that the virulent phylotypes are not more active in
multiplying than the neutral phylotype. However, this was different when analyzing the biofilm-forming activity of the 4 phylotypes. Phylotypes HM-552 and HM- 523 showed to be much more active in producing biofilm than the other virulent and the neutral phylotype of C. acnes. This behavior nicely illustrates their virulence. The results obtained with LPM Complex showed interesting activity too. Whereas the growth of HM-552 was relatively hard to stop, its biofilm-forming activity was fully abolished with LPM Complex. Altogether, from these results it can be concluded that LPM Complex has clear benefits in reducing the microbial aspects of acne lesions, such as papules and pustules.
Conclusion The above results show that LPM Complex can provide the skin of the male skin care consumer with essential support in a daily skin care regime. Other in vitro and in vivo studies were performed with LPM Complex. These are not included in this article but illustrated the potency of LPM Complex. LPM Complex reduced the number of comedones, papules and pustules. Volunteers in the in vivo study also reported on LPM Complex’ perceivable and visible results, even improving Quality of Life of the test subjects. Being effective, natural and safe, LPM
Complex is an active ingredient which fits the needs of the modern-day male skin care consumer.
PC October 2020
‘Neutral’
Associated with acne
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