Effects of cosmetics containing purified honeybee venom on acne vulgaris

Transcription

1 43 rd APIMONDIA 2013 Effects of cosmetics containing purified honeybee venom on acne vulgaris Sang Mi Han National Academy of Agricultural Science,

2 Collecting bee venom by electric shock device Bee venom collector made in Korea Placing the collector frames at entrance Collect for 30 min 2/25

3 Harvest bee venom Off wires and glass plates Dried bee venom on the glass Scraped off using a sharp scraperand collected bee venom 3/25

4 Purified bee venom Before purification After purification Commercial purified bee venom 4/25

5 Purified bee venom - Electron microscopy Before 5/75/13

6 Purified bee venom - Electron microscopy After 6/75

7 Results skin schematic Skin Schematic 7/25

8 Acne pathogenesis

9 Antibacterial effect-mic & EC 50 Bacteria MIC (µg /ml) EC 50 (µg /ml) P.acnes clindamycin-resistant P. acnes S.epidermidis S. pyrogenes /25

10 Time-kill curves for bee venom P. acnes clindamycin-resistant P. acnes S.epidermidis S. pyrogenes Control ( ), 2 MIC ( ), 10 MIC ( ) 10/25

11 CFU assay for bee venom Purified bee venom (PBV) has antimicrobial activity against P. acnes. (A) Various concentrations of PBV were incubated with P. acnes for 4 hr at 37 C and tested for antimicrobial activity using the CFU assay. (B) The antimicrobial activity of PBV in a qualitative CFU assay which involved plating PBV with P. acnes on solid media and counting individual colonies to determine the CFU.

12 Electron micrographs (A) SEM analysis demonstrated the effect of PBV on the surface of P. acnes as compared to the surfaces of untreated control bacteria. (B) TEM revealed that PBV treatment disrupted the sharply layered surface architecture of P. acnes, making the cell wall less distinct and more permeable to fluid influx. 12/32

13 Cytotoxicity in human cells human epidermal keratinocyte (HEK) human monocytic THP-1 Cytotoxicity of BV on human epidermal keratinocyte (HEK) and human monocytic (THP-1). Cells were cultured with the indicated concentrations of BV in media at 37 for 24 or 48 h. The viability of cells was determined by the MTT assay

14 Antiinflammation effects BV inhibits P.acnes induced secretion of proinflammatory cytokines. Doesdependent effect of BV on P. acnes-induced TNF-α release. THP-1 cells were stimulated with or without heat-killed P. acnes, and then the supernatants were harvested.

15 Antiinflammation effects PBV inhibits P.acnes induced secretion of proinflammatory cytokines. Doesdependent effect of BV on P. acnes-induced IL-8 release. THP-1 cells were stimulated with or without heat-killed P. acnes, and then the supernatants were harvested.

16 Skin safety Dermal toxicity Sex number Fluid of treatment (ml/kg) Concentration of BV (mg/kg) Vehicle M Control F T1 M F /25

17 Skin safety Skin irritation 17/25

18 Skin safety Eye irritation 18/25

19 Skin safety Toxic test in rats with single transdermal injection Skin irritation test in rabbits Skin sensitization test in guinea pigs Eye irritation test in rabbits Non-toxic Non-toxic Non-toxic Non-toxic OECD: Principles of Good Laboratory Practice 19/25

20 Sensory test & bee venom cosmetics Korean acne grading system Grade Description 1 Papules* 10 2 papules papules 31, nodules** 10 4 nodules 11-20, ± mild ongoing scars 5 nodules 21-30, ± moderate ongoing scars 6 nodules 30, ± severe ongoing scars ± sinus tract 20/25

21 Sensory test & bee venom cosmetics The change of Korean Acne Grading System (KAGS) based on numbers of inflammatory and noninflammatory lesions KAGS Control group (n=6) PBV group (n=6) Comparison of (grade) Week 0 Week 2 Week 0 Week 2 group, P value* P value** /25

22 Sensory test & bee venom cosmetics The average decrement of skin microorganism level (RLU) Class Control group (n=6) PBV group (n=6) Comparison of group P value* Week ± 3, ± 3,911 Week ± 2, ± 6, P value** The average decrement (%) 4.7% 57.5% 22/25

23 Sensory test & bee venom cosmetics BEFORE AFTER BEFORE AFTER [Subject: woman in her 20 s] Usage Period: 14 days Frequency: 3~4 times/day [Subject: man in his 20 s] Usage Period: 14 days Frequency: 3~4 times/day BEFORE AFTER BEFORE AFTER Usage Period: 14 days Frequency: 3~4 times/day [Subject: man in his 20 s] Usage Period: 21 days Frequency: 3~4 times/day 23/25

24 PBV benefits Helps treatment and prevention of acne Cumulative effect: Result build up over period of continual use This is because PBV has function of; Antibacterial Anti-inflammatory 24/25

25 Thank you!!!