Effects of cosmetics containing purified honeybee venom on acne vulgaris

Sang Mi Han National Academy of Agricultural Science,

43 rd APIMONDIA 2013

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Bee venom collector made in Korea

Collect for 30 min

Placing the collector frames at entrance

Collecting bee venom by electric shock device

Dried bee venom on the glass

Off wires and glass plates

Scraped off using a sharp scraperand

collected bee venom

Harvest bee venom

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Purified bee venom

Before purification After purification Commercial purified bee venom

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Purified bee venom - Electron microscopy

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Before

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Purified bee venom - Electron microscopy

After

Results – skin schematic

Skin Schematic

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Acne pathogenesis

Antibacterial effect-MIC & EC50

Bacteria MIC (µg /ml) EC50 (µg /ml)

P.acnes 0.086 0.097

clindamycin-resistant P. acnes 0.067 0.086

S.epidermidis 0.104 0.156

S. pyrogenes 0.121 0.2

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Time-kill curves for bee venom

Control (●), 2 × MIC (▲), 10 × MIC (○)

P. acnes clindamycin-resistant P. acnes

S. pyrogenes S.epidermidis

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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.

CFU assay for bee venom

Electron micrographs

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(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.

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

Antiinflammation effects

BV inhibits P.acnes–induced secretion of proinflammatory cytokines. Does-dependent 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.

PBV inhibits P.acnes–induced secretion of proinflammatory cytokines. Does-dependent 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.

Antiinflammation effects

Skin safety

Sex number Fluid of treatment (ml/kg)

Concentration of BV (mg/kg)

Vehicle M 5 3 0

Control F 5 3 0

T1 M 5 3 1500

F 5 3 1500

Dermal toxicity

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Skin irritation

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Skin safety

Eye irritation

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Skin safety

Non-toxic

Skin sensitization test in guinea pigs

Eye irritation test in rabbits

Non-toxic

Non-toxic

Non-toxic

Toxic test in rats with single transdermal injection

Skin irritation test in rabbits

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OECD: “Principles of Good Laboratory Practice”

Skin safety

Sensory test & bee venom cosmetics

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Grade Description

1 Papules* ≤ 10

2 papules 11-30

3 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

Korean acne grading system

Sensory test & bee venom cosmetics

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The change of Korean Acne Grading System (KAGS) based on numbers of inflammatory and noninflammatory lesions

KAGS

(grade)

Control group (n=6) PBV group (n=6) Comparison of

group, P value* Week 0 Week 2 Week 0 Week 2

0 0 0 0 3

0.027 1 2 2 2 2

2 2 3 2 1

3 2 1 2 0

4 0 0 0 0

5 0 0 0 0

6 0 0 0 0

P value** 0.363 0.001

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 0 17.539 ± 3,202 17.825 ± 3,911

0.012 Week 2 16.708 ± 2,268 7.566 ± 6,082

P value** 0.448 0.007

The average decrement (%) 4.7% 57.5%

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Sensory test & bee venom cosmetics

[Subject: woman in her 20’s]

BEFORE AFTER

[Subject: man in his 20’s]

BEFORE AFTER

BEFORE AFTER

[Subject: man in his 20’s]

BEFORE AFTER

Usage Period: 14 days Frequency: 3~4 times/day

Usage Period: 21 days Frequency: 3~4 times/day

Usage Period: 14 days Frequency: 3~4 times/day

Usage Period: 14 days Frequency: 3~4 times/day

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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

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Thank you !!!