Archives of
RDeermatological
Arch. Dermatol. Res. 264, 369-371 (1979)
searcn
9 Springer-Verlag 1979
Investigations on the Bacteriostatic Effect of Benzoyl Peroxide D. Fanta, H. Bardach, and Ch. Poitscheck 2nd Department of Dermatology, University of Vienna (Head: Prof. Dr. G. Niebauer), AlserstraBe 4, A-1090 Wien, Austria
Benzoyl peroxide (BP), a potent oxidizing agent, used for a long time by surgeons to improve wound healing and by dermatologists to treat ulcers and pyodermas, lately was also found to be effective in the management of acne vulgaris [1, 7, 8, 10]. The bacteriostatic effect is believed to be due to reduction of bacterial amino acids to pantothionic acid. The importance of this for the treatment of acne vulgaris was first recognized by Fulton [3], who not only found a significant reduction in the microbial flora of the follicular infundibulum following topical use of BP, but also a distinct decrease in free fatty acids as measured by infrared spectroscopy, the latter serving as a criterion for the presence of Propionibacterium acnes (P.a.). This was supported by G l o o r et al. [5], who performed thin layer chromatography of skin surface lipids. Moreover, local application of BP also produces a keratolytic effect, probably due to liberation, by contact with organic material, of benzoic acid, which like salicylic acid [6] causes lysis of the intercellular substance in the stratum corneum. In addition, the authors could show by autoradiography and quantitative determination of skin surface lipids, a decrease in the production of sebum [2, 3], so that benzoyl peroxide may also be endowed with true sebostatic properties, so beneficial in acne vulgaris. In the present study it has been our objective to elucidate further the bacteriostatic effect of BP. For this purpose the inhibitory action of BP on colonies of P.a. was tested in comparison to the vehicle alone.
Material and Methods Following thorough cleaning of the facial skin with 70 % ethanol seborrheic filaments were expressed with a comedone expressor, the exprimate transferred into sterile blood-isotonic saline and homogenized. With a small metal loop, the homogenizedmaterial was inoculated into brain-heart agar with 1 ~o dextrose for isolation of P,a. and anaerobically cultured at 37~ for 5 days. Pure cultures of P.a. were then densely inoculated into Petri dishes covered with 10 ml of this selective agar. 7 mm holes were punched out with a cork piercer and filled with the followingagents: Offprint requests to: Dr. H. Bardach (address see above)
0340-3696/79/0264/0369/$1.00
370
D. Fanta et al.
Table 1. Baeteriostatic activity of benzoyl peroxide on Propionibacterium aches Test material
Inhibition area (mm)
Benzoyl peroxide 5% - ointment Benzoyl peroxide 10 ~ - ointment Benzoyl peroxide 5 % - in ethanol Benzoyl peroxide 10 % - in ethanol Gel vehicle Gel vehicle Erythromycin (aqueous solution)
45 45 - 50 15 20 45 40 55 46 24 35 18 18
Terramycin hydrochloride (aqueous solution)
2% 0.02% 0.0002 % 5% o.o5 % o.oo5%
5 % and i0 % BP preparation in a gel base 1 consisting of colloidal magnesium aluminum silicate, hydroxypropylmethyl-cellulose, polyoxyethylen lauryl ether, citric acid, absolute alcohol, and distilled water; 5 % and 10 % suspension of BP in 70 % ethanol; the BP free vehicle; the BP free vehicle without absolute alcohol. 0.5 ml of each test substance was put into the punched out holes. After anaerobic culturing for 5 days at 37~ the areas of inhibition were measured (Table 1).
Results The 5 ~ and 10 % BP preparation p r o d u c e d inhibitory areas o f about the same size after 5 days culturing, i.e., 45 and 50 mm, respectively. Similar areas o f inhibition were measured with the isolated gel vehicle as well as the alcohol free vehicle, i.e., 45 and 40 m m respectively. Significantly smaller areas o f inhibition occurred with the alcoholic suspension o f BP.
Discussion T o d a y the use o f antibiotics in the treatment o f acne vulgaris is generally accepted. The development and p r o p a g a t i o n o f this type o f therapy is mainly based on the rationale, that P.a. plays a significant role in the pathogenesis o f acne by p r o d u c t i o n o f epitheliotoxic and irritating enzymes and metabolites as well as by its antigenic properties. Reduction o f P.a. in the microbial flora o f the follicular contents correlates well with improvement o f this disease. A l t h o u g h the bacteria are sensitive in vitro to a variety o f antibiotics and chemotherapeutic agents, only the systemic administration o f low doses o f tetracyclines and erythromycin has gained wide acceptance because o f their beneficial effect and low toxicity. Topical application o f these antibiotics has also been quite effective. Reduction o f free acids in skin surface lipids is used as a measure o f successful bacteriostatic therapy in ache vulgaris [9]. This was also observed to various degrees 1 Panoxyl 5 & 10, Stiefel Lab. Offenbach/Main
Bacteriostatic Effect of Benzoyl Peroxide
371
following application o f b e n z o y l peroxide in different vehicles, although the vehicle o f the preparations employed in this study had itself a similar, albeit less significant effect. Similar findings were reported by G l o o r [5]. Since examination o f the bacteriostatic properties o f BP itself is a p r o b l e m because of its instability as well as p o o r solubility in solvents suitable for in vitro m e t h o d s - in this respect the areas o f inhibition p r o d u c e d by BP in ethanol are to be interpreted with c a u t i o n - w e decided to answer this question by comparing the areas o f inhibition using a commercial preparation and a BP-free vehicle. The results of the present study indicate that the vehicle contributes significantly to the bacteriostatic effect. This get vehicle, which has proved to be optimal for dermatologic use, p r o d u c e d areas o f inhibition o f nearly the same size as those elicited by the BP preparation alone. A p a r t f r o m the absolute alcohol present in the vehicle, the surfactant polyoxyethylen lauryl ether m a y also play a causative role in regard to this effect. The above mentioned sebostatic effect, however, was only observed with the BP containing preparation, but not with the vehicle itself [2, 3].
Conclusion Until recently, the effectivity o f BP in the treatment o f acne vulgaris was considered to be primarily a function o f its bacteriostatic action. However, in vitro investigation using pure colonies o f P.a. showed that the areas o f inhibition p r o d u c e d by a 5 % and 10 % commercial BP preparation and the isolated vehicle were o f a b o u t equal size.
References 1. Fanta, D., Bardach, H., Jurecka, W. : Klinische Erfahrungen mit Benzoylperoxidin der Behandlung der Acne vulgaris. Wien. Klim Wsehr. 89, 748-751 (1977) 2. Fanta, D., Jurecka, W. : Autoradiographie investigations in benzoyl peroxide-treated skin. Acta Dermatol-Venereol. (in press) 3. Fanta, D., Mtiller, M. M. : Effect of benzoyl peroxide on skin surface lipids. Dermatologica (in press) 4. Fulton, J. E., Farzad-Bakshandeh, A., Bradley, S. : Studies on the mechanism of action of topical benzoyl peroxide and vitamin A acid in acne vulgaris. J. Cutan. Pathol. 1, 191- 200 (1974) 5. Gloor, M., Hummel, A., Friedrich, H. G.: Experimentelle Untersuchungen zur Benzoylperoxidtherapie der Ache vulgaris. Z. H + G 5, 657-663 (1975) 6_ Huber, C., Christophers, E. : "Keratolytic" effect of salicylic acid. Arch. Dermatol. Res. 257, 293 297 (1977) 7. Meigel, W. N., Prtickner, H. : Therapie der Akne vulgaris mit Panoxyl. Z. Hautkr. 52, 725-732 (1977) 8. Pace, W. E. : A benzoyl peroxide-sulfur cream for ache vulgaris. Canad. Med. An. J. 93, 252-254 (1965) 9. Puhvel, S. M., Reisner, R. M. : Effect of antibiotics on the lipases of Corynebacterium aeries in vitro. Arch. Dermatol. 106, 45-49 (1972) 10. Vasarinsh, D. : Benzoyl-peroxid-sulfur lotion. Arch. Dermatol. 98, 113-186 (1968)
Received September 18, 1978
RDeermatological
Arch. Dermatol. Res. 264, 369-371 (1979)
searcn
9 Springer-Verlag 1979
Investigations on the Bacteriostatic Effect of Benzoyl Peroxide D. Fanta, H. Bardach, and Ch. Poitscheck 2nd Department of Dermatology, University of Vienna (Head: Prof. Dr. G. Niebauer), AlserstraBe 4, A-1090 Wien, Austria
Benzoyl peroxide (BP), a potent oxidizing agent, used for a long time by surgeons to improve wound healing and by dermatologists to treat ulcers and pyodermas, lately was also found to be effective in the management of acne vulgaris [1, 7, 8, 10]. The bacteriostatic effect is believed to be due to reduction of bacterial amino acids to pantothionic acid. The importance of this for the treatment of acne vulgaris was first recognized by Fulton [3], who not only found a significant reduction in the microbial flora of the follicular infundibulum following topical use of BP, but also a distinct decrease in free fatty acids as measured by infrared spectroscopy, the latter serving as a criterion for the presence of Propionibacterium acnes (P.a.). This was supported by G l o o r et al. [5], who performed thin layer chromatography of skin surface lipids. Moreover, local application of BP also produces a keratolytic effect, probably due to liberation, by contact with organic material, of benzoic acid, which like salicylic acid [6] causes lysis of the intercellular substance in the stratum corneum. In addition, the authors could show by autoradiography and quantitative determination of skin surface lipids, a decrease in the production of sebum [2, 3], so that benzoyl peroxide may also be endowed with true sebostatic properties, so beneficial in acne vulgaris. In the present study it has been our objective to elucidate further the bacteriostatic effect of BP. For this purpose the inhibitory action of BP on colonies of P.a. was tested in comparison to the vehicle alone.
Material and Methods Following thorough cleaning of the facial skin with 70 % ethanol seborrheic filaments were expressed with a comedone expressor, the exprimate transferred into sterile blood-isotonic saline and homogenized. With a small metal loop, the homogenizedmaterial was inoculated into brain-heart agar with 1 ~o dextrose for isolation of P,a. and anaerobically cultured at 37~ for 5 days. Pure cultures of P.a. were then densely inoculated into Petri dishes covered with 10 ml of this selective agar. 7 mm holes were punched out with a cork piercer and filled with the followingagents: Offprint requests to: Dr. H. Bardach (address see above)
0340-3696/79/0264/0369/$1.00
370
D. Fanta et al.
Table 1. Baeteriostatic activity of benzoyl peroxide on Propionibacterium aches Test material
Inhibition area (mm)
Benzoyl peroxide 5% - ointment Benzoyl peroxide 10 ~ - ointment Benzoyl peroxide 5 % - in ethanol Benzoyl peroxide 10 % - in ethanol Gel vehicle Gel vehicle Erythromycin (aqueous solution)
45 45 - 50 15 20 45 40 55 46 24 35 18 18
Terramycin hydrochloride (aqueous solution)
2% 0.02% 0.0002 % 5% o.o5 % o.oo5%
5 % and i0 % BP preparation in a gel base 1 consisting of colloidal magnesium aluminum silicate, hydroxypropylmethyl-cellulose, polyoxyethylen lauryl ether, citric acid, absolute alcohol, and distilled water; 5 % and 10 % suspension of BP in 70 % ethanol; the BP free vehicle; the BP free vehicle without absolute alcohol. 0.5 ml of each test substance was put into the punched out holes. After anaerobic culturing for 5 days at 37~ the areas of inhibition were measured (Table 1).
Results The 5 ~ and 10 % BP preparation p r o d u c e d inhibitory areas o f about the same size after 5 days culturing, i.e., 45 and 50 mm, respectively. Similar areas o f inhibition were measured with the isolated gel vehicle as well as the alcohol free vehicle, i.e., 45 and 40 m m respectively. Significantly smaller areas o f inhibition occurred with the alcoholic suspension o f BP.
Discussion T o d a y the use o f antibiotics in the treatment o f acne vulgaris is generally accepted. The development and p r o p a g a t i o n o f this type o f therapy is mainly based on the rationale, that P.a. plays a significant role in the pathogenesis o f acne by p r o d u c t i o n o f epitheliotoxic and irritating enzymes and metabolites as well as by its antigenic properties. Reduction o f P.a. in the microbial flora o f the follicular contents correlates well with improvement o f this disease. A l t h o u g h the bacteria are sensitive in vitro to a variety o f antibiotics and chemotherapeutic agents, only the systemic administration o f low doses o f tetracyclines and erythromycin has gained wide acceptance because o f their beneficial effect and low toxicity. Topical application o f these antibiotics has also been quite effective. Reduction o f free acids in skin surface lipids is used as a measure o f successful bacteriostatic therapy in ache vulgaris [9]. This was also observed to various degrees 1 Panoxyl 5 & 10, Stiefel Lab. Offenbach/Main
Bacteriostatic Effect of Benzoyl Peroxide
371
following application o f b e n z o y l peroxide in different vehicles, although the vehicle o f the preparations employed in this study had itself a similar, albeit less significant effect. Similar findings were reported by G l o o r [5]. Since examination o f the bacteriostatic properties o f BP itself is a p r o b l e m because of its instability as well as p o o r solubility in solvents suitable for in vitro m e t h o d s - in this respect the areas o f inhibition p r o d u c e d by BP in ethanol are to be interpreted with c a u t i o n - w e decided to answer this question by comparing the areas o f inhibition using a commercial preparation and a BP-free vehicle. The results of the present study indicate that the vehicle contributes significantly to the bacteriostatic effect. This get vehicle, which has proved to be optimal for dermatologic use, p r o d u c e d areas o f inhibition o f nearly the same size as those elicited by the BP preparation alone. A p a r t f r o m the absolute alcohol present in the vehicle, the surfactant polyoxyethylen lauryl ether m a y also play a causative role in regard to this effect. The above mentioned sebostatic effect, however, was only observed with the BP containing preparation, but not with the vehicle itself [2, 3].
Conclusion Until recently, the effectivity o f BP in the treatment o f acne vulgaris was considered to be primarily a function o f its bacteriostatic action. However, in vitro investigation using pure colonies o f P.a. showed that the areas o f inhibition p r o d u c e d by a 5 % and 10 % commercial BP preparation and the isolated vehicle were o f a b o u t equal size.
References 1. Fanta, D., Bardach, H., Jurecka, W. : Klinische Erfahrungen mit Benzoylperoxidin der Behandlung der Acne vulgaris. Wien. Klim Wsehr. 89, 748-751 (1977) 2. Fanta, D., Jurecka, W. : Autoradiographie investigations in benzoyl peroxide-treated skin. Acta Dermatol-Venereol. (in press) 3. Fanta, D., Mtiller, M. M. : Effect of benzoyl peroxide on skin surface lipids. Dermatologica (in press) 4. Fulton, J. E., Farzad-Bakshandeh, A., Bradley, S. : Studies on the mechanism of action of topical benzoyl peroxide and vitamin A acid in acne vulgaris. J. Cutan. Pathol. 1, 191- 200 (1974) 5. Gloor, M., Hummel, A., Friedrich, H. G.: Experimentelle Untersuchungen zur Benzoylperoxidtherapie der Ache vulgaris. Z. H + G 5, 657-663 (1975) 6_ Huber, C., Christophers, E. : "Keratolytic" effect of salicylic acid. Arch. Dermatol. Res. 257, 293 297 (1977) 7. Meigel, W. N., Prtickner, H. : Therapie der Akne vulgaris mit Panoxyl. Z. Hautkr. 52, 725-732 (1977) 8. Pace, W. E. : A benzoyl peroxide-sulfur cream for ache vulgaris. Canad. Med. An. J. 93, 252-254 (1965) 9. Puhvel, S. M., Reisner, R. M. : Effect of antibiotics on the lipases of Corynebacterium aeries in vitro. Arch. Dermatol. 106, 45-49 (1972) 10. Vasarinsh, D. : Benzoyl-peroxid-sulfur lotion. Arch. Dermatol. 98, 113-186 (1968)
Received September 18, 1978
