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Histological evaluation of hair follicle due to papain's effect
Abstract
Histological alterations in the skin and hair follicle of mice were evaluated as a result of the application of gel and cream formulas containing papain as a harmless treatment for hirsutism. Papain is a proteolytic enzyme and it has been used in pharmaceutical, cosmetic and nutrition areas. The purpose of this study was to evaluate the efficacy of a depilatory product, through histological analysis using light microscopy. Gel and cream formulas containing papain were developed and daily applied on the back of two groups of mice for 31 days. The depilatory effect of the gel formula applied on the first group was less evident. The second group treated with the cream formula presented an intensive depilatory effect; the morphometrical analysis showed dilation of about 55% of the hair follicle lumen and an increase of the thickness of epidermis. Papain cream had a significantly higher depilatory effect than the papain gel.
... É uma enzima que possui aplicação em diversos segmentos industriais, incluindo o da cosmética, onde seu emprego está bastante propagado no Brasil (MIÚRA, 2012). Capaz de promover a hidrólise de ligações peptídicas tanto do colágeno quanto da queratina no extrato córneo da pele (SIM et al., 2000;TRAVERSA;MACHADO-SANTELLI;VELASCO, 2007), a papaína é inserida em formulações cosméticas com ação esfoliante e depilatória (TRAVERSA; MACHADO-SANTELLI;VELASCO, 2007, MIÚRA, 2012, além de ser usada em peelings (AZEVEDO, 2018), visto que a mesma atua a nível celular da epiderme, renovando-a (BERLINCK, 2015). ...
... É uma enzima que possui aplicação em diversos segmentos industriais, incluindo o da cosmética, onde seu emprego está bastante propagado no Brasil (MIÚRA, 2012). Capaz de promover a hidrólise de ligações peptídicas tanto do colágeno quanto da queratina no extrato córneo da pele (SIM et al., 2000;TRAVERSA;MACHADO-SANTELLI;VELASCO, 2007), a papaína é inserida em formulações cosméticas com ação esfoliante e depilatória (TRAVERSA; MACHADO-SANTELLI;VELASCO, 2007, MIÚRA, 2012, além de ser usada em peelings (AZEVEDO, 2018), visto que a mesma atua a nível celular da epiderme, renovando-a (BERLINCK, 2015). ...
... É uma enzima que possui aplicação em diversos segmentos industriais, incluindo o da cosmética, onde seu emprego está bastante propagado no Brasil (MIÚRA, 2012). Capaz de promover a hidrólise de ligações peptídicas tanto do colágeno quanto da queratina no extrato córneo da pele (SIM et al., 2000;TRAVERSA;MACHADO-SANTELLI;VELASCO, 2007), a papaína é inserida em formulações cosméticas com ação esfoliante e depilatória (TRAVERSA; MACHADO-SANTELLI;VELASCO, 2007, MIÚRA, 2012, além de ser usada em peelings (AZEVEDO, 2018), visto que a mesma atua a nível celular da epiderme, renovando-a (BERLINCK, 2015). ...
Sendo possível a obtenção mediante processos biotecnológicos, as enzimas são biocatalisadoras orgânicas de origem proteica responsáveis por acelerar reações químicas, onde sua utilização abrange diversas áreas, dentre estas, a cosmética. O presente trabalho teve como objetivo avaliar o potencial para bioprospecção de enzimas com aplicabilidade no setor cosmético. Para elaboração deste artigo foi realizado um estudo de revisão bibliográfica nas plataformas Google Acadêmico, Scientific Eletronic Library Online (SciELO) e PubMed, bem como acesso em um livro disponível na biblioteca da Universidade São Francisco, na qual a coleta de dados se baseou em critérios de inclusão e exclusão, a partir do tema proposto. Após a realização do levantamento bibliográfico, discutiu-se acerca do crescimento dos produtos intitulados com naturais, a utilização da biodiversidade como fonte para a bioprospecção de enzimas, além da abordagem de três classes de enzimas (superóxido-dismutase, lipases e proteases) e suas respectivas aplicações na cosmética.
... The cosmetic, medical and pharmaceutical segments have started to use papain to improve product quality due to the increase in the use of bio-derived and eco-friendly ingredients [6,7]. Papain has been applied as a debridement agent of wounds [8][9][10], exfoliation agent [11,12], depilatory agent [13] and transdermal drug permeation enhancer [14], for example. The chemical stability of papain, which may decrease depending on the storage and presence of other ingredients in the formulation, is the most evaluated effect in terms of skin application, principally related to the complexation of the enzyme with polymers [2,13,[15][16][17]. ...
... Papain has been applied as a debridement agent of wounds [8][9][10], exfoliation agent [11,12], depilatory agent [13] and transdermal drug permeation enhancer [14], for example. The chemical stability of papain, which may decrease depending on the storage and presence of other ingredients in the formulation, is the most evaluated effect in terms of skin application, principally related to the complexation of the enzyme with polymers [2,13,[15][16][17]. ...
Objective:
This work aims to evaluate the effects of enzyme concentration, pH, temperature, and time course degree of hydrolysis (DH) of papain regarding further development of pharmaceutical and cosmetic formulations.
Methods:
The hydrolysis of casein, collagen, keratin, and porcine skin at pH and temperature ranges of the human skin was evaluated. Also, low contact times of enzyme-substrate were studied. The incorporation of 3 mM of cysteine improved the caseinolytic (PU), collagenolytic (CU), and keratinolytic (KU) activities of papain.
Results:
In general, the increase from 0.1 to 1.0 or 2.0 mg mL-1 of papain improved PU, CU, and KU. When 2.0 mg mL-1 of papain was used, the highest DH of casein, collagen, and keratin were obtained at 240 min (14, 35, and 6%, respectively). The decrease in pH and temperature reduced all proteolytic activities, but papain maintained at least 50 and 40% of its activity at 26 °C and pH 4.5, respectively. Scanning electron micrographs of the surface of the skin showed that papain application had exfoliating activity.
Conclusion:
This pre-formulation study demonstrated that papain concentration, time of application, and pH of the product should be evaluated when developing a product to promote the hydrolysis of the proteins of the skin.
... Sections were stained with hematoxylin and eosin. Finally examined under a light microscope for structures [12]. ...
... Prolonged use of reishi can also give hirsutism. This study gave effective results on mice as mice or any other animal has a higher number of terminal hair than in human Also, the skin of mice is more permeable than that of human sp So it is possible that a smaller amount penetrated [12]. That means lesser actives reaching the follicle that can even mean a lesser effect. ...
Objective: The aim of this work to formulate, evaluate and compare the effectiveness of herbal creams containing extract of reishi and tea tree oil for treating hirsutism. Methods: Herbal ingredients were authenticated. Cream base was initially formulated. Three formulations of herbal cream were prepared. Reishi ethanolic extract, tea tree oil, and combination of tea tree oil and reishi extract were added to the cream base and formulated cream were named as RHC, THC and RTC respectively. In vitro evaluations on herbal creams were done for the physicochemical characteristics. In vivo studies were carried out on female Swiss Albino mice for the activity against hair growth by topical application of cream to shaved skin. The histological and morphometric evaluation was carried out. Skin irritancy study was conducted. Results: The herbal creams showed desirable physicochemical properties like pH, viscosity and spreadability. Statistical analysis for the length of hair was performed by using one way ANOVA followed by DUNNET’S post hoc test where THC and RTC were found to be significant whereas RHC showed no significant reduction of hair growth compared to control. RTC showed a significant effect at p<0.05 and hair growth reduction was significant for THC at p<0.001 compared to the control group. RTC and THC showed mild to moderate reduction in the size of the hair follicles with a reduction of sebaceous gland size in the histological analysis. Conclusion: Topical application of herbal creams to mice showed that hair growth was fastest in group RHC and was slowest in group THC and intermediate with RTC. It can be concluded that these herbal actives can be used as an effective treatment against hirsutism. Within the study period, tea tree oil was found to be more effective than reishi extract and the combination product. Further formulation studies and in vivo studies need to be carried out on reishi to assess its effectiveness against hirsutism.
... Therefore, management approaches that are as effective as possible need to be constantly explored. Follicular iontophoretic delivery of proteolytic enzymes has been examined and shown to induce profound depilatory effects [75,76]. In fact, an early study evaluating the effect of the proteolytic enzymes papain and chymotrypsin on hair follicles of transgenic mice demonstrated that their iontophoretic application produced intense degenerative effects on structures of the hair follicle down to the bulb region, resulting in a long-lasting depilatory effect [77]. ...
Objective:
Iontophoresis is defined as the use of electric current to drive molecules across cell membranes through an electrolyte solution. In therapeutic context, it is used to facilitate the administration of bioactive substances, either systemically or locally. The technique presents various advantages and that is why it has been successfully used by a plethora of medical sciences. The constantly developing field of dermato-cosmetic science has also taken advantage of the possibilities offered by iontophoresis, aiming to enhance the delivery of the applied active ingredients and, thus, induce the desired cosmetic effects.
Methods:
Available literature was examined for evidence-based reports of safe and successful iontophoresis of pharmaceutical and cosmetic substances, in order to explore different iontophoretic applications in the field of dermato-cosmetic and dermato-aesthetic sciences.
Conclusion:
Iontophoresis can be safely and successfully used in the treatment of ageing, photoageing, hyperpigmentation, oxidative stress, hair loss, hair removal, acne, acne sequelae and cellulite, providing many possibilities for enhanced treatment results.
... Papain has better depilatory efficacy in creams than in gel formulations (Traversa et al., 2007). Two of the peel products we investigated (No. 3 and 6) were in gel form (Table 1). ...
Our goal was to verify the proteolytic mode of action and activity levels among several commercial cosmetic facial peels advertised by manufacturers as “enzymatic”. Eleven enzyme peels were analyzed for their proteolytic activity against casein as a generic substrate and compared to the activity found in pineapple and papaya fruits. The highest specific protease activity was observed in the flesh of a pineapple (5.88 U/g). Only two products demonstrated sufficient activity (0.924 and 0.238 U/g) to be called “enzyme peels”. Three products showed trace activity (0.023–0.125 U/g), albeit too low to exert a significant exfoliating effect. Six preparations had no detectable enzyme activity.
... 14 Some studies have already studied the effects of stabilizers with proteolytic enzymes. Traversa et al. 40 45 showed that the encapsulation of papain improved stability, enhanced activity, and reduced skin irritation in wound management. Moreover, immobilization may also limit or reduce the digestion that the own proteases promote in themselves. ...
Background
Proteolytic enzymes are biological catalysts that can compose cosmetic formulations: These enzymes are capable of mimicking the desquamation process of the skin, acting as exfoliants. Although enzymatic exfoliation is not new and commercial products were easily found, there is a lack of scientific literature about this topic.
Methods
A search was carried out until 2021 in different scientific databases (Web of Science, Scopus, Scielo, PubMed, etc.). In vitro and in vivo studies that evaluated the application of enzymes aiming to exfoliate the skin or with a similar cosmetic or dermatological application were selected.
Results
Only 11 articles were found, and, among them, few studies applied enzymes as exfoliants in clinical trials. Nevertheless, the results demonstrate that the enzymes can exfoliate the skin and improve some desired characteristics of the organ. Papain, bromelain, keratinases, and microbial proteases are some enzymes already applied as exfoliants. The study of pH, temperature, and stabilization of the enzymes in cosmetic formulations were also demonstrated to be important aspects to be evaluated, principally in preventing loss of enzyme activity and possible allergens/irritations on the skin.
Conclusion
This literature review showed the main aspects that should be evaluated before considering producing or applying proteolytic enzymes in exfoliation products/procedures. The use of enzymatic exfoliation has potential in the cosmetic industry. Hence, further robust in vivo studies are needed before the enzymatic exfoliation can be recommended with safety as a treatment modality in the current conditions.
... Others have used bacterial keratinases [6] which do not degrade collagen, thereby preserving the length and integrity of the collagen α-chains [7]. The enzyme papain, which has both collagenolytic and keratinolytic activities, has been shown to have a depilatory effect [8]. Acid pre-treatment is invariably used for raw materials (pig skin, and fish skin and bone) that have fewer inter or intra-molecular collagen cross-links to extract gelatin [9,10], but it is not usually used in the preparation of raw materials from bovines slaughtered close to maturity which have collagen cross-links that are resistant to acid solubilization. ...
PurposeHides are the by-product of slaughter houses which are mostly used for leather production. In Canada, the hides are either disposed of with other slaughter waste or sold at a very low price. Dehairing of hides is a prerequisite for either leather or gelatin production from it. Therefore, the effect of hide dehairing method on subsequent gelatin extraction and quality was investigated.Methods
Bovine hides (BH) were dehaired using either 5% acetic acid (AA), 10% calcium hydroxide (CH), 0.02% keratinase (KTN), 2.5% papain (PP), or not at all (control; CT), with control BH subsequently treated with 5% AA (CTAA).ResultsMean bovine hide gelatin (BHG) yields (dry basis) were 11.37%, 54.25%, 45.07%, 18.88% and 55.02% for CT, AA, CTAA, CH, and KTN, respectively. Gel strength was highest in AA followed by the CTAA, CT and CH and KTN treatments. The molecular weight (MW) distribution pattern showed that dehairing of BH with enzymes degraded the collagen extensively, increased proportions of low MW peptides that translated into low gel strength.Conclusions
Acetic acid, which is extensively using in food industry, can be used to dehair BH as pre-treatment to extracting high quality gelatin.Graphic Abstract
... Others have used bacterial keratinases [3] which do not degrade collagen, thereby preserving the length and integrity of the collagen αchains [4]. The enzyme papain, which has both collagenolytic and keratinolytic activities, has been shown to have a depilatory effect [5]. Acid pre-treatment is invariably used for raw materials (pig skin, and sh skin and bone) that have fewer inter or intra-molecular collagen cross-links to extract gelatin [6,7], but it is not usually used in the preparation of raw materials from bovine animals slaughtered close to maturity which have collagen cross-links that are resistant to acid solubilization. ...
Purpose Hides are the by product of slaughter houses which are mostly used for leather production. In Canada, the hides are either disposed of with other slaughter waste or sold at a very low price. Dehairing of hides is a prerequisite for either leather or gelatin production from it. Therefore, the effect of hide dehairing method on subsequent gelatin extraction and quality was investigated.
Methods Bovine hides (BH) were dehaired using either 5% acetic acid (AA), 10% calcium hydroxide (CH), 0.02% keratinase (KTN), 2.5% papain (PP), or not at all (control; CT), with control BH subsequently treated with 5% AA (CTAA).
Results Mean bovine hide gelatin (BHG) yields (dry basis) were 11.37%, 54.25%, 45.07%, 18.88% and 55.02% for CT, AA, CTAA, CH, and KTN, respectively. Gel strength was highest in AA followed by the CTAA, CT and CH and KTN treatments. The molecular weight (MW) distribution pattern showed that dehairing of BH with enzymes degraded the collagen extensively, increased proportions of low MW peptides that translated into low gel strength.
Conclusions Acetic acid, which is extensively using in food industry, can be used to dehair BH as pre-treatment to extracting high quality gelatin.
... Topical enzymes, mainly keratinases, are also used to increase the activity of topically applied drugs by enhancing skin and nail penetration (FixaFungus Inc., 2015a, b). Further, enzymes are investigated for the prevention and treatment of biofilm-related oral diseases (Pleszczynska et al., 2017) and as active agents in personal care products, mainly for depilation (Sanghvi et al., 2016;Traversa et al., 2007), skin cleansing, desquamation and lightening (Del Rosso, 2013;Draelos, 2015;Zhong et al., 2015) as well as prevention of ultravioletinduced damage (Emanuele et al., 2013) with several enzymebased products on the market. In earlier publications, high level production of alkaline protease (APr) with considerable keratinolytic activity has been achieved using recombinant B. subtilis DB 100 (p5.2) cells, harboring the multi-copy p5.2 plasmid that carries the complete alkaline protease (aprE) gene (Zaghloul et al., 2000(Zaghloul et al., , 2011(Zaghloul et al., , 2010. ...
Enzymes may offer great potentials in topical pharmaceutical applications provided that treatment conditions are controlled for efficacy and safety. In this study, the effect of alkaline protease produced by recombinant Bacillus subtilis cells on the ex-vivo permeability of rabbit ear skin was investigated under different conditions of enzyme activity (5-60 units) and exposure time (15–60 min). Data for transepidermal water loss (TEWL) and permeation of a hydrophilic dye, rhodamine B (Rb), indicated biphasic activity-dependent and exposure time-dependent skin permeability. Maximum effects were obtained at 20 proteolytic units and 30 min exposure. Findings proved consistent with histopathological changes indicating progressive stratum corneum (SC) loss and disruption of the dermo-epidermal junction at 20 units and up to 30 min exposure time followed by dermal hyalinization at longerexposure. This was associated with progressive loss of skin hair. Applying the identified pretreatment conditions to transdermal delivery of vardenafil in a gel base across dorsal rat skin indicated a significant increase in plasma levels at 30 and 60 min with minimal histopathological changes 5 days post enzyme treatment. Accordingly, the recombinant B. subtilis alkaline protease offers promise as a pharmaceutical enzyme for transdermal drug delivery bioenhancement and dermatological applications.
... These characteristics make the enzyme a promising alternative for culinary and industrial applications (Kim and Lee, 1995). In the pharmaceutical field, papain, collagenase and other protease activities may be used for wound healing, necrotic tissue removal and reepithelialization (Traversa et al., 2007;Saarialho et al., 1993;Thompson et al., 1973;Choi and Laursen, 2000;Su et al., 2009). These enzymes may also be used in the debridement of necrotic tissue in burn patients (Kim, 2007). ...
The rhizomes of common ginger (Zingiber officinale
Roscoe) contain substances with antimicrobial activity
and proteolytic enzymes and thus may have various
pharmaceutical applications. The aim of the present
study was to prepare Zingiber officinale Roscoe
rhizome extracts for pharmaceutical use, preserving the
proteolytic enzyme activity and testing the antimicrobial
activity, in order to develop topical formulations.
Two extracts were obtained - aqueous and glycolic
– and assessed for their physical, physicochemical
and organoleptic characteristics. To measure their
proteolytic activity, the extracts were assayed for
enzymatic hydrolysis of 1.2% casein solution, at pH
6.0 and 37°C; papain was used for comparison. The
antimicrobial activity of the glycolic extract of Zingiber
officinale Roscoe was tested by microdilution; the
inoculants were prepared from cultures of Escherichia
coli, Staphylococcus aureus, Staphylococcus epidermidis
and Pseudomonas aeruginosa. There was no growth
of S. aureus and S. epidermidis at concentrations of
150 mg/mL or more of extract, whereas P. aeruginosa
was inhibited from 100 mg/mL and E.coli from 75 mg/
mL. Emulsion formulations were prepared as vehicles
for the extracts and their stability was tested. The
results showed proteolytic activity in both Z. officinale
rhizome extracts, the glycolic extract having 691.68
PU/g juice and the aqueous extract, 338.14 PU/g juice.
The formulations were stable, especially the one that
contained the glycolic extract. In sum, the formulations
showed satisfactory stability and the Z. officinale extract
showed bactericidal activity against the cultures tested;
the results are promising for the use of the extract in
foods, medicine and cosmetics.
... We developed seventeen O/W emulsions containing 0.8 % (w/w) papain (obtained from Wallerstein, Brazil) (Traversa et al., 2007). They were composed of: emulsifying wax NF (purchased from Chemyunion -Brazil); paraffinum liquidum (purchased from Mapric -Brazil); myristyl lactate and isopropyl palmitate (gifts from Croda, Brazil); ethylparaben, isobutylparben, methylparaben, propylparaben (and) phenoxyethanol; decyl oleate and hydroxylated milk glycerides (gift from Ionquímica, Brazil); dibutyl adipate (gift from Cognis, Brazil); ammonium acryloyldimethyltaurate/VP copolymer (gift from Clariant, Brazil); carbomer and acrylates/C10-30 alkyl acrylate crosspolymer (purchased from Noveon Consumer Specialties Lubrizol, Brazil); hydroxyethylcellulose (purchased from Galena, Brazil); propylene glycol (purchased from Cosmotec, Brazil); and cyclopentasiloxane (gift from Dow Corning -Brazil). ...
A papaína é uma enzima utilizada em formulações tópicas como agente proteolítico debridante no tratamento de lesões abertas de grande extensão e queimaduras. É, também, empregada na pele íntegra como agente promotor da permeação cutânea de princípios ativos, peeling químico e como agente depilatório progressivo. A estabilidade de formulações contendo enzimas não é facilmente alcançada. No presente trabalho realizou-se a modificação da enzima com polietilenoglicol, visando maior estabilidade das formulações. A realização do Teste Estabilidade Normal comparativo entre as formulações contendo as formas da enzima não modificada e modificada demonstrou que a última apresentou um perfil de estabilidade diferenciado, nas diferentes condições (5,0 ± 1,0 °C; 22,0 ± 2,0 °C; 40,0 ± 2,0 °C). A condição de 5,0 ± 1,0 °C foi a mais adequada para a formulação contendo papaína não modificada enquanto a 22,0 ± 2,0 °C foi indicada para aquela contendo a forma modificada. Estes resultados confirmaram o aumento da estabilidade da papaína modificada comparada com a livre e seu potencial de aplicação em formulações de uso tópico.
... These characteristics make the enzyme a promising alternative for culinary and industrial applications (Kim and Lee, 1995). In the pharmaceutical field, papain, collagenase and other protease activities may be used for wound healing, necrotic tissue removal and reepithelialization (Traversa et al., 2007; Saarialho et al., 1993; Thompson et al., 1973; Choi and Laursen, 2000; Su et al., 2009). These enzymes may also be used in the debridement of necrotic tissue in burn patients (Kim, 2007). ...
The rhizomes of common ginger (Zingiber officinale Roscoe) contain substances with antimicrobial activity and proteolytic enzymes and thus may have various pharmaceutical applications. The aim of the present study was to prepare Zingiber officinale Roscoe rhizome extracts for pharmaceutical use, preserving the proteolytic enzyme activity and testing the antimicrobial activity, in order to develop topical formulations. Two extracts were obtained-aqueous and glycolic – and assessed for their physical, physicochemical and organoleptic characteristics. To measure their proteolytic activity, the extracts were assayed for enzymatic hydrolysis of 1.2% casein solution, at pH 6.0 and 37°C; papain was used for comparison. The antimicrobial activity of the glycolic extract of Zingiber officinale Roscoe was tested by microdilution; the inoculants were prepared from cultures of Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. There was no growth of S. aureus and S. epidermidis at concentrations of 150 mg/mL or more of extract, whereas P. aeruginosa was inhibited from 100 mg/mL and E.coli from 75 mg/ mL. Emulsion formulations were prepared as vehicles for the extracts and their stability was tested. The results showed proteolytic activity in both Z. officinale rhizome extracts, the glycolic extract having 691.68 PU/g juice and the aqueous extract, 338.14 PU/g juice. The formulations were stable, especially the one that contained the glycolic extract. In sum, the formulations showed satisfactory stability and the Z. officinale extract showed bactericidal activity against the cultures tested; the results are promising for the use of the extract in foods, medicine and cosmetics.
... Principles in Animal Research adopted by the Brazilian College of Animal Experimentation. 10 After asepsis of the dorsal region with ethanol, the treated area had hair removal by stainless steel blade, and it was divided into superior and inferior left and right dorsal areas. During 21 consecutive days, each area of 2.0 cm 2 received individually, twice a day (morning and evening), 0.5 g of formulations (emulsion or gel) with caffeine, caffeine + sodium benzoate, SAC, and control (formulation without active substance). ...
Cellulite is a physiological condition that presents etiologic plurality. Caffeine and its derivatives are used in anticellulite cosmetics due to their lipolytic activity on fatty cells. Siloxanetriol alginate caffeine (SAC) is a silanol derived from organic silicon. Radicals primarily from SAC are caffeine and the mannuronic acid.
This study aims to analyze the effects of caffeine and siloxanetriol alginate caffeine on fatty tissue by histological evaluation.
Formulations were developed with caffeine, caffeine + sodium benzoate or SAC and were applied topically for 21 days on Wistar female mice. The study regarded the histological aspects by determination of diameter and number of fatty cells with a light microscope.
Emulsion with caffeine caused a reduction of 17% on the diameter of the fatty cells compared with the control. The emulsion with caffeine + sodium benzoate did not cause alterations on cell diameter. Emulsion with SAC provoked reduction on fatty cell diameters of 16%. No significant alterations were observed on the diameter of the fatty cells treated with gels, although it was noticed that gel with SAC promoted a reduction of 26% on the number of fatty cells.
Emulsion with SAC was considered more indicated to promote the lipolytic action on fatty tissue, acting as a complement to treat cellulite. When sodium benzoate was added to the preparations, it inhibited the caffeine efficiency. Gel was not an adequate vehicle to be incorporated with caffeine and SAC.
... Principles in Animal Research adopted by the Brazilian College of Animal Experimentation. 10 After asepsis of the dorsal region with ethanol, the treated area had hair removal by stainless steel blade, and it was divided into superior and inferior left and right dorsal areas. During 21 consecutive days, each area of 2.0 cm 2 received individually, twice a day (morning and evening), 0.5 g of formulations (emulsion or gel) with caffeine, caffeine + sodium benzoate, SAC, and control (formulation without active substance). ...
Background Cellulite is a physiological condition that presents etiologic plurality. Caffeine and its derivatives are used in anticellulite cosmetics due to their lipolytic activity on fatty cells. Siloxanetriol alginate caffeine (SAC) is a silanol derived from organic silicon. Radicals primarily from SAC are caffeine and the mannuronic acid.Aims This study aims to analyze the effects of caffeine and siloxanetriol alginate caffeine on fatty tissue by histological evaluation.Methods Formulations were developed with caffeine, caffeine + sodium benzoate or SAC and were applied topically for 21 days on Wistar female mice. The study regarded the histological aspects by determination of diameter and number of fatty cells with a light microscope.Results Emulsion with caffeine caused a reduction of 17% on the diameter of the fatty cells compared with the control. The emulsion with caffeine + sodium benzoate did not cause alterations on cell diameter. Emulsion with SAC provoked reduction on fatty cell diameters of 16%. No significant alterations were observed on the diameter of the fatty cells treated with gels, although it was noticed that gel with SAC promoted a reduction of 26% on the number of fatty cells.Conclusions Emulsion with SAC was considered more indicated to promote the lipolytic action on fatty tissue, acting as a complement to treat cellulite. When sodium benzoate was added to the preparations, it inhibited the caffeine efficiency. Gel was not an adequate vehicle to be incorporated with caffeine and SAC.
Skin is the ultimate barrier between body and environment and prevents water loss and penetration of pathogens and toxins. Internal and external stressors, such as ultraviolet radiation (UVR), can damage skin integrity and lead to disorders. Therefore, skin health and skin ageing are important concerns and increased research from cosmetic and pharmaceutical sectors aims to improve skin conditions and provide new anti‐ageing treatments. Biomolecules, compared to low molecular weight drugs and cosmetic ingredients, can offer high levels of specificity. Topically applied enzymes have been investigated to treat the adverse effects of sunlight, pollution and other external agents. Enzymes, with a diverse range of targets, present potential for dermatological use such as antioxidant enzymes, proteases and repairing enzymes. In this review, we discuss enzymes for dermatological applications and the challenges associated in this growing field.
Although the loss of scalp hair is distressing and many medical treatments focus on its restoration, the removal of body hair has been adopted since ancient times. Beauty standards, which r eflect the culture of each society, have been presenting the depilated body as absolutely desirable. Through the ages various methods of hair removal have been used depending on the requirements of the individuals. In recent years, Laser and Intense Pulse Light devices have been considered as the most promising solution for excess hair growth, without excluding the efficacy of other methods to induce satisfactory epilatory results. The enzyme-based hair removal method has received little recognition even though experimental and clinical data support its efficacy to provide long term or even permanent epilation. The present review presents these data and examines the likelihood of considering the aforementioned method as ideal.
In pH 4.0 Britton-Robinson buffer medium, PdCl(2) was able to react with enzymes (EZ) such as lysozyme (LYSO) and papain (PAP) to form a coordination complex (EZ-PdCl(2) ), which further reacted with MoO(4) (2-) to form a ternary complex (MoO(4) (2-) -EZ-PdCl(2) ). As a result, the absorption and fluorescence spectra changed; new spectra of resonance Rayleigh scattering (RRS), second-order scattering (SOS) and frequency-doubling scattering (FDS) appeared and their intensities were enhanced greatly. The maximum RRS, SOS and FDS wavelengths of two ternary complexes were located at 310, 560 and 350 nm, respectively. The increments of scattering intensity were directly proportional to the concentrations of EZ within certain ranges. The detection limits (3σ) of LYSO and PAP were 4.5 and 14.0 ng/mL (RRS method), 9.6 and 57.8 ng/mL (SOS method), and 5.2 and 106.0 ng/mL (FDS method). Taking the MoO(4) (2-) -LYSO-PdCl(2) system, which was more sensitive, as an example, the effects of coexisting substances were evaluated. The methods showed excellent selectivity. Accordingly, new rapid, convenient, sensitive and selective scattering methods for the determination of LYSO and PAP were proposed and applied to determine LYSO in egg white with satisfactory results. The reaction mechanism and basis of the enhancement of scattering were discussed. Copyright © 2012 John Wiley & Sons, Ltd.
Acid unfolding of non-inhibited papain at pH 2 was studied by means of spectroscopic and electrophoresis techniques as well as activity assays. We found a molten globule like species (A state) similar to that previously reported for bromelain and S-carboxy-methyl-papain. We demonstrated that this A state is not thermodynamically stable but a metastable conformer which decays into an unfolded conformation in a few hours. The mechanism of acid unfolding to the A state proved to be completely irreversible, with a biphasic time evolution of spectroscopic signals characteristic of the existence of a kinetic intermediate. This latter species showed properties in-between native and A state such as secondary structure, exposition of hydrophobic area and tryptophan environment, but a native like hydrodynamic radius. Native papain seems to unfold at acid pH through at least two kinetic barriers, being its pro-region mandatory to conduct and stabilize its active structure. Computer simulations of acid unfolding, followed by ANS docking, identified three regions of cavity formation induced by acid media which might be used as regions to be fortified by protein engineering in the quest for extreme-resistant proteases or as hot-spots for protease inactivation.
To compare the effectiveness of finasteride and flutamide in the treatment of hirsutism in patients with polycystic ovary syndrome (PCOS) and with idiopathic hirsutism.
Randomized study.
One hundred and ten hirsute patients were selected: 64 women with PCOS and 46 with idiopathic hirsutism.
Patients were assigned randomly to receive 5mg finasteride once daily or 250mg of flutamide twice daily, for 12 consecutive months. Hirsutism was evaluated at 12 months of therapy, with the Ferriman-Gallwey score and with measurement of the terminal hair diameters (microm) taken from four different body areas. Blood samples were taken for assessment of endocrine and hematochemical parameters. Side effects were monitored during the treatment.
Both finasteride and flutamide induced a significant decrease in the hirsutism scores and hair diameters at the end of 12 months. Finasteride reduced the Ferriman-Gallwey score by 31.4% in the PCOS cases and by 34.2% in the idiopathic hirsutism cases, and hair diameter by 27.0-34.1% in PCOS and by 29.6-37.9% in idiopathic hirsutism. Flutamide reduced the Ferriman-Gallwey score by 56.7% in PCOS and by 50.9% in idiopathic hirsutism, and hair diameter by 50. 3-60.0% in PCOS and by 47.7-56.5% in idiopathic hirsutism. Flutamide did not induce hormone variations, while finasteride increased testosterone levels by 40% in PCOS and by 60% in idiopathic hirsutism and decreased 3alpha-androstanediol glucuronide (3alpha-diolG) by 66.7% in PCOS and by 69.5% in idiopathic hirsutism. No important side effects or changes in the hematochemical parameters were observed with finasteride, while two patients (3.6%) in the flutamide group expressed abnormal transaminase levels after 6 months of treatment. Dry skin also appeared significantly more with flutamide (67.3%) than with finasteride (23.6%).
Both drugs are effective in the treatment of hirsutism but flutamide is more effective than finasteride.
Nearly 50 years ago, Chase published a review of hair cycling in which he detailed hair growth in the mouse and integrated hair biology with the biology of his day. In this review we have used Chase as our model and tried to put the adult hair follicle growth cycle in perspective. We have tried to sketch the adult hair follicle cycle, as we know it today and what needs to be known. Above all, we hope that this work will serve as an introduction to basic biologists who are looking for a defined biological system that illustrates many of the challenges of modern biology: cell differentiation, epithelial-mesenchymal interactions, stem cell biology, pattern formation, apoptosis, cell and organ growth cycles, and pigmentation. The most important theme in studying the cycling hair follicle is that the follicle is a regenerating system. By traversing the phases of the cycle (growth, regression, resting, shedding, then growth again), the follicle demonstrates the unusual ability to completely regenerate itself. The basis for this regeneration rests in the unique follicular epithelial and mesenchymal components and their interactions. Recently, some of the molecular signals making up these interactions have been defined. They involve gene families also found in other regenerating systems such as fibroblast growth factor, transforming growth factor-beta, Wnt pathway, Sonic hedgehog, neurotrophins, and homeobox. For the immediate future, our challenge is to define the molecular basis for hair follicle growth control, to regenerate a mature hair follicle in vitro from defined populations, and to offer real solutions to our patients' problems.
Four topical drugs were studied with regard to their ability to promote healing of open wounds in 60 Wistar rats. Five study groups were defined. Four of the five groups received one of the following substances: papain, sugar, minoxidil, or glucan. The fifth group or control group received saline solution (NaCl, 0.9%). An experimental model with wound standardization (6 × 3.5 cm) was used. The raw area and new epithelium were quantified in each rat by taking photographs from which planimetry was performed. The closure of the first lesion occurred on the 49th postoperative day. Quantitative assessment of collagen deposition was performed using image analyzing software on slides stained with picrosirius and observed under polarized light. Epithelium formation was greatest in the minoxidil group, followed (in decreasing order) by the sugar, papain, control, and glucan groups. Significant differences in collagen deposition were observed among all the groups. The greatest amount of collagen was quantified in the glucan group, followed (in decreasing order) by the sugar, minoxidil, papain, and control groups. This data and possible mechanisms for the interference of each substance in wound healing are considered.
For cosmetic applications, papain or lysozyme was conjugated to a soluble biopolymer produced by Schizophyllum commune. Stability of the conjugated enzymes were significantly enhanced, such that more than 90% of the initial activity remained after a month storage at 45C, even in a cosmetic formulation including various oils and surfactants. Cosmetic lotion containing 1% papain conjugate was more effective in exfoliating stratum corneum of skin than the lotion containing 5% lactic acid, one of the popular exfoliating agents.
A spin-labeled p-chloromercuribenzoate (SL-PMB) and a fluorescence probe, 6-acryloyl-2-dimethylaminonaphthalene (Acrylodan), both of which bind to the single SH group located in the active site of papain, were used to investigate the interaction of papain (EC 3.4.22.2) with two protein denaturants. It was found that the active site of papain was highly stable in urea solution, but underwent a large conformational change in guanidine hydrochloride solution. Electron paramagnetic resonance and fluorescence results were in agreement and both paralleled enzymatic activity of papain with respect to both the variation in pH and denaturation. These results strongly suggest that SL-PMB and Acrylodan labels can be used to characterize the physical state of the active site of the enzyme.
The percutaneous absorption of compounds through the skin of selected animal species was compared to that occurring with human skin, using in vitro diffusion cell techniques. The permeability of back skin of the following animals was examined: Hormel miniature pigs, Osborne-Mendel rats, NIH hairless mice, and Swiss mice. Benzoic acid, acetylsalicylic acid, and urea, dissolved in a petrolatum vehicle, were applied and permeability constants were determined. With the faster penetrating compounds, benzoic acid and acetylsalicylic acid, mouse skin and hairless mouse skin were similar and were the most permeable. Little difference was observed in acetylsalicylic acid absorption of human, pig, and rat skin. Pig skin and hairless mouse skin were the best animal models for the slower penetrating urea. The thicknesses of the stratum corneum, epidermis, and whole skin were determined from microtome sections prepared from frozen skin. In addition, the density of hair follicles in each type of skin was determined. In general terms, thickness of the stratum corneum was found to be important: the thickest stratum corneum was that of pig skin and the thinnest was that of the mouse. Often however, differences in permeability and stratum corneum thickness did not correlate on a 1:1 basis, which can be indicative of differences in structure of the different types of skin. The animal model of choice is dependent on the compound. For benzoic acid and acetylsalicylic acid, the pig and rat are good models for human skin. In the case of slow absorbing compounds, such as urea, diffusion through appendages in the skin may make undesirable the use of skin of densely haired animals.
Human exposure to papain, a cysteine proteinase, is associated with hypersensitivity reactions. We demonstrate in mice that enzymatically active papain preferentially induces an IgG1 response and results in mast cell degranulation, both features typical of an allergic reaction. Inactive papain, blocked with E-64, appears to desensitize mice to subsequent challenge by active enzyme. These results suggest that the enzymatic activity of papain is critical in inducing an allergic response and that the use of inactive allergens may be a possible strategy for desensitizing allergic individuals.
To compare the effectiveness of finasteride and flutamide in the treatment of idiopathic hirsutism.
Randomized study.
Department of Gynecological Endocrinology, University of Brescia, Italy.
Forty-six women with idiopathic hirsutism were selected.
Patients were assigned randomly to receive 5 mg of finasteride once daily or 250 mg of flutamide twice daily for 12 consecutive months.
Hirsutism was evaluated at 6 and 12 months of therapy by measuring the Ferriman-Gallwey score and the terminal-hair diameters (microm) taken from different body areas. Blood samples were taken and side effects were monitored during the treatment.
Both finasteride and flutamide induced a statistically significant decrease in hirsutism scores and hair diameters at the end of 12 months. Finasteride reduced the Ferriman-Gallwey score by 20.5% at 6 months and by 34.2% at 12 months, and hair diameter by 18.9%-23.6% at 6 months and by 29.6%-37.9% at 12 months. Flutamide reduced the Ferriman-Gallwey score by 26.6% at 6 months and by 50.9% at 12 months, and hair diameter by 22.3%-28.2% at 6 months and by 47.7%-56.5% at 12 months. Flutamide did not induce hormonal variations, whereas finasteride increased T levels by 60% and decreased 3alpha-androstanediol glucuronide by 69.5% at 12 months.
Both drugs were effective in the treatment of idiopathic hirsutism, but flutamide was more effective than finasteride.
N(6)-Cyclopentyladenosine (CPA), a structural analog of adenosine, is a vasodilator with extensive pharmacological effects. However, little is known about the effect of CPA on wound healing and hair growth.
Cellular responses to CPA were measured in vitro by tetrazolium dye reduction and in vivo by bromodeoxyuridine (BrdU) uptake. The effect of CPA on healing of incisional and excisional wounds on the dorsum of diabetic (db/db, n = 94) and nondiabetic (db/+, n = 20) mice and hair growth along the wound margin was evaluated with wound breaking strength, wound closure rate, and quantitative histology.
CPA stimulated proliferation of BALB/3T3 fibroblasts and human dermal microvascular endothelial cells in both quiescent and nonquiescent phases. Wounds treated with CPA at 10 microM showed a significant increase in the number of BrdU-labeled cells, including keratinocytes, fibroblasts, endothelial cells, and cells in sebaceous glands and the outer root sheath of hair follicles, compared with controls (P < 0.05). CPA application (5.1 microg/daily for 12 days) significantly increased the breaking strength of incisional wounds at day 24 postwound (P < 0.05). Excisional wound closure rate in the CPA-treated group (3.4 microg/daily for 15 days) was accelerated starting at day 10 postwound compared with controls (P < 0.01). Tissue sections from CPA-treated wounds showed a sevenfold increase in hair follicle number, compared with controls (P < 0.01). Enhanced hair growth along the wound margin was revealed in CPA-treated groups.
CPA stimulated proliferation of many cell types in vivo and in vitro and enhanced wound healing and hair growth. Therefore, CPA could be an interesting candidate for clinical application.
To study the effects of proteolytic enzymes on mice hair follicles, particularly on cells of the bulge area regarded as follicle stem cells.
Previous application by iontophoresis of proteolytic enzymes on guinea pig skin resulted in degenerative effects on hair follicles and the hypothesis was proposed that some of the affected cells could be stem cells.
To mark putative stem cells transgenic mice were produced carrying the lac-Z gene fused to the Upstream Regulatory Region (URR) of Human Papilloma Virus 11 (HPV11), as they express this gene specifically in the cells of the bulge area. Chymotrypsin and papain were applied on skin by iontophoresis, trypsin in the form of liposomes.
Enzyme application, both by electrophoresis and as liposomes, led to intense degenerative effects of the hair follicle, such as detachment of the inner root sheath, cystic dilation of the hair shaft and presence of epithelial cells within the lumen. Some of these cells represent hair follicle stem cells expressing beta-galactosidase (beta-gal), having been detached from the bulge area as a result of enzyme treatment, implying impairment of their function.
The purpose of this study was to test the ability of topical formulations of finasteride and flutamide to re-enlarge hair follicles in male-pattern baldness. This was evaluated by an experimental model of human scalp skin graft transplanted onto SCID mice. A comparison was made between formulations containing finasteride and flutamide, and a vehicle formulation in terms of the mean hairs per graft, length, diameter of the shafts, and structures of the growth stages of the hair. Flutamide and finasteride had a significantly higher effect (P<0.05) than the placebo in all the tested parameters, but flutamide demonstrated more hair per graft and longer hair shafts than finasteride (P<0.05). The number of hairs per graft for flutamide and finasteride groups were 1.22+/-0. 47 and 0.88+/-0.95 hairs/0.5 mm2 graft, respectively, versus 0. 35+/-0.6 hairs/graft for vehicle-treated graft. Similarly, hair lengths for flutamide and finasteride were 5.82+/-0.50 and 4.50+/-0. 32 mm, respectively, versus 2.83+/-0.18 mm for the vehicle-treated grafts. An in vitro diffusion study of flutamide gel using hairless mouse skin demonstrated the beneficial effect of the vehicle composition in comparison with a hydroalcoholic solution or a gel containing no penetration enhancer. It is therefore suggested that this topical composition containing flutamide or finasteride may effectively result in regression of male-pattern baldness.
Objective:
To compare the long-term (1 year) effects of flutamide (250 mg/d) and finasteride (5 mg/d) for the treatment of hirsutism in women.
Design:
Randomized, prospective clinical study.
Setting:
Departments of Gynecology and Obstetrics and Endocrinology at Erciyes University Medical Faculty, Kayseri, Turkey.
Patient(s):
Seventy patients with hirsutism were studied.
Intervention(s):
Thirty-five patients were treated with flutamide (250 mg/d) and 35 patients with finasteride (5 mg/d) for 12 months. Hirsutism score, hormone levels, and multiscreen blood chemistry were measured at 3-month intervals.
Main outcome measure(s):
Reduction in hair growth.
Result(s):
The modified Ferriman-Gallwey scores for hirsutism decreased significantly at months 6 and 12 from a mean +/- SD of 17. 8 +/- 5.8 to 6.0 +/- 3.4 and 17.8 +/- 5.8 to 4.8 +/- 3.2, respectively, in group 1; and from 19.1 +/- 6.1 to 14.2 +/- 4.9 and 19.1 +/- 6.1 to 11.3 +/- 5.0 in group 2, respectively. There were no statistically significant differences in any of the hormonal indices in group 1, but in group 2, E(2) and sex hormone-binding globulin increased significantly while DHEAS decreased significantly at 12 months of therapy.
Conclusion(s):
This study shows that flutamide (250 mg/d) is more effective than finasteride (5 mg/d) in reducing hair growth. We conclude that flutamide (250 mg/d) may represent a more effective and well tolerated treatment for patients.
The location of follicular and epidermal stem cells in mammalian skin is a crucial issue in cutaneous biology. We demonstrate that hair follicular stem cells, located in the bulge region, can give rise to several cell types of the hair follicle as well as upper follicular cells. Moreover, we devised a double-label technique to show that upper follicular keratinocytes emigrate into the epidermis in normal newborn mouse skin, and in adult mouse skin in response to a penetrating wound. These findings indicate that the hair follicle represents a major repository of keratinocyte stem cells in mouse skin, and that follicular bulge stem cells are potentially bipotent as they can give rise to not only the hair follicle, but also the epidermis.
The hair follicle possesses progenitor cells for continued hair follicle cycling and for epidermal keratinocytes, melanocytes and Langerhans cells. These different cell types can be targeted by topical gene delivery to mouse skin. Using a combination of liposomes and DNA, we demonstrated the feasibility of targeting hair follicle cells in human scalp xenografts as well. We defined liposome composition and stage of the hair cycle as important parameters influencing transfection of human hair follicles. Transfection occurred only during anagen onset. Considerations and obstacles for using gene therapy to treat alopecias and skin disease are discussed. A theoretical framework for future gene therapy treatments for cutaneous and systemic disorders is presented.
Optimisation of drug delivery through human skin is important in modern therapy. This review considers drug-vehicle interactions (drug or prodrug selection, chemical potential control, ion pairs, coacervates and eutectic systems) and the role of vesicles and particles (liposomes, transfersomes, ethosomes, niosomes). We can modify the stratum corneum by hydration and chemical enhancers, or bypass or remove this tissue via microneedles, ablation and follicular delivery. Electrically assisted methods (ultrasound, iontophoresis, electroporation, magnetophoresis, photomechanical waves) show considerable promise. Of particular interest is the synergy between chemical enhancers, ultrasound, iontophoresis and electroporation.
The hair follicle possesses progenitor cells required for continuous hair follicle cycling and for epidermal keratinocytes, melanocytes and Langerhans cells. These different cell types can be the target of topical gene delivery in the skin of the mouse. Using a combination of liposomes and DNA, we demonstrate the feasibility of targeting hair follicle cells in human scalp xenografts. We consider liposome composition and stage of the hair cycle as important parameters influencing transfection of human hair follicles. Transfection is possible only during the early anagen phase. Factors and obstacles for the use of gene therapy in treating alopecia and skin diseases are discussed. A theoretical framework for future treatment of cutaneous and systemic disorders using gene therapy is presented.
One long-standing approach for improving transdermal drug delivery uses penetration enhancers (also called sorption promoters or accelerants) which penetrate into skin to reversibly decrease the barrier resistance. Numerous compounds have been evaluated for penetration enhancing activity, including sulphoxides (such as dimethylsulphoxide, DMSO), Azones (e.g. laurocapram), pyrrolidones (for example 2-pyrrolidone, 2P), alcohols and alkanols (ethanol, or decanol), glycols (for example propylene glycol, PG, a common excipient in topically applied dosage forms), surfactants (also common in dosage forms) and terpenes. Many potential sites and modes of action have been identified for skin penetration enhancers; the intercellular lipid matrix in which the accelerants may disrupt the packing motif, the intracellular keratin domains or through increasing drug partitioning into the tissue by acting as a solvent for the permeant within the membrane. Further potential mechanisms of action, for example with the enhancers acting on desmosomal connections between corneocytes or altering metabolic activity within the skin, or exerting an influence on the thermodynamic activity/solubility of the drug in its vehicle are also feasible, and are also considered in this review.
Influência do método de depilação na espessura da epiderme de animais tratados com papaína
- E Traversa
- M V R Velasco
- G M Machado-Santelli
- A T B Ottolenghi
- D C P Menegotto
Traversa, E., Velasco, M.V.R., Machado-Santelli, G.M., Ottolenghi, A.T.B.,
Menegotto, D.C.P., 2002. Influência do método de depilação na espessura da epiderme de animais tratados com papaína. Cosmet. Toiletries 14,
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- J D Bancroft
- A Stevens
- W B Barry
Bancroft, J.D., Stevens, A., 1996. Theory and Practice of Histological Tech-niques. Churchill Livingstone, Edinburgh. Barry, W.B., 2001. Novel mechanisms and devices to enable successful trans-dermal drug delivery. Eur. J. Pharm. Sci. 14, 101–114.
Involvement of follicular stem cells in forming not only the follicle but also the epidermis
- Taylor
Influência do método de depilação na espessura da epiderme de animais tratados com papaína
- Traversa