Chemical Profile and Biological Effects of an Herbal Mixture for the Development of an Oil-in-Water Cream.

Three individual hydroalcoholic extracts derived from Hamamelis virginiana leaves, Krameria lappacea root, Salix alba bark, and the resulting herbal mixture (HM) were assessed for the phytochemical profile as well as for antibacterial and cytotoxic potential. The chemical composition of the individual extracts and of their mixture was analyzed by chromatographical (LC-MS) and spectrophotometrical methods. The antimicrobial properties were evaluated by using the agar-well diffusion and the broth microdilution assays, whereas the potential cytotoxicity was investigated on human keratinocyte cell line by MTT method and apoptosis test. The HM composition revealed important amounts of valuable polyphenolic compounds provided from the individual extracts, having synergistic biological effects. All tested extracts displayed in vitro antimicrobial properties, with a significantly higher efficacy noticed for the HM when tested against Staphylococcus aureus. Moreover, none of the tested extracts was responsible for in vitro cytotoxicity against the human keratinocytes in the selected concentration range. Furthermore, the HM was included in an oil-in-water cream for the nonpharmacological treatment of seborrheic dermatitis, developed and optimized by using a QbD approach. A D-optimal experimental plan with four factors that varied on two levels was used to investigate the effect of the quantitative variation of the formulation factors (emulsifier, co-emulsifier, thickening agent, oily phase ratio) on the characteristics of the cream in terms of firmness, consistency, adhesiveness, stringiness, spreadability, and viscosity. Based on the experimental results, an optimal formulation containing 2.5% emulsifier and 20% oily phase was prepared and analyzed. The obtained results showed appropriate quality characteristics of this novel cream, which may be used in the future to manage the associated symptoms of seborrheic dermatitis.

Unveiling the Ability of Witch Hazel (Hamamelis virginiana L.) Bark Extract to Impair Keratinocyte Inflammatory Cascade Typical of Atopic Eczema.

Hamamelis virginiana L. bark extract is a traditional remedy for skin affections, including atopic dermatitis/eczema (AD). Hamamelis preparations contain tannins, including hamamelitannin (HT), although their pharmacological role in AD is still unknown. This study aimed to study the rational for its topical use by considering the impact of crucial biomarkers on AD pathogenesis. A standardized extract (HVE) (0.5−125 µg/mL) was compared to hamamelitannin (HT), its main compound (0.5−5 µg/mL), in a model of human keratinocytes (HaCaTs), challenged with an AD-like cytokine milieu (TNF-α, IFN-γ, and IL-4). HVE inhibited the release of mediators involved in skin autoimmunity (IL-6 and IL-17C) and allergy (TSLP, IL-6, CCL26, and MMP-9) with a concentration-dependent fashion (IC50s < 25 µg/mL). The biological mechanism was ascribed, at least in part, to the impairment of the NF-κB-driven transcription. Moreover, HVE counteracted the proliferative effects of IL-4 and recovered K10, a marker of skin differentiation. Notably, HT showed activity on well-known targets of IL-4 pathway (CCL26, K10, cell proliferation). To the best of our knowledge, this work represents the first demonstration of the potential role of Hamamelis virginiana in the control of AD symptoms, such as itch and skin barrier impairment, supporting the relevance of the whole phytocomplex.

Extratos de romã e hamamélis: existe ação antimicrobiana sobre cepas clínicas de Acinetobacter baumannii e Klebsiella pneumoniae multirresistentes? / Pomegranate and witch hazel extracts: is there antimicrobial action on clinical strains of multiresistant Acinetobacter baumannii and Klebsiella pneumoniae?

Acinetobacter baumannii e Klebsiella pneumoniae são patógenos prioritários globais resistentes a antibióticos, portanto, é relevante encontrar métodos alternativos para controlá-los, e os fitoterápicos têm sido investigados para essa finalidade. Em vista disso, o objetivo desse estudo foi avaliar a atividade antimicrobiana e antibiofilme do extrato glicólico de Hamamelis virginiana (hamamélis) e do extrato hidroalcóolico de Punica granatum (romã) isolados e em combinação sobre cepas ATCC e clínicas de A. baumannii e K. pneumoniae. Primeiramente, foi determinada a Concentração Inibitória Mínima (CIM) e a Concentração Microbicida Mínima (CMM) pelo teste de microdiluição em caldo, segundo Clinical and Laboratory Standards Institute (CLSI), após, o potencial antimicrobiano da combinação entre esses extratos foi analisado sobre esses micro-organismos por meio da técnica "checkerboard". Posteriormente, a redução da atividade metabólica dos biofilmes foi avaliada pelo teste de MTT (contato de 5 min e 24h), para tanto a CMM e as duas concentrações acima dela foram utilizadas. A concentração correspondente a 2X a CMM dos extratos combinados foi utilizada no teste de MTT para o tratamento dos biofilmes no tempo de 5 min e 24h. Os extratos isolados e combinados apresentaram ação microbicida contra A. baumannii e K. pneumoniae na forma planctônica. O extrato de romã apresentou CMM de 1,71 a 3,43 mg/mL para A. baumannii e de 3,43 a 13,75 mg/mL para K. pneumoniae, enquanto que Hamamélis exibiu CMM de 12,5 a 25 mg/mL para A. baumannii e de 25 a 50 mg/mL para K. pneumoniae. O uso combinado dos extratos resultou em treze concentrações aditivas para A. baumannii e quinze para K. pneumoniae. A redução de viabilidade em biofilmes de A. baumannii chegou a 64,94% para o extrato de romã, e 83,45% para o extrato de Hamamélis. K. pneumoniae apresentou redução máxima de 75,66% com extrato de Romã e de 76,56% com o extrato de Hamamelis. A redução do biofilme variou conforme a concentração e cepa quando tratado com a combinação dos extratos. Com isso, pode-se concluir que os extratos apresentam importante ação antibacteriana e antibiofilme sobre os patógenos avaliados, podendo ser considerados potenciais agentes terapêuticos para o combate destes patógenos.

Pomegranate and witch hazel extracts: is there antimicrobial action on clinical strains of multiresistant Acinetobacter baumannii and Klebsiella pneumoniae? [dissertation]. São José dos Campos (SP): São Paulo State University (Unesp), Institute of Science and Technology; 2021. ABSTRACT Acinetobacter baumannii and Klebsiella pneumoniae are global priority antibiotic resistant pathogens, therefore, it is relevant to find alternative methods to control them, and herbal medicines have been investigated for this purpose. In view of this, the aim of this study was to evaluate the antimicrobial and antibiofilm activity of the glycolic extract of Hamamelis virginiana (witch hazel) and the hydroalcoholic extract of Punica granatum (pomegranate) isolated and in combination on ATCC and clinical strains of A. baumannii and K. pneumoniae. First, the Minimum Inhibitory Concentration (MIC) and the Minimum Microbicide Concentration (MMC) were determined by the broth microdilution test, according to the Clinical and Laboratory Standards Institute (CLSI), after which the antimicrobial potential of the combination of these extracts was analyzed on these microorganisms through the "checkerboard" technique. Subsequently, the reduction of the metabolic activity of biofilms was evaluated by the MTT test (5 min and 24h contact), for both the CMM and the two concentrations above it were used. The concentration corresponding to 2X the CMM of the combined extracts was used in the MTT test for the treatment of biofilms in a time of 5 min and 24h. The isolated and combined extracts showed microbicidal action against A. baumannii and K. pneumoniae in the planktonic form. The pomegranate extract showed MMC from 1.71 to 3.43 mg/mL for A. baumannii and from 3.43 to 13.75 mg/mL for K. pneumoniae, while Witch Hazel exhibited MMC from 12.5 to 25 mg µg/ml for A. baumannii and from 25 to 50 mg/ml for K. pneumoniae. The combined use of the extracts resulted in thirteen additive concentrations for A. baumannii and fifteen for K. pneumoniae. The viability reduction in A. baumannii biofilms reached 64.94% for the pomegranate extract, and 83.45% for the Witch Hazel extract. K. pneumoniae showed a maximum reduction of 75.66% with Pomegranate extract and 76.56% with Hamamelis extract. Biofilm reduction varied according to concentration and strain when treated with the combination of extracts. Thus, it can be concluded that the extracts have an important antibacterial and antibiofilm action on the evaluated pathogens, and can be considered potential therapeutic agents to combat these pathogens.

Avaliação in vitro da atividade antimicrobiana dos extratos de Calendula officinalis, Capsicum annuum L e Hamamelis virginiana L. sobre cepas clínicas multirresistentes de Klebsiella pneumoniae e Pseudomonas aeruginosa / In vitro Evaluation of antimicrobial activity of extracts from Calendula officinalis, Capsicum annuum L and Hamamelis virginiana L. on multiresistant clinical straines of Klebsiella pneumoniae and Pseudomonas aeruginosa

O conhecimento dos efeitos dos extratos naturais de plantas vem cada vez mais se mostrando um campo promissor. Extratos de Calendula officinalis, Capsicum annuum e Hamamelis virginiana apresentam propriedades analgésicas, anti-inflamatórias, antioxidantes e potencial antimicrobiano que deve ser aprofundado. Assim, o objetivo foi avaliar ação antimicrobiana dos extratos glicólicos em culturas planctônicas e biofilmes de cepas padrão e clínicas de Klebsiella pneumoniae e Pseudomonas aeruginosa. Foi realizado o teste de microdiluição em caldo segundo Clinical and Laboratory Standards Institute, para determinação da Concentração Inibitória Mínima (CIM) e Concentração Microbicida Mínima (CMM) dos extratos. Foram realizados biofilmes monotípicos, nos tempos de contato de 5 min e 24 h, utilizando o teste de MTT. A CIM e CMM variaram de 1,56 a 50 mg/mL para todas as cepas avaliadas. C. annuum (50 mg/mL) apresentou redução de 0,25% entre 8,60% para cepa ATCC de P. aeruginosa (5 min) e em 24 h redução de 99,89% entre 99,99%, destacando-se os extratos de C. annuum (100 mg/mL) e os de C. officinalis. C. annuum (200 mg/mL) apresentou redução de 18% de K. pneumoniae (ATCC 4352). Em 24 h houve redução de 8,86%-75,74%, para o extrato de C. annuum (100 mg/mL). Os extratos apresentaram resultados mais satisfatórios quando expostos aos tratamentos de 24 horas. As cepas clínicas K1, K2 e K3 responderam ao tratamento de C. annuum (50 e 100 mg/mL) (p>0,05%). Para cepa clínica K4, os extratos de C. annuum (50 e 100 mg/mL) e C. officinalis (12,5 mg/mL) promoveram redução de viabilidade semelhante a clorexidina 2% (p>0,05%). O extrato de C. annuum (50 e 100 mg/mL), promoveu a redução de viabilidade de P1 e P2, enquanto o extrato de C. officinalis (25 e 50 mg/mL), reduziu P1 (p>0,05%). Para as cepas P3 e P4 o extrato de 50 e 100 mg/mL de C. annuum, apresentaram resultados semelhantes a clorexidina. Conclui-se que todos os extratos apresentaram ação antimicrobiana em cultura planctônica, no entanto, o extrato de C. annuum foi o que apresentou importante ação antibiofilme (24 h) sobre cepas clínicas multirresistentes de K. pneumoniae e P. aeruginosa, podendo ser considerado um potencial agente antimicrobiano.

Empirical knowledge of natural plant extracts is increasingly proving to be a promising field. Extracts of Calendula officinalis, Capsicum annuum and Hamamelis virginiana have analgesic, anti-inflammatory, antioxidant and antimicrobial potential that should be further developed. This research objective was to evaluate the antimicrobial action of glycolic extracts in planktonic cultures and biofilms of standard and clinical strains of Klebsiella pneumoniae and Pseudomonas aeruginosa. The broth microdilution test was performed according to the Clinical and Laboratory Standards Institute, to determine the Minimum Inhibitory Concentration (MIC) and Minimum Microbicide Concentration (MMC) of the extracts. Tests were performed on monotypic biofilms, at contact times of 5 min and 24 h, using the MTT test. The MIC and CMM of the extracts ranged from 1.56 to 50 mg/mL for all strains evaluated. C. annuum (50 mg/mL) showed a reduction of 0.25% between 8.60% for the ATCC strain of P. aeruginosa (5 min) and in 24 h a reduction of 99.89% between 99.99%, highlighting extracts from C. annuum (100 mg/mL) and from C. officinalis. C. annuum (200 mg/mL) showed an 18% reduction in K. pneumoniae (ATCC 4352). In 24 h there was a reduction of 8.86% between 75.74% for the extract of C. annuum (100 mg/mL). The extracts showed more satisfactory results when exposed to 24-hour treatments. Clinical strains K1, K2 and K3 responded to the treatment of C. annuum (50 and 100 mg/mL) (p>0.05%). For clinical strain K4, extracts of C. annuum (50 and 100 mg/mL) and C. officinalis (12.5 mg/mL) promoted a reduction in viability similar to chlorhexidine 2% (p>0.05%). The extract of C. annuum (50 and 100 mg/mL), reduced the viability of P1 and P2, while the extract of C. officinalis (25 and 50 mg/mL), reduced P1 (p>0.05% ). For strains P3 and P4, the extract of 50 and 100 mg/mL of C. annuum showed similar results to chlorhexidine. It is concluded that all extracts showed antimicrobial action in planktonic cultures, however, the extract of C. annuum was the one that showed an important antibiofilm action (24 h) on multiresistant clinical strains of K. pneumoniae and P. aeruginosa and can be considered a potential antimicrobial agent

Clinical trial shows lasting function of a new moisturizing cream against vaginal dryness.

Vaginal dryness and associated symptoms may occur in women of any age and are a frequent burden after menopause. The North American Menopause Society recommends long-acting non-hormonal vaginal creams as first-line therapy. A new type of such creams was developed (Remifemin® FeuchtCreme in Austria and Germany). This hormone-free cream contains hamamelis virginiana distillate and well-established vaginal cream ingredients. We explored its physical function and tolerability in an open interventional clinical trial. 20 postmenopausal women (54 to 76 years, median 60) treated their vaginal dryness using this cream once daily for 7 days and reported about their symptoms before, 4 to 8 h after first and 14 to 22 h after last application. A physician assessed tolerability and local physical function. All patients completed the study. Local physical function significantly improved from dryness at baseline (mean 4.0 ± SD 1.8) to a normal moisture level, on average (6.3 ± 2.1 after first, 6.7 ± 2.1 after last application, p = 0.0001). Subjective assessment of a feeling vaginal dryness showed a significant improvement at both times (p = 0.0001). Onset and duration of feeling moisturized were reported to be 0 to 2 min and 11.3 ± 6.9 h after application. All women reported vaginal dryness at baseline. 55 and 80% of patients reported no dryness after first application and at the end of the investigation. The cream was seen at the application site for up to 21 h. Tolerability assessments did not reveal any relevant change over time. There were four adverse events in 4 patients, all not serious and of mild intensity: urinary urgency (2), diarrhoea (1) and mild spotting after first application (1). The latter was caused by the dry surface of the applicator and was avoided by moisturizing the surface of the applicator at subsequent applications. In conclusion, these study results indicate a well-tolerated and long-acting function of this new vaginal moisturizing cream. Further clinical research in more patients will follow.

Contact dermatitis as an adverse reaction to some topically used European herbal medicinal products - part 2: Echinacea purpurea-Lavandula angustifolia.

This review focuses on contact dermatitis as an adverse effect of a selection of topically used herbal medicinal products for which the European Medicines Agency has completed an evaluation up to the end of November 2013 and for which a Community herbal monograph has been produced. Part 2: Echinacea purpurea Moench-Lavandula angustifolia Mill.