Identification of Flavonoids, Antioxidant and Antiproliferative Activity of Aqueous Infusions of Calendula officinalis L., Chelidonium majus L., Teucrium chamaedrys L. and Alchemilla vulgaris L.

Research background: The current changes in the global economy, characterised by the climate crisis and the economic and health impact of the COVID-19 pandemic, have led to a significant demand for medicinal herbs. This trend is expected to increase significantly by 2050. In this study, we investigated the biopotential of aqueous infusions of four medicinal plants: Calendula officinalis, Chelidonium majus, Teucrium chamaedrys and Alchemilla vulgaris. Experimental approach: The flavonoid analysis of the aqueous infusions of the selected plants was carried out using the RP-HPLC technique. The antiproliferative activity of the prepared aqueous plant infusions was analysed against three human cancer cell lines (MDA-MD-231, T24 and A549), while the antioxidant potential was measured using three antioxidant methods (DPPH, FRAP and Rancimat assay). Results and conclusions: T. chamaedrys had the highest total phenolics (expressed as GAE (2061±42) mg/L), free radical scavenging activity (IC50=1.9 mg/mL) and Fe(III) reducing antioxidant power (expressed as FeCl2 (9798±27) mg/L). At a concentration of 1 mg/mL, the antiproliferation of T24 by C. majus was 96 % and of MDA-MD-231 cells by A. vulgaris was 75 % after 72 h. After principal component analysis, T. chamaedrys and C. majus were grouped together. Quercetin glucoside and antioxidant capacity (DPPH) contributed the most to differentiate these infusions from the other two. Novelty and scientific contribution: This study represents a comparative analysis of the biopotential of four medicinal plants. A new RP-HPLC method was developed to separate the flavonoids in the herbal infusions. This is the first report on the presence of kaempferol-3-O-rutinoside in C. officinalis and isorhamnetin-3-O-rutinoside in A. vulgaris aqueous infusion. For the first time, C. majus has been shown to contribute to the oxidative stability of edible oil. Furthermore, this is the first comparative study on the antiproliferative activity of selected medicinal plants against the cell lines MDA-MD-231, T24 and A549.

Satureja hortensis L. and Calendula officinalis L., Two Romanian Plants, with In Vivo Antiparasitic Potential against Digestive Parasites of Swine.

Internal parasitic diseases of swine constitute a major welfare and health concern in low-input livestock farming. Due to an increase in chemical resistance, phytotherapeutic remedies have become an alternative for the prophylaxis and therapy of digestive parasitosis, albeit few remedies have been subjected to scientific validation. Low-input swine farming in Romania has adopted the traditional use of phytotherapy for controlling pathogens in livestock. The current study aimed to assess the antiparasitic potential of Calendula officinalis and Satureja hortensis against digestive parasites of swine in two low-input farms. The fecal samples were collected from sows, fatteners, and weaners, and were tested using the following coproparasitological methods: centrifugal sedimentation, flotation (Willis, McMaster egg counting technique), Ziehl-Neelsen stain modified by Henricksen, modified Blagg method, and in vitro nematode larvae/protozoan oocyst cultures. Six species of digestive parasites were diagnosed, namely Ascaris suum, Trichuris suis, Oesophagostomum spp., Balantioides coli, Eimeria spp., and Cryptosporidium spp., in various combinations, dependent on the swine category. A dose of 140 mg/kg bw/day of C. officinalis and 100 mg/kg bw/day of S. hortensis powders administered for 10 consecutive days revealed a strong antiprotozoal and anthelmintic activity on the aforementioned parasites. The curative efficacy can be attributed to the presence of polyphenols, sterols, tocopherols, and methoxylated flavones. In conclusion, our results indicate that S. hortensis and C. officinalis are promising alternatives to the commercially available antiparasitics, enabling their use as natural antiparasitic products against gastrointestinal parasites in pigs.

Analysis of the Impact of Ethanol Extract of Calendula officinalis L. on Human Fibroblast Cell Cultures Using the PANsys 3000 Device for Breeding and Visualization of Cells.

Calendula officinalis L. promotes wound healing and might be effective in gingival fibroblast stimulation. The influence of different concentrations of Calendula officinalis L. ethanol extract on human gingival fibroblast was visualized using PANsys 3000-a fully automated cell culture device used for in vitro culture to study cells under conditions similar to in vivo. The human fibroblast cells were isolated from gingival tissue. The 100% brew of Calendula officinalis L., as well as 7% and 20% Calendula officinalis L. ethanol extract, were added to the cultured cells and observed for 72 h. The qualitative and quantitative composition of the volatile compounds of marigold Calendula officinalis L. flowers are presented in this study. The essential oil compounds of the decoction were isolated with solid-phase microextraction (SPME) and analyzed with gas chromatography coupled with tandem mass spectrometry (GC-MS/MS). The presence of terpenoids, flavonoids, and other compounds was demonstrated. The composition was correlated with the fragrance properties. Observation of gingival fibroblast showed that there were no changes in cell morphology and proliferation after 100% Calendula officinalis L. brew stimulation. The growth and cell division were not inhibited. Likewise, the addition of 7% or 20% ethanol in water extract of Calendula officinalis L. stimulation did not inhibit the fibroblast proliferation. Overall, ethanol extracts of Calendula officinalis L. decrease the alcohol cytotoxic influence on gingival fibroblasts.

Effects of Calendula officinalis and Capsicum annum glycolic extracts on planktonic cells and biofilms of multidrug-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa.

Empirical knowledge of natural plant extracts is increasingly proving to be a promising field. The effect of Calendula officinalis L. (CO) and Capsicum annum (CA) glycolic extracts (GlExt) have potential that should be further developed in microbial tests. The effect of CO-GlExt and CA-GlExt was evaluated on eight multidrug-resistant clinical strains of Klebsiella pneumoniae and Pseudomonas aeruginosa, as well as collection strains for each bacterial. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined in comparison with 0.12% chlorhexidine. The tests were performed on single species biofilms, at 5 min and 24 h, using the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. The MIC and MBC of the extract ranged from 1.56 to 50 mg mL-1 for all strains evaluated. Analysis of the MTT assay revealed a strong antimicrobial potential of CA-GlExt, comparable to chlorhexidine. The findings suggest that CA-GlExt is effective against multidrug-resistant strains of K. pneumoniae and P. aeruginosa in planktonic state and biofilms.

Effect of Ethylene and Abscisic Acid on Steroid and Triterpenoid Synthesis in Calendula officinalis Hairy Roots and Saponin Release to the Culture Medium.

Phytohormones (plant growth regulators) can be applied as efficient elicitors to enhance the productivity of plant in vitro cultures, due to their significance in regulating the plant metabolism and strong influence on plant defense responses. In the present study, the effects of exogenous ethylene (ETY, applied in the form of ethephon as an ethylene-generating agent) and abscisic acid (ABA) on the synthesis of triterpenoids and steroids in Calendula officinalis hairy roots were investigated. ABA appeared to be an efficient elicitor of the biosynthesis of triterpenoid oleanolic acid (almost two-fold) and the release of its glycosides (saponins) to the culture medium (up to 6.6-fold). ETY had only a slight effect on triterpenoid metabolism; instead, it strongly influenced steroid metabolism, leading to profound modifications of the quantitative profiles of these compounds, particularly the ratio of stigmasterol to sitosterol. Both the applied phytohormones influenced the interplay between steroid and triterpenoid biosynthetic pathways, revealing the symptoms of their competition.

Efectividad terapéutica de la jalea de caléndula al 1 % en pacientes con estomatitis aftosa recurrente / Therapeutic effectiveness of calendula jelly at 1% in patients with aphthous recurrent stomatitis

Introducción: Para el tratamiento de pacientes con estomatitis aftosa recurrente suele emplearse el extracto fluido y la tintura de Caléndula officinalis L. como colutorio disuelto en agua. Teniendo en cuenta los inconvenientes que generan estas formulaciones se elaboró una jalea al 1 % contentiva de los principios activos de la referida planta. Objetivo: Evaluar la efectividad terapéutica de la jalea de caléndula al 1 % en pacientes con estomatitis aftosa recurrente. Métodos: Se realizó una intervención terapéutica en 30 pacientes con estomatitis aftosa recurrente, atendidos en el Policlínico Docente 30 de Noviembre de Santiago de Cuba, desde marzo hasta diciembre de 2019. La muestra se dividió en 2 grupos: control y de estudio, que recibieron tratamiento convencional y jalea de caléndula al 1 %, respectivamente. Se tuvieron en cuenta algunas variables de interés, a saber: edad, sexo, tiempo de remisión del dolor y de cicatrización, respuesta al tratamiento y efectividad terapéutica. Se empleó el porcentaje como medida de resumen y el estadígrafo de Ji al cuadrado con un nivel de significación de 0,05. Resultados: En la serie predominaron el grupo etario de 15- 24 años de edad (33,3 %) y el sexo femenino (60,0 %). En los tratados con la jalea de caléndula, la remisión del dolor y la cicatrización de las lesiones ulcerativas se evidenciaron en las primeras 24 y 48 horas, respectivamente, y la mayoría de estos habían mejorado a las 72 horas. Conclusiones: La jalea de caléndula al 1 % es efectiva como analgésico y cicatrizante en pacientes con estomatitis aftosa recurrente.

Introduction: The officinalis L Calendula fluid extract and tincture is usually used as mouthwash dissolved in water for the treatment of patients with aphthous recurrent stomatitis. Taking into account the inconveniences that generate these formulations a jelly at 1 % was elaborated with the active principles of the plant above mention. Objective: To evaluate the therapeutic effectiveness of the calendula jelly at 1% in patients with aphthous recurrent stomatitis. Methods: A therapeutic intervention in 30 patients with aphthous recurrent stomatitis, assisted in 30 de Noviembre Teaching Polyclinic was carried out in Santiago de Cuba, from March to December, 2019. The sample was divided in 2 groups: control and study that received conventional treatment and calendula jelly at 1%, respectively. Some variables of interest were taken into account, such as: age, sex, pain regression and scaring time, response to the treatment and therapeutic effectiveness. The percentage was used as summary measure and the chi-squared test with a significance level of 0.05. Results: In the series there was a prevalence of the 15 - 24 age group (33.3 %) and female sex (60.0 %). In the patients treated with the calendula jelly, pain regression and scaring of the ulcerative lesions were evidenced in the first 24 and 48 hours, respectively, and most of them had improved at 72 hours. Conclusions: The calendula jelly at 1% is effective as analgesic and healing in patients with aphthous recurrent stomatitis.

Effects and Characterization of Some Topical Ointments Based on Vegetal Extracts on Incision, Excision, and Thermal Wound Models.

The present paper aims to formulate and characterize four phytotherapeutic ointments based on Hippophae fructus, Calendulae flos, Bardanae folium, and Millefolii herba, which are included in a novel ointment base. In order to investigate the healing properties of the ointments, in vivo experimental wound models of linear incision, circular excision, and thermal burn were performed on Wistar rats. Topical treatment was performed daily for 21 days. Determination of the wound contraction rate (WCR), the period of reepithelization, and histopathological examination were achieved. Additionally, for the tested ointments, oscillatory and rotational rheological tests were carried out, and for the extracts, HPLC analysis was performed. The results demonstrate that the tested novel ointments are safe for use and the most effective ointment proved to be the one based on Arctium lappa, followed by that of Calendulae flos.

Hydrogel Delivery System Containing Calendulae flos Lyophilized Extract with Chitosan as a Supporting Strategy for Wound Healing Applications.

Calendulae flos is a valued plant material with known anti-inflammatory and antimicrobiological properties. The limitation for its use in the treatment of chronic wounds is the lack of adhesion to the required site of action. Obtaining the Calendulae flos lyophilized extract from water-ethanolic extract allowed to prepare valuable material whose biological activity in the wound healing process was confirmed by a positive result of the scratch test. The Calendulae flos lyophilized extract was standardized for the contents of the chlorogenic acid and the narcissin. The significant potency of the Calendulae flos pharmacological activity has become the reason for studies on its novel applications in combination with the multifunctional chitosan carrier, to create a new, valuable solution in the treatment of chronic wounds. The use of chitosan as a carrier allowed for the controlled release of the chlorogenic acid and the narcissin. These substances, characterized by prolonged release from the chitosan delivery system, were identified as well permeable, based on the results of the studies of the parallel artificial membrane permeability assay (PAMPA Skin) a model simulating permeability through membrane skin. The combination of the Calendulae flos lyophilized extract and the chitosan allowed for synergy of action towards hyaluronidase inhibition and effective microbiological activity. Optimization of the hypromellose hydrogel preparation ensuring the required rheological properties necessary for the release of the chlorogenic acid and the narcissin from the chitosan delivery system, as well as demonstrated antimicrobial activity allows indicating formulations of 3% Calendulae flos lyophilized extract with chitosan 80/500 in weight ratio 1:1 and 2% or 3% hypromellose as an important support with high compliance of response and effectiveness for patients suffering from chronic wounds.

Effects of EDTA, citric acid, and tartaric acid application on growth, phytoremediation potential, and antioxidant response of Calendula officinalis L. in a cadmium-spiked calcareous soil.

The improved efficiency of cadmium (Cd) phytoextraction potential of Calendula officinalis L. was evaluated in Cd-spiked calcareous soil, using various chelating agents. In a greenhouse study, three chelating agents, including EDTA, citric acid (CA), and tartaric acid (TA), were applied to Cd-spiked soils (50 and 100 mg kg-1) under C. officinalis L. cultivation. According to the results, C. officinalis grew normally without any toxicity signs at various Cd levels of the soil; however, with increasing the Cd levels, the plant dry weight biomass decreased, and activities of antioxidant enzymes (AOEs) increased. The application of CA and TA in Cd-spiked soils improved the physiologic traits of plants and mitigated the Cd stress since the activities of AOEs decreased. Oppositely, due to increasing the Cd excessive permeability to the root of the plant, EDTA application diminished the physiologic traits and increased the activities of AOEs. The results also showed that all the chelators, especially EDTA, markedly increased the Cd mobility factor (from 58.80% to 65.20-89.60%) in Cd-spiked soils. The bioconcentration factor (BCF = 1.3-2.90) and translocation factor (TF = 1.28-1.58) of Cd, which were >1 in all treated and untreated plant samples, as well as the accumulated Cd >100 mg kg-1, demonstrated that C. officinalis is a Cd-hyperaccumulator plant which could remediate Cd by the phytoextraction process. Regarding the biodegradation of CA, as well as the increased TF efficiency of Cd and plant biomass of CA treatments (by decreasing oxidative stress), compared to EDTA and TA treatments, it is recommended that CA be used as a superior chelating agent to enhance the efficiency of Cd phytoremediation in C. officinalis.

Evaluation of Calendula officinalis L. (marigold) flower as a natural growth promoter in comparison with an antibiotic growth promoter on growth performance, carcass traits and humoral immune responses of broilers.

The objective of this experiment was to investigate the effects of dried powder of Calendula officinalis L. (marigold) flower as an antibiotic growth promoter substitute on growth performance, organ weights, and immunological parameters in broilers. A total of 240 mixed sex broilers (Ross 308) at 1 d of age were individually weighed and randomly assigned to 4 treatments with 4 replicates of 15 birds for 6 wk. The dietary treatments were: 1) a basal diet (control); 2) control plus 4.5 mg flavophospholipol/kg of diet; 3) control plus 5 g marigold/kg of diet; 4) control plus 10 g marigold/kg of diet. Antibody responses against Newcastle (NDV), influenza (AI) viruses, and sheep red blood cells (SRBC) were measured. Supplementing 4.5 mg flavophospholipol/kg of diet enhanced body weight (BW) of broilers at 14 and 28 d of age (P < 0.05), but final BW at 42 d was not markedly affected. At 14 and 28 d of age, broilers fed diets supplemented with 10 g marigold/kg of diet had significantly lower BW compared with broilers fed the basal diet supplemented with antibiotic or 5 g marigold/kg of diet. During the starter period, broilers fed diets supplemented with antibiotic had significantly (P < 0.05) better FCR compared with broilers supplemented with 10 g marigold/kg of diet, but did not differ from broilers fed the basal diet or basal diet supplemented with 5 g marigold/kg of diet. There were no significant differences in FCR between treatments, during grower, finisher phases as well as for the whole experiment. Carcass yield was significantly (P < 0.05) higher in broilers supplemented with 5 g marigold/kg of diet compared with broilers in other groups. The treatments failed to induce any marked effect on immune parameters. In conclusion, the results of this experiment showed that supplementation of 5 and 10 g dried powder of marigold/kg of diet has no affirmative influence on growth performance of broilers.

Medicinal plants as therapeutic options for topical treatment in canine dermatology? A systematic review.

BACKGROUND: Medicinal plants have been used traditionally since centuries for wound care and treatment of skin diseases both in human and animals. Skin diseases are one of the most common reasons for owners to take their dog to the veterinarian. The demands for treatment and prophylaxis of these diseases are broad. A wide range of bacteria including antibiotic-resistant bacteria can be involved, making the treatment challenging and bear an anthropo-zoonotic potential. The aim of this review is to systematically evaluate based on recent scientific literature, the potential of four medicinal plants to enrich the therapeutic options in pyoderma, canine atopic dermatitis, otitis externa, wounds and dermatophytosis in dogs. RESULTS: Based on four books and a survey among veterinarians specialized in phytotherapy, four medicinal plants were chosen as the subject of this systematic review: Calendula officinalis L. (Marigold), Hypericum perforatum L. agg. (St. John's Wort), Matricaria chamomilla L. (syn. Matricaria recutita L., Chamomile) and Salvia officinalis L. (Sage). According to the PRISMA statement through literature research on two online databases a total of 8295 publications was screened and narrowed down to a final 138 publications for which full-text documents were analyzed for its content resulting in a total of 145 references (21 clinical, 24 in vivo and 100 in vitro references). CONCLUSIONS: All four plants were proven to have antibacterial and antifungal effects of a rather broad spectrum including antibiotic-resistant bacteria. This makes them an interesting new option for the treatment of pyoderma, otitis externa, infected wounds and dermatophytosis. Marigold, St. John's Wort and Chamomile showed wound-healing properties and are thus promising candidates in line to fill the therapeutic gap in canine wound-healing agents. St. John's Wort and Chamomile also showed anti-inflammatory and other beneficial effects on healthy skin. Due to the wide range of beneficial effects of these medicinal plants, they should be taken into account for the treatment of dermatologic diseases in dogs at least in future clinical research.

La efectividad de la crema Calendula officinalis L como tratamiento de la estomatitis aftosa recurrente / Calendula officinalis L cream effective treatment for recurrent aphthous stomatitis

RESUMEN Introducción: la estomatitis aftosa recurrente es una enfermedad habitual en las urgencias de Periodontología. Objetivo: evaluar la efectividad de la crema de Calendula officinalis L. al 10%, en el tratamiento de la estomatitis aftosa recurrente, en una clínica estomatológica. Método: se realizó un ensayo no controlado- no aleatorizado, en una población de 15 a 34 años de edad; que acudieron a la consulta de periodoncia de la Clínica Estomatológica Docente "Manuel Angulo Farrán" en Holguín, Cuba, en el período comprendido del año 2015 al 2017. La muestra fue de 100 pacientes, tratada durante 6 días, con consultas cada 48 horas. Resultados: el 59% de los pacientes fueron del sexo femenino, el 63% entre los 25 y 34 años, y en el 97% predominaron las aftas menores. La crema en estudio mejoró los síntomas en el 97% de los pacientes, y el 94% se curó. Conclusiones: la crema de Calendula officinalis L al 10% parece eficaz en el tratamiento de la estomatitis aftosa recurrente, aunque se requieren ensayos clínicos controlados y aleatorizados que lo demuestren de forma definitiva.

ABSTRACT Introduction: recurrent aphthous stomatitis is a frequent disease in periodontology emergencies. Objective: to evaluate the Calendula officinalis L at 10% cream effectiveness, as treatment for recurrent aphthous stomatitis, in a stomatological clinic. Method: non-control, non-randomized trial was conducted in a population between 15 and 34 years old, who attended "Manuel Angulo Farrán" Stomatological Teaching Clinic in Holguín, Cuba, from 2015 to 2017. Sample was 100 patients, treated in consultations every 48 hours, for 6 days. Results: the 59% of the patients were female, 63% between 25 to 34 years, and minor aphthae predominated in the 97%. The studied cream improved symptoms in 97% of patients and 94% were completely cured. Conclusions: Calendula officinalis L at 10% cream seems effective in the treatment of recurrent aphthous stomatitis, although controlled and randomized clinical trials would definitively demonstrate it.

Plant cell culture as emerging technology for production of active cosmetic ingredients.

Plants have always been the main source for active cosmetic ingredients, having proven health beneficial effects on human, such as anti-aging, antioxidant, anti-inflammatory, UV-protective, anti-cancer, anti-wrinkle, skin soothing, whitening, moisturizing, etc. Extracts from herbal, aromatic and/or medicinal plants have been widely used as effective active ingredients in cosmeceuticals or nutricosmetics, especially in products for topical application and skin-care formulations. However, over the past decade, there has been an increasing interest to plant cell culture - derived active cosmetic ingredients. These are "new generation" of high quality natural products, produced by the modern plan biotechnology methods, which usually showed stronger activities than the plant extracts obtained by the classical methods. In this review, the advantages and the current progress in plant cell culture technology for the production of active cosmetic ingredients have been summarized, and discussed in details within a presented case study for calendula stem cell product development.

Lettuce and marigold intercropping: crops productivity and marigold's flavonoid content / Consórcio entre alface e calêndula: produtividade das culturas e teor de flavonóides em calêndula

ABSTRACT: Intercropping of vegetables and medicinal plants might produce favorable interactions for both crops, resulting in increased production and profit per unit area. It is known that active compounds can change because of several factors, including the cropping system. Therefore, the goal of this study was to evaluate the effect of intercropping of lettuce and marigold on the productivity of these crops and on the flavonoid content of marigold flowers. The experiment was conducted in the experimental area of EPAMIG in Oratorios-MG. Treatments consisted of lettuce + marigolds intercropping between rows, lettuce + marigold intercropping between plants, and monocultures of each species. Spacing was 0.3x0.3m. The experimental design consisted of randomized blocks with ten repetitions. Harvest of lettuce and marigold flowers started 45 days after transplantation (DAT) and extended up to 72 DAT for marigolds. Lettuce and marigold intercropping appears feasible because lettuce production did not differ between the monoculture and intercropped cultivations, and marigold productivity was higher when intercropped with lettuce. No change in the flavonoid (active chemicals of medicinal interest) content in the floral capitula of marigolds was observed.

RESUMO: O cultivo consorciado entre hortaliças e plantas medicinais pode promover interação benéfica entre as duas culturas, resultando em aumento na produção e no lucro por unidade de área. Sabe-se também que os princípios ativos podem ser alterados por vários fatores, incluindo o sistema de cultivo. Dessa forma, o objetivo do presente trabalho foi avaliar o efeito do consórcio entre alface e calêndula na produtividade destas culturas e no teor de flavonoides em flores de calêndula. O experimento foi conduzido na área experimental da EPAMIG, em Oratórios-MG. Os tratamentos consistiram em: alface + calêndula entre linhas, alface + calêndula entre plantas e os monocultivos de alface e de calêndula. O espaçamento utilizado foi de 0,3x0,3m. O delineamento experimental foi em blocos casualizados com dez repetições. A colheita de alface e das flores de calêndula iniciou-se 45 dias após o transplantio (DAT), sendo a colheita de calêndula estendida até os 72 DAT. O consórcio entre alface e calêndula é viável, pois a produção da alface é semelhante entre sistemas consorciados e cultivo solteiro, e a produtividade da calêndula é maior quando consorciada com a alface, não havendo alteração no teor de flavonoides (princípio ativo de interesse medicinal) dos capítulos florais de calêndula.

Optimization of chemometric approaches for the extraction of isorhamnetin-3-O-rutinoside from Calendula officinalis L.

The application of Design of Experiments (DoE) to the determination of optimum conditions for an extraction process relies on the correct selection of mathematical models. The linear model is the one typically used; however, in some cases it does not always have superior performance, ignoring the real nature of the data and its appropriate descriptive model. In order to evaluate the extraction efficiency of isorhamnetin-3-O-rutinoside from flowers of Calendula officinalis L. a multivariate factorial analysis was used. Simulations were conducted using linear, quadratic, full cubic and special cubic models. A Simplex-Centroid design was chosen as it delivered greater precision with only minor errors versus other models tested. Analyses were performed by capillary zone electrophoresis using sodium tetraborate buffer (40mmolL(-1), pH 9.4) containing 10% methanol. The detection was linear over a range of 8.0-50.0mgL(-1) (r(2)=0.996), and the limits of detection (LOD) and quantification (LOQ) for isorhamnetin-3-O-rutinoside were 3.44mgL(-1) and 11.47mgL(-1), respectively. The full cubic model showed the best extraction results, with an error of 3.40% compared to analysis of variance, and a determination coefficient of 0.974. The difference between the responses at the reference point, calculated by the model, and the experimental response, varies around 2.72% for full cubic model. Comparison of the four models showed the full cubic model was the most appropriate one, allowing greater efficiency in the extraction of isorhamnetin-3-O-rutinoside. Selection of the model made it possible to obtain a 60% increase in sensitivity compared to the linear model.

Preclinical evaluation of collagen type I scaffolds, including gelatin-collagen microparticles and loaded with a hydroglycolic Calendula officinalis extract in a lagomorph model of full-thickness skin wound.

Previously, we have developed collagen type I scaffolds including microparticles of gelatin-collagen type I (SGC) that are able to control the release of a hydroglycolic extract of the Calendula officinalis flower. The main goal of the present work was to carry out the preclinical evaluation of SGC alone or loaded with the C. officinalis extract (SGC-E) in a lagomorph model of full-thickness skin wound. A total of 39 rabbits were distributed in three groups, of 13 animals each. The first group was used to compare wound healing by secondary intention (control) with wound healing observed when wounds were grafted with SGC alone. Comparison of control wounds with wounds grafted with SGC-E was performed in the second group, and comparison of wounds grafted with SGC with wounds grafted with SGC-E was performed in the third group. Clinical follow-ups were carried in all animals after surgery, and histological and histomorphometric analyses were performed on tissues taken from the healed area and healthy surrounding tissue. Histological and histomorphometric results indicate that grafting of SGC alone favors wound healing and brings a better clinical outcome than grafting SGC-E. In vitro collagenase digestion data suggested that the association of the C. officinalis extract to SGC increased the SGC-E cross-linking, making it difficult to degrade and affecting its biocompatibility.

Análise de amostras de flores de Calêndula (Calendula officinalis L, Asteraceae) comercializadas na grande Curitiba / Analysis of samples of flowers Calendula (Calendula officinalis L. ) Asteraceae commercially in Curitiba, PR, Brazil

A espécie Calendula officinalis L. é uma planta herbácea pertencente à família Asteraceae, suas flores apresentam propriedades antimicrobianas, emolientes, antiinflamatórias e tonificantes da pele. Os flavonoides são utilizados como marcadores para avaliar a qualidade da droga vegetal. O objetivo do presente trabalho foi avaliar a identidade e qualidade de seis amostras de flores de C. officinalis L. comercializadas na Grande Curitiba (PR). Foram realizados os ensaios de identificação macroscópica e de pureza, determinação de flavonoides, perfil do extrato etanólico em cromatografia em camada delgada, avaliação das atividades antioxidante e atividade antibacteriana sobre Staphylococcus aureus ATCC 25923. Todas as seis amostras analisadas foram identificadas como C. officinalis L. e foram aprovadas no perfil cromatográfico, porém não atenderam aos requisitos farmacopeicos de pureza e teor de flavonoides, fatores que contribuíram para a diminuição da atividade antioxidante e antibacteriana. A má qualidade de plantas medicinais, decorrente de processos produtivos inadequados, pode ocasionar ausência ou diminuição do efeito terapêutico esperado, além de possíveis efeitos tóxicos.(AU)

Calendula officinalis L., is an herbaceous plant belonging to the family Asteraceae, its flowers have antimicrobial, emollient, anti-inflammatory and toning of the skin properties. Flavonoids are used as chemical markers to evaluate the quality of plant drug. The aim of this study was to evaluate the identity and quality of six samples of C. officinalis L. flowers commercialized in Curitiba City and Metropolitan Area (State of Paraná, Brazil). Tests of macroscopic identification, purity, determination of flavonoids were performed, in addition to the profile of the ethanolic extract in thin layer chromatography, evaluation of antioxidant activity and antibacterial activity against Staphylococcus aureus ATCC 25923. All six samples analyzed were identified as C. officinalis L. and were approved in chromatographic profile, but samples were in discordance with parameters established by Brazilian Pharmacopoeia of purity and content of flavonoids, factors that have contributed to the decline of antioxidant activity and antibacterial. The poor quality of medicinal plants as a result of productive processes inadequate can cause absence or reduction of the therapeutic effect expected, in addition to possible toxic effects.(AU)

Hyperaccumulator oilcake manure as an alternative for chelate-induced phytoremediation of heavy metals contaminated alluvial soils.

The ability of hyperaccumulator oilcake manure as compared to chelates was investigated by growing Calendula officinalis L for phytoremediation of cadmium and lead contaminated alluvial soil. The combinatorial treatment T6 [2.5 g kg(-1) oilcake manure+5 mmol kg(-1) EDDS] caused maximum cadmium accumulation in root, shoot and flower up to 5.46, 4.74 and 1.37 mg kg(-1) and lead accumulation up to 16.11, 13.44 and 3.17 mg kg(-1), respectively at Naini dump site, Allahabad (S3). The treatment showed maximum remediation efficiency for Cd (RR=0.676%) and Pb (RR=0.202%) at Mumfordganj contaminated site (S2). However, the above parameters were also observed at par with the treatment T5 [2.5 g kg(-1) oilcake manure +2 g kg(-1) humic acid]. Applied EDDS altered chlorophyll-a, chlorophyll-b, and carotene contents of plants while application of oilcake manure enhanced their contents in plant by 3.73-8.65%, 5.81-17.65%, and 7.04-17.19%, respectively. The authors conclude that Calendula officinalis L has potential to be safely grown in moderately Cd and Pb-contaminated soils and application of hyperaccumulator oilcake manure boosts the photosynthetic pigments of the plant, leading to enhanced clean-up of the cadmium and lead-contaminated soils. Hence, the hyperaccumulator oilcake manure should be preferred over chelates for sustainable phytoremediation through soil-plant rhizospheric process.

Physical chemistry evaluation of stability, spreadability, in vitro antioxidant, and photo-protective capacities of topical formulations containing Calendula officinalis L. leaf extract

Calendula is used widely in cosmetic formulations that present phenolic compounds in their chemical constitution. The objective of our research was to develop and evaluate the stability of topical formulations containing 5% hydro-ethanolic extract of calendula leaves, including spreadability, and in vitro photo-protective, and antioxidant capacity. To evaluate the stability, we used organoleptic characteristics, pH, and viscosity parameters. Antioxidant capacity was measured by the DPPH (2,2-diphenyl-1-picrylhydrazyl) method, and the photo-protective capacity by SPF spectrophotometric measure. All formulations were stable. The calendula extract formulations in gel and cream showed no significant variations in pH, and the cream formulations presented lower viscosity variations than gel formulations. The spreadability of the gel formulations was superior to those in cream. The formulations also presented good antioxidant capacities and an FPS of around 1.75. In accordance with the results, the formulations can be used as antioxidants, but considering the low SPF obtained, calendula cannot be considered as a stand-alone sunscreen, yet may well be tested in future studies towards verifying enhancement of synthetic sunscreens.

A calêndula é amplamente utilizada em formulações cosméticas, apresentando compostos fenólicos em sua constituição química. Desta forma, o objetivo desta pesquisa foi desenvolver e avaliar a estabilidade de formulações tópicas contendo 5% de extrato hidroetanólico das folhas de calêndula, bem como a espalhabilidade, capacidade antioxidante e fotoprotetora in vitro nas mesmas. Para a avaliação da estabilidade, foram usados parâmetros como a verificação das características organolépticas, pH e viscosidade. A capacidade antioxidante foi verificada pelo método do DPPH (2,2-difenil,1- picril-hidrazila) e a capacidade fotoprotetora pela medida espectrofotométrica do FPS. Para as formulações testadas, observou-se que apresentaram uma boa estabilidade. As formulações de creme e gel com extrato de calêndula não apresentaram variações significativas nos valores de pH e o creme apresentou as menores variações de viscosidade em relação ao gel. A espalhabilidade das formulações de gel foi superior à do creme. As formulações também apresentaram uma boa capacidade antioxidante e um FPS em torno de 1.75. De acordo com os resultados, a formulação pode ser utilizada com ação antioxidante, porém com o FPS obtido, a calêndula não pode ser considerada um filtro solar isolado, mas poderá ser testada em estudos futuros para verificar a potencialização de filtros solares sintéticos.

Acción antiedemagénica del extracto de Caléndula officinalis L. de producción nacional / Evaluation of the anti-edematogenic action of Calendula officinalis L. extract of domestic production

Introducción: la Caléndula officinalis L., es una planta conocida y utilizada por sus cualidades terapéuticas. A los extractos de sus flores se le atribuye, entre sus funciones, ser protector de los vasos sanguíneos. Objetivo: evaluar la acción antiedemagénica del extracto de hojas de Caléndula officinalis de producción nacional.Métodos: se utilizaron 120 ratas Wistar divididas en cuatro grupos de 30 animales, un grupo control y el resto con concentración de extracto de C. officinalis de 50, 100 y 200 mg/kg de peso, respectivamente; administrado por vía oral durante 3, 7 y 14 días. Se utilizaron como agentes edemagénicos la histamina y la dextrana grado A. Se midieron los edemas producidos en el lomo de las ratas y se calculó su área.Resultados: se halló una disminución (p < 0,05) en el área del edema inducido por la histamina y la dextrana en los tres grupos tratados con el extracto de C. officinalis. Para ambos agentes edemagénicos los resultados fueron similares y no significativos para las dosis de 50 y 100 mg/kg de peso, aunque la disminución fue mayor (p < 0,05) con la dosis de 200 mg/kg de peso. La reducción del edema fue dependiente del tiempo de tratamiento.Conclusiones: se demostró la acción antiedemagénica del extracto de C. officinalis, la dosis de 200 mg/Kg de peso fue la mejor y 14 días el tiempo de mejor efecto. Se recomienda su uso como flebotónico en el tratamiento de los trastornos vasculares venosos(AU)

Introduction: Calendula officinalis is a well-known plant used by its therapeutic qualities. Blood vessel protection is attributed to the extracts from its flowers. Objective: to evaluate the anti- edematogenic action of the extract from national Calendula officinalis leaves. Methods: one hundred twenty Wistar rats divided into four groups of 30 animals were used; one group control and three with C. Officinalis extract concentrations of 50, 100 and 200 mg/kg of weight, respectively, which were orally administered for 3,7 and 14 days. Histamine and the Dextrane A were the edematogenic agents. The edemas appeared in the rats' back were measured to estimate the area.Results: there was a decrease (p < 0.05) in the edema area induced by histamine and the dextrane A in three groups treated with C. officinalis extract. For both a edematogenic agents, the results were similar and not significant for 50 and 100 mg/kg of weight, although the decrease was substantial (p < 0.05) with the dose of 200 mg/kg. The reduction of the edema depended on the length of treatment. Conclusions: The anti-edematogenic action of the C. officinalis extract was shown, being the 200 mg/kg dose the best and the 14-day length of treatment the most effective. It was recommended to use it as phlebotonic agent in the treatment of the venous vascular disorders(AU)