Thesis--Novi Sad, Yugoslavia. / Title also in English: The rhizosphere microflora of matricaria chamomilla in solonetz and chernozem.

The fungus <i>Colletotrichum truncatum</i> is a potential biocontrol agent (BA) against the noxious weed scentless chamomile (<i>Metricaria perforata</i> Mérat; syn.: <i>Tripleurospermum perforatum</i> (Mérat) Lainz) in western Canada. This potential BA, however, is taxonomically related to the anthracnose pathogen on lentil, raising questions about crop safety. Ribosomal DNA (rDNA) internal transcribed space (ITS) regions of <i>C. truncatum</i> isolates collected from different plant hosts were examined, and compared with additional Colletotrichum species. Sequences were amplified with the universal primers its4 and its5, and <i>C. truncatum</i> isolates from scentless chamomile and selected legume crops were differentiated consistently. All scentless chamomile isolates fell within a single cluster in phylogenetic trees, regardless of their geographic origins. These isolates were more closely related to lentil isolates of <i>C. truncatum</i> than to isolates from the other host species. Soybean isolates, with more falcate and slender conidia and slightly bigger appressoria, were distinguishable from other <i>C. truncatum</i> isolates, while the isolates from scentless chamomile, lentil and pea were morphologically more similar. Based on sequence information, strain-specific PCR primers were designed for <i>C. truncatum</i> isolates from these hosts and used to amplify specific DNA bands (markers) from isolates of <i>C. truncatum</i>. This technique may be used for rapid detection and differentiation of <i>C. truncatum,</i> from scentless chamomile and designated legume species, as well as for tracking the BA after release. Inoculation trials were conducted using detached leaves and whole plants to determine potential cross infection of these <i>C. truncatum</i> isolates. Isolates from scentless chamomile caused disease only on their original host, but not on lentil, pea, soybean or alfalfa. In contrast, lentil isolates caused severe disease on lentil and pea, light symptoms on alfalfa, but no disease on the other hosts tested. Potential penetration of lentil leaves by scentless chamomile isolates was tested, with 2-23% incidence of the fungus from inoculated detached, senescence leaves but disease symptoms were not observed on either detached leaves or whole plants. Examination of the infection process revealed that scentless chamomile and lentil isolates had a similar pattern of infection and disease development on their respective hosts; infection vesicles were produced 24 h after inoculation, both primary and secondary infection hyphae were present, and the onset of disease symptoms tended to coincide with the development of secondary hyphae. The current study provided molecular and pathological evidence that differentiates the potential BA of scentless chamomile from <i>C. truncatum</i> isolates from lentil, pea and soybean.

The fungus <i>Colletotrichum truncatum</i> is a potential biocontrol agent (BA) against the noxious weed scentless chamomile (<i>Metricaria perforata</i> Mérat; syn.: <i>Tripleurospermum perforatum</i> (Mérat) Lainz) in western Canada. This potential BA, however, is taxonomically related to the anthracnose pathogen on lentil, raising questions about crop safety. Ribosomal DNA (rDNA) internal transcribed space (ITS) regions of <i>C. truncatum</i> isolates collected from different plant hosts were examined, and compared with additional Colletotrichum species. Sequences were amplified with the universal primers its4 and its5, and <i>C. truncatum</i> isolates from scentless chamomile and selected legume crops were differentiated consistently. All scentless chamomile isolates fell within a single cluster in phylogenetic trees, regardless of their geographic origins. These isolates were more closely related to lentil isolates of <i>C. truncatum</i> than to isolates from the other host species. Soybean isolates, with more falcate and slender conidia and slightly bigger appressoria, were distinguishable from other <i>C. truncatum</i> isolates, while the isolates from scentless chamomile, lentil and pea were morphologically more similar. Based on sequence information, strain-specific PCR primers were designed for <i>C. truncatum</i> isolates from these hosts and used to amplify specific DNA bands (markers) from isolates of <i>C. truncatum</i>. This technique may be used for rapid detection and differentiation of <i>C. truncatum,</i> from scentless chamomile and designated legume species, as well as for tracking the BA after release. Inoculation trials were conducted using detached leaves and whole plants to determine potential cross infection of these <i>C. truncatum</i> isolates. Isolates from scentless chamomile caused disease only on their original host, but not on lentil, pea, soybean or alfalfa. In contrast, lentil isolates caused severe disease on lentil and pea, light symptoms on alfalfa, but no disease on the other hosts tested. Potential penetration of lentil leaves by scentless chamomile isolates was tested, with 2-23% incidence of the fungus from inoculated detached, senescence leaves but disease symptoms were not observed on either detached leaves or whole plants. Examination of the infection process revealed that scentless chamomile and lentil isolates had a similar pattern of infection and disease development on their respective hosts; infection vesicles were produced 24 h after inoculation, both primary and secondary infection hyphae were present, and the onset of disease symptoms tended to coincide with the development of secondary hyphae. The current study provided molecular and pathological evidence that differentiates the potential BA of scentless chamomile from <i>C. truncatum</i> isolates from lentil, pea and soybean.

The Asteraceae family is frequently used to describe several medicinal plants which contain various phytochemical compounds including phenols, flavonoids and terpenoids. Among the Asteraceae family German chamomile (Matricaria chamomilla L.) and yarrow (Achillea millefolium L.) plants are extant species used in contemporary medicine. These phytochemical compounds have been traditionally used since ancient times in health care systems worldwide as a source of medicines. The use of micropropagation is essential to improve and increase these active compounds via plant tissue culture within a short period of time using the application of key plant growth regulators (PGRs). Furthermore, quantitative and qualitative analysis using high performance liquid chromatography- ultraviolet detector (HPLC-UV) and gas chromatography- flame ionisation detector (GC-FID) of potential medicinal compounds expressed by both chamomile and yarrow are important points. The protocol of in vitro shoots, roots and callus formation of chamomile and yarrow seeds culture were investigated using Murashige and Skoog (MS) medium supplemented with different concentrations of plant growth regulators (PGRs). MS culture medium containing 0.5 mgL-1 IAA and 1.0 mgL-1 of GA3 were found to be the best culture medium for chamomile and yarrow seeds. In this project in vitro and in vivo growth rates of selected plant species were also investigated. In the earlier growth stages yarrow plants were found to grow much quicker than chamomile, while the yield of chamomile flowers was significantly (p ≤ 0.001) more than yarrow flowers. The phenolic, flavonoid and terpenoid compounds content of leaves and flowers of plants produced from both cultures were also studied. HPLC-UV analysis showed that chlorogenic acid, apigenin-7-O-glucoside and luteolin dominated as the main phenol and flavonoid compounds recovered in both in vitro and in vivo chamomile and yarrow cultures. However, GC-FID analysis indicated that farnesene and nerolidol were detected as the main terpenoid compounds present in the two culture conditions used to grow chamomile and yarrow plants. Moreover, this research examines how chamomile and yarrow plants can produce and improve their phytochemical compounds content not only under well-watered conditions but also under drought stress conditions. The main phenol and flavonoid compounds of chlorogenic acid, caffeic acid, apig-7-glucoside, umbelliferon and luteolin were found in chamomile and yarrow varieties grown under both well-watered and drought stress conditions using (HPLC-UV), however farnesene, nerolidol, chamazulene, α-(-)- bisabol and bisabolol oxide A were observed in the plant essential oils (EOs) using Soxhlet extraction and GC-FID analysis. The antibacterial activity of plant EOs was also investigated using disc diffusion and 96 well plates. In vivo chamomile EO showed the highest antibacterial activity against gram-positive and gram-negative bacteria strains. In addition, in vitro yarrow EO showed the greatest effect on the death of bacteria strains.

As propriedades dos extratos vegetais vêm sendo cada vez mais investigadas, suscitando o interesse em aliar o potencial destes para minimizar efeitos do fotoenvelhecimento da pele decorrentes da ação dos radicais livres à fotoproteção proporcionada pelos filtros solares. Apesar do uso difundido de vários extratos vegetais, alguns deles, como a Matricaria chamomilla, ainda carecem de estudos acerca de suas propriedades e efeitos biológicos na pele, especialmente quando este é veiculado em formulações dermocosméticas. Assim, esse trabalho teve por objetivo a avaliação da estabilidade e da eficácia pré-clínica de formulações fotoprotetoras contendo o extrato de Matricaria chamomilla ou dos componentes isolados do vegetal, a-bisabolol e apigenina, na proteção de danos causados pela radiação ultravioleta (UV). Para tal, verificou-se a estabilidade dos ativos veiculados em formulação com FPS previamente determinado, bem como outras formulações de protetor solar foram desenvolvidas e a estas adicionadas extrato de camomila ou alfa-bisabolol, sendo testados por centrifugação, pH e análise visual. A estabilidade física das formulações mais estáveis foi analisada por estudos reológicos. A estabilidade química de formulações contendo extrato de camomila, apigenina e bisabolol foi determinada por Cromatografia Líquida de Alta Eficiência (HPLC). Estas formulações acrescidas de apigenina, extrato de camomila, ou alfa-bisabolol finalmente foram avaliadas no que tange à eficácia pré-clínica. Neste estudo, camundongos foram submetidos à radiação UV e o efeito protetor das formulações foi avaliado através de técnicas de biofísica nãoinvasiva e análise de imagem, que forneceram medidas de eritema, o teor de água do estrato córneo, a perda de água transepidérmica, ecogenicidade e espessura da derme. Os resultados permitiram selecionar as formulações fisicamente estáveisl para ambas as combinações de filtros UV associados contendo as substâncias ativas em estudo. No estudo de estabilidade química, a apigenina foi mais estável quando comparada às outras substâncias ativas em estudo e o bisabolol mostrou estabilidade química baixa. Todas as formulações foram eficazes na redução da perda de água transepidérmica. Os efeitos destas formulações na pele foram muito positivos, pois todas elas levaram a aumentos ecogenicidade e valores menores de espessura quando comparada às regiões irradiadas. Esses resultados mostraram efeitos significativos na melhoria do processo inflamatório causado pela radiação. Finalmente, as formulações desenvolvidas foram estáveis e eficazes na proteção da pele. Além disso, quando o extrato de camomila ou apigenina esteve presente na formulação fotoprotetora os efeitos foram mais pronunciados. / Particular plant extracts are being increasingly studied for their properties, in order to combine their potential to minimize skin photoaging effects from the action of free radicals with photoprotection provided by sunscreens. Despite of the widespread use of various plant extracts, some of them as it is the case of Matricaria chamomilla still need mored detailed studies especially on their properties and biological effects on skin, especially in dermocosmetic formulations. The aim of the present study was to assess the stability and the preclinical efficacy of sunscreen formulations containing M. chamomilla extract or its isolated components, alpha-bisabolol and apigenin, in the protection from damage caused by ultraviolet radiation. For this purpose, the physical stability in a pre-determined SPF formulation added of the active substances under study was determinated. Other sunscreen formulations were developed and supplemented with chamomile extract or alpha-bisabolol, and these were tested for centrifugation stability, pH and visual analysis. Physical stability of the most stable formulations was estimated by rheological measurements. The chemical stability of the formulations containing chamomile extract, apigenin, and bisabolol was determinated by High Efficiency Liquid Chromatography (HPLC). These formulations with chamomile extract, apigenin or alpha-bisabolol were finally evaluated in terms of preclinical efficacy. In this study, hairless mice were submitted to UV radiation and the protective effect of the formulations was evaluated using noninvasive biophysics techniques and image analysis, in which we obtained measures of erythema, the water content of stratum corneum, transepidermal water loss, the echogenicity and the thickness of the dermis. The results allowed selecting physicallystable formulations for both the combinations of UV-filters associated containing the actives substances under study. In the chemical stability study, the apigenin present in the extract was more stable when compared to the other active substances under study and the bisabolol showed a low chemical stabilitity. All the formulations were effective in the transepidermic water-loss reduction. The effects of these formulations on skin were very positive since all of them led to ecogenicity increases and lower thickness values when compared to irradiated regions. These results showed significant effects in the improvement of the inflammatory process caused by radiation. Finally, the formulations developed were stable and effective in the skin protection. In addition, when the chamomile extract or apigenin was present in the formulation the photoprotective effects were more pronounced.

A Chamomilla recutita L. é uma das plantas medicinais mais cultivadas no Brasil e no mundo. Os extratos da C. recutita são de interesse para as indústrias farmacêuticas e cosméticas, visto que estes apresentam atividades anti-inflamatória, antioxidante e adstringente. A ação terapêutica do extrato pode ser mais pronunciada que a ação terapêutica de um de seus ativos isolados. No entanto, a incorporação de um extrato em uma formulação pode ser difícil devido à baixa estabilidade dos extratos, bem como à possibilidade de gerarem instabilidade das formulações. Microencapsulando o extrato com um carreador é possível aumentar estabilidade do extrato quanto evitar instabilidade na formulação. Além disso, a microencapsulação é capaz de fornecer outras vantagens, como uma liberação controlada. Dois processos foram estudados como alternativas para a microencapsulação do óleo essencial e do extrato hidroalcoólico da C. recutita usando quitosana como carreador: o spray drying e o spray freeze drying. Planejamentos fatorais foram utilizados para determinar os fatores que mais influenciaram no diâmetro médio das micropartículas, eficiência de encapsulação e teor dos marcadores e rendimento do processo. A apigenina e a apigenina-7-glicosídeo foram usadas como marcadores do extrato hidroalcoólico e o óxido de bisabolol A foi usado como marcador do óleo essencial. Os processos de spray drying e spray freeze drying dos dois extratos foram otimizados e as micropartículas resultantes foram caracterizadas com relação ao diâmetro médio, rendimento do processo, teor e eficiência de encapsulação dos marcadores, atividade antioxidante in vitro, densidade, índice de Carr, fator de Hausner, umidade, morfologia, perfil de liberação n vitro e estabilidade. Os resultados mostraram que o processo de spray drying apresentou os melhores resultados para eficiência de encapsulação, com valores de aproximadamente 98%, 95% e 80% para apigenina, apigenina-7-glicosídeo e óxido de bisabolol A, respectivamente. As eficiências de encapsulação obtidas no processo de spray freeze drying foram de aproximadamente 59%, 58% e 38% para os mesmos marcadores, respectivamente. As micropartículas produzidas por spray freeze drying apresentaram formato irregular e poroso, enquanto as produzidas por spray drying apresentaram formato esférico e superfícies mais lisas, sem poros ou fissuras. Ao contrário do que ocorreu com o extrato hidroalcoólico, a perda do marcador do óleo foi elevada no processo de spray drying, com teor final de 35%. Os teores dos marcadores ficaram acima de 80% para o processo de spray freeze drying do óleo e acima de 90% para o extrato hidroalcoólico. As micropartículas produzidas por spray drying do extrato hidroalcoólico e do óleo e por spray freeze drying do extrato hidroalcoólico e do óleo apresentaram diâmetro médio de 5,1 ?m, 5,0 ?m, 31,0 ?m e 96,4 ?m, respectivamente. Ensaios de liberação in vitro mostraram que as micropartículas foram capazes de sustentar a liberação dos respectivos marcadores. Os estudos de permeação in vitro das micropartículas produzidas por spray drying do extrato hidroalcoólico também mostraram que estas foram capazes de sustentar a liberação. A microencapsulação proporcionou em todos os casos um aumento considerável da estabilidade. As micropartículas produzidas por spray drying do extrato hidroalcoólico apresentaram teores de marcadores no mínimo 50% maiores que o extrato puro após 90 dias. O spray freeze drying se mostrou como a melhor alternativa para produção de micropartículas de quitosana contendo o óleo essencial de C. recutita, enquanto o processo de spray drying se mostrou como uma ótima alternativa para microencapsulação do extrato hidroalcoólico da C. recutita. / Chamomilla recutita L. is one of the most cultivated medicinal plants in Brazil and around the world. Its extracts are important to both the pharmaceutical and cosmetics industries due to its therapeutic applications, such as an anti-inflammatory, antioxidant, and astringent. The therapeutic effects of an extract may be more pronounced than those of an isolated active compound. However, the incorporation of an extract in a formulation is difficult due to the low stability of extracts and the potential instabilities they may cause in formulations. Microencapsulating an extract in a carrier is a potential way of increasing the stability of an extract and avoiding instabilities in a formulation. Compound microencapsulation also brings other advantages, such as controlled release rates. Two processes were studied as alternatives to microencapsulating C. recutita essential oil and C. recutita hydroalcoholic extract using chitosan as a carrier: spray drying and spray freeze drying. Factorial designs were used to determine which process factors most influence the mean diameter, encapsulation efficiency and content of the chemical markers, and process yield. Apigenin and apigenin-7-glucoside were used as chemical markers for the hydroalcoholic extract and bisabolol oxide A was used as the chemical marker for the essential oil. The spray drying and spray freeze drying processes for both the oil and hydroalcoholic extract were optimized and the resulting microparticles were further characterized to determine mean diameter, process yield, marker encapsulation efficiency and content, in vitro antioxidant activity, density, Carr index, Hausner factor, water content, morphology, in vitro release profiles and stability. The results showed spray drying had the best encapsulation efficiency results, with about 98%, 95% e 80% of the apigenin, apigenin-7-glucoside and bisabolol oxide A content, respectively, inside the microparticles. The encapsulation efficiencies obtained in the spray freeze drying process were about 59%, 58% e 38% for the same chemical markers, respectively. Microparticles produced by spray freeze drying were irregular and porous, whereas microparticles produced by spray drying were spherical and fairly smooth, without porous or cracks. Contrary to what happened with the hydroalcoholic extract, oil marker content was low for spray dried microparticles, with final content at 35%. Chemical markers contents were above 80% for the oil and above 90% for the hydroalcoholic extract in spray freeze dried microparticles. Spray dried microparticles containing extract and oil and spray freeze dried microparticles containing extract and oil had mean diameter of 5.1 ?m, 5.0 ?m, 31.0 ?m and 96.4 ?m, respectively. In vitro release profiles showed all microparticles were able to sustain their respective marker release rates. In vitro permeation studies of spray dried microparticles containing hydroalcooholic extract also showed sustained release rates for the corresponding markers. Microencapsulation also provided considerable increase in C. recutita hydroalcoholic extract stability and C. recutita essential oil stability. After 90 days spray dried microparticles containing hydroalcoholic extract presented marker content 50% higher than the pure hydroalcoholic extract. Spray freeze drying was the best alternative to produce chitosan microparticles containing C. recutita essential oil, while spray drying was shown to be an excellent way to microencapsulate C. recutita hydroalcoholic extract in chitosan.

O presente estudo teve como objetivo avaliar a influência da administração do chá de camomila durante 21 dias sobre parâmetros antioxidantes e estresse oxidativo das glândulas parótida e submandibular. Bem como o efeito do consumo do chá no transporte de glicose do fígado e possíveis alterações sistêmicas. Os grupos estudados foram divididos em controle não tratado (C), controle tratado com chá de camomila (CC), diabético não tratado (DM) e diabético tratado com chá de camomila (DMC). A indução do diabetes nos grupos DM e DMC foi realizada com injeção intraperitoneal de estreptozotocina (60 mg/Kg de peso corporal). No 7º dia experimental, os animais do grupo CC e DMC iniciaram o tratamento com o chá de camomila. Foram avaliados os sistemas antioxidantes enzimáticos por meio da atividade das enzimas superóxido dismutase (SOD), catalase (CAT) e glutationa peroxidase (GPx) e da via não enzimática pelo total antioxidante (TAS). O estresse oxidativo foi avaliado por meio da quantificação de malondialdeído (MDA). Além disso foram analisados os níveis de insulina séricos, glicemia, peso e consumo hídrico e alimentar. A via de transporte de glicose foi estudada por meio da avaliação da expressão e quantificação das proteínas AKT e AMPK. Os resultados obtidos confirmam o potencial hipoglicemiante do chá de camomila, bem como sua ação no controle da polidipsia. O chá não alterou os níveis de insulina além disso provocou uma redução da expressão da p-AMPK. O tratamento com chá ocasionou um aumento nos valores da atividade da GPx e CAT tanto na glândula parótida como na submandibular. No entanto, para o TAS o chá provocou aumento apenas na glândula submandibular. Na glândula parótida o chá diminuiu a peroxidação lipídica. Os resultados obtidos confirmam o potencial antioxidante do chá de camomila na glândula parótida dos animais diabéticos após tratamento. Conclusão: O tratamento com o chá de camomila aponta como sendo promissor na prevenção dos danos oxidativos presentes no diabetes mellitus, tanto nas glândulas salivares através da redução do MDA, quanto sistemicamente pelo seu potencial hipoglicemiante. / The present study aimed to evaluate the influence of the administration of chamomile tea for 21 days on antioxidant parameters and oxidative stress of the parotid and submandibular glands. As well as the effect of tea consumption on the transport of glucose from the liver and possible systemic alterations. The groups studied were divided into untreated control (C), control treated with chamomile tea (CC), untreated diabetic (DM) and diabetic treated with chamomile tea (DMC). The induction of diabetes in the DM and DMC groups was performed with intraperitoneal injection of streptozotocin (60 mg / kg body weight). On the 7th experimental day, the CC and DMC animals started treatment with chamomile tea. The enzymatic antioxidant systems were evaluated through the activity of the enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) and non-enzymatic pathway by total antioxidant (SAT). Oxidative stress was evaluated by the quantification of malondialdehyde (MDA). Serum insulin levels, glycemia, weight, and water and food intake were also analyzed. The glucose transport pathway was studied by evaluating the expression and quantification of AKT and AMPK proteins. The results confirm the hypoglycemic potential of chamomile tea, as well as its action in the control of polydipsia. The tea did not alter insulin levels, in addition it caused a reduction of p-AMPK expression. Treatment with tea caused an increase in the values of GPx and CAT activity in both the parotid gland and the submandibular gland. However, for TAS the tea caused only increase in the submandibular gland. In the parotid gland the tea decreased lipid peroxidation. The results confirm the antioxidant potential of chamomile tea in the parotid gland of diabetic animals after treatment. Conclusion: Treatment with chamomile tea is shown to be promising in preventing the oxidative damage present in diabetes mellitus, both in the salivary glands through the reduction of MDA and systemically due to its hypoglycemic potential.

The traditional herbal medicinal product Myrrhinil-Intest® is a fixed herbal combination, which is marketed in Germany since 1959 and applied in medical practice for the treatment of gastrointestinal disorders such as functional diarrhea, irritable bowel syndrome and inflammatory bowel disease. It contains myrrh, which is described as the oleo-gum resin from mainly Commiphora molmol Engler (Burseraceae), coffee charcoal, which are the milled roasted to blackening outer seed parts of green dried Coffea Arabica Linné (Rubiaceae) fruits and chamomile flowers - the flower heads of Matricaria recutita Linné (Asteraceae). The clinical effectiveness of Myrrhinil-Intest® for the treatment of various gastrointestinal disorders was demonstrated in several clinical studies and is described in various experience reports, however its pharmacological profile is not fully elucidated. Within the present study the spasmolytic and anti-inflammatory potential of the components myrrh, chamomile and coffee charcoal was investigated. Therefore pharmacological, histological and molecular biological methods were utilised. Spasmolytic activity was characterised using isometric tension measurement with rat isolated small intestinal preparations. Anti-inflammatory potential was assessed with different methods using isolated rat small intestinal preparations and immune cell lines. Inflammation was induced with TNBS and LPS respectively. Additionally, the influence of the herbal components on the gene expression profile of native human macrophages after LPS/IFNγ stimulation was determined by microarray gene expression analysis. Chamomile flower and myrrh exerted spasmolytic effects, whereby the more pronounced spasmolytic effects of myrrh were mediated via calcium channel blockade. Myrrh and chamomile flower exerted anti-inflammatory effects.

A Chamomilla recutita L. é uma das plantas medicinais mais cultivadas no Brasil e no mundo. Os extratos da C. recutita são de interesse para as indústrias farmacêuticas e cosméticas, visto que estes apresentam atividades anti-inflamatória, antioxidante e adstringente. A ação terapêutica do extrato pode ser mais pronunciada que a ação terapêutica de um de seus ativos isolados. No entanto, a incorporação de um extrato em uma formulação pode ser difícil devido à baixa estabilidade dos extratos, bem como à possibilidade de gerarem instabilidade das formulações. Microencapsulando o extrato com um carreador é possível aumentar estabilidade do extrato quanto evitar instabilidade na formulação. Além disso, a microencapsulação é capaz de fornecer outras vantagens, como uma liberação controlada. Dois processos foram estudados como alternativas para a microencapsulação do óleo essencial e do extrato hidroalcoólico da C. recutita usando quitosana como carreador: o spray drying e o spray freeze drying. Planejamentos fatorais foram utilizados para determinar os fatores que mais influenciaram no diâmetro médio das micropartículas, eficiência de encapsulação e teor dos marcadores e rendimento do processo. A apigenina e a apigenina-7-glicosídeo foram usadas como marcadores do extrato hidroalcoólico e o óxido de bisabolol A foi usado como marcador do óleo essencial. Os processos de spray drying e spray freeze drying dos dois extratos foram otimizados e as micropartículas resultantes foram caracterizadas com relação ao diâmetro médio, rendimento do processo, teor e eficiência de encapsulação dos marcadores, atividade antioxidante in vitro, densidade, índice de Carr, fator de Hausner, umidade, morfologia, perfil de liberação n vitro e estabilidade. Os resultados mostraram que o processo de spray drying apresentou os melhores resultados para eficiência de encapsulação, com valores de aproximadamente 98%, 95% e 80% para apigenina, apigenina-7-glicosídeo e óxido de bisabolol A, respectivamente. As eficiências de encapsulação obtidas no processo de spray freeze drying foram de aproximadamente 59%, 58% e 38% para os mesmos marcadores, respectivamente. As micropartículas produzidas por spray freeze drying apresentaram formato irregular e poroso, enquanto as produzidas por spray drying apresentaram formato esférico e superfícies mais lisas, sem poros ou fissuras. Ao contrário do que ocorreu com o extrato hidroalcoólico, a perda do marcador do óleo foi elevada no processo de spray drying, com teor final de 35%. Os teores dos marcadores ficaram acima de 80% para o processo de spray freeze drying do óleo e acima de 90% para o extrato hidroalcoólico. As micropartículas produzidas por spray drying do extrato hidroalcoólico e do óleo e por spray freeze drying do extrato hidroalcoólico e do óleo apresentaram diâmetro médio de 5,1 ?m, 5,0 ?m, 31,0 ?m e 96,4 ?m, respectivamente. Ensaios de liberação in vitro mostraram que as micropartículas foram capazes de sustentar a liberação dos respectivos marcadores. Os estudos de permeação in vitro das micropartículas produzidas por spray drying do extrato hidroalcoólico também mostraram que estas foram capazes de sustentar a liberação. A microencapsulação proporcionou em todos os casos um aumento considerável da estabilidade. As micropartículas produzidas por spray drying do extrato hidroalcoólico apresentaram teores de marcadores no mínimo 50% maiores que o extrato puro após 90 dias. O spray freeze drying se mostrou como a melhor alternativa para produção de micropartículas de quitosana contendo o óleo essencial de C. recutita, enquanto o processo de spray drying se mostrou como uma ótima alternativa para microencapsulação do extrato hidroalcoólico da C. recutita. / Chamomilla recutita L. is one of the most cultivated medicinal plants in Brazil and around the world. Its extracts are important to both the pharmaceutical and cosmetics industries due to its therapeutic applications, such as an anti-inflammatory, antioxidant, and astringent. The therapeutic effects of an extract may be more pronounced than those of an isolated active compound. However, the incorporation of an extract in a formulation is difficult due to the low stability of extracts and the potential instabilities they may cause in formulations. Microencapsulating an extract in a carrier is a potential way of increasing the stability of an extract and avoiding instabilities in a formulation. Compound microencapsulation also brings other advantages, such as controlled release rates. Two processes were studied as alternatives to microencapsulating C. recutita essential oil and C. recutita hydroalcoholic extract using chitosan as a carrier: spray drying and spray freeze drying. Factorial designs were used to determine which process factors most influence the mean diameter, encapsulation efficiency and content of the chemical markers, and process yield. Apigenin and apigenin-7-glucoside were used as chemical markers for the hydroalcoholic extract and bisabolol oxide A was used as the chemical marker for the essential oil. The spray drying and spray freeze drying processes for both the oil and hydroalcoholic extract were optimized and the resulting microparticles were further characterized to determine mean diameter, process yield, marker encapsulation efficiency and content, in vitro antioxidant activity, density, Carr index, Hausner factor, water content, morphology, in vitro release profiles and stability. The results showed spray drying had the best encapsulation efficiency results, with about 98%, 95% e 80% of the apigenin, apigenin-7-glucoside and bisabolol oxide A content, respectively, inside the microparticles. The encapsulation efficiencies obtained in the spray freeze drying process were about 59%, 58% e 38% for the same chemical markers, respectively. Microparticles produced by spray freeze drying were irregular and porous, whereas microparticles produced by spray drying were spherical and fairly smooth, without porous or cracks. Contrary to what happened with the hydroalcoholic extract, oil marker content was low for spray dried microparticles, with final content at 35%. Chemical markers contents were above 80% for the oil and above 90% for the hydroalcoholic extract in spray freeze dried microparticles. Spray dried microparticles containing extract and oil and spray freeze dried microparticles containing extract and oil had mean diameter of 5.1 ?m, 5.0 ?m, 31.0 ?m and 96.4 ?m, respectively. In vitro release profiles showed all microparticles were able to sustain their respective marker release rates. In vitro permeation studies of spray dried microparticles containing hydroalcooholic extract also showed sustained release rates for the corresponding markers. Microencapsulation also provided considerable increase in C. recutita hydroalcoholic extract stability and C. recutita essential oil stability. After 90 days spray dried microparticles containing hydroalcoholic extract presented marker content 50% higher than the pure hydroalcoholic extract. Spray freeze drying was the best alternative to produce chitosan microparticles containing C. recutita essential oil, while spray drying was shown to be an excellent way to microencapsulate C. recutita hydroalcoholic extract in chitosan.

As propriedades dos extratos vegetais vêm sendo cada vez mais investigadas, suscitando o interesse em aliar o potencial destes para minimizar efeitos do fotoenvelhecimento da pele decorrentes da ação dos radicais livres à fotoproteção proporcionada pelos filtros solares. Apesar do uso difundido de vários extratos vegetais, alguns deles, como a Matricaria chamomilla, ainda carecem de estudos acerca de suas propriedades e efeitos biológicos na pele, especialmente quando este é veiculado em formulações dermocosméticas. Assim, esse trabalho teve por objetivo a avaliação da estabilidade e da eficácia pré-clínica de formulações fotoprotetoras contendo o extrato de Matricaria chamomilla ou dos componentes isolados do vegetal, a-bisabolol e apigenina, na proteção de danos causados pela radiação ultravioleta (UV). Para tal, verificou-se a estabilidade dos ativos veiculados em formulação com FPS previamente determinado, bem como outras formulações de protetor solar foram desenvolvidas e a estas adicionadas extrato de camomila ou alfa-bisabolol, sendo testados por centrifugação, pH e análise visual. A estabilidade física das formulações mais estáveis foi analisada por estudos reológicos. A estabilidade química de formulações contendo extrato de camomila, apigenina e bisabolol foi determinada por Cromatografia Líquida de Alta Eficiência (HPLC). Estas formulações acrescidas de apigenina, extrato de camomila, ou alfa-bisabolol finalmente foram avaliadas no que tange à eficácia pré-clínica. Neste estudo, camundongos foram submetidos à radiação UV e o efeito protetor das formulações foi avaliado através de técnicas de biofísica nãoinvasiva e análise de imagem, que forneceram medidas de eritema, o teor de água do estrato córneo, a perda de água transepidérmica, ecogenicidade e espessura da derme. Os resultados permitiram selecionar as formulações fisicamente estáveisl para ambas as combinações de filtros UV associados contendo as substâncias ativas em estudo. No estudo de estabilidade química, a apigenina foi mais estável quando comparada às outras substâncias ativas em estudo e o bisabolol mostrou estabilidade química baixa. Todas as formulações foram eficazes na redução da perda de água transepidérmica. Os efeitos destas formulações na pele foram muito positivos, pois todas elas levaram a aumentos ecogenicidade e valores menores de espessura quando comparada às regiões irradiadas. Esses resultados mostraram efeitos significativos na melhoria do processo inflamatório causado pela radiação. Finalmente, as formulações desenvolvidas foram estáveis e eficazes na proteção da pele. Além disso, quando o extrato de camomila ou apigenina esteve presente na formulação fotoprotetora os efeitos foram mais pronunciados. / Particular plant extracts are being increasingly studied for their properties, in order to combine their potential to minimize skin photoaging effects from the action of free radicals with photoprotection provided by sunscreens. Despite of the widespread use of various plant extracts, some of them as it is the case of Matricaria chamomilla still need mored detailed studies especially on their properties and biological effects on skin, especially in dermocosmetic formulations. The aim of the present study was to assess the stability and the preclinical efficacy of sunscreen formulations containing M. chamomilla extract or its isolated components, alpha-bisabolol and apigenin, in the protection from damage caused by ultraviolet radiation. For this purpose, the physical stability in a pre-determined SPF formulation added of the active substances under study was determinated. Other sunscreen formulations were developed and supplemented with chamomile extract or alpha-bisabolol, and these were tested for centrifugation stability, pH and visual analysis. Physical stability of the most stable formulations was estimated by rheological measurements. The chemical stability of the formulations containing chamomile extract, apigenin, and bisabolol was determinated by High Efficiency Liquid Chromatography (HPLC). These formulations with chamomile extract, apigenin or alpha-bisabolol were finally evaluated in terms of preclinical efficacy. In this study, hairless mice were submitted to UV radiation and the protective effect of the formulations was evaluated using noninvasive biophysics techniques and image analysis, in which we obtained measures of erythema, the water content of stratum corneum, transepidermal water loss, the echogenicity and the thickness of the dermis. The results allowed selecting physicallystable formulations for both the combinations of UV-filters associated containing the actives substances under study. In the chemical stability study, the apigenin present in the extract was more stable when compared to the other active substances under study and the bisabolol showed a low chemical stabilitity. All the formulations were effective in the transepidermic water-loss reduction. The effects of these formulations on skin were very positive since all of them led to ecogenicity increases and lower thickness values when compared to irradiated regions. These results showed significant effects in the improvement of the inflammatory process caused by radiation. Finally, the formulations developed were stable and effective in the skin protection. In addition, when the chamomile extract or apigenin was present in the formulation the photoprotective effects were more pronounced.