Made available in DSpace on 2014-06-11T19:32:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-16Bitstream added on 2014-06-13T19:02:38Z : No. of bitstreams: 1 piffer_rc_dr_botfm.pdf: 1433858 bytes, checksum: 0b4876bf7f03faeb50a5129b53d87275 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A corticoterapia vem sendo muito utilizada na prática obstétrica, em gestantes com risco de parto prematuro, no intuito de promover a aceleração da maturidade pulmonar fetal. Entretanto, altos níveis de corticosteróides podem interferir com o pico de testosterona perinatal, importante para a diferenciação sexual do hipotálamo. Este processo é mediado por hormônios gonadais durante o período perinatal e responsável pelas diferenças fisiológicas, morfológicas, comportamentais e neuroanatômicas entre machos e fêmeas adultos. O hipotálamo de mamíferos, antes da diferenciação sexual, está organizado do tipo feminino . No macho, o hipotálamo precisa ser masculinizado e defeminizado para que apareça o padrão tônico de secreção de gonadotrofinas e o comportamento de monta. Desta maneira, objetivouse investigar através de parâmetros reprodutivos, metabólicos, farmacológicos e de biologia molecular, possíveis alterações resultantes da terapia com betametasona no período pré-natal em ratos, avaliando o desenvolvimento dos descendentes machos, do nascimento até a vida adulta reprodutiva. Ratas Wistar prenhes receberam 0,1mg/kg de betametasona (grupo Betametasona) ou salina (grupo Controle) no 12º, 13º, 18º e 19º dias de prenhez (IM). A exposição pré-natal à betametasona reduziu ao nascer a massa corporal e a distância anogenital; o peso úmido da glândula adrenal e a corticosterona plasmática também estavam reduzidos ao nascer em relação ao grupo Controle, indicando comprometimento do eixo hipotálamo-hipófise-adrenal. No entanto, esta terapia não alterou o desenvolvimento físico da prole, exceto pelo atraso na descida testicular. Na vida adulta, a exposição pré-natal à betametasona reduziu o peso úmido do testículo e da secreção da vesícula seminal, bem como a... / Corticosteroids are used widely in obstetric clinical practice in pregnant women at risk for preterm delivery. This therapy promotes fetal lung maturation, thus reducing the incidence of respiratory distress syndrome, which in turn decreases neonate mortality and morbidity. In spite of this, much less is known about the interference of high corticosteroid levels at the testosterone peak during the critical period of brain sexual differentiation. Thus, the aim this study was to investigate reproductive, metabolic, pharmacologycal, and molecular biology parameters in male rats exposed prenatally to betamethasone. Pregnant rats received 0.1mg/kg of betamethasone (Betamethasone group) or saline (Control group) on the 12th, 13th, 18th, and 19th days of pregnancy (IM). The prenatal treatment with betamethasone reduced the body weight, the anogenital distance, the wet weight of adrenal, and plasmatic corticosterone level of male pups at birth. However, this terapy did not alter the physical development, but there was retardation in the age of testicular descent. In adulthood, there were reductions in the wet weight of testis, in the seminal vesicle secretion, in the adrenal wet weight and as well as plasmatic corticosterone level. The fertility reduction observed in the Betamethasone group probably was caused by decreased sperm quality, sperm production, and plasmatic testosterone level, and not for alterations in contraction response of seminal vesicle and vas deferens, to autonomic drugs, so important for ejaculatory process. Prenatal exposure to betamethasone also damaged sexual behavior. There was a delay in the latency to first ejaculation, as well as a decrease in the numbers of postejaculatory intromissions, total intromissions and total ejaculations. Although 80% of the betamethasone-treated animals exhibited male sexual behavior, ...(Complete abstract click electronic access below)

Orientador: Oduvaldo Câmara Marques Pereira / Banca: Daniela Cristina Ceccatto Gerardin / Banca: Angelina Zanesco / Banca: Célia Regina Nogueira / Banca: Marilza Vieira Cunha Rudge / Resumo: A corticoterapia vem sendo muito utilizada na prática obstétrica, em gestantes com risco de parto prematuro, no intuito de promover a aceleração da maturidade pulmonar fetal. Entretanto, altos níveis de corticosteróides podem interferir com o pico de testosterona perinatal, importante para a diferenciação sexual do hipotálamo. Este processo é mediado por hormônios gonadais durante o período perinatal e responsável pelas diferenças fisiológicas, morfológicas, comportamentais e neuroanatômicas entre machos e fêmeas adultos. O hipotálamo de mamíferos, antes da diferenciação sexual, está organizado do tipo feminino . No macho, o hipotálamo precisa ser masculinizado e defeminizado para que apareça o padrão tônico de secreção de gonadotrofinas e o comportamento de monta. Desta maneira, objetivouse investigar através de parâmetros reprodutivos, metabólicos, farmacológicos e de biologia molecular, possíveis alterações resultantes da terapia com betametasona no período pré-natal em ratos, avaliando o desenvolvimento dos descendentes machos, do nascimento até a vida adulta reprodutiva. Ratas Wistar prenhes receberam 0,1mg/kg de betametasona (grupo Betametasona) ou salina (grupo Controle) no 12º, 13º, 18º e 19º dias de prenhez (IM). A exposição pré-natal à betametasona reduziu ao nascer a massa corporal e a distância anogenital; o peso úmido da glândula adrenal e a corticosterona plasmática também estavam reduzidos ao nascer em relação ao grupo Controle, indicando comprometimento do eixo hipotálamo-hipófise-adrenal. No entanto, esta terapia não alterou o desenvolvimento físico da prole, exceto pelo atraso na descida testicular. Na vida adulta, a exposição pré-natal à betametasona reduziu o peso úmido do testículo e da secreção da vesícula seminal, bem como a ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Corticosteroids are used widely in obstetric clinical practice in pregnant women at risk for preterm delivery. This therapy promotes fetal lung maturation, thus reducing the incidence of respiratory distress syndrome, which in turn decreases neonate mortality and morbidity. In spite of this, much less is known about the interference of high corticosteroid levels at the testosterone peak during the critical period of brain sexual differentiation. Thus, the aim this study was to investigate reproductive, metabolic, pharmacologycal, and molecular biology parameters in male rats exposed prenatally to betamethasone. Pregnant rats received 0.1mg/kg of betamethasone (Betamethasone group) or saline (Control group) on the 12th, 13th, 18th, and 19th days of pregnancy (IM). The prenatal treatment with betamethasone reduced the body weight, the anogenital distance, the wet weight of adrenal, and plasmatic corticosterone level of male pups at birth. However, this terapy did not alter the physical development, but there was retardation in the age of testicular descent. In adulthood, there were reductions in the wet weight of testis, in the seminal vesicle secretion, in the adrenal wet weight and as well as plasmatic corticosterone level. The fertility reduction observed in the Betamethasone group probably was caused by decreased sperm quality, sperm production, and plasmatic testosterone level, and not for alterations in contraction response of seminal vesicle and vas deferens, to autonomic drugs, so important for ejaculatory process. Prenatal exposure to betamethasone also damaged sexual behavior. There was a delay in the latency to first ejaculation, as well as a decrease in the numbers of postejaculatory intromissions, total intromissions and total ejaculations. Although 80% of the betamethasone-treated animals exhibited male sexual behavior, ...(Complete abstract click electronic access below) / Doutor

Orientador: Gilberto de Nucci / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-12T13:53:54Z (GMT). No. of bitstreams: 1 Oliveira_LinaSayuriOdoBuenode_M.pdf: 1845448 bytes, checksum: 4fea923152025c316ca4c99487b21970 (MD5) Previous issue date: 2008 / Resumo: Betametasona é um corticosteróide sintético designado para exercer a função de um glicocorticóide ativo. Como um álcool livre, betametasona apresenta uma vasta aplicação clínica com atividade antiinflamatória e imunossupressora. A betametasona foi extraída com 0,5mL de plasma humano por extração líquido-líquido (ELL) utilizando Cloranfenicol como padrão interno. O método utilizou uma corrida cromatográfica de 2,5min, com a coluna analítica C18 (100mm×2.1mm i.d.), a calibração da curva linear de 0,05ng/mL a 50ng/mL (r2 > 0,993). A exatidão inter-corrida dos controles de qualidade foi 92.3% (CQA 0.15ng/mL), 90.7% (CQB 4.0ng/mL) e 97.2% (CQC 40ng/mL). A precisão inter-corrida dos controles de qualidade foi 8.7% (CQA 0.15ng/mL), 9.0% (CQB 4.0ng/mL) e 9.8% (CQC 40ng/mL). O método aqui descrito foi empregado em estudo de biodisponibilidade relativa de duas formulações contendo dexclorfeniramina/betametasona 2mg/0.25mg comprimido. A razão da média geométrica e dos respectivos intervalos de confiança (IC) de Betametasona/Celestamine® foram 107.61% (101.62-113.95%) para AUClast, 106.93% 102.08-112.00% para AUC0-inf e 105.06% (96.56-114.31%) para Cmax. O intervalo deconfiança de 90% calculado para a razão individual das médias de Cmax, ASC0-72h, ASClast e ASC0-inf para betametasona/celestamine® estavam dentro do intervalo de 80- 125% definido pela Agência Vigilância Sanitária (ANVISA). No presente estudo um método, rápido, sensível e robusto foi desenvolvido para a determinação e quantificação de betametasona em plasma humano através da cromatografia líquida acoplada a espectrometria de massas usando fotoionização à pressão atmosférica em modo negativo. / Abstract: Betamethasone is a synthetic corticosteroid designed to exert a marked glucocorticoid activity. As the free alcohol, betamethasone finds widespread clinical applications related to its anti-inflammatory and immunosuppressant activity. Betamethasone was extracted from 0.5 ml human plasma by liquid-liquid extraction (LLE) using chloramphenicol as internal standard. The method has a chromatographic run of 2.5 min using a C18 analytical column (100mm×2.1mm i.d.) and the linear calibration curve over the range was linear from 0.05 to 50 ng ml-1 (r2 > 0.993). The between-run accuracy, based on the relative standard deviation replicate quality controls was 92.3% (0.15 ng ml-1), 90.7% (4.0 ng ml-1) and 97.2% (40 ng ml-1). The between-run precision for the above-mentioned concentrations was 8.7, 9.0 and 9.8%, respectively. The method herein described was employed in a bioequivalence study of two formulations of dexchlorpheniramine/betamethasone 2 mg/0.25 mg tablets. The geometric mean and respective 90% confidence interval (Cl) of betamethasone/Celestamine® percent ratios were 107.61% (101.62-113.95%) for AUClast, 06.93% 102.08-112.00%for AUC0-inf and 105.06% (96.56-114.31%) for Cmax. In adition, the calculated 90% Cl for mean Cmax, AUClast and AUCinf betamethasone/celestamine® individual ratios were within the 80-125% interval defined by the Agência Vigilância Sanitária - ANVISA. In the present study, a fast, sensitive, robust method was developed for the determination and quantification of betamethasone in human plasma by liquid chromatography coupled with tandem mass spectrometry, using photospray ionization in negative mode. / Mestrado / Mestre em Farmacologia

Magister Pharmaceuticae - MPharm / Background/Introduction: Atopic dermatitis (AD) is one of the most prevalent diseases worldwide. It is a rapidly growing field of study with several research avenues to explore its pathophysiology and to find innovative treatment and management regimens. Clinically, it is classified as a non-contagious, intensely pruritic, inflammatory, chronic skin disorder mediated by abnormalities associated with atopy. Symptoms include inflammation, redness, pain and a negative impact on the patient‘s overall quality of life. Chronic itching often leads to the formation of lichenified skin, which may increase the thickness of the epidermis and exacerbate the barrier function of the skin. AD is treated with topical corticosteroids which help to decrease inflammation. However, lichification of the skin may decrease the efficacy of topical dosage forms. Nanomedicine is a rapidly developing field where advances have been made using ethosomes for topical delivery. As such, corticosteroid loaded ethosomal formulations containing hydrocortiosone acetate (HCA) and betamethasone valerate (BMV) were developed and characterised to develop novel tools for topical drug delivery. Aim: This study aimed at developing corticosteroid loaded ethosomes as a pre-formulation component for inclusion in a topical dosage form. To date, no ethosmal formulation with HCA and BMV has been investigated for topical drug delivery. Method: Ethosomes were synthesised using the hot method and the cold method, a modified version of a double emulsion (o/w/o), solvent evaporation technique, as developed by Touitou et al, 2007.1 Ethosomes were prepared using fixed concentrations of either BMV or HCA (10 mg/ml), ethanol (30% v/v) and purified water (70% v/v) and were comminuted using bath sonication or mini-extrusion. Centrifugation and centrifugal drying were used to purify and isolate the ethosomes for solid state characterisation. The morphology was determined using Scanning electron microscopy (SEM). Ethosomes were characterised using: dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FTIR), hot stage microscopy (HSM), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). The encapsulation efficiency (EE) and drug loading (DL) were determined using validated HPLC methods. Finally, the drug release was determined using Franz diffusion cells and mathematical models were fitted to the % cumulative release data to determine the release kinetics. Results: Ethosomes were assessed according to the following criteria for topical drug delivery which were determined using dynamic light scattering (DLS): Hydrodynamic diameter (HdD), ~ 200 nm, polydispersity index (PdI) < 0.5 and zeta potential (ζp) ± 30 mV. The optimum formulations contained phosphatidylcholine (PC) 50 mg/ml. Extrusion was found to be the best method for particle reduction based on the reproducibility of the results. The HdD was 163.8±31.99 and 147.7±19.91 for BMV loaded ethosomes and HCA loaded ethosomes respectively and both formulations had an acceptable PdI of 0.049 and 0.111, respectively. SEM analyses indicated that the ethosomes had a spherical shape. Encapsulation of the APIs was verified by the thermoanalyses and possible intermolecular interactions were identified using FTIR. BMV loaded and HCA loaded ethosomes had a respective EE of 74.57 % and 37.30 %, and a DL of 14.91 % and 7.46 %, respectively. The release kinetics best fit the Peppas-Sahlin model indicative of an anomalous non-Fickian diffusion coupled with polymer relaxation and zero order release. Conclusions: BMV and HCA loaded ethosomes for topical drug delivery were successfully synthesised and characterised. These novel nanoparticles have provided an array of avenues for further investigation and application in the topical delivery of corticosteroids

Os glicocorticóides são muito utilizados na clínica veterinária por serem potentes inibidores da resposta inflamatória. São encontrados em muitas preparações tópicas, sozinhos ou combinados com outros medicamentos e estão presentes, principalmente, em produtos destinados ao uso otológico. Mesmo quando aplicados topicamente, sabe-se que existe certo grau de absorção. Com a disponibilidade de novos glicocorticóides sintéticos mais potentes, vem sendo sugerido recentemente, a ocorrência de hiperadrenocorticismo iatrogênico em pequenos animais. Com a finalidade de se testar a hipótese que a utilização de preparações otológicas contendo glicocorticóides podem ocorrer em efeitos sistêmicos, o presente estudo avaliou três antiinflamatórios esteroidais comumente utilizados no tratamento de otite em cães e gatos, quais sejam a hidrocortisona, a betametasona e a triancinolona. Para tanto, ratos foram tratados tópicamente, via auricular, a cada 12 horas, durante 7, 14 e 21 dias, com os fármacos descritos acima. Utilizamos o edema de pata induzido pela injeção de carragenina como forma de avaliar a existência de níveis efetivos de antiinflamatórios circulantes. Ainda, foram avaliados vários parâmetros tais como variação de peso corpóreo, peso relativo de baço e adrenais, hemograma completo, índice de proliferação de linfócitos e fagocitose e burst oxidativo realizados por neutrófilos circulantes. Além disso, foi avaliado o nível de corticosterona plasmática nos animais tratados com glicocorticóides tópicos. O tratamento tópico com antiinflamatórios esteroidais levou a redução no volume de edema inflamatório, mais evidente com a triancinolona. De forma geral, houve queda no ganho, ou mesmo perda de peso corpóreo durante o tratamento tópico com os antiinflamatórios esteroidais empregados. Em alguns casos, essa redução de peso corpóreo foi acompanhada de diminuição do peso relativo do baço e adrenais, particularmente com a betametasona. A principal alteração observada no hemograma foi leucopenia, mais evidente com a triancinolona. Houve redução dos níveis plasmáticos de corticosterona, mais duradoura com a betametasona. Os tratamentos tópicos causaram discreta redução na intensidade e percentual de fagocitose realizada por neutrófilos circulantes, bem como na produção de espécies reativas de oxigênio. O estudo aponta que glicocorticóides tópicos comumente empregados na terapêutica veterinária podem ser absorvidos, na dependência da molécula empregada e de seu tempo de utilização, causando alterações sistêmicas relevantes já após 7 dias de tratamento. Sugerimos uma maior cautela e acompanhamento veterinário dos pacientes nos casos que haja necessidade de emprego de tais medicamentos por períodos prolongados. / Glucocorticoids are commonly employed in veterinary practice because they are potent inhibitors of the inflammatory response. They are present in many topical formulations, alone or combined with other active compounds, particularly in otic ointments. Even when applied topically, certain degree of absortion is known to occur. Following the availability of novel, more potent synthetic steroidal anti-inflammatory drugs, cases of iatrogenic hyperadrenocorticism have been suggested. Aiming on testing the hypothesis that glucocorticoid-containing otic ointments may lead to systemic effects, we employed three different synthetic glucocorticoids widely used for the treatment of otitis in small animals (hydrocortisone, betamethasone, and triamcinolone) our study Rats were treated topically on their ears, every 12 hours, for 7, 14, or 21 days with the drugs described above. We first employed carrageenan-induced paw edema in order to evaluate the presence of effective levels of circulating anti-inflammatory drugs. We also evaluated several parameters such as change in body weight, relative weight of spleen and adrenal glands, complete hemogram, proliferation index of lymphocytes, and phagocytosis and oxidative burst performed by circulating neutrophils. Moreover, we assessed the plasmatic levels of corticosterone in rats treated topically with anti-inflammatory steroids. Topical treatment led to a reduction on the volume of the inflammatory edema induced by carrageenan, more evident following triamcinolone. Altogether, rats treated with these molecules ceased to gain, or lost weight on the course of the experiment. In some cases, such reduction was accompanied by a decrease on the relative weight of the spleen and adrenal glands, particularly after betamethasone. The most commonly observed change in the hemogram was leucopenia, evident following triamcinolone. We found reduced levels of circulating corticosterone, more persistent after betamethasone. Topically applied glucocorticoids caused a discrete reduction in the intensity and percentage of phagocytosis by circulating neutrophils, and a reduction on the oxidative burst performed by these cells. This study shows that topical steroidal anti-inflammatory drugs, so commonly employed in veterinary practice, can be absorbed, depending on the molecule and duration of treatment, leading to systemic effects, which are already relevant after 7 days. Based on our findings, we suggest caution and a close follow-up by a qualified veterinarian whenever a long-term therapy based on these molecules and route of administration is required.

Os glicocorticóides são muito utilizados na clínica veterinária por serem potentes inibidores da resposta inflamatória. São encontrados em muitas preparações tópicas, sozinhos ou combinados com outros medicamentos e estão presentes, principalmente, em produtos destinados ao uso otológico. Mesmo quando aplicados topicamente, sabe-se que existe certo grau de absorção. Com a disponibilidade de novos glicocorticóides sintéticos mais potentes, vem sendo sugerido recentemente, a ocorrência de hiperadrenocorticismo iatrogênico em pequenos animais. Com a finalidade de se testar a hipótese que a utilização de preparações otológicas contendo glicocorticóides podem ocorrer em efeitos sistêmicos, o presente estudo avaliou três antiinflamatórios esteroidais comumente utilizados no tratamento de otite em cães e gatos, quais sejam a hidrocortisona, a betametasona e a triancinolona. Para tanto, ratos foram tratados tópicamente, via auricular, a cada 12 horas, durante 7, 14 e 21 dias, com os fármacos descritos acima. Utilizamos o edema de pata induzido pela injeção de carragenina como forma de avaliar a existência de níveis efetivos de antiinflamatórios circulantes. Ainda, foram avaliados vários parâmetros tais como variação de peso corpóreo, peso relativo de baço e adrenais, hemograma completo, índice de proliferação de linfócitos e fagocitose e burst oxidativo realizados por neutrófilos circulantes. Além disso, foi avaliado o nível de corticosterona plasmática nos animais tratados com glicocorticóides tópicos. O tratamento tópico com antiinflamatórios esteroidais levou a redução no volume de edema inflamatório, mais evidente com a triancinolona. De forma geral, houve queda no ganho, ou mesmo perda de peso corpóreo durante o tratamento tópico com os antiinflamatórios esteroidais empregados. Em alguns casos, essa redução de peso corpóreo foi acompanhada de diminuição do peso relativo do baço e adrenais, particularmente com a betametasona. A principal alteração observada no hemograma foi leucopenia, mais evidente com a triancinolona. Houve redução dos níveis plasmáticos de corticosterona, mais duradoura com a betametasona. Os tratamentos tópicos causaram discreta redução na intensidade e percentual de fagocitose realizada por neutrófilos circulantes, bem como na produção de espécies reativas de oxigênio. O estudo aponta que glicocorticóides tópicos comumente empregados na terapêutica veterinária podem ser absorvidos, na dependência da molécula empregada e de seu tempo de utilização, causando alterações sistêmicas relevantes já após 7 dias de tratamento. Sugerimos uma maior cautela e acompanhamento veterinário dos pacientes nos casos que haja necessidade de emprego de tais medicamentos por períodos prolongados. / Glucocorticoids are commonly employed in veterinary practice because they are potent inhibitors of the inflammatory response. They are present in many topical formulations, alone or combined with other active compounds, particularly in otic ointments. Even when applied topically, certain degree of absortion is known to occur. Following the availability of novel, more potent synthetic steroidal anti-inflammatory drugs, cases of iatrogenic hyperadrenocorticism have been suggested. Aiming on testing the hypothesis that glucocorticoid-containing otic ointments may lead to systemic effects, we employed three different synthetic glucocorticoids widely used for the treatment of otitis in small animals (hydrocortisone, betamethasone, and triamcinolone) our study Rats were treated topically on their ears, every 12 hours, for 7, 14, or 21 days with the drugs described above. We first employed carrageenan-induced paw edema in order to evaluate the presence of effective levels of circulating anti-inflammatory drugs. We also evaluated several parameters such as change in body weight, relative weight of spleen and adrenal glands, complete hemogram, proliferation index of lymphocytes, and phagocytosis and oxidative burst performed by circulating neutrophils. Moreover, we assessed the plasmatic levels of corticosterone in rats treated topically with anti-inflammatory steroids. Topical treatment led to a reduction on the volume of the inflammatory edema induced by carrageenan, more evident following triamcinolone. Altogether, rats treated with these molecules ceased to gain, or lost weight on the course of the experiment. In some cases, such reduction was accompanied by a decrease on the relative weight of the spleen and adrenal glands, particularly after betamethasone. The most commonly observed change in the hemogram was leucopenia, evident following triamcinolone. We found reduced levels of circulating corticosterone, more persistent after betamethasone. Topically applied glucocorticoids caused a discrete reduction in the intensity and percentage of phagocytosis by circulating neutrophils, and a reduction on the oxidative burst performed by these cells. This study shows that topical steroidal anti-inflammatory drugs, so commonly employed in veterinary practice, can be absorbed, depending on the molecule and duration of treatment, leading to systemic effects, which are already relevant after 7 days. Based on our findings, we suggest caution and a close follow-up by a qualified veterinarian whenever a long-term therapy based on these molecules and route of administration is required.

Vesicular delivery systems, like liposomes, represent an attractive approach to improving skin delivery of drugs. Liposomes are composed of one or several lipid bilayers surrounding an aqueous compartment. They are able to encapsulate hydrophilic compounds into their inner compartment and lipophilic compounds into their lipid bilayers. In the first part of this thesis, we developed a new dermal delivery system combining the advantages of cyclodextrin inclusion complexes and those of deformable liposomes. This system was called drugs-in-cyclodextrin-in-deformable liposome. Betamethasone was chosen as the model drug. These systems were successfully developed and characterised. In vitro diffusion studies using Franz diffusion cells and polycarbonate membranes gave promising results. In order to better mimic the in vivo conditions, ex vivo penetration studies using pig ear skin and Franz diffusion cells were developed. The tape stripping method was used to determine the amount of drug in the stratum corneum. The amount of drug in the epidermis, dermis and receptor medium was also assayed. These methods were successfully validated. Unfortunately, ex vivo penetration studies of the developed skin delivery system could not confirm the promising in vitro results. Our research was then directed towards studies allowing a better understanding of liposome skin penetration mechanisms. In the second part of this thesis, the influence of several parameters on the skin penetration behaviour of liposomes was investigated. The formulation importance on skin penetration efficiency was highlighted by the comparison of two liposomal systems. In the first system, betamethasone is encapsulated by the help of cyclodextrin inclusion complexes in the aqueous compartment. In the second system, betamethasone is encapsulated alone in the lipid bilayer. The second system shows high membrane permeability of the encapsulated betamethasone. Despite this decreased drug retention, the second system enhances the skin penetration of betamethasone. This high membrane permeability and the better penetration of the second formulation support the mechanism of penetration as a free drug substance. In addition, no difference in penetration is observed between liposomes containing betamethasone alone and a non liposomal dispersion. Several surfactants or edge activators were incorporated in the lipid bilayer of liposomes in order to obtain deformable liposomes. Unfortunately, these deformable liposomes are unable to enhance skin penetration compared to classical non deformable liposomes. The addition of charged lipids in the lipid bilayer of liposomes encapsulating either betamethasone or betamethasone dipropionate was investigated. The addition of negatively charged lipids enhances the penetration of the encapsulated drug. The use of negatively charged liposomes enhanced the epidermis accumulation of betamethasone 9.3 times compared with the ethanolic solution. This improvement is not observed with the corresponding non liposomal dispersion, indicating that the vesicle form is of high importance. In the case of the ester, the use of negatively charged liposomes enhances the epidermis accumulation 5.5 times compared with the Diprosone® lotion and only 1.6 times compared with the ethanolic solution. The penetration of a more lipophilic drug, with a high intrinsic penetration, is less enhanceable when incorporated in liposomes. Confocal microscopy was performed to visualise skin penetration of fluorescently labelled liposomes. The lipophilic dye rhodamine B encapsulated in the lipid bilayer as betamethasone penetrates the epidermis at the same level as NBD-phosphatidylcholine. On the other hand, the hydrophilic dye calcein encapsulated in the aqueous compartment as betamethasone-cyclodextrin complexes does not penetrate the epidermis. These observations seem to confirm the ex vivo results. In addition, the skin penetration of rhodamine B seems to be improved by the incorporation in negatively charged liposomes compared with classical ones. Finally, the efficiency of the negatively charged vesicles was evaluated in vivo on volunteers by the skin blanching assay. Carbomer gel containing the negatively charged vesicles seems to enhance the blanching response of encapsulated betamethasone in comparison with the ethanolic hydroxypropylcellulose gel. However, results are not significantly different. This could be explained by the small number of volunteers. On the other hand, in the case of formulations containing betamethasone dipropionate, no tendency could be highlighted. These results seem to confirm that the use of negatively charged liposomes is more attractive for improving the efficiency of a lesser lipophilic drug, betamethasone, in comparison with the more lipophilic ester, betamethasone dipropionate. These results need to be confirmed using a higher number of volunteers./ Les systèmes vésiculaires, comme les liposomes, représentent une approche intéressante pour améliorer ladministration cutanée de médicaments. Les liposomes sont des vésicules sphériques composées dune ou plusieurs bicouches lipidiques entourant une cavité aqueuse. Ils sont capables dencapsuler des molécules hydrophiles dans leur cavité aqueuse, ainsi que de retenir des molécules hydrophobes dans leur bicouche lipidique. Dans la première partie de la thèse, nous nous sommes intéressés au développement dun nouveau système dadministration cutanée combinant les avantages des complexes dinclusion dans les cyclodextrines et ceux des liposomes déformables, aboutissant à un nouveau concept appelé : « drugs-in-cyclodextrin-in-deformable liposome». La bétaméthasone a été utilisée comme molécule modèle. Ces nouveaux systèmes ont été mis au point et caractérisés avec succès. Les premières études de diffusion réalisées in vitro, à travers des membranes synthétiques et au moyen des cellules de diffusion de Franz, étaient prometteuses. Afin de se rapprocher des conditions in vivo, des études de pénétration des liposomes ont alors été mises au point, ex-vivo, au moyen de cellules de diffusion et à travers des peaux doreilles de cochons. La méthode du « tape stripping » a permis de déterminer la quantité de bétaméthasone dans le stratum corneum. La quantité de substance active dans lépiderme, le derme et le mileu récepteur des cellules de Franz a également été déterminée. Ces méthodes ont été validées avec succés. Malheureusement, les études sur peaux doreilles de cochons nont pas permis de confirmer nos premières conclusions. Les recherches ont alors été orientées vers des études permettant de mieux comprendre les mécanismes de pénétration cutanée des liposomes. Dans la seconde partie de la thèse, linfluence de différents paramètres sur la pénétration cutanée a été étudiée. La comparaison de la pénétration cutanée de deux systèmes liposomiaux différents a mis en évidence limportance de la formulation des liposomes sur leur efficacité. Dans le premier, la bétaméthasone est encapsulée comme précédemment à laide de cyclodextrines dans la cavité aqueuse des liposomes déformables. Dans le second système, la bétaméthasone est encapsulée seule au niveau de la bicouche lipidique des liposomes. Ce second système a montré une plus grande perméabilité de la molécule modèle encapsulée. Malgré cette plus faible rétention, le deuxième système a permis daugmenter la pénétration de la molécule modèle. La plus grande perméabilité et la meilleure pénétration du deuxième système permettent de conclure que la bétaméthasone pénétrerait sous une forme non encapsulée dans les liposomes. De plus, aucune différence na été observée entre la pénétration cutanée des liposomes encapsulant la bétaméthasone dans leur bicouche et une dispersion non liposomale. Plusieurs tensio-actifs ont également été incorporés au niveau de la bicouche des liposomes afin dobtenir des liposomes déformables. Malheureusement, la faible augmentation de pénétration observée par rapport aux liposomes classiques non déformables nest pas significative. Lajout de lipides chargés au niveau de la bicouche lipidique de liposomes encapsulant soit la bétaméthasone soit le dipropionate de bétaméthasone a été investigué. Laddition de lipides chargés négativement a permis daugmenter la pénétration de la molécule encapsulée. Lutilisation de liposomes chargés négativement a permis daugmenter la quantité de bétaméthasone au niveau de lépiderme dun facteur 9,3 par rapport à une solution éthanolique de bétaméthasone. Cette augmentation na pas été observée avec la dispersion non liposomale correspondante, indiquant que la formulation sous forme de vésicules est très importante. En ce qui concerne le dipropionate de bétaméthasone, lencapsulation dans les liposomes chargés négativement a augmenté la quantité dester dans lépiderme dun facteur 5,5 par rapport à la spécialité Diprosone® lotion et seulement dun facteur 1,6 par rapport à la solution éthanolique. Lencapsulation dans les liposomes chargés négativement semble donc moins avantageuse dans le cas dune molécule plus lipophile, comme le dipropionate de bétaméthasone et qui possède déjà une bonne pénétration cutanée intrinsèque par rapport à une molécule plus hydrophile comme la bétamethasone. La pénétration de liposomes rendus fluorescents a été visualisée grâce à la microscopie confocale. Un marqueur lipophile, la rhodamine B, encapsulé dans la bicouche comme la bétaméthasone, pénètre bien dans la peau de la même façon que la phosphatidylcholine fluorescente. Par contre, un marqueur hydrophile, la calcéine, encapsulé dans la cavité des liposomes comme les complexes bétaméthasone-cyclodextrines, ne pénètre pas dans lépiderme. Ces observations confirment les résultats de dosage ex vivo au moyen des cellules de Franz. De plus, la pénétration dans la peau de la rhodamine B semble être plus importante lorsquelle est encapsulée dans les liposomes chargés négativement par rapport aux liposomes non chargés. Enfin, lefficacité des liposomes négatifs a été évaluée in vivo sur des volontaires grâce à lévaluation du pouvoir de blanchiement des corticostéroïdes. Le gel de carbomère contenant les liposomes chargés négativement semble augmenter la réponse de la bétaméthasone par rapport au gel éthanolique dhydroxypropylcellulose. Cependant, les résultats ne sont pas significatifs. Une explication pourrait être le faible nombre de volontaires. Par contre, dans le cas des liposomes contenant le dipropionate de bétaméthasone, il a été plus difficile de dégager une tendance. Ces résultats semblent confirmer que lutilisation des liposomes chargés négativement est plus intéressante dans le cas dune molécule moins lipophile comme la bétamethasone par rapport à une molécule plus lipophile comme le dipropionate de bétaméthasone. Ces résultats nécessitent cependant dêtre confirmés sur un plus grand nombre de volontaires.

Approximately one in four men will have surgery for ingunial hernia in their lifetime. In Sweden, 16 000 procedures are performed each year. To investigate the possible link between handling of nerves and sensory disturbance, 97 groins in 92 patients were examined one year after inguinal hernia surgery. Sensory disturbances were found to be common after open surgery (29 %), but were not seen after the laparoscopic procedures. No significant relationship between sensory disturbance and handling of nerves or pain was seen. The risk for recurrence has been significantly reduced due to the use of prosthetic meshes, but continued surveillance of this important outcome will always be necessary. In that context, the time frame in which recurrence develops in relation to possible risk factors can help our understanding of the underlying mechanisms. To explore such temporal relationships, 142,578 patients were included in a register study. A relative over-risk for early recurrence was seen after suture repair, laparoscopic repair, after postoperative complications, and after surgery for previous recurrence. Corticosteroids are known to decrease pain and nausea after several surgical procedures. In a randomised trial on open hernia surgery, 398 patients were randomised to treatment with 12 mg of betamethasone or placebo. Decreased levels of pain were seen on the day of surgery, the next day and after one month. No difference was seen on days 2-7 and after one year. Nausea was not common and did not differ between the groups. Reoperation is sometimes performed to correct a presumed structural defect thought to cause the long-term pain. In order to evaluate the result of such treatment, 111 cases were analysed based on register data, questionnaires and medical records. Sixty-two per-cent of the patients reported an improvement compared to before the reoperation, but a high level of pain remaining (42 %), and impaired quality of life was seen. There was no clear advantage for any surgical intervention over the other.

A utilização do corticosteróide antenatal objetivando induzir artificialmente a maturação fetal e garantir a sobrevivência de neonatos críticos é bem estabelecido e rotineiramente utilizado em Medicina. Por outro lado, a utilização pós-natal de corticóide, como garantia de melhores condições clínicas do neonato, ainda figura como expressivo desafio tanto em neonatologia humana quanto veterinária. Deste modo, este estudo tem como objetivo avaliar a eficácia da corticoterapia pré ou pós-natal em cordeiros prematuros extremos na melhora da condição clínica, pulmonar, metabólica e hemodinâmica; necessidade de assistência ventilatória e na garantia de sobrevivência neonatal. Para tal, cordeiros prematuros, nascidos aos 135 dias de gestação, foram aleatoriamente alocados nos grupos: corticoterapia materna pré-natal (CORT PRÉ; n=8), corticoterapia pós-natal (CORT PÓS; n=9) e controle (CONT; n=5). A corticoterapia foi realizada com betametasona em dose única de 0,5 mg/Kg, por via de aplicação intra muscular, aos 133 dias de gestação ou aos 10 minutos após o nascimento para os grupos CORT PRÉ e CORT PÓS, respectivamente. O parto foi induzido utilizando-se o fármaco aglepristona às 49 horas prévias à data estimada para o parto. Os neonatos foram avaliados quanto ao escore Apgar, condição clínica geral (freqüência cardíaca, freqüência respiratória, tônus muscular e irritabilidade reflexa), temperatura corpórea, hemogasometria arterial, glicemia, lactatemia, concentrações séricas das enzimas antioxidantes (superoxido dismutase SOD e glutationa peroxidase GPx) e marcador do estresse oxidativo (TBARS) em 10 momentos pontuais durante os 3 dias subsequentes ao parto. Para os cordeiros destinados ao suporte ventilatório, avaliamos o tempo (em minutos) necessário para o início do protocolo ventilatório a partir do nascimento e o tempo de permanência na ventilação. Os dados foram analisados por meio de testes paramétricos e não-paramétricos, com nível de significância de 95%. As variáveis também foram submetidas ao teste de correlação de Spearman. Como resultados, observamos melhor condição de vitalidade e tônus muscular nos neonatos tratados em relação ao grupo controle. Quanto à avaliação cardiogênica, a betametasona pré-natal apresentou efeito bradicárdico, enquanto a administração pós-natal resultou em efeito taquicárdico. Tanto os resultados de frequência respiratória como temperatura corpórea foram estatisticamente superiores nos neonatos tratados em relação ao controle. Todos os cordeiros nasceram bradicárdicos e bradipneicos, com valores normais atingidos a partir de 10 minutos. Observamos hiperglicemia nos grupos tratados, com valor estatisticamente superior no grupo CORT PÓS. O valor de lactato foi estatisticamente superior no grupo CORT PÓS em relação ao controle. Quanto aos componentes do equilíbrio ácido-básico, os neonatos do grupo CORT PRÉ tiveram valores significativamente maiores de bicarbonato, enquanto menores pressões de CO2 foram observadas a partir de 60 minutos, embora sem diferença quanto ao pH e BE. Os tratamentos não determinaram alterações nos valores de PO2 e SO2. Entretanto, os diferentes protocolos corticoterápicos influenciaram no perfil hematológico neonatal, com valores significativamente maiores de hematócrito no grupo CORT PÓS, seguido pelo grupo CORT PRÉ. Observamos também atuação do tratamento pós-natal nos valores de SOD e correlação positiva com os valores de hematócrito e hemoglobina. Comparando-se o percentual de cordeiros submetidos à assistência ventilatória, verificou-se maior necessidade no grupo controle. Os neonatos do grupo CORT PRÉ permaneceram por tempo menor sob assistência ventilatória, em relação ao grupo controle Com base nos resultados obtidos, concluímos que a corticoterapia pós-natal ou pré-natal favorece a condição clínica neonatal (vitalidade, tônus muscular e funções vitais), a função pulmonar (trocas gasosas e compensação aos desequilíbrios ácido-básicos) e a atividade metabólica (controle glicêmico). O tratamento pós-natal aumentou a atividade da enzima SOD, reduzindo os riscos de danos pulmonares. Ainda, o tratamento com betametasona pós-natal ou materna pré-natal diminui a necessidade de assistência ventilatória em cordeiros prematuros extremos / The use of corticosteroid aiming artificially fetal lung maturation and survival of critical neonates is used as a routine in medicine. On the other hand, the postnatal steroid therapy used to improve clinical conditions of the newborn, It\'s still a therapeutic challenge both in human as veterinary neonatology. Thus, the aims this study is to assess the effectiveness of corticosteroid, used in the pre or postnatal period, in extremely preterm lambs to improve clinical, pulmonary, metabolic and hemodynamic condition; the need for ventilatory support and neonatal survival rate. For it, preterm lambs, born with 135 days of gestation, were randomly allocated into three groups: prenatal maternal corticosteroid therapy (CORT PRE; n = 8), postnatal corticosteroid therapy (CORT POST; n = 9) and control (CONT; n = 5). The steroid therapy was performed with betamethasone in a single dose of 0.5 mg/ kg, by intramuscular route of administration, at 133 days of pregnancy our after 10 minutes from the birth in the groups CORT PRÉ and CORT PÓS, respectively. The labor was induced with aglepristone administrated 49 hours prior to the estimated date of birth. Newborns were assessed by Apgar score, general medical condition (heart rate, respiratory rate, muscle tone and reflex irritability), body temperature, blood gas analysis, blood glucose, blood lactate concentration, serum concentrations of antioxidant enzymes (superoxide dismutase - SOD and glutathione peroxidase - GPx) and marker of oxidative stress (TBARS). The assessments were performed in 10 moments during the three days from delivery. For ventilated newborns, we assessed the needed time (in minutes) between birth and the beginning of ventilatory protocol, as well as length of stay on ventilatory support. Data were analyzed using parametric and nonparametric tests, with 95% of significance level. The variables were assessed from Spearman correlation test. As results, we noted better condition of vitality and muscle tone in newborns treated when compared with control group. Regarding cardiogenic values, antenatal betamethasone resulted in bradycardic effect, while its postnatal administration was related with tachycardia. Both results of respiratory and body temperature rates were significantly higher in newborns treated compared to the control. All lambs born whith bradycardia and bradypnea, reached normal values from 10 minutes of life. Hyperglycemia was observed in the treated groups, with statistically higher values in CORT PÓS group. The lactate value was statistically higher in CORT PÓST group compared to the control. Concerning the components of the acid-base balance, the CORT PRÉ showed significantly higher values of bicarbonate, while lower pressures of CO2 were observed from 60 minutes, although there were no differences in pH and BE values. The treatments did not determine changes in PO2 and SO2 values. However, the different protocols of steroid therapy influenced on neonatal blood profile, with significantly higher hematocrit values in CORT POST group, followed by CORT PRÉ. We also observed influence of postnatal treatment in SOD values as well as positive correlation between SOD and the hematocrit and hemoglobin values. Comparing the percentage of lambs submitted to mechanical ventilation, there were most ventilated neonates in the control group. Lower ventilation time was observed in the CORT PRÉ group. Then we conclude that, the postnatal or antenatal steroids improve neonatal clinical condition (vitality, muscle tone and vital functions), lung function (gas exchange and compensation to acid-base imbalances) and metabolic activity (glycemic control). The postnatal treatment increased the activity of SOD by reducing the risks of pulmonary damage. Moreover, treatment with postnatal betamethasone or prenatal maternal reduces the need for mechanical ventilation in extremely preterm lambs

Corticosteroide por via espinal é uma das alternativas no processo de tratamento da dor radicular quando o tratamento conservador falhou. Sua administração por via epidural vem sendo reavaliada devido ao risco de sequelas, provenientes da obstrução de vasos sanguíneos, associadas a outros efeitos adversos próprios dos corticosteroides. O presente estudo visou avaliar o efeito analgésico da administração de diferentes corticosteroides, exemplificados pela dexametasona e pela betametasona, administrados por via epidural. Vinte e seis pacientes portadores de dor neuropática secundária à hérnia de disco participaram do estudo. A punção sacral foi realizada com combinação de 40 mg de lidocaína, 30 µg de clonidina e 10 mg de dexametasona ou 10 mg de betametasona, diluídos para volume final 10 ml, com solução fisiológica a 0,9%, após comprovação com contraste não-iodado do perfeito posicionamento da agulha no espaço epidural. Este procedimento foi repetido uma vez por semana por 2 semanas seguidas, e após um intervalo de 3 semanas foi realizado mais duas vezes, uma vez por semana, sendo o total de 4 procedimentos por paciente. Treze pacientes iniciaram com dexametasona epidural durante as primeiras duas semanas, e prosseguiram com betametasona após período de descanso de três semanas, sendo que cada paciente atuou como seu próprio controle. Os pacientes foram avaliados em relação à analgesia, incidência de efeitos adversos e alteração das concentrações plasmáticas de íons, cortisol, adrenocorticotrofina e glicemia. Vinte e três pacientes completaram o estudo. Cada paciente foi o seu próprio controle e, no mesmo paciente, o efeito analgésico da dexametasona foi superior ao da betametasona. (p<0,05). Cortisol e adrenocorticotrofina plasmáticos reduziram no sétimo dia após realização do bloqueio (p<0,05), As demais medidas de concentrações plasmáticas de íons Na+, K+, Ca++, glicemia inicial, pós-prandial e hemoglobina glicada foram semelhantes entre os grupos, e sem alteração quando comparados com valores iniciais (p>0,05). Ambos grupos relataram insônia relativa nos primeiros dias pós-bloqueio (p<0,05). Não houve alteração de peso corporal e da pressão arterial durante o tratamento entre os fármacos avaliados (p>0,05). Entretanto, 3 pacientes que receberam dexametasona e 2 que receberam betametasona inicialmente, apresentam medida de pressão ocular aumentada (p<0,05). Como conclusões, a analgesia da dexametasona foi superior quando comparada à da betametasona, (p<0,05). Entretanto ambos grupos apresentaram aumento da pressão ocular e prejuízo do sono noturno, não relevante este último. Finalmente, uma vez que a dexametasona foi superior à betametasona em relação à analgesia e demonstrada ser mais segura em relação à formação de agregados, sugere-se a dexametasona como escolha. / Although the primarily indication of epidural corticosteroids as part of the treatment of acute neuropathic pain when the conservative treatment have failed, recently there have been concerns related to which corticosteroid would be the best indication, as there is the risk of particulate aggregation and serious adverse effects secondary to their own pharmacological properties or intra vessel aggregation and obliteration, as well their efficacy as analgesics. This actual study was designed to evaluate two different corticosteroids, i.e., dexamethasone and betamethasone, as coajuvants in epidural management of acute radicular pain. Twenty-six patients with history of neuropathic pain secondary to disc herniation acted as their own control related to the epidural administration of dexamethasone and betamethasone. Sacral puncture was performed with a combination of 40 mg of lidocaine, 30 µg of clonidine and 10 mg of dexamethasone or 10 mg of betamethasone, diluted to final volume 10 ml with 0.9% physiological solution after non-iodinated contrast of the perfect positioning of the needle in the epidural space.Thirteen patients have started with dexamethasone during the first two weekly procedures, and after 3 weeks of wash-out were submitted to two weekly sequence of sacral betamethasone. The other patients have started with epidural betamethasone, followed by dexamethasone after 3-week rest. Each patient acted as its own control. Patients were evaluated related to analgesia, blood pressure, weight gain, adverse effects and plasmatic measurements of ions, glicemia, ACTH and cortisol. Twenty-three patients completed the study. The analgesic effect was higher when dexamethasone was used in the same patient (p <0.05) Plasma cortisol and ACTH reduced on the 7th day after the block (p<0.05). The plasmatic concentrations of the ions Na+, K+, Ca++, control and post-prandial glicemia, blood pressure, weight were similar between groups and did no differ from initial control values (p>0.05). Both groups reported relative insomnia in the first post-blockade days (P <0.05). However, 3 patients that received dexamethasone and 2 that received betamethasone had ocular pressure increase (p<0.05). As conclusions, both drugs resulted in still unaware increase in ocular pressure and sleep disturbance. Because dexamethasone analgesia was superior and there were no differences regarding adverse effects between either epidural betamethasone or dexamethasone, based on the literature, it would be more appropriate the utilization of dexamethasone.