The blood-brain barrier restricts the access of many compounds, including therapeutic agents, to the brain. Several human studies indicate that nasal administration of hydrophilic compounds, such as peptides, can bypass the blood-brain barrier. The aims of this thesis were to develop and refine models for this direct nose-to-brain transfer. In a mouse model, [3H]-dopamine was given as a unilateral nasal dose. The resulting radioactivity in the ipsilateral olfactory bulb was significantly higher than that in the contralateral bulb and peaked at 4 h. Tape section autoradiography showed that the radioactivity was concentrated in the olfactory nerve layer and the glomerular layer of the olfactory bulb. The olfactory transfer of dopamine was also studied in vitro. At a lower donor concentration, the mucosal-to-serosal dopamine permeability was higher than the serosal-to-mucosal permeability, but at a higher concentration, the permeability coefficients were similar. Together, these results suggest that the olfactory transfer of dopamine has an active component. Olfactory transfer of fluorescein-labeled dextran through the epithelium and deeper tissues was studied in a rat model, which enabled visualization of the transfer using fluorescence microscopy. Although the epithelial transfer appeared to be mainly intracellular, transfer in the following deeper tissues was extracellular. Without altering the route of uptake, a gellan gum formulation enhanced the uptake of fluorescein dextran. The enhancing effect was considered likely to be the result of an increased residence time in the nasal cavity. In conclusion, dopamine and fluorescein-labeled dextran were identified as suitable model compounds for the study of olfactory drug transfer mechanisms and the influence of drug formulation. Two new in vitro models of olfactory transfer were compared. Also, a rat model, which enabled the visualization of the entire nose-to-brain transfer, was developed.

Trichobilharzia regenti is a nasal avian schistosome which has during the initial phase of infection an affinity to the nervous system. Larvae migrate through the central nervous system to the nasal mucosa of waterfowl, where they mature and reproduce. Until now this infection phase has been described only marginally. Adults are located in the nasal mucosa approximately from 13th to 24th day post infection. During this life phase, they migrate through the vascularized connective tissue and lay eggs, the presence of which has been detected in the tissue only. Maturation and hatching of miracidia occur in the tissue (unlike human schistosomes). The parasite causes inflammation, and the tissue is infiltrated with immune cells. Lymphocytes, granulocytes, macrophages, plasma cells and giant multi-nuclear cells were described by histological methods. The thesis is focused on a more detailed description of cellular immune response and histopathological changes of the tissue by means of histological stains, and antibody/lectin probes. The flukes were observed more frequently in the blood vessel lumen, together with a higher number of immune cells compared to the healthy duck. Infiltration by a high number of lymphocytes occurred in the tissue, the macrophages were frequently observed in clusters around the...

Luffa operculata é uma planta medicinal popularmente usada para tratamento de rinites e rinossinusites. A infusão de seu fruto seco é usada no nariz, liberando secreção mucosa profusa, mas pode causar irritação, epistaxe ou anosmia. Avaliamos os efeitos de diferentes concentrações da infusão do fruto seco da Luffa operculata na velocidade de transporte mucociliar (VTM), freqüência de batimento ciliar (FBC), diferença de potencial transepitelial (DPT) e morfologia do epitélio (microscopia de luz e eletrônica de transmissão), no modelo do palato isolado de rã. Os resultados apontam para dano epitelial dose-dependente no epitélio mucociliar, sugerindo que ela seja potencialmente nociva à mucosa nasal humana / Luffa operculata is a medicinal plant popularly used for treatment of rhinitis and rhinosinusitis. Its dry fruit infusion is used into the nose, delivering profuse mucous secretion, but may cause nasal mucosa irritation, epistaxis or anosmia. We evaluated the effects of different concentrations of Luffa operculata dry fruit infusion on mucociliary transport velocity (MTV), ciliary beat frequency (CBF), transepithelial potential difference (TPD) and epithelial morphology (light and electron transmission microscopy) of the isolated frog palate preparation. Results pointed to dose-dependent epithelial damage on mucociliary epithelium, suggesting that it is potentially noxious to the human nasal mucosa

In dieser Arbeit wurden die beiden Membranen Schweinekornea und Rindernasenmukosa hinsichtlich ihrer Permeabilität für den lipophilen Arzneistoff Androstendion (AD), der sowohl zur ophthalmologischen Anwendung als auch für die nasale Applikation mit systemischer Wirkung von Interesse ist, verglichen. Zusätzlich wurden identische Versuche mit der synthetischen Membran, Nephrophan(r), durchgeführt. Neben der gepufferten Arzneistofflösung, für die aufgrund des differenzierten Membranaufbaus Permeationskoeffizienten (Peff) für AD im Verhältnis von 3:1:4 (Mukosa : Kornea : Nephrophan(r)) resultierten, standen zwei entwickelte wasserkontinuierliche, nicht-ionische Mikroemulsionen (ME) und ihre Einzelkomponenten als Trägerformulierungen bei den Permeationsstudien im Vordergrund. Darüber hinaus wurde ein ME-System mit einem kationischen Kotensid entwickelt, charakterisiert und in die Untersuchungen einbezogen. Die getesteten Trägerformulierungen führten an den einzelnen Membranen zu unterschiedlichen Ergebnissen. Um auch den "hydrophilen Permeationsweg" einzuschließen, wurde parallel Fluorescein-Na (FSC) als hydrophile Modellsubstanz getestet. Als mögliche Ursache für diese differenzierten Ergebnisse wurde ein Einfluss der Additiva und Formulierungen auf das Verteilungsverhalten des lipophilen AD zwischen Donatorlösung und Membran in Betracht gezogen und daher in einem weiteren Versuchsblock die Penetrationsrate untersucht. Darüber hinaus wurde parallel dazu der metabolische Abbau, den AD während des Membrandurchtritts durch die vorhandenen Enzyme erfahren kann, berücksichtigt. Die Resultate zeigten Übereinstimmung mit den Permeationsergebnissen indem die Additiva und Formulierungen die Penetration in das Gewebe und den Metabolismus mehr oder weniger herabsetzten. Zur Charakterisierung der systemischen Verfügbarkeit von AD nach nasaler Applikation wurden im Anschluss an die Permeations- und Penetrationsversuche In-vivo-Studien an Kaninchen durchgeführt, die hier allerdings lediglich mit orientierendem Charakter einbezogen werden konnten. Um schließlich die Verhältnisse am Auge bzw. den mehrschichtigen Tränenfilm hinsichtlich einer Applikation der ME-Systeme modellhaft zu simulieren, wurden biophysikalische Untersuchungen in einem Langmuir-Trog mit Meibom''schen Drüsensekret als Oberflächenfilm durchgeführt, die über mögliche Interaktionen der Formulierungen bzw. ihrer Bestandteile mit der Tränenlipidschicht des Auges Auskunft geben sollten. Hier zeigten sich günstige Einflüsse der ME auf die Tränenlipidschicht, die vor allem bei einer Anwendung der ME bei Trockenem Auge von Vorteil sein können. / In these studies in vitro permeability of porcine cornea and bovine nasal mucosa was investigated and compared to each other using the lipophilic drug androstenedione (AD), which is of interest for ocular use as well as nasal, systemical administration. Additionally, the artificial membrane, Nephrophan(r), was used for identical investigations. Because of the differentiated membrane structure AD-permeation behaviour out of buffer solution resulted in a ratio of permeability coefficients (Peff) of 3:1:4 (nasal mucosa : cornea : Nephrophan(r)). Furthermore, two water-continuous, non-ionic microemulsions (ME) and their isolated components were investigated as carrier formulations. Additionally, a new ME containing a cationic co-surfactant was developed, characterized and included in the permeability studies as well. Permeation out of these carrier formulations also resulted in different Peff in case of all tissues. For including studies of the hydrophilic transport way flourescein-sodium (FSC) was investigated as well representing a hydrophilic model substance. Influence of the additives and formulations on the partition behaviour of AD between membrane and donor solution was considered to partially cause these results. Therefore, penetration of AD was investigated together with the metabolic conversion of AD caused by enzymes located in the biological membranes. The additives and formulations decreased penetration into the tissue as well as metabolism of the drug. These findings corresponded with and could therefore explain the results of the permeability studies to some extend. For characterizing systemical availability of AD after nasal administration and improving the results of the permeability and penetration investigations in vivo studies using rabbits were carried out. However, these studies could give but marginal information and therefore be incorporated for orientation only. Furthermore, biophysical investigations were carried out using a Langmuir trough with Meibomian gland secrete (MGS) as the surface layer in order to simulate the multiple layer tear film. These studies were supposed to give some information about interactions between the ME or their isolated components, respectively, and the lipid layer of the tear film, regarding ocular administration of these formulations. The results showed suitable influence of the ME on the MGS, which can especially be advantageous for a use in Dry eye syndrome.

INTRODUÇÃO: A toxina botulínica tipo A (TXB-A) tem sido testada no tratamento da rinite, principalmente nos casos de rinite idiopática. Sugere-se que um estado de hiper-reatividade do nervo trigêmeo esteja envolvido na fisiopatologia da rinite idiopática. O nervo trigêmeo possui fibras sensitivas não mielinizadas tipo C (FSNMT-C) que contém os neuropeptídeos substância P (SP) e peptídeo relacionado ao gene da calcitonina (CGRP). O óxido nítrico (NO) produzido pelas enzimas óxido nítrico sintase (NOS) também está envolvido nesse processo de neurorregulação nasal. O transporte mucociliar, mecanismo primário de defesa do sistema respiratório, é formado pelo batimento ciliar e muco nasal, e esses componentes podem ser influenciados por diferentes neuropeptídeos e neurotransmissores presentes na mucosa nasal. OBJETIVO: O objetivo deste estudo foi avaliar o efeito da TXB-A sobre a expressão da SP, CGRP e óxido nítrico sintase neural (nNOS), além de sua influência sobre o transporte mucociliar nasal em coelhos. MÉTODOS: Coelhos machos saudáveis da linhagem Nova Zelândia foram divididos em dois grupos: o grupo tratamento recebeu TXB-A (25UI) na concha nasomaxilar (CNM) do lado direito e soro fisiológico a 0,9% (SF0,9%) na CNM esquerda. O grupo controle recebeu SF0,9% na CNM direita e nenhuma intervenção na CNM esquerda. Foram investigados os efeitos da TXB-A sobre a expressão da SP, CGRP e nNOS no tecido de CNM por meio da imuno-histoquímica. Para esta análise, dividiu-se o tecido em camada externa (CE, acima da membrana basal) e camada interna (CI, abaixo da membrana basal). Avaliou-se também a presença de apoptose celular, a frequência de batimento ciliar (FBC), o perfil histoquímico do muco nasal (glicoproteínas ácidas e neutras) e a espessura do epitélio (ESP-CE). RESULTADOS: Foi observado um aumento significativo na quantidade de células apoptóticas na CNM do grupo tratamento que recebeu TXB-A em comparação aos controles (p <= 0,001). A CNM do grupo tratamento que recebeu SF0,9% exibiu um aumento na quantidade de células apoptóticas na CI ao comparar com os controles (CNM SF0,9%, p=0,035) (CNM sem intervenção, p=0,022), e também um aumento da expressão de SP na CE em comparação aos controles (CNM SF0,9%, p=0,021) (CNM sem intervenção, p=0,040). A expressão de CGRP apresentou um aumento na CNM do grupo tratamento que recebeu TXB-A em comparação à CNM sem intervenção (p=0,008). A FBC, expressão de nNOS, perfil histoquímico do muco nasal e ESP-CE não apresentaram diferenças significativas. DISCUSSÃO: O aumento da expressão de CGRP e SP pode ter sido causado por inibição de sua exocitose vesicular pela TXB-A, levando ao seu acúmulo intracelular. Não foram observadas diferenças significativas na FBC ou perfil histoquímico do muco nasal, indicando que o aumento da expressão de CGRP e SP não foi associado à inflamação. O aumento da quantidade de células apoptóticas e da expressão de SP na CNM SF0,9% do grupo tratamento pode ter sido causado por um efeito central da TXB-A. CONCLUSÃO: A administração nasal de TXB-A aumentou a expressão de CGRP e SP, possivelmente por acúmulo intracelular por causa da inibição da sua exocitose, mas sem alterar a FBC e o perfil histoquímico do muco nasal / INTRODUCTION: Botulinum toxin type A (BoNT-A) has been assessed in the treatment of rhinitis, especially in cases of idiopathic rhinitis. Trigeminal hyper-responsiveness appears to be involved in the pathological process of idiopathic rhinitis. Trigeminal nociceptive type C unmyelinated sensory fibers contain the neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). Nitric oxide (NO) produced by the enzyme nitric oxide synthase (NOS) are also involved in this nasal neurorregulation process. The mucociliary clearance, primary defense system of the respiratory system, is composed by the ciliary beat and nasal mucus. These components can be influenced by different nasal neuropeptides and neurotransmitters. OBJECTIVE: The aim of this study was to evaluate the effect of BoNT-A on the expression of SP, CGRP and neural nitric oxide synthase (nNOS), and its influence on nasal mucociliary clearance in rabbits. METHODS: Healthy New Zealand male rabbits were divided into two groups: the treatment group was challenged with BoNT-A (25UI) in the right nasomaxillary turbinate (NMT) and saline (SF0.9%) in the left NMT. The control group received SF0.9% in the right NMT and no-intervention in the left NMT. We investigated the effects of BoNT-A on SP, CGRP and nNOS expression in the NMT tissue by immunohistochemistry. Each area of interest was subdivided into an internal layer (IL: below the basement membrane) and outer layer (OL: above the basement membrane) for analysis. It was also assessed signs of cellular apoptosis, ciliary beat frequency (CBF), mucus histochemical profile (acidic and neutral glycoproteins) and epithelial thickness (EP-TH). RESULTS: It was observed a significant increase in the amount of apoptotic cells in the BoNT-A-challanged NMT compared with controls (p <= 0.001). The NMT of treatment group which received only SF0.9% showed an increase in the amount of apoptotic cells in the IL compared with controls (NMT SF0.9%, p = 0.035) (NMT no-intervention, p = 0.022), and also an increase in the SP expression in the OL compared with controls (NMT SF0.9%, p = 0.021) (NMT no-intervention, p = 0.040). CGRP expression showed higher expression in the BoNT-A-challanged NMT compared with no-intervention NMT (p=0.008). The CBF, nNOS expression, mucus histochemical profile and EP-TH did not show significant differences. DISCUSSION: The increased CGRP and SP expression could be due to inhibition of vesicular exocytosis by BoNT-A, leading to CGRP and SP intracellular accumulation. No significant differences in CBF or mucus histochemical profile were observed, indicating that the increased CGRP and SP expression was not associated with inflammation. The increase in the amount of apoptotic cells and SP expression in the SF0.9% NMT of treatment group may be due to a central effect of BoNT-A. CONCLUSION: Nasal administration of BoNT-A increased SP and CGRP expression, possibly via inhibition of their release, but did not change the CBF or mucus profile

INTRODUÇÃO: A toxina botulínica tipo A (TXB-A) tem sido testada no tratamento da rinite, principalmente nos casos de rinite idiopática. Sugere-se que um estado de hiper-reatividade do nervo trigêmeo esteja envolvido na fisiopatologia da rinite idiopática. O nervo trigêmeo possui fibras sensitivas não mielinizadas tipo C (FSNMT-C) que contém os neuropeptídeos substância P (SP) e peptídeo relacionado ao gene da calcitonina (CGRP). O óxido nítrico (NO) produzido pelas enzimas óxido nítrico sintase (NOS) também está envolvido nesse processo de neurorregulação nasal. O transporte mucociliar, mecanismo primário de defesa do sistema respiratório, é formado pelo batimento ciliar e muco nasal, e esses componentes podem ser influenciados por diferentes neuropeptídeos e neurotransmissores presentes na mucosa nasal. OBJETIVO: O objetivo deste estudo foi avaliar o efeito da TXB-A sobre a expressão da SP, CGRP e óxido nítrico sintase neural (nNOS), além de sua influência sobre o transporte mucociliar nasal em coelhos. MÉTODOS: Coelhos machos saudáveis da linhagem Nova Zelândia foram divididos em dois grupos: o grupo tratamento recebeu TXB-A (25UI) na concha nasomaxilar (CNM) do lado direito e soro fisiológico a 0,9% (SF0,9%) na CNM esquerda. O grupo controle recebeu SF0,9% na CNM direita e nenhuma intervenção na CNM esquerda. Foram investigados os efeitos da TXB-A sobre a expressão da SP, CGRP e nNOS no tecido de CNM por meio da imuno-histoquímica. Para esta análise, dividiu-se o tecido em camada externa (CE, acima da membrana basal) e camada interna (CI, abaixo da membrana basal). Avaliou-se também a presença de apoptose celular, a frequência de batimento ciliar (FBC), o perfil histoquímico do muco nasal (glicoproteínas ácidas e neutras) e a espessura do epitélio (ESP-CE). RESULTADOS: Foi observado um aumento significativo na quantidade de células apoptóticas na CNM do grupo tratamento que recebeu TXB-A em comparação aos controles (p <= 0,001). A CNM do grupo tratamento que recebeu SF0,9% exibiu um aumento na quantidade de células apoptóticas na CI ao comparar com os controles (CNM SF0,9%, p=0,035) (CNM sem intervenção, p=0,022), e também um aumento da expressão de SP na CE em comparação aos controles (CNM SF0,9%, p=0,021) (CNM sem intervenção, p=0,040). A expressão de CGRP apresentou um aumento na CNM do grupo tratamento que recebeu TXB-A em comparação à CNM sem intervenção (p=0,008). A FBC, expressão de nNOS, perfil histoquímico do muco nasal e ESP-CE não apresentaram diferenças significativas. DISCUSSÃO: O aumento da expressão de CGRP e SP pode ter sido causado por inibição de sua exocitose vesicular pela TXB-A, levando ao seu acúmulo intracelular. Não foram observadas diferenças significativas na FBC ou perfil histoquímico do muco nasal, indicando que o aumento da expressão de CGRP e SP não foi associado à inflamação. O aumento da quantidade de células apoptóticas e da expressão de SP na CNM SF0,9% do grupo tratamento pode ter sido causado por um efeito central da TXB-A. CONCLUSÃO: A administração nasal de TXB-A aumentou a expressão de CGRP e SP, possivelmente por acúmulo intracelular por causa da inibição da sua exocitose, mas sem alterar a FBC e o perfil histoquímico do muco nasal / INTRODUCTION: Botulinum toxin type A (BoNT-A) has been assessed in the treatment of rhinitis, especially in cases of idiopathic rhinitis. Trigeminal hyper-responsiveness appears to be involved in the pathological process of idiopathic rhinitis. Trigeminal nociceptive type C unmyelinated sensory fibers contain the neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). Nitric oxide (NO) produced by the enzyme nitric oxide synthase (NOS) are also involved in this nasal neurorregulation process. The mucociliary clearance, primary defense system of the respiratory system, is composed by the ciliary beat and nasal mucus. These components can be influenced by different nasal neuropeptides and neurotransmitters. OBJECTIVE: The aim of this study was to evaluate the effect of BoNT-A on the expression of SP, CGRP and neural nitric oxide synthase (nNOS), and its influence on nasal mucociliary clearance in rabbits. METHODS: Healthy New Zealand male rabbits were divided into two groups: the treatment group was challenged with BoNT-A (25UI) in the right nasomaxillary turbinate (NMT) and saline (SF0.9%) in the left NMT. The control group received SF0.9% in the right NMT and no-intervention in the left NMT. We investigated the effects of BoNT-A on SP, CGRP and nNOS expression in the NMT tissue by immunohistochemistry. Each area of interest was subdivided into an internal layer (IL: below the basement membrane) and outer layer (OL: above the basement membrane) for analysis. It was also assessed signs of cellular apoptosis, ciliary beat frequency (CBF), mucus histochemical profile (acidic and neutral glycoproteins) and epithelial thickness (EP-TH). RESULTS: It was observed a significant increase in the amount of apoptotic cells in the BoNT-A-challanged NMT compared with controls (p <= 0.001). The NMT of treatment group which received only SF0.9% showed an increase in the amount of apoptotic cells in the IL compared with controls (NMT SF0.9%, p = 0.035) (NMT no-intervention, p = 0.022), and also an increase in the SP expression in the OL compared with controls (NMT SF0.9%, p = 0.021) (NMT no-intervention, p = 0.040). CGRP expression showed higher expression in the BoNT-A-challanged NMT compared with no-intervention NMT (p=0.008). The CBF, nNOS expression, mucus histochemical profile and EP-TH did not show significant differences. DISCUSSION: The increased CGRP and SP expression could be due to inhibition of vesicular exocytosis by BoNT-A, leading to CGRP and SP intracellular accumulation. No significant differences in CBF or mucus histochemical profile were observed, indicating that the increased CGRP and SP expression was not associated with inflammation. The increase in the amount of apoptotic cells and SP expression in the SF0.9% NMT of treatment group may be due to a central effect of BoNT-A. CONCLUSION: Nasal administration of BoNT-A increased SP and CGRP expression, possibly via inhibition of their release, but did not change the CBF or mucus profile

Luffa operculata é uma planta medicinal popularmente usada para tratamento de rinites e rinossinusites. A infusão de seu fruto seco é usada no nariz, liberando secreção mucosa profusa, mas pode causar irritação, epistaxe ou anosmia. Avaliamos os efeitos de diferentes concentrações da infusão do fruto seco da Luffa operculata na velocidade de transporte mucociliar (VTM), freqüência de batimento ciliar (FBC), diferença de potencial transepitelial (DPT) e morfologia do epitélio (microscopia de luz e eletrônica de transmissão), no modelo do palato isolado de rã. Os resultados apontam para dano epitelial dose-dependente no epitélio mucociliar, sugerindo que ela seja potencialmente nociva à mucosa nasal humana / Luffa operculata is a medicinal plant popularly used for treatment of rhinitis and rhinosinusitis. Its dry fruit infusion is used into the nose, delivering profuse mucous secretion, but may cause nasal mucosa irritation, epistaxis or anosmia. We evaluated the effects of different concentrations of Luffa operculata dry fruit infusion on mucociliary transport velocity (MTV), ciliary beat frequency (CBF), transepithelial potential difference (TPD) and epithelial morphology (light and electron transmission microscopy) of the isolated frog palate preparation. Results pointed to dose-dependent epithelial damage on mucociliary epithelium, suggesting that it is potentially noxious to the human nasal mucosa