Chitosan has proven through the years as a versatile biomaterial to be used in pharmaceutical applications. Its mucoadhesive properties as well as its ability to manipulate the tight junctions in epithelium membranes have qualified it as an effective drug carrier in controlled drug delivery systems. Microparticles or beads as they are forward called in this study have advantages over conventional drug dosage forms because of a large surface to volume ratio and have the ability to target a specific site for drug release. Indomethacin is an anti-inflammatory drug that causes gastrointestinal side effects in conventional immediate-release dosage forms. The goal is to manipulate the drug delivery vehicle to target the intestines/colon as the site for drug delivery and to minimize this side effect. Thus chitosan beads have been chosen as a drug delivery system for indomethacin in this study. Chitosan beads have been prepared through the ionotropic gelation method using tripolyphophate (TPP) as a cross-linking agent. To prepare the most effective bead to encapsulate indomethacin different formulation and system variables (pH of the TPP solution, the concentration of the TPP solution as well as the indomethacin concentration) have been evaluated according to the following parameters: morphology, drug loading capacity and swelling capability. The ideal pH of the TPP solution was determined at 8.7 and the most effective TPP and indomethacin concentration were 5% w/v and 4% w/v respectively. The chitosan concentration was kept at 3% w/v throughout the study. These concentrations were used to examine the effect of pharmaceutical excipients on the indomethacin release from chitosan beads. The effect of the different excipients namely, ExplotabⒽ(0.25% w/v), Ac-Di-SolⓀ (0.5% w/v) and Vitamin C (0.25% w/v), on the morphology, drug loading capacity, swelling capability as well as the drug release of indomethacin chitosan beads (ICB's) were also studied. The excipients were used in the individually above mentioned concentrations and in combination with each other in the same concentrations. These formulations were used in dissolution studies over a period of 6 hours in PBS pH 7.4 solutions. The indomethacin release rate increased when an excipient was added to the formulation and it dramatically increased when the excipients were added in their various combinations, compared to the formulation that did not contain excipients. / Contents: Chitosan -- Controlled drug delivery -- Indomethacin -- Inotropic gelation -- Tripolyphosphate (TPP) -- Explotab® -- Ac-Di-Sol® -- Vitamin C / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2007.

Controlled release formulations offer many advantages over conventional dosage forms. These include reduced plasma fluctuations and improved patient comp1i:nce. Complex controlled release formulations such as those with enteric release properties, often require additional steps in the production phase. The costs and economic impact associated with these complex controlled release dosage formulations often outweigh the immediate benefits. Thus the development of an economic method to produce controlled release particles is of great importance especially in third world countries. In controlled release formulations, the drug is generally dispersed throughout a polymer matrix. The rate of drug release is often determined by the viscosity or complexity of the polymer matrix through which the drug needs to diffuse in order to be released. With enteric release the polymer coating, insoluble in an acidic environment is often applied in the final phase of production. Chitosan is a versatile polymer of natural origin with many favourable characteristics. These include its safety, biocompatibility, and biodegradability. Simple methods can be applied and modified to produce controlled release particles form chitosan. The effect of modern controlled release polymers such as Aqoat AS-HF, Eudragit SlOO and Kollidon SR was investigated. Chitosan beads and chitosan-polymer beads, as well as chitosan granules and chitosan-polymer granules, were prepared and investigated as possible controlled release formulations. Ketoprofen was chosen as the model drug. Chitosan beads and chitosan-polymer beads were prepared by inotropic gelation in tripolyphosphate. Chitosan granules and chitosan-polymer matrix granules were prepared by binding chitosan with an acetic acid solution as a granulating system. The beads and granules appeared differed in appearance as well as in the results obtained from various experiments. Granules prepared in the study did not appear to be effective with regards to enteric and controlled release. Beads prepared form Kollidon SR appeared to be effective with regards to enteric and controlled release, with Kollidon 1% and 5% w/v chitosan beads achieving good drug loading of up to 73.13% and releasing less than 15 % of the total drug content in 0.1 M HCI after 60 minutes. Drug release continued steadily for up to 360 minutes in pH 7.2. It was concluded that Kollidon SR loaded chitosan beads nay be a viable controlled release dosage form with enteric release properties, and that future experiments, possibly with lower polymer concentrations, are worthwhile / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2006.

Orientador: Rosana Almada Bassani / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-09T08:29:36Z (GMT). No. of bitstreams: 1 Penna_LarissaBarretta_M.pdf: 693702 bytes, checksum: 8dc2d8f7e3689a2bd6879211f15ccf07 (MD5) Previous issue date: 2007 / Resumo: Tem sido descrita supersensibilidade ao efeito cronotrópico do isoproterenol (ISO) em átrios direitos de ratos submetidos a choque nas patas, mas há pouca informação com relação à resposta inotrópica, principalmente ventricular. O objetivo deste trabalho foi estudar os efeitos inotrópico e pró-arrítmico de catecolaminas em miócitos ventriculares esquerdos de ratos nesta condição de estresse, em comparação a animais controles. Ratos Wistar machos adultos foram submetidos a sessões diárias de choque nas patas (30 min de duração) por 3 dias. Foram medidos o encurtamento celular de pico e a freqüência de contrações espontâneas das células. Obteve-se curvas concentração-efeito para noradrenalina (NA), ISO, 3-isobutil-1-metilxantina (IBMX) e CaCl2, com a determinação dos parâmetros Rmax (resposta máxima) e pD2 (-log da concentração do agonista que produz uma resposta= 0,5 Rmax). A população de receptores adrenérgicos (RA) _ foi analisada pelo método de Schild. Observamos, em células de animais do grupo choque, redução da Rmax inotrópica à NA e ao ISO. Estas diferenças permaneceram durante bloqueio de RA_1, mas foram abolidas pelo bloqueio de RA_1 (aumento da Rmax ao ISO apenas no grupo choque) ou _2 (redução da Rmax no apenas no grupo controle). O valor pA2 do metoprolol e a resposta inotrópica a IBMX e CaCl2 foram equivalentes entre os grupos. As células de ratos do grupo choque se mostraram mais reativas à ação pró-arrítmica da NA e do ISO, mas não aos demais agonistas. Todos os procedimentos que aboliram a redução de Rmax inotrópica a catecolaminas em miócitos de animais estressados também atenuaram as diferenças na resposta arrítmica a estes agonistas, o que pode indicar que a resposta inotrópica anormal no grupo choque seja conseqüência da maior atividade espontânea induzida pelas catecolaminas. Em conclusão, os resultados indicam que o estresse por choque nas patas causa aumento da reatividade de miócitos ventriculares de rato ao efeito pró-arrítmico de catecolaminas e reduz a eficácia aparente destes agonistas em estimular o inotropismo / Abstract: Supersensitivity to isoproterenol (ISO) chronotropic effect has been described in right atria from footshock-stressed rats, but information on inotropic responsiveness, particularly of the ventricles, is scarce. The goal of this study was to analyze the inotropic and pro-arrhythmic effect of catecholamines on left ventricular myocytes from footshockstressed rats, in comparison with cells from control animals. Adult male Wistar rats were submitted to 30-min long daily footshock sessions for 3 days. Peak cell shortening and the rate of spontaneous contractions during rest were measured in myocytes at 23 oC. Concentration-effect curves were obtained for noradrenaline (NA), ISO, 3-isobutyl-1- methylxanthine (IBMX) e CaCl2, for estimation of the following parameters Rmax (maximum response) and pD2 (-log of the agonist molar concentration that evokes halfmaximum response). The _ adrenoceptor (AR) population was analyzed by the Schild¿s method. In the footshock group, Rmax values to NA and ISO were decreased. This difference was unaffected by _1AR blockade, but was abolished by blockade of _1AR (which increased Rmax to ISO in the footshock group only) or _2-AR (which decreased Rmax to ISO solely in the control group). Metoprolol pA2 value and the inotropic responsiveness to IBMX and CaCl2 were similar in both groups. Cells from footshockstressed rats were more responsive to NA and ISO pro-arrhythmic action. All procedures that abolished the difference in inotropic response between footshock and control groups also diminished the differences in arrhythmic responses. These results indicate that the inotropic hyporesponsiveness to catecholamines in the footshock group may be the result of increased spontaneous activity induced by these agonists. We conclude that footshock stress enhances catecholamine pro-arrhythmic effect in rat ventricular myocytes, while simultaneously decreases the inotropic apparent efficacy of these agonists / Mestrado / Fisiologia / Mestre em Biologia Funcional e Molecular

Secretin and VIP stimulate cardiac adenylyl cyclase activity and exert a positive inotropic action in several mammalian species. This study examined positive inotropic activity and cAMP levels in rat papillary muscle. Isoproterenol and secretin increased contractions by 150 ± 31% and 129 ± 27%, respectively. VIP increased contraction by 30 ± 21% only at 10 μM. Isoproterenol significantly increased cAMP levels by 82%, whereas increases by secretin (58%) and VIP (56%) were not significant. These results are consistent with reports that secretin and VIP stimulate cardiac adenylyl cyclase in the rat, but suggest that cAMP tissue levels cannot totally explain the positive inotropic responses to secretin and VIP.

The parmacological effects of guinea pig vasoactive intestinal peptide (VIP) were studied in isolated perfused guinea pig hearts. Bolus injections of VIP produced a dose-dependent tachycardia that was not affected by atenolol. A decrease in amplitude of ventricular contractions occurred in response to all doses of VIP. This response was preceded by a small increase in amplitude in 3 of 6 hearts at the highest dose. VIP produced a decrease in perfusion pressure which was prominent after coronary tone was elevated with [Arg8]-vasopressin. The present findings support speculation that VIP may have a role in the regulation of heart rate and coronary blood flow.

Kardialen Strukturen, wie dem koronaren oder endokardialen Endothel, dem Myokard und auch dem Perikard werden unter physiologischen und pathophysiologischen Bedingungen zunehmend autokrine oder parakrine Funktionen zugesprochen. Es ist gut belegt, dass das Herz durch Freisetzung von löslichen Mediatoren nach Myokardischämie einen entscheidenden Anteil an der postischämischen Regulation der Vasomotion hat. Allerdings weniger bekannt ist die Bedeutung einer Mediator-vermittelten kardialen Autoregulation bei postischämischen Veränderungen der Myokardkontraktilität. In dieser Arbeit wird eine neue negativ inotrope Substanz(en) (NIS) beschrieben, die nach myokardialer Ischämie aus isolierten Herzen freigesetzt wird und die an sequentiell perfundierten Herzen, die als Bioassay eingesetzt werden, einen deutlichen kardiodepressiven Effekt hervorruft. In isolierten Feld-stimulierten Rattenkardiomyozyten reduziert NIS dosisabhängig die systolische Zellverkürzung und den Ca2+-Transienten (Konfokale Laser Scan Mikroskopie). Der negativ inotrope Effekt setzt sowohl in isolierten Herzen als auch Kardiomyozyten schnell ein und ist reversibel. Katecholamine maskieren und überspielen den negativ inotropen Effekt in Abhängigkeit von der Ischämiedauer. Voltage clamp Untersuchungen auf Einzelzellebene zeigten, dass NIS den Ca2+-Einstrom Ica über die L-Typ Ca2+-Kanäle reduziert. Somit scheint NIS die Myokardkontraktilität und Zellverkürzung über eine Verminderung der intrazellulären systolischen Ca2+-Konzentrationen durch Blockade der L-Typ Ca2+-Kanäle zu reduzieren und nicht etwa über eine Ca2+-Desensitivierung. Derzeit ist noch nicht geklärt, über welchen Mechanismus NIS den Ca2+-Einstrom blockiert. Da weder die Gewebsspiegel von cGMP und cAMP noch die PKA Aktivität durch NIS moduliert werden, ist es unwahrscheinlich, dass eine Dephosphorylierung von Untereinheiten des L-Typ Ca2+-Kanals der Funktionsweise von NIS zu Grunde liegt. Die Ergebnisse legen nahe, dass NIS direkt mit dem Ca2+-Kanal interagiert, z.B. durch Bindung an ein Kanalprotein. Die chemische Struktur von NIS ist derzeit noch ungeklärt, allerdings gibt es Hinweise, dass es sich nicht um ein Protein handelt. Die Substanz(en) ist stabil, Hitze-resistent (56°) und ein dialysierbares Molekül mit einem geringen Molekulargewicht (< 0.5 kDa). Die kardiodepressorische Substanz(en) wird nicht vom Koronarendothel freigesetzt. Eine abschließende Bewertung, ob NIS durch Aggravation der kontraktilen Dysfunktion myokardschädigend oder durch Senkung des myokardialen Sauerstoffverbrauches kardioprotektiv wirkt, ist derzeit noch nicht möglich. / Autocrine and paracrine functions are increasingly being attributed under physiological and pathophysiological conditions to cardiac structures such as the coronary or endocardial endothelium, the myocardium, and the pericardium as well. Reliable evidence exists to confirm that the heart, through the release of soluble mediators after myocardial ischemia, plays a decisive role in post-ischemic regulation of vasomotion. Less well-known, however, is the significance of mediator-effected cardiac autoregulation in cases of post-ischemic changes of myocardial contractility. This study describes a new negative inotropic substance or substances, NIS, released from isolated hearts after myocardial ischemia. NIS elicits marked cardiodepressive effects on sequentially perfused hearts used as bioassays. In isolated field-stimulated rat cardiomyocytes, NIS reduces, as a function of dose, systolic cell shortening and Ca2+ transients (as detected by confocal laser-scan microscopy). The negative inotropic effect occurs quickly both in isolated hearts as well as in cardiomyocytes, and is reversible. Catecholamines counteract the negative inotropic effect, as a function of ischemia duration. Voltage-clamp investigations on the single-cell level have disclosed that NIS reduces Ca2+ inflow Ica via L-type Ca2+ channels. NIS appears to decrease myocardial contractility and cell shortening through reduction of intracellular systolic Ca2+ concentration, by blockade of L-type Ca2+ channels: and not, say, by Ca2+ desensitization. It has not yet been definitely established by which mechanism NIS blocks Ca2+ inflow. Since NIS modulates neither the tissue level of cGMP and cAMP, nor PKA activity, it is improbable that NIS acts by dephosphorization of sub-units of the L-type Ca2+ channel. The results of this study imply that NIS directly interacts with the Ca2+ channel, perhaps by binding to a channel protein. Although the chemical structure of NIS has not yet been elucidated, there are indications that it is not a protein. The substance(s) is/are stable, heat resistant (up to 56°C), and dialyzable as a molecule with low molecular weight (< 0.5 kDa). It is not yet possible to provide conclusive evaluation of whether NIS acts to damage the myocardium by aggravation of the contractile dysfunction, or whether it exerts cardioprotective action by diminishing myocardial oxygen consumption.

No description available.