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Development of a novel co-culture based in vitro model system to study the wound healing processAbraham, Suraj 07 September 2010
Drug development research on wound repair is challenging and inefficient due to the complex nature of wound healing and scarring processes and the limitations of available in vitro or in vivo models used for preclinical drug testing. Many patients who undergo elective back surgery develop post-surgical complications resulting from excess peridural scarring in and around the site of operation. We tested the effects of two anti-inflammatory compounds, quercetin and L-2-oxothiazolidine-4-carboxylate (OTC), in ameliorating peridural scar formation following spinal laminectomy surgery in laboratory rats. Western blot and immunocytochemical analyses indicated that the peridural scar tissue contained MyoD-positive myoblast cells and expressed prolyl-4-hydroxylase (P4H), a fibroblast marker. Treatment with 1 mM OTC reduced activation of ERK1/2 and p38 mitogen-activated protein kinases (MAPK) at 21 days post-surgery suggesting potential anti-scarring mechanism. However, large animal to animal variation in the expression levels of collagen biosynthesis markers made it difficult to demonstrate any efficacy of quercetin or OTC in reducing peridural scar formation. The shortcomings of this live animal approach led us to develop a novel three-dimensional (3-D) <i>in vitro</i> wound repair model for evaluating quercetin and OTC effects. High-density micromass co-cultures seeded at a 1:3 ratio of FR 3T3 fibroblast cells and L8 myoblast cells formed 3-D microtissues <i>in vitro</i> that expressed MyoD, P4H, and á-smooth muscle actin. The micromass tissue layer remained adherent to the culture plate when inflicted with a single laceration injury, which allowed monitoring of cell migration into the wound site. Wounded cultures were treated with quercetin, OTC and other agents (TGF- â1, mitomycin, p38 inhibitor SB202190, ERK inhibitor PD184352) to determine their effects on collagen accumulation, wound closure rates, MAPK activation, and gene transcript expression. Both OTC and quercetin treatments reduced collagen biosynthesis in dose-dependent manner. In addition, 1.5 mM OTC accelerated wound closure and significantly reduced p38 MAPK activation without affecting ERK1/2. In contrast, 40 µM quercetin delayed wound closure in micromass co-cultures and reduced ERK1/2 activation. Our in vitro findings suggest that OTC might have potential as an anti-scarring agent. Importantly, our novel micromass co-culture system shows promise as an improved 3-D scaffold-free in vitro model for use in preclinical drug development research.
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Development of a novel co-culture based in vitro model system to study the wound healing processAbraham, Suraj 07 September 2010 (has links)
Drug development research on wound repair is challenging and inefficient due to the complex nature of wound healing and scarring processes and the limitations of available in vitro or in vivo models used for preclinical drug testing. Many patients who undergo elective back surgery develop post-surgical complications resulting from excess peridural scarring in and around the site of operation. We tested the effects of two anti-inflammatory compounds, quercetin and L-2-oxothiazolidine-4-carboxylate (OTC), in ameliorating peridural scar formation following spinal laminectomy surgery in laboratory rats. Western blot and immunocytochemical analyses indicated that the peridural scar tissue contained MyoD-positive myoblast cells and expressed prolyl-4-hydroxylase (P4H), a fibroblast marker. Treatment with 1 mM OTC reduced activation of ERK1/2 and p38 mitogen-activated protein kinases (MAPK) at 21 days post-surgery suggesting potential anti-scarring mechanism. However, large animal to animal variation in the expression levels of collagen biosynthesis markers made it difficult to demonstrate any efficacy of quercetin or OTC in reducing peridural scar formation. The shortcomings of this live animal approach led us to develop a novel three-dimensional (3-D) <i>in vitro</i> wound repair model for evaluating quercetin and OTC effects. High-density micromass co-cultures seeded at a 1:3 ratio of FR 3T3 fibroblast cells and L8 myoblast cells formed 3-D microtissues <i>in vitro</i> that expressed MyoD, P4H, and á-smooth muscle actin. The micromass tissue layer remained adherent to the culture plate when inflicted with a single laceration injury, which allowed monitoring of cell migration into the wound site. Wounded cultures were treated with quercetin, OTC and other agents (TGF- â1, mitomycin, p38 inhibitor SB202190, ERK inhibitor PD184352) to determine their effects on collagen accumulation, wound closure rates, MAPK activation, and gene transcript expression. Both OTC and quercetin treatments reduced collagen biosynthesis in dose-dependent manner. In addition, 1.5 mM OTC accelerated wound closure and significantly reduced p38 MAPK activation without affecting ERK1/2. In contrast, 40 µM quercetin delayed wound closure in micromass co-cultures and reduced ERK1/2 activation. Our in vitro findings suggest that OTC might have potential as an anti-scarring agent. Importantly, our novel micromass co-culture system shows promise as an improved 3-D scaffold-free in vitro model for use in preclinical drug development research.
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Epidural blockade and the catabolic response to surgery : an integrated analysis of perioperative protein and glucose metabolism using stable isotope kinetics in the fasted and fed stateLattermann, Ralph January 2002 (has links)
The present project investigated the effect of epidural blockade with local anesthetic on the catabolic stress response during and immediately after abdominal surgery in fasting patients and during infusion of glucose at 2 mg·kg-1·min-1. The kinetics of glucose and protein metabolism were assessed by the stable isotope tracers [6,6-2H2]glucose and L-[1-13C]-leucine. / Epidural blockade was associated with a lower plasma glucose concentration and glucose production when compared to control subjects in the fasted state. Whole body protein breakdown, amino acid oxidation and protein synthesis were suppressed during surgery, and epidural blockade had no modifying effect on perioperative protein metabolism. The suppression of endogenous glucose production by exogenous glucose was more pronounced in the presence of epidural blockade. Perioperative protein metabolism, however, was not influenced by epidural blockade during glucose infusion. / Although epidural blockade suppressed glucose metabolism both in the fasted state and during glucose administration, it failed to exert a modifying effect on perioperative protein metabolism.
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Analgesia after total hip replacement epidural versus psoas compartment block /Bosch, Johannes Coenraad. January 2005 (has links)
Thesis (MMed.(Anaesthesiology-Faculty of Health Sciences))--University of Pretoria, 2005. / Includes bibliographical references.
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Efeitos analgésicos da neostigmina e morfina, isoladas ou associadas, pela via peridural em cães submetidos a cirurgia ortopédica nos membros pélvicos / Analgesic effect of epidural neostigmine and/or morphine after canine orthopedic surgery on a pelvic limbRodrigo Luiz Marucio 10 December 2012 (has links)
Agonistas colinérgicos (neostigmina), administrados por via espinhal, potencializam a analgesia dos opióides por aumentar a concentração de acetilcolina no líquido cérebro-espinhal. O objetivo deste trabalho foi verificar a duração e a eficácia analgésica, assim como a ocorrência de efeitos adversos, da neostigmina e morfina, isoladas ou associadas, administradas por via peridural em cães submetidos à cirurgia ortopédica nos membros pélvicos. Foram utilizados 30 cães de diferentes raças, machos ou fêmeas, de comportamento dócil submetidos à cirurgia ortopédica. Os cães foram prémedicados com meperidina (4 mg/kg IM); após 30 minutos, indução anestésica com propofol (5 mg/kg) e manutenção da anestesia com isofluorano. Após estabilização da anestesia, um cateter peridural era introduzido e a anestesia peridural foi realizada com lidocaína 2% (5 mg/kg). No final da cirurgia, os animais eram distribuídos aleatoriamente em 3 grupos de 10 e recebiam tratamento analgésico pelo cateter peridural como segue: grupo MOR, 0,1 mg/kg de morfina; grupo NEO, 5 µg/kg de neostigmina; e grupo MOR+NEO, associação de 0,1 mg/kg de morfina e 5 µg/kg de neostigmina. Soluções ajustadas com solução NaCl 0,9% até um volume total de 0,4 ml/kg, sendo o estudo caracterizado como prospectivo, clínico, tipo cego. Variáveis paramétricas mensuradas: frequência cardíaca (FC), frequência respiratória (f), temperatura retal (T°C) e as pressões arteriais sistólica, média e diastólica (PAS, PAM e PAD). Analgesia pós-operatória verificada por meio da escala analógica visual (EAV) e escala numérica descritiva (END). Tempos de avaliação: 1, 2, 3, 4, 6, 8, 12, 16 e 24 horas após o final da cirurgia. Caso o animal recebesse nota maior ou igual a quatro para EAV ou END, era realizado resgate analgésico com morfina 0,2 mg/kg (IV), morfina 0,1 mg/kg (peridural) mais meloxican 0,2 mg/kg (IV). Não houve diferenças significativas entre os grupos quanto aos valores demográficos e às variáveis paramétricas. Realizou-se resgate analgésico em 7 animais do grupo NEO, 4 do grupo MOR e 2 do grupo MOR+NEO. O grupo MOR+NEO apresentou menores valores nos escores de dor (EAV) no tempo 1 h em relação ao grupo NEO e no tempo 4 h em relação ao grupo MOR. Quanto aos efeitos adversos, não houve diferenças entre os grupos. A neostigmina como agente isolado não foi eficaz para o tratamento da dor pósoperatória, e a associação de morfina e neostigmina apresentou benefícios sem aumentar a incidência dos efeitos adversos comumente observados quando comparado ao uso isolado da morfina. / The epidural administration of cholinesterase inhibitor drug (nesotigmine) improves morphine analgesia for increased acetylcholine concentration in the cerebrospinal fluid. The aim of this study was to evaluate the possible analgesic effects of neostigmine and the possible potentiation of morphine analgesia in dogs undergoing orthopaedic pelvic limb surgery. Thirty healthy dogs, males or females, from several breeds were selected. They were sedated with meperidine (4 mg/kg IM). 30 minutes later, anestesia was induced with propofol (5 mg/kg IV) and anesthesia was maintained with isoflurane. An epidural catheter was inserted and local anaesthesia was performed with lidocaine 2% (5 mg/kg). At the end of surgical operation, the animals were randomly distributed into three groups of 10 animals each and received the analgesic treatment via epidural catheter using a factorial design: MOR group received 0.1 mg/kg morphine, while NEO group received 5 µg/kg neostigmine and MOR+NEO group received the combination of 0.1 mg/kg of morphine plus 5 µg/kg of neostigmine. In all cases, drug administration was completed with 0.4 ml of 0,9% NaCl. The study was characterized as a prospective, double-blind, randomized clinical trial. Parametric variables measured were heart rate (HR), respiratory rate (f), rectal temperature (T °C) and noninvasive estimation of systolic, diastolic and mean blood pressure (SBP, DBP, and MAP). Postoperative analgesia was evaluated on a visual analogue scale (VAS) and a descriptive numerical scale (DNS) at 1, 2, 3, 4, 6, 8, 10, 12, 16 and 24 hours after the end of the surgery. When the VAS and DNS were equal or greater than four, postoperative analgesia was supplied with morphine 0.2 mg/kg (IV), morphine 0.1 mg/kg (epidural) plus meloxicam 0.2 mg/kg (IV). There were no statistically significant differences in demographic and parametric variables between the groups. Supplemental analgesia were administered in 7 animals of NEO group, 4 animals of MOR group and 2 animals of MOR+NEO group. Animals of MOR+NEO group showed lower values in pain scores (VAS) than animals of NEO group at time 1 hour and animals of MOR group at time 4 hours. The incidence of side effects was similar between the three treatment groups. In short, neostigmine alone was not effective in treatment postoperative pain in dogs undergoing orthopaedic surgery. The analgesics effects of neostigmine plus morphine showed benefits without increasing the incidence of adverses events commonly observed when compared to the use of morphine alone.
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The Relationship between Epidural Analgesia during Childbirth and Childbirth OutcomesRamstad, Marsha January 2004 (has links)
Epidural analgesia has increased in usage dramatically in the United States as a means
of comfort for labor pain. Prior studies have connected epidural analgesia to an increase in
cesarean birth rate, an increase in use of instrumentation, an increase in length of labor,
episiotomy rate, and maternal fever. Epidural analgesia has produced additional costs to
the patient and society. The purpose of this study is to examine the relationship between
epidural analgesia during childbirth and childbirth outcomes. The data for this study were obtained from a retrospective patient record review of 200
systematically selected labor patients who delivered in 2002 at a midwestern hospital. The
epidural analgesia rate was 72% at this facility in 2002, a significant increase from the
previous 5 years. Using the Chi-square test of independence, 3 relationship was established
between epidural analgesia and four of the variables examined. A statistically significant
relationship was found to exist between epidural analgesia and cesarean birth rate, pitocin
augmentation, and the first and second stages of labor with the total sample. The results of
the study are important for healthcare providers who are relaying influential wellness
information to childbearing women and their partners. The results indicate a need for further education for healthcare providers on alternative methods of pain relief for their patients during childbirth.
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Variabilidad del medio interno inducida por infusión rápida de solución salina fisiológica en las gestantes programadas para cesárea electiva previo a la cirugía en el Hospital San Bartolomé de Lima Perú 2018Rojas Maturano, César Augusto January 2019 (has links)
Evalúa la variabilidad del medio interno de la gestante que va a ser sometida a cesárea electiva luego de infundir solución salina fisiológica, de manera rápida a un volumen de 12 mL/kg de peso corporal, en el Hospital San Bartolomé. Se seleccionaron al azar y por conveniencia a 15 gestantes a término que iban a ser sometidas a operación cesárea y, como parte del procedimiento anestésico, al inicio se le infundió 12 mL/kg de cloruro de sodio al 0,9 %. Se tomaron dos muestras venosas, la primera antes de la anestesia regional y la segunda al inicio de la extracción fetal. En ambos casos las muestras fueron analizadas con una demora máxima de 15 minutos. El equipo usado fue Rapid Lab 1265 marca SIEMENS. El pH, la cloremia y la bicarbonatemia tuvieron una variabilidad significativa (p< 0,05) no así el sodio. Se demostró la variabilidad significativa del medio interno por la rápida administración endovenosa de cloruro de sodio al 0,9 %. / Tesis
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Epidural blockade and the catabolic response to surgery : an integrated analysis of perioperative protein and glucose metabolism using stable isotope kinetics in the fasted and fed stateLattermann, Ralph January 2002 (has links)
No description available.
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Nivel de satisfacción materna con la analgesia epidural para el control del dolor del trabajo de parto. Hospital Nacional Dos de Mayo. marzo-mayo del 2012Luque Pinto, Luis Alex January 2014 (has links)
Publicación a texto completo no autorizada por el autor / El documento digital no refiere asesor / Determina el nivel de satisfacción de las pacientes con la analgesia epidural para el control del dolor del trabajo de parto. Estudio de tipo descriptivo, observacional y transversal, en el Hospital Nacional Dos de Mayo, entre marzo y mayo del 2012. Se estudia a 40 pacientes obstétricas para conocer el nivel de satisfacción de la analgesia epidural para el control del dolor del trabajo de parto. Se estima las frecuencias absolutas y relativas para las variables cualitativas y las medidas de tendencia central y de dispersión para las variables cuantitativas. Entre las características clínicas de las pacientes con analgesia epidural que participan en el presente estudio, tienen como edad promedio 21.5±5.1 años, donde se observa que el 35% son gestantes adolescentes. Asimismo, el 77.5% en su mayoría son primigesta. Además, el 65% de gestantes tienen sobrepeso y el 89.5% tiene anemia leve o moderada. Respecto a las características de la práctica de la analgesia epidural en trabajo en parto el 78.9% de las gestantes con analgesia epidural tienen un buen pujo, siendo el 86.9% mayores a 39 semanas, la mayoría de los RN son de sexo masculino (73.7%) con Apgar normal al minuto de 92.5% y a los 5 minutos normal en todos los pacientes. La cesárea es necesaria en dos casos. Sobre el nivel de insatisfacción respecto al dolor antes de la analgesia epidural es “Severo” para todas las pacientes (100%), pero después de la analgesia sólo el 35% de pacientes menciono haber tenido un dolor “Leve” o “Moderado”, inclusive el 100% recomendaría el procedimiento y además lo volvería a aceptar. En resumen, las gestantes mencionan que el nivel de satisfacción es “Bueno” (97.5%) con la analgesia epidural, solo un caso califica de regular su satisfacción (2.5%). Adicionalmente se obtiene alguna información comparando el nivel de satisfacción según gestación: solo las primigestas califican como regular el nivel de satisfacción (2.5%) y respecto a la calificación del dolor después de la analgesia el 25% menciona como “Leve” y 5% como Moderado. Concluye que la mayoría de las gestantes califica de “Buen” nivel de satisfacción de la analgesia epidural. Además todas las gestantes recomendarían el procedimiento y en un parto posterior, opinan que también lo volverían a utilizar. / Trabajo académico
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Perioperative protein sparing in diabetes mellitus type 2 patients : an integrated analysis of perioperative protein and glucose metabolism using stable isotope kineticsKopp Lugli, Andrea. January 2006 (has links)
The potential effects of nutritional support with amino acids or dextrose and epidural blockade on the catabolic response to surgery were investigated in diabetic patients undergoing colorectal surgery. Protein and glucose metabolism were assessed with a stable isotope infusion technique using the two stable isotopes L-[1-13C]leucine and [6,6-2H2 ]glucose. / 1. The first intervention of a postoperative infusion of amino acids avoided pronounced hyperglycaemia in diabetic patients after colorectal surgery and achieved a positive protein balance compared to dextrose. / 2. The second intervention of a short term infusion of amino acids postoperatively blunted protein breakdown and stimulated protein synthesis. This resulted in a positive protein balance in patients with epidural blockade compared to patient controlled analgesia with intravenous morphine. With regard to glucose metabolism, amino acid supply after surgery decreased glucose clearance and endogenous glucose production independent from type of analgesia.
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