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1.
Acta cir. bras ; 37(1): e370103, 2022. tab
Artículo en Inglés | LILACS, VETINDEX | ID: biblio-1364251

RESUMEN

Introduction: Portal hypertension still represents an important health problem worldwide. In the search for knowledge regarding this syndrome, experimental studies with animal models have proven to be useful to point the direction to be taken in future randomized clinical trials. Purpose: To validate the experimental model of portal hypertension and esophagogastric varices in a medium-sized animal. Methods: This study included five minipigs br1. Midline laparotomy with dissection of the portal vein and production of a calibrated stenosis of this vein was performed. Measurement of pressure in the portal venous and digestive endoscopic were performed before and five weeks after the production of a stenosis. Results: All animals were 8 months old, average weight of 17 ± 2.5 kg. The mean pressure of the portal vein immediately before the partial ligation of the portal vein was 8.9 + 1.6 mm Hg, with 26.6 + 5.4 mm Hg in the second measurement five weeks later (p < 0.05). No gastroesophageal varices or hypertensive portal gastropathy were seen at endoscopy procedures in our sample at any time in the study. Conclusion: Portal vein ligation in minipigs has been validated in the production of portal hypertension, but not in the formation of esophageal varices.


Asunto(s)
Animales , Porcinos Enanos/cirugía , Várices Esofágicas y Gástricas/cirugía , Hipertensión Portal/cirugía
2.
Sci Rep ; 11(1): 6281, 2021 03 18.
Artículo en Inglés | MEDLINE | ID: mdl-33737695

RESUMEN

Pathological changes after third-generation drug-eluting stent implantation remain unclear. We compared the tissue responses of coronary arteries after the implantation of third-generation abluminal biodegradable-polymer everolimus-eluting stent (3rd EES) and second-generation durable-polymer EES (2nd EES) using autopsy specimens and an atherosclerotic porcine model. We compared the histology of stented coronary arteries obtained by autopsy performed 1-10 months after 3rd EES (n (number of cases) = 4, stent-implanted period of 3-7 months) and 2nd EES (n (number of cases) = 9, stent-implanted period of 1-10 months) implantations. The ratio of covered stent struts was higher with 3rd EESs than with 2nd EESs (3rd; 0.824 ± 0.032 vs. 2nd; 0.736 ± 0.022, p = 0.035). Low-density lipoprotein receptor knockout minipigs were stented with 3rd or 2nd EES in the coronary arteries and the stented regions were investigated. The fibrin deposition around the 2nd EES was more prominent. Additionally, higher density of smooth muscle cells was confirmed after the 3rd EES implantation. Pathological examination after the 3rd EES demonstrated a combination of less fibrin deposition and more rapid acquisition of well-developed neointima as compared to the 2nd EES at autopsy and the atherosclerotic porcine model.


Asunto(s)
Enfermedad de la Arteria Coronaria/cirugía , Modelos Animales de Enfermedad , Stents Liberadores de Fármacos/efectos adversos , Stents Liberadores de Fármacos/clasificación , Everolimus/administración & dosificación , Neointima/etiología , Intervención Coronaria Percutánea/efectos adversos , Porcinos Enanos/cirugía , Porcinos/cirugía , Implantes Absorbibles/efectos adversos , Anciano , Anciano de 80 o más Años , Animales , Animales Modificados Genéticamente , Autopsia , Vasos Coronarios/patología , Vasos Coronarios/cirugía , Femenino , Fibrina/metabolismo , Técnicas de Inactivación de Genes , Humanos , Masculino , Persona de Mediana Edad , Neointima/metabolismo , Placa Aterosclerótica/cirugía , Diseño de Prótesis , Receptores de LDL/genética , Resultado del Tratamiento
3.
Lab Anim ; 54(6): 576-587, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32063097

RESUMEN

Swine models had been popular in paediatric oesophageal surgery in the past. Although being largely replaced by rodent models, swine experienced a revival with the establishment of minipig models. However, none of them has ever been investigated for similarity to humans. We conducted a pilot study to determine whether three-week old Pietrain piglets and three-month old Aachen Minipigs are suitable for experimental paediatric oesophageal atresia surgery. We tested the operation's feasibility, performed a necropsy, weighed organs, measured organ length and calculated relative weights and lengths, and measured laboratory parameters. We used multidimensional scaling to assess the similarity of the swine breeds with previously published human data. Pietrain piglets had a higher a priori bodyweight than Aachen Minipigs (Δ = 1.31 kg, 95% confidence interval (CI): 0.37-2.23, p = 0.015), while snout-to-tail length was similar. Pietrain piglets had higher absolute and relative oesophageal lengths (Δ = 5.43 cm, 95% CI: 2.2-8.6; p = 0.0062, q1* = 0.0083 and Δ = 11.4%, 95% CI: 5.1-17.6; p = 0.0025, q3* = 0.0053). Likewise, absolute and relative small intestinal lengths were higher in Pietrains, but all other parameters did not differ, with the exception of minor differences in laboratory parameters. Multidimensional scaling revealed three-week old Pietrain piglets to be similar to two-month old humans based on their thoracoabdominal organ weights. This result indicates three-week old Pietrain piglets are a suitable model of paediatric oesophageal atresia surgery, because clinically many procedures are performed at around eight weeks age. Three-month old Aachen Minipigs were more dissimilar to eight-week old humans than three-week old Pietrain piglets.


Asunto(s)
Modelos Animales de Enfermedad , Atresia Esofágica/cirugía , Análisis de Escalamiento Multidimensional , Sus scrofa/cirugía , Animales , Femenino , Humanos , Lactante , Masculino , Tamaño de los Órganos , Proyectos Piloto , Porcinos , Porcinos Enanos/cirugía
4.
Nat Commun ; 10(1): 2244, 2019 05 21.
Artículo en Inglés | MEDLINE | ID: mdl-31113942

RESUMEN

Before they are used in the clinical setting, the effectiveness of artificially produced human-derived tissue-engineered medical products should be verified in an immunodeficient animal model, such as severe combined immunodeficient mice. However, small animal models are not sufficient to evaluate large-sized products for human use. Thus, an immunodeficient large animal model is necessary in order to properly evaluate the clinical efficacy of human-derived tissue-engineered products, such as artificial grafts. Here we report the development of an immunodeficient pig model, the operational immunodeficient pig (OIDP), by surgically removing the thymus and spleen, and creating a controlled immunosuppressive protocol using a combination of drugs commonly used in the clinical setting. We find that this model allows the long-term accommodation of artificial human vascular grafts. The development of the OIDP is an essential step towards a comprehensive and clinically relevant evaluation of human cell regeneration strategies at the preclinical stage.


Asunto(s)
Órganos Bioartificiales , Prótesis Vascular , Huésped Inmunocomprometido , Modelos Animales , Ingeniería de Tejidos , Animales , Implantación de Prótesis Vascular/métodos , Línea Celular , Fibroblastos , Humanos , Masculino , Impresión Tridimensional , Bazo/inmunología , Bazo/cirugía , Porcinos , Porcinos Enanos/inmunología , Porcinos Enanos/cirugía , Timo/inmunología , Timo/cirugía , Factores de Tiempo
5.
J Vet Sci ; 15(3): 417-22, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24962406

RESUMEN

A purely laparoscopic four-port approach was created for left hepatectomy in pigs. A polyethylene loop was placed on the left two hepatic lobes for traction and lift. Next, penetrating ligation of the lobes using of a double row of silk sutures was performed to control bleeding. A direct hepatic transection was completed using a monopolar hook electrode without meticulous dissection of the left hepatic vein. The raw surface of the liver was coagulated and sealed with fibrin glue. Lobes were retrieved through an enlarged portal. Laparoscopic hepatic lobectomy was completed in all pigs without the use of specialized instruments and with a mean operative time of 179 ± 9 min. No significant perioperative complications were observed. The average weight of each resected lobe was 180 ± 51 g. Complete blood count as well as serum organics and enzyme levels normalized after about 2 weeks. During necropsy, adhesion of the hepatic raw surface to the gastric wall and omentum were observed. No other abnormalities were identified. This minimally invasive left hepatectomy technique in swine could serve as a useful model for investigating liver diseases and regeneration, and offer preclinical information to improve hepatobiliary surgical procedures.


Asunto(s)
Hepatectomía/veterinaria , Porcinos/cirugía , Animales , Femenino , Hepatectomía/métodos , Laparoscopía/métodos , Laparoscopía/veterinaria , Hígado/cirugía , Masculino , Cuidados Posoperatorios/métodos , Cuidados Posoperatorios/veterinaria , Porcinos Enanos/cirugía
6.
Artículo en Inglés | WPRIM (Pacífico Occidental) | ID: wpr-194856

RESUMEN

A purely laparoscopic four-port approach was created for left hepatectomy in pigs. A polyethylene loop was placed on the left two hepatic lobes for traction and lift. Next, penetrating ligation of the lobes using of a double row of silk sutures was performed to control bleeding. A direct hepatic transection was completed using a monopolar hook electrode without meticulous dissection of the left hepatic vein. The raw surface of the liver was coagulated and sealed with fibrin glue. Lobes were retrieved through an enlarged portal. Laparoscopic hepatic lobectomy was completed in all pigs without the use of specialized instruments and with a mean operative time of 179 +/- 9 min. No significant perioperative complications were observed. The average weight of each resected lobe was 180 +/- 51 g. Complete blood count as well as serum organics and enzyme levels normalized after about 2 weeks. During necropsy, adhesion of the hepatic raw surface to the gastric wall and omentum were observed. No other abnormalities were identified. This minimally invasive left hepatectomy technique in swine could serve as a useful model for investigating liver diseases and regeneration, and offer preclinical information to improve hepatobiliary surgical procedures.


Asunto(s)
Animales , Femenino , Masculino , Hepatectomía/métodos , Laparoscopía/métodos , Hígado/cirugía , Cuidados Posoperatorios/métodos , Porcinos/cirugía , Porcinos Enanos/cirugía
7.
J Invest Surg ; 26(2): 63-71, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23281597

RESUMEN

The authors have extensive experience in surgery, anesthesia, and perioperative care in both miniature and domestic swine in the research setting. This manuscript describes the techniques that have been developed over the last four decades while developing porcine surgical models and their use in translational research. Swine are conditioned and socialized during a seven-day period prior to entering into a survival surgical protocol. Housing is designed to be nonstressful and accommodate the needs of the species. Induction of anesthesia is generally performed in the animal's pen. Injectable agents are administered sc in the neck using a butterfly catheter technique. The anesthetic protocol is designed to consider the physiologic effects of the agents and their potential effects on the research being performed. Complete aseptic technique is performed and includes final application of adhesive iodine impregnated drapes over the surgical site. Preemptive analgesia is performed using parenteral analgesics, local anesthetics, and/or epidural analgesics. Animals are monitored intensively intraoperatively and during the postoperative period with frequent recordings of physiologic parameters. Observation during the postoperative period is continuous until the animal fully recovers the righting reflex. Observations of the animals in the postoperative period are performed at least daily until surgical incisions are healed. In protocols which create defects or impede physiologic function daily monitoring may have to be performed in conjunction with specific therapies or tests throughout the duration of the protocol.


Asunto(s)
Porcinos/cirugía , Analgesia/veterinaria , Anestesia/veterinaria , Crianza de Animales Domésticos , Animales , Vivienda para Animales , Dolor Postoperatorio/prevención & control , Dolor Postoperatorio/veterinaria , Atención Perioperativa/veterinaria , Porcinos Enanos/cirugía
8.
Clin Exp Immunol ; 167(3): 556-64, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22288599

RESUMEN

Despite recent tissue-engineering advances, there is no effective way of replacing all the functions of the larynx in those requiring laryngectomy. A recent clinical transplant was a success. Using quantitative immunofluorescence targeted at immunologically relevant molecules, we have studied the early (48 h and 1 week) immunological responses within larynxes transplantated between seven pairs of National Institutes of Health (NIH) minipigs fully homozygous at the major histocompatibility complex (MHC) locus. There were only small changes in expression of some molecules (relative to interindividual variation) and these were clearest in samples from the subglottic region, where the areas of co-expression of CD25(+) CD45RC(-) CD8(-) and of CD163(+) CD172(+) MHC-II(-) increased at 1 week after transplant. In one case, infiltration by recipient T cells was analysed by T cell receptor (TCR) Vß spectratype analysis; this suggested that changes in the T cell repertoire occur in the donor subglottis mucosal tissues from day 0 to day 7, but that the donor and recipient mucosal Vß repertoires remain distinct. The observed lack of strong immunological responses to the trauma of surgery and ischaemia provides encouraging evidence to support clinical trials of laryngeal transplantation, and a basis on which to interpret future studies involving mismatches.


Asunto(s)
Laringe/trasplante , Porcinos Enanos/inmunología , Porcinos Enanos/cirugía , Animales , Antígenos CD/metabolismo , Femenino , Genes Codificadores de la Cadena beta de los Receptores de Linfocito T , Laringe/inmunología , Laringe/patología , Complejo Mayor de Histocompatibilidad , Masculino , Modelos Animales , Porcinos , Porcinos Enanos/genética , Factores de Tiempo , Inmunología del Trasplante , Trasplante Homólogo
9.
Lab Anim ; 46(1): 59-64, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22056891

RESUMEN

Swine are a frequent species of choice for testing new surgical procedures and for transplantation studies. However, information concerning best practice to prepare pigs for surgery and postoperative treatment and monitoring is limited, despite a perception that preoperative socialization is beneficial. Therefore we examined the effect of preoperative visits by project personnel on compliance of 26 National Institutes of Health (NIH) minipigs subject to corneal transplantation. We briefly describe sedation and anaesthesia protocols developed for surgery and multiple postoperative interventions in order to facilitate interpretation of data relating to pig compliance. Preoperative visit variables and measures of preoperative socialization were correlated with postoperative outcome. Principal component analysis (PCA) of postoperative outcome variables identified a factor accounting for 53.5% of the variance that was significantly associated with two factors derived from PCA of preoperative factors (accounting, respectively, for 54.7% and 26.0% of the variance; P = 0.019 for the overall model, P = 0.041 and 0.040 for factors 1 and 2, respectively), such that more time spent with pigs before surgery and higher socialization scores were associated with less postoperative stress and difficulty of eye medication. Moreover, two of the preoperative visit variables, time spent with only one person in the pen and time spent with two or more people in the pen, contributed predominantly to PCA factors 1 and 2, respectively, indicating that they were fulfilling two qualitatively different requirements for socialization. We conclude that NIH minipigs are fully compliant with anaesthetic and postoperative experimental procedures provided they are well-socialized to project personnel before surgery.


Asunto(s)
Bienestar del Animal , Conducta Social , Porcinos Enanos/fisiología , Porcinos Enanos/cirugía , Porcinos/fisiología , Porcinos/cirugía , Crianza de Animales Domésticos , Animales , Análisis Químico de la Sangre , Trasplante de Córnea , Femenino , Ciencia de los Animales de Laboratorio/normas , Masculino , Periodo Posoperatorio , Periodo Preoperatorio , Estrés Fisiológico , Porcinos/genética , Porcinos Enanos/genética
10.
Lab Anim ; 46(1): 77-80, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22072625

RESUMEN

We describe the placement of a left ventricular assist device (LVAD) in a pig with spontaneously occurring atrial septal defect (ASD) (incidental finding) that created a right-left cardiac shunt, with subsequent severe hypoxaemia. Early diagnosis was critical in order to prevent end-organ damage due to hypoxaemia. Adequate monitoring alerted us to the deterioration in oxygenation, haemodynamics and cerebral oxygen metabolism. This forced us to change the level of assistance provided by the pump, and thus dramatically correct this impairment. Necropsy revealed an ostium secundum ASD. In conclusion, if hypoxaemia presents after implementation of an LVAD, the presence of a right-left shunt must be ruled out. The first step must be a judicious reduction in assist device flow to minimize intracardiac shunting. Subsequently, atrial septal closure of the defect should be considered. We report an experimental model of severe hypoxaemia after placement of an LVAD as part of a larger research project.


Asunto(s)
Modelos Animales de Enfermedad , Defectos del Tabique Interatrial/fisiopatología , Corazón Auxiliar/efectos adversos , Hipoxia/etiología , Animales , Defectos del Tabique Interatrial/complicaciones , Defectos del Tabique Interatrial/diagnóstico , Tabiques Cardíacos/patología , Ventrículos Cardíacos/fisiopatología , Hemodinámica , Hipoxia/diagnóstico , Masculino , Porcinos/cirugía , Porcinos Enanos/cirugía
11.
J Vis Exp ; (52)2011 Jun 26.
Artículo en Inglés | MEDLINE | ID: mdl-21730947

RESUMEN

The emergence of the Göttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects the close resemblance of these animals to human anatomy and physiology (1-6).The size of the Göttingen minipig permits the use of surgical equipment and advanced imaging modalities similar to those used in humans (6-8). The aim of this instructional video is to increase the awareness on the value of minipigs in biomedical research, by demonstrating how to perform tracheal intubation, transurethral bladder catheterization, femoral artery and vein catheterization, as well as transcardial perfusion. Endotracheal Intubation should be performed whenever a minipig undergoes general anesthesia, because it maintains a patent airway, permits assisted ventilation and protects the airways from aspirates. Transurethral bladder catheterization can provide useful information about about hydration state as well as renal and cardiovascular function during long surgical procedures. Furthermore, urinary catheterization can prevent contamination of delicate medico-technical equipment and painful bladder extension which may harm the animal and unnecessarily influence the experiment due to increased vagal tone and altered physiological parameters. Arterial and venous catheterization is useful for obtaining repeated blood samples and monitoring various physiological parameters. Catheterization of femoral vessels is preferable to catheterization of the neck vessels for ease of access, when performing experiments involving frame-based stereotaxic neurosurgery and brain imaging. When performing vessel catheterization in survival studies, strict aseptic technique must be employed to avoid infections(6). Transcardial perfusion is the most effective fixation method, and yields preeminent results when preparing minipig organs for histology and histochemistry(2,9). For more information about anesthesia, surgery and experimental techniques in swine in general we refer to Swindle 2007. Supplementary information about premedication and induction of anesthesia, assisted ventilation, analgesia, pre- and postoperative care of Göttingen minipigs are available via the internet at http://www.minipigs.com(10). For extensive information about porcine anatomy we refer to Nickel et al. Vol. 1-5(11).


Asunto(s)
Procedimientos Quirúrgicos Cardíacos/métodos , Cateterismo Periférico/métodos , Arteria Femoral/cirugía , Intubación Intratraqueal/métodos , Porcinos Enanos/cirugía , Vejiga Urinaria/cirugía , Cateterismo Urinario/métodos , Animales , Catéteres de Permanencia , Perfusión/métodos , Porcinos
12.
J Neurosci Methods ; 200(1): 41-6, 2011 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-21723320

RESUMEN

BACKGROUND: Deep brain stimulation (DBS) in experimental animals has promoted new indications and refined existing treatments. Implantation of downscaled clinical DBS leads directly compatible with commercially available implantable pulse generators can however be challenging. Accordingly, we have developed a lead implantation device (LID) and technique for minimally invasive implantation of experimental multicontact DBS leads in large research animals. MATERIALS AND METHODS: The LID carries a small-diameter biocompatible polyimide guide tube which is first used for inserting the stimulating end of the lead and then implanted subcutaneously with the rest of the lead. The functionality of the device was tested by implanting 2 different designs of experimental DBS leads in 12 Göttingen minipigs for up to 12 weeks. The brains were histologically analyzed in order to assess implantation accuracy and local tissue reaction. RESULTS AND CONCLUSIONS: The LID was easy to handle and capable of accurate stereotaxic implantation of downscaled experimental DBS leads in the predetermined target brain structures with minimal surrounding tissue reaction. The device may benefit future large animal DBS research as it allows for precise implantation of DBS leads and may have implications for further refinement of clinical DBS leads.


Asunto(s)
Estimulación Encefálica Profunda/instrumentación , Electrodos Implantados/normas , Procedimientos Quirúrgicos Mínimamente Invasivos/instrumentación , Implantación de Prótesis/instrumentación , Instrumentos Quirúrgicos/normas , Porcinos Enanos/cirugía , Animales , Tamaño Corporal/fisiología , Encéfalo/fisiología , Encéfalo/cirugía , Estimulación Encefálica Profunda/métodos , Modelos Animales de Enfermedad , Femenino , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Procedimientos Neuroquirúrgicos/instrumentación , Procedimientos Neuroquirúrgicos/métodos , Implantación de Prótesis/métodos , Porcinos
13.
Invest Radiol ; 46(8): 495-503, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21427591

RESUMEN

OBJECTIVES: : To investigate the effects of immediate intramyocardial implantation of autologous bone marrow-derived mononuclear cells (BMDMNCs) in minipig myocardium after coronary artery ligation with magnetic resonance imaging (MRI), histopathologic, and immunochemical studies. MATERIALS AND METHODS: : Of the 12 minipigs subjected to permanent left anterior descending coronary artery ligation, 6 were immediately treated with intramyocardial BMDMNCs (3 × 10 cells) implantation in the infarct area, whereas the other 6 were treated with intramyocardial injection of an equal amount of saline served as controls. Cardiovascular MRIs, including cine, first-pass, and late gadolinium enhancement (LGE) sequences, were performed on postoperative days 3 and 90. Postmortem infarct size and the degree of fibrosis on histopathologic examination were compared between 2 groups. The degree of BMDMNC differentiation was assessed with flow cytometry, whereas engraftment of BMDMNCs and vascular density was evaluated with confocal immunofluorescence microscopy and immunohistochemical staining, respectively. RESULTS: : There were no significant differences in cardiac function, first-pass dynamics, and LGE between the BMDMNC-treated group and the control group on day 3. On day 90, the BMDMNC-treated group had significantly higher left ventricular (LV) mass/body-weight ratio, lower end-diastolic or end-systolic LV volume/body-weight ratios, higher ejection fraction, better contractility, greater upslope and peak enhancement of the infarct areas, smaller hypoperfused area on first-pass study, and smaller enhanced area and infarct transmurality on LGE MRI than the control group. Flow cytometry revealed high cellular positivity of mesenchymal stem cell surface markers (CD90 and CD271) of the in vitro expanded cells on day 21 after cell culture. In the infarct and peri-infarct areas of the BMDMNC-treated group, there was limited myogenic-like cell differentiation, some engrafted undifferentiated cells, but prominent CD31-positive endothelial cells. On the other hand, a significantly higher number of α-smooth muscle actin-positive small vessel (≤ 25 µm) was noted in the BMDMNC-treated group compared with that in the controls. CONCLUSIONS: : After myocardial infarction in a swine model, immediate intramyocardial BMDMNCs implantation may promote neovascularization with resultant improvement in LV function, perfusion, and myocardial viability as demonstrated on cardiovascular MRI.


Asunto(s)
Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Vasos Coronarios/cirugía , Imagen por Resonancia Cinemagnética/instrumentación , Infarto del Miocardio/terapia , Miocardio/patología , Porcinos Enanos/cirugía , Adapaleno , Animales , Células de la Médula Ósea , Trasplante de Médula Ósea , Vasos Coronarios/lesiones , Modelos Animales de Enfermedad , Selectina E , Fibrosis/patología , Fibrosis/terapia , Citometría de Flujo , Inmunohistoquímica , Técnicas In Vitro , Ligadura , Miocardio/citología , Naftalenos , Molécula-1 de Adhesión Celular Endotelial de Plaqueta , Volumen Sistólico , Porcinos , Antígenos Thy-1 , Factores de Tiempo , Función Ventricular Izquierda
14.
J Am Assoc Lab Anim Sci ; 50(1): 65-72, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21333166

RESUMEN

Threats of nuclear and other radiologic exposures have been increasing, but no countermeasure for acute radiation syndrome has been approved by regulatory authorities. Because of their similarity to humans in regard to physiology and anatomy, we are characterizing Gottingen minipigs as a model to aid the development of radiation countermeasures. Irradiated minipigs exhibit immunosuppression, severe thrombocytopenia, vascular leakage, and acute inflammation. These complications render serial acquisition of blood samples problematic. Vascular access ports (VAP) facilitate serial sampling, but their use often is complicated by infections and fibrin deposition. We demonstrate here the successful use of VAP for multiple blood samplings in irradiated minipigs. Device design and limited postoperative prophylactic antimicrobial therapy before irradiation were key to obtaining serial sampling, reducing swelling, and eliminating infection and skin necrosis at the implantation site. Modifications of previous protocols included the use of polydioxanone sutures instead of silk; eliminating chronic port access; single-use, sterile, antireflux prefilled syringes for flushing; strict aseptic weekly maintenance of the device, and acclimating animals to reduce stress. VAP remained functional in 19 of 20 irradiated animals for as long as 3 mo. The remaining VAP failed due to a small leak in the catheter, leading to clot formation. VAP-related sepsis occurred in 2 minipigs. Blood sampling did not cause detectable stress in nonanesthetized sham-irradiated animals, according to leukograms and clinical signs.


Asunto(s)
Catéteres de Permanencia/veterinaria , Traumatismos Experimentales por Radiación/cirugía , Porcinos Enanos/cirugía , Bienestar del Animal , Animales , Profilaxis Antibiótica/veterinaria , Catéteres de Permanencia/efectos adversos , Diseño de Equipo/veterinaria , Masculino , Cuidados Posoperatorios/veterinaria , Traumatismos Experimentales por Radiación/sangre , Estrés Fisiológico , Porcinos , Porcinos Enanos/sangre , Porcinos Enanos/microbiología
15.
Cell Reprogram ; 12(6): 689-97, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21108536

RESUMEN

Research into transplantation strategies to treat spinal cord injury (SCI) is frequently performed in rodents, but translation of results to clinical patients can be poor and a large mammalian model of severe SCI is needed. The pig has been considered an optimal model species in which to perform preclinical testing, and the Yucatan minipig can be cloned successfully utilizing somatic cell nuclear transfer (SCNT). However, induction of paralysis in pigs poses significant welfare and nursing challenges. The present study was conducted to determine whether Yucatan SCNT clones could be used to develop an SCI animal model for cellular transplantation research. First, we demonstrated that transection of the sacrocaudal spinal cord in Yucatan SCNT clones produces profound, quantifiable neurological deficits restricted to the tail. We then established that neurospheres could be isolated from brain tissue of green fluorescence protein (GFP) transfected SCNT clones. Finally, we confirmed survival of transplanted GFP-expressing neural stem cells in the SCI lesion and their differentiation into glial and neuronal lineages for up to 4 weeks without immunosuppression. We conclude that this model of sacrocaudal SCI in Yucatan SCNT clones represents a powerful research tool to investigate the effect of cellular transplantation on axonal regeneration and functional recovery.


Asunto(s)
Modelos Animales , Traumatismos de la Médula Espinal/patología , Médula Espinal/cirugía , Trasplante de Células Madre/métodos , Porcinos Enanos/cirugía , Animales , Conducta Animal , Cauda Equina/patología , Cauda Equina/cirugía , Técnicas de Cultivo de Célula , Células Cultivadas , Clonación de Organismos , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Regeneración Nerviosa/fisiología , Técnicas de Transferencia Nuclear , Recuperación de la Función/fisiología , Sacro , Médula Espinal/patología , Porcinos
17.
Xenotransplantation ; 16(1): 11-8, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19243556

RESUMEN

BACKGROUND: Islet cell transplantation is a novel therapeutic modality for the cure of diabetes. Pig islet cells are an attractive substitute for human islet cells; however, they are known to be particularly difficult to isolate because of a weak islet capsule and a tendency to be fragmented during enzymatic digestion. Therefore, parameters favoring successful pig islet isolation were investigated using specific-pathogen-free (SPF) miniature pigs. METHODS: Sixty-eight SPF miniature pigs were used for islet isolation. Birth weight, body weight, age, sex, pregnancy history, and the fasting blood glucose levels of each pig were determined. Each pig's general condition was assessed with regard to feeding status and physical activity. Pancreas procurement was performed by one surgical team. Anesthesia duration, operation duration, procedure quality, and perfusate type were recorded. After pancreatectomy, a biopsy was performed for islet density analysis. Decapsulation, cannulation duration, degree of distension, and cold ischemic time were assessed. During islet isolation, pancreas weight, digestion time, and digested tissue proportion were recorded. Isolation results were evaluated by total islet equivalents (IEQ), islet equivalents per gram of pancreas (IEQ/g), isolation index, islet recovery rate, purity, and visual grade. To identify the predictors of higher islet isolation yield, we performed binary logistic regression analysis with significant (P < 0.05) variables from the univariate analysis. RESULTS: The pigs were categorized into high (n = 34) and low yield (n = 34) groups according to the median IEQ/g or total IEQ values. Body weight and age were significantly different between the two groups. Being male or a positive history of pregnancy in females was factors favoring successful islet isolation. General condition assessments failed to estimate islet isolation results. Long anesthesia duration, which might have caused ischemic injury to the pancreas, negatively affected islet isolation results. Decapsulation, cannulation duration, and subsequent pancreas distension were significantly important in successful islet isolation. Inter-lot variability of Liberase was not observed because of screening processes performed before purchase. Isolation index and islet recovery rate correlated well with islet yields. CONCLUSIONS: Multivariate analysis using total IEQ and IEQ/g as outcome variables indicated that age older than 2, being male and moderate distension by Liberase injection are major determinants influencing successful islet isolation.


Asunto(s)
Trasplante de Islotes Pancreáticos , Islotes Pancreáticos/cirugía , Porcinos Enanos/cirugía , Recolección de Tejidos y Órganos/métodos , Animales , Biopsia , Colagenasas/farmacología , Femenino , Humanos , Islotes Pancreáticos/efectos de los fármacos , Islotes Pancreáticos/patología , Masculino , Análisis Multivariante , Análisis de Regresión , Caracteres Sexuales , Organismos Libres de Patógenos Específicos , Porcinos , Termolisina/farmacología , Trasplante Heterólogo
18.
J Am Assoc Lab Anim Sci ; 45(6): 27-34, 2006 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17089988

RESUMEN

Vascular access ports (VAPs) for studies requiring intermittent or continuous infusion and frequent sampling are well accepted and widely used in large animal species. However, the use of medical devices such as VAPs to facilitate sample collection can lead to complications. Noninfectious complications of VAP implantation can result from thrombotic or mechanical obstructions, other mechanical problems, and animal-associated complications. To facilitate our research, we surgically implanted VAPs in the right external jugular vein of 6 adult male and 3 female Yucatan miniswine (age, 12 mo) to enable collection of blood samples every 30 min for 8 h and then every 8 h for as long as 60 d. The VAPs were operational an average of 35.6 d (range, 29 to 56 d) and had an overall success rate of 77.8% with 7 of 9 VAPs functional. In these 7 animals, 53.1 samples on average (range, 28 to 95 samples) were collected from each VAP. Rates of noninfectious complications were 60% for thrombotic events and 40% for nonthrombotic events over the course of this study.


Asunto(s)
Catéteres de Permanencia/veterinaria , Complicaciones Posoperatorias/veterinaria , Enfermedades de los Porcinos/etiología , Porcinos Enanos/cirugía , Animales , Catéteres de Permanencia/efectos adversos , Infarto Cerebral/etiología , Infarto Cerebral/veterinaria , Femenino , Venas Yugulares/cirugía , Masculino , Porcinos/cirugía
19.
J Craniomaxillofac Surg ; 34(2): 93-9, 2006 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-16423531

RESUMEN

BACKGROUND: Obliteration of the frontal sinus is a generally accepted management for patients with severe craniofacial trauma in whom the sinus cannot be repaired or adequately drained. Effective obliteration requires complete removal of mucosa, sufficient filling of the sinus cavity as well as effective occlusion of the nasofrontal duct. The use of various filling materials for sinus obliteration has been described and include autologous fat, bone, muscle and lyophilized cartilage. MATERIAL AND METHODS: In order to appreciate the advantages of the water-jet scalpel an animal study in 10 juvenile (4 months old) and 10 adult (18 months) minipigs was performed. In 5 juvenile and 5 adult animals the frontal sinus mucosa was removed using the water jet, while in the others bone curettes were used. No occlusion of the frontonasal duct was carried out and no filling material was implanted. All animals were followed-up for 40 days. After sacrifice, radiological, histological and computerized histomorphometrical examinations were carried out. RESULTS: Removal of the mucosa from the sinus was simplified when using the water jet. Previous studies have shown that the selected tissue structures were less traumatized when compared with conventional surgical techniques. In young animals (only) and exclusively following the use of a water-jet scalpel osseous occlusion of the frontal sinus was found at follow-up. Histomorphometric findings revealed a significant increase of bone formation in the frontal sinus area of these animals. CONCLUSION: Based on these experimental results in a small series of juvenile minipigs the water-jet technique is a faster and less traumatic technique (less blood loss) for osteoplastic surgery of the frontal sinus. Further long-term studies are needed to evaluate the possibilities for applying this technique in post-traumatic craniofacial surgery in man.


Asunto(s)
Seno Frontal/cirugía , Mucosa Nasal/cirugía , Osteogénesis , Porcinos Enanos/cirugía , Animales , Estudios de Casos y Controles , Seno Frontal/lesiones , Instrumentos Quirúrgicos , Porcinos/cirugía , Irrigación Terapéutica/métodos , Agua
20.
Lab Anim ; 39(3): 280-3, 2005 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-16004686

RESUMEN

A pilot study was performed to investigate whether the Göttingen minipig is a suitable animal model for creating and closing oroantral communications (OACs) and to test whether these defects can be closed with a biodegradable polyurethane (PU) foam. In three adult minipigs, an OAC was created on both sides of the maxilla. The left side was closed by a standard surgical buccal flap procedure, the right side by applying a PU foam. The pigs were killed after two weeks, one month and three months, respectively. Postmortem and histological examination showed that an OAC was created in only one of six cases. In the remaining cases, the infraorbital canal was perforated instead of the floor of the maxillary sinus. It was concluded that the Göttingen minipig is not a suitable animal model for OAC investigations. As a result, the closure of OACs with a biodegradable PU could not be evaluated.


Asunto(s)
Modelos Animales de Enfermedad , Fístula Oroantral/terapia , Poliuretanos/uso terapéutico , Porcinos Enanos/cirugía , Animales , Estudios de Evaluación como Asunto , Seno Maxilar/patología , Seno Maxilar/cirugía , Fístula Oroantral/cirugía , Porcinos
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