Feasibility of biodegradable PLGA common bile duct stents: An in vitro and in vivo study

The current study investigates the feasibility of using a biodegradable polymeric stent in common bile duct (CBD) repair and reconstruction. Here, poly(l-lactide-co-glycolide) (PLGA, molar ratio LA/GA = 80/20) was processed into a circular tube- and dumbbell-shaped specimens to determine the in vitro degradation behavior in bile. The morphology, weight loss, and molecular weight changes were then investigated in conjunction with evaluations of the mechanical properties of the specimen. Circular tube-shaped PLGA stents with X-ray opacity were subsequently used in common bile duct exploration (CBDE) and primary suturing in canine models. Next, X-ray images of CBD stents in vivo were compared and levels of serum liver enzymes and a histological analysis were conducted after stent transplantation. The results showed that the PLGA stents exhibited the required biomedical properties and spontaneously disappeared from CBDs in 4-5 weeks. The degradation period and function match the requirements in repair and reconstruction of CBDs to support the duct, guide bile drainage, and reduce T-tube-related complications.

Polymeric photosensitizer-embedded self-expanding metal stent for repeatable endoscopic photodynamic therapy of cholangiocarcinoma

Photodynamic therapy (PDT) is a new therapeutic approach for the palliative treatment of malignant bile duct obstruction. In this study, we designed photosensitizer-embedded self-expanding nonvascular metal stent (PDT-stent) which allows repeatable photodynamic treatment of cholangiocarcinoma without systemic injection of photosensitizer. Polymeric photosensitizer (pullulan acetate-conjugated pheophorbide A; PPA) was incorporated in self-expanding nonvascular metal stent. Residence of PPA in the stent was estimated in buffer solution and subcutaneous implantation on mouse. Photodynamic activity of PDT-stent was evaluated through laserexposure on stent-layered tumor cell lines, HCT-116 tumor-xenograft mouse models and endoscopic intervention of PDT-stent on bile duct of mini pigs. Photo-fluorescence imaging of the PDT-stent demonstrated homogeneous embedding of polymeric Pheo-A (PPA) on stent membrane. PDT-stent sustained its photodynamic activities at least for 2 month. And which implies repeatable endoscopic PDT is possible after stent emplacement. The PDT-stent after light exposure successfully generated cytotoxic singlet oxygen in the surrounding tissues, inducing apoptotic degradation of tumor cells and regression of xenograft tumors on mouse models. Endoscopic biliary in-stent photodynamic treatments on minipigs also suggested the potential efficacy of PDT-stent on cholangiocarcinoma. In vivo and in vitro studies revealed our PDT-stent, allows repeatable endoscopic biliary PDT, has the potential for the combination therapy (stent plus PDT) of cholangiocarcinoma. © 2014 Elsevier Ltd.

A comparative study of antibody expressions in primary biliary cirrhosis and autoimmune cholangitis using phage display

Primary biliary cirrhosis (PBC) and autoimmune cholangitis (AIC) are serologic expressions of an autoimmune liver disease affecting biliary ductular cells. Previously we screened a phage-displayed random peptide library with polyclonal IgG from 2 Australian patients with PBC and derived peptides that identified a single conformational (discontinuous) epitope in the inner lipoyl domain of the E2 subunit of the pyruvate dehydrogenase complex (PDC-E2), the characteristic autoantigen in PBC. Here we have used phage display to investigate the reactivity of PBC sera from 2 ethnically and geographically distinct populations, Japanese and Australian, and the 2 serologic expressions, PBC and AIC. Random 7-mer and 12-mer peptide libraries were biopanned with IgG from 3 Japanese patients with PBC and 3 with AIC who did not have anti-PDC-E2. The phage clones (phagotopes) obtained were tested by capture enzyme-linked immunosorbent assay (ELISA) for reactivity with affinity-purified anti-PDC-E2, and compared with those obtained from Australian patients with PBC. Peptide sequences of the derived phagotopes and sequences derived by biopanning with irrelevant antisera were aligned to develop a guide tree based on physicochemical similarity. Both Australian and Japanese PBC-derived phagotopes were distributed in branches of the guide tree that contained the peptide sequences MH and FV previously identified as part of an immunodominant conformational epitope of PDC-E2, indicating that epitope selection was not influenced by the racial origin of the PBC sera. Biopanning with either PBC or AIC-derived IgG yielded phagotopes that reacted with anti-PDC-E2 by capture ELISA, further establishing that there is a similar autoimmune targeting in PBC and AIC.

Effect of Dietary Unsaturated Oils on the Biosynthesis of Cholesterol, and on Biliary and Fecal Excretion of Cholesterol and Bile Acids in Rats1

The aim of the present work was to investigate whether the hypocholesterolemic effect of polyunsaturated oils is due to inhibition of cholesterol synthesis or increased excretion of cholesterol and bile acids through the bile and feces of animals. Separate groups of rats were fed diets containing 10% safflower oil, coconut oil or hydrogenated vegetable oils for 30 days, after which the hepatic cholesterol and bile acid synthesis and their excretion through the bile and feces were studied. As compared to the rats in the other two groups, those given the diet containing 10% safflower oil showed markedly increased rates of bile flow and excreted through their bile and feces markedly higher amounts of cholesterol and bile acids. At the same time incorporation of [1-14C] acetate and [2-14C] mevalonate into the liver cholesterol and conversion of [4-14C] cholesterol into 14C-bile acids were also higher in the same rats. In the light of these observations it has been discussed that in the animals given polyunsaturated oils, biliary and fecal loss of cholesterol and bile acids far outweighs the activation of cholesterol synthesis and thereby effectively lowers the serum cholesterol levels.

Epidemiology and Treatment Options of Primary Biliary Cirrhosis

Primary biliary cirrhosis (PBC) is caused by an autoimmune inflammation of the small bile ducts. It results to destruction of bile ducts, accumulation of the bile in the liver, and cirrhosis. The prevalence and incidence of PBC is increasing in the Western world. The prevalence is highest in the USA (402 per million) and incidence in Scotland (49/million/year). Our aim was to assess the epidemiology of PBC in Finland. Patients for the epidemiological study were searched from the hospital discharge records from year 1988 to 1999.The prevalence rose from 103 to 180/million from 1988 to 1999, an annual increase of 5.1%. The incidence rose from 12 to 17 /million/year, an annual increase of 3.5%. The age at death increased markedly from 65 to 76 years. The risk of liver related deaths diminished over time. The treatment of PBC is based on Ursodeoxycholic acid (UDCA). During 20 years 50% of patients end up with cirrhosis. Our treatment option was to combine budesonide, a potent corticosteroid with a high first pass metabolism in the liver, to UDCA and evaluate the liver effects and systemic effects such as bone mass density (BMD) changes. Our aim was to find out if combination of laboratory tests would serve as a surrogate marker for PBC and help reducing the need for liver biopsy. Non-cirrhotic PBC patients were randomized to receive budesonide 6 mg/day combined to UDCA 15 mg /kg/day or UDCA alone for three years. The combination therapy with UDCA and budesonide was effective: stage improved 22%, fibrosis 25%, and inflammation 32%. In the UDCA group the changes were: 20% deterioriation in stage and 70% in fibrosis, but a 10% improvement in inflammation. BMD in femoral neck decreased by 3.6% in the combination group and by 1.9% in the UDCA group. The reductions in lumbar spine were 2.8% and 0.7%. Pharmacokinetics did not differ between the stages of PBC. HA, PIIINP, bile acids, and AST were significantly different within stages I-III and could differentiate the mild fibrosis (F0F1) from the moderate (F2F3). The combination of these individual markers (PBC-score) further improved the accuracy. The area under the ROC of the PBC score, using a cut of value 66, had a sensitivity of 81.4% and a specificity of 65.2% to classify the stage of PBC. The prevalence of PBC in Finland increases, which results from increasing incidence and improved survival. The combination of budesonide and UDCA improves liver histology compared to UDCA alone in non-cirrhotic stages of PBC. The treatment may reduce BMD. Hyaluronic acid, PIIINP, AST, and bile acids may serve as tools to monitor the treatment response in the early stages of PBC. The budesonide and UDCA combination therapy is an option for those patients who do not receive full response from UDCA and are still at the non-cirrhotic stage of PBC.

Valor nutritivo del pasto pangola (Digitaria decumbens stent.) vr. transvala a distintos niveles de fertilización y diversas edades de corte

Este estudio Se desarrolló en el periodo de julio a diciembre de 1990 en el Centro de Investigación Zootecnia “La Polvosa” de la UCA (Universidad Centroamericana), situada en el km 23 112 de la carretera nueva a León, depto. de Managua. El centro se ubica a una elevación de 40 msnrn a 1 2 grados 12 minutos, Latitud norte y 86 grados 22 minutos, longitud oeste registra temperaturas promedio anuales de 32 grados centígrados los y unos 8OO mm de precipitación media por año, por lo cual, es posible calificar su zona agroecológica como de trópico seco se evaluó la respuesta en términos de valor nutritivo del pasto Pangola (Digitaria decumbens Stent.) vr. Transvala, utilizándose un diseño experimental en bloques completos al azar (BCA) con arreglo factorial, se estudió el efecto de dos factores (Niveles de fertilizantes-edades de cortes) con cuatro repeticiones, formándose un total de 16 tratamientos El ensayo contó con un área experimental total de 357.75 mt2 para la toma de muestras, se empleó el método del metro cuadrado realizándose en las parcelas un muestreo sin reemplazo para cada frecuencia en estudio. Previo al momento experimental, se efectúo una poda de control procediéndose después a aplicar de una sola vez los respectivos niveles de fertizante. El estudio estadístico contempló el uso del análisis de varianza (ANDEVA), se hizo una separción de medias por Tukey, y se midió la influencia porcentual (Individual y asociada) de 4 niveles de fertilizante (0, 50,100 y150 kg urea (46%N2) / mz) y 4 edades de corte (15, 30.. 45y 60 días), sobre nueve variables o componentes bromatológicos (MS, PB,FC, ELN, EE, EB (Kcal/ 100g), Cz, Ca y P). Todos los componentes bromatológicos evaluados excepto el EE.. presentaron una variación significativa (P<0.05), bajo el efecto de los factores en estudio. Los niveles de fertilizante influyeron ios valores de mayor mérito con la dosis •150 kg urea / rnz, observando la PB.. 9.76% ;el ELN, 56.54 y la EB,190.97 Kcal ,' 100 g. En tanto que los 50 kg1mz mostraron !os mayores índices en FC (28.88), EE (2.75) y Cz. (11.:39) las parcelas no fertilizadas brindaron los porcentajes más altos de MS (23.78) y Ca (0.60),y el nivel 100 kg/ mz El mejor en P (0.39) y el más bajo en FC (26.61). A su vez con O kg / mz_. Observaron sus níveles minimos la PB (6.91), el ELN (53.58). el valor energético (175.45 Kcal/ 100 g) y el P (O.28), y con 100 kg/ mz los menores en MS y Cz (25.08 y 10.38 respectivamente), a. los 150kg/mz el EE (2.29) y el Ca (0.49), rindieron sus índices más bajos. En cuanto a la frecuencia de corte, la edad 15 días presentó los máximos contenidos en PH (11.95), ELN (56.45),EB (202.40 Kcal/ 100g), EE (2.65) y el mínimo en FC (27.09). Observando el corte a los 60 días, las mayores proporciones en MS (30.91) Cz (12.43) )'Ca (0.58),en tanto que con 30 y 45 días se dieron los más altos valores de P (0.34) FC (28.55) respectivamente los índices más bajos de PB (6.48),ELN (52.63), Ca (0.47)y P (0.30), se obtuvieron a los 45 días de madurez mientras que la MS (22.88) y la Cz (9.56),alcanzaron sus valores más pequeños con 15 días de corte a los 60 días, la EB (175. 42 Kcal ./ 100g) y el EE (2.20) respondieron con sus por cientos más bajos. Para los tratamientos resultantes de la combinación de un nivel de fertilizante y una edad de corte, la interacción 50 kg/mz-60 días reportó el mayor contenido en MS (32.72), sobresaliendo el tratamiento 150 kg / mz-15 días al presentar los valores más altos en EB (214.39 Kcal/100g), PB (14.30) y ELN (59.02), aunque la combinación 50 kg/mz-30 días brindó igual valor en ELN y el máximo en Cz (13.10) con aplicaciones de 100 kg/mz y frecuencia de 15 días se obtuvieron los menores índices en FC (24.82) y Cz (9.37). Consiguiéndose en O kg / mz-15 días el valor más alto en Ca (0.76) y el menor en P (0.22), observando la MS su índice más bajo (18.37) con el tratamiento 100 kg / mz-15 días. Finalmente, en las interacciones que incluyen la frecuencia 45 días, se encontró que las parcelas no fertilizadas presentaron los menores contenidos en PB (5.02), ELN (48.95) y EB (165.44 Kcal/ 1oo g). en cambio al aplicar 100 kg /rnz, se obtuvieron (para la misma interacción) los valores más bajos en EE (1.72), Ca (0.43) y el mayor en P (0.44),en tanto los niveles 50 y 150 kg / mz influyeron los índices más altos en EE (3.85) y FC (30.75) respectivamente.

Novel nanostructured biomaterials: implications for coronary stent thrombosis.

BACKGROUND: Nanomedicine has the potential to revolutionize medicine and help clinicians to treat cardiovascular disease through the improvement of stents. Advanced nanomaterials and tools for monitoring cell-material interactions will aid in inhibiting stent thrombosis. Although titanium boron nitride (TiBN), titanium diboride, and carbon nanotube (CNT) thin films are emerging materials in the biomaterial field, the effect of their surface properties on platelet adhesion is relatively unexplored. OBJECTIVE AND METHODS: In this study, novel nanomaterials made of amorphous carbon, CNTs, titanium diboride, and TiBN were grown by vacuum deposition techniques to assess their role as potential stent coatings. Platelet response towards the nanostructured surfaces of the samples was analyzed in line with their physicochemical properties. As the stent skeleton is formed mainly of stainless steel, this material was used as reference material. Platelet adhesion studies were carried out by atomic force microscopy and scanning electron microscopy observations. A cell viability study was performed to assess the cytocompatibility of all thin film groups for 24 hours with a standard immortalized cell line. RESULTS: The nanotopographic features of material surface, stoichiometry, and wetting properties were found to be significant factors in dictating platelet behavior and cell viability. The TiBN films with higher nitrogen contents were less thrombogenic compared with the biased carbon films and control. The carbon hybridization in carbon films and hydrophilicity, which were strongly dependent on the deposition process and its parameters, affected the thrombogenicity potential. The hydrophobic CNT materials with high nanoroughness exhibited less hemocompatibility in comparison with the other classes of materials. All the thin film groups exhibited good cytocompatibility, with the surface roughness and surface free energy influencing the viability of cells.