RESUMEN
The limited supply of cartilage tissue with appropriate sizes and shapes needed for reconstruction and repair has stimulated research in the area of hydrogels as scaffolds for cartilage tissue engineering. In this study we demonstrate that poly(ethylene glycol) (PEG)-based semi-interpenetrating (sIPN) network hydrogels, made with a crosslinkable poly(ethylene glycol)-dimethacrylate (PEGDM) component and a non-crosslinkable interpenetration poly(ethylene oxide) (PEO) component, and seeded with chondrocytes support cartilage construct growth having nominal thicknesses of 6 mm and relatively uniform safranin-O stained matrix when cultured statically, unlike constructs grown with prefabricated macroporous scaffolds. Even though changing the molecular weight of the PEO from 100 to 20 kDa reduces the viscosity of the precursor polymer solution, we have demonstrated that it does not appear to affect the histological or biochemical characteristics of cartilaginous constructs. Extracellular matrix (ECM) accumulation and the spatial uniformity of the ECM deposited by the embedded chondrocytes decreased, and hydrogel compressive properties increased, as the ratio of the PEGDM:PEO in the hydrogel formulation increased (from 30:70 to 100:0 PEGDM:PEO). Total collagen and glycosaminoglycan contents per dry weight were highest using the 30:70 PEGDM:PEO formulation (24.4+/-3.5% and 7.1+/-0.9%, respectively). The highest equilibrium compressive modulus was obtained using the 100:0 PEGDM:PEO formulation (0.32+/-0.07 MPa), which is similar to the compressive modulus of native articular cartilage. These results suggest that the versatility of PEG-based sIPN hydrogels makes them an attractive scaffold for tissue engineering of cartilage.
RESUMEN
PURPOSE: To examine students' performances on Step 1 and Step 2 of the United States Medical Licensing Examination (USMLE) following the implementation of a problem-based learning curriculum. METHOD: Performances on Step 1 of the USMLE for four classes at the University of Missouri-Columbia School of Medicine that completed a new problem-based learning curriculum (1997, 1998, 1999, and 2000) were compared with those of the last two classes to learn in the traditional curriculum (1995 and 1996). Performances on Step 2 of the USMLE for the classes of 1997, 1998, and 1999 were also compared with those of the classes of 1995 and 1996. The authors analyzed matriculation data (GPAs and MCAT scores) for all six classes. They compared all data with those of U.S. and Canadian first-time USMLE takers. RESULTS: The mean scores were higher on USMLE Step 1 for classes in the problem-based learning curriculum than for classes in the traditional curriculum. The mean scores for Step 2 were above the national mean for classes in the revised curriculum and below the national mean for classes in the traditional curriculum. The admission profiles of these classes were essentially the same before and after the change in curriculum. CONCLUSIONS: Major PBL revisions of the curriculum did not compromise the performances of medical students on the licensing examinations; in fact, they may have contributed to higher scores.
Asunto(s)
Curriculum , Educación Médica , Evaluación Educacional , Licencia Médica , Aprendizaje Basado en Problemas , Estudiantes de Medicina , Canadá , Distribución de Chi-Cuadrado , Humanos , Missouri , Facultades de Medicina , Estados UnidosRESUMEN
The effect of polymer chemistry on adhesion, proliferation, and morphology of human articular cartilage (HAC) chondrocytes was evaluated on synthetic degradable polymer films and tissue culture polystyrene (TCPS) as a control. Two-dimensional surfaces of poly(glycolide) (PGA), poly(L-lactide) (L-PLA), poly(D,L-lactide) (D,L-PLA), 85:15 poly(D,L-lactide-co-glycolide) (D,L-PLGA), poly(epsilon-caprolactone) (PCL), 90:10 (D,L-lactide-co-caprolactone) (D,L-PLCL), 9:91 D,L-PLCL, 40:60 L-PLCL, 67:33 poly(glycolide-co-trimethylene carbonate) (PGTMC), and poly(dioxanone) (PDO) were made by spin-casting into uniform thin films. Adhesion kinetics were studied using TCPS and PCL films and revealed that the rate of chondrocyte adhesion began to level off after 6 h. Degree of HAC chondrocyte adhesion was studied on all the substrates after 8 h, and ranged from 47 to 145% of the attachment found on TCPS. The greatest number of chondrocytes attached to PGA and 67:33 PGTMC polymer films, and attachment to PCL and L-PLA films was statistically lower than that found on PGA (p < 0.05). There was no correlation between amount of chondrocyte attachment to the substrates and the substrates' water contact angle. Chondrocytes proliferated equally well on all the substrates resulting in equivalent cell numbers on all the substrates at both day 4 and day 7 of the culture. However, these total cell numbers were reached as a result of a 88- and 42-fold expansion on PDO and PLA, respectively, which was significantly higher than the 11-fold expansion found on TCPS (p < 0.05). The greater fold expansion of the cells on PDO and L-PLA films may be attributed to the availability of space for cells to grow, since their numbers at the start of culture were fewer following the 8 h attachment period. This suggests that regardless of initial seeding density on these degradable polymer substrates (i.e., if some minimum number of cells are able to attach), they will eventually populate the surfaces of all these polymers given sufficient space and time.
Asunto(s)
Cartílago Articular/citología , Adhesión Celular , Condrocitos/citología , Polímeros/metabolismo , División Celular , Células Cultivadas , Dioxanos/metabolismo , Humanos , Cinética , Proteínas de la Membrana/metabolismo , Microscopía Electrónica de Rastreo , Poliésteres/metabolismo , Polímeros/química , Factores de TiempoRESUMEN
Neonatal rat calvarial osteoblasts were cultured in 90% porous, 75:25 poly(DL-lactic-co-glycolic acid) (PLGA) foam scaffolds for up to 56 days to examine the effects of the cell seeding density, scaffold pore size, and foam thickness on the proliferation and function of the cells in this three-dimensional environment. Osteoblasts were seeded at either 11.1 x 10(5) or 22.1 x 10(5) cells per cm2 onto PLGA scaffolds having pore sizes in the range of 150-300 or 500-710 microm with a thickness of either 1.9 or 3.2 mm. After 1 day in culture, 75.6 and 68.6% of the seeded cells attached and proliferated on the 1.9 mm thick scaffolds of 150-300 microm pore size for the low and high seeding densities, respectively. The number of osteoblasts continued to increase throughout the study and eventually leveled off near 56 days, as indicated by a quantitative DNA assay. Osteoblast/foam constructs with a low cell seeding density achieved comparable DNA content and alkaline phosphatase (ALPase) activity after 14 days, and mineralization results after 56 days to those with a high cell seeding density. A maximum penetration depth of osseous tissue of 220+/-40 microm was reached after 56 days in the osteoblast/foam constructs of 150-300 microm pore size initially seeded with a high cell density. For constructs of 500-710 microm pore size, the penetration depth was 190+/-40 microm under the same conditions. Scaffold pore size and thickness did not significantly affect the proliferation or function of osteoblasts as demonstrated by DNA content, ALPase activity, and mineralized tissue formation. These data show that comparable bone-like tissues can be engineered in vitro over a 56 day period using different rat calvarial osteoblast seeding densities onto biodegradable polymer scaffolds with pore sizes in the range of 150-710 microm. When compared with the results of a previous study where similar polymer scaffolds were seeded and cultured with marrow stromal cells, this study demonstrates that PLGA foams are suitable substrates for osteoblast growth and differentiated function independent of cell source.
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Materiales Biocompatibles , Matriz Ósea/citología , Ácido Láctico , Osteoblastos/citología , Ácido Poliglicólico , Polímeros , Fosfatasa Alcalina/metabolismo , Animales , Desarrollo Óseo/fisiología , Adhesión Celular/fisiología , Recuento de Células , División Celular/fisiología , Células Cultivadas , Osteoblastos/enzimología , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas , Ratas Sprague-DawleyRESUMEN
Porous biodegradable poly(DL-lactic-co-glycolic acid) foams were seeded with rat marrow stromal cells and implanted into the rat mesentery to investigate in vivo bone formation at an ectopic site. Cells were seeded at a density of 6.83 x 10(5) cells/cm2 onto polymer foams having pore sizes ranging from either 150 to 300 to 710 microns and cultured for 7 days in vitro prior to implantation. The polymer/cell constructs were harvested after 1, 7, 28, or 49 days in vivo and processed for histology and gel permeation chromatography. Visual observation of hematoxylin and eosin-stained sections and von Kossa-stained sections revealed the formation of mineralized bonelike tissue in the constructs within 7 days postimplantation. Ingrowth of vascular tissue was also found adjacent to the islands of bone, supplying the necessary metabolic requirements to the newly formed tissue. Mineralization and bone tissue formation were investigated by histomorphometry. The average penetration depth of mineralized tissue in the construct ranged from 190 +/- 50 microns for foams with 500-710-microns pores to 370 +/- 160 microns for foams with 150-300-microns pores after 49 days in vivo. The mineralized bone volume per surface area and total bone volume per surface area had maximal values of 0.28 +/- 0.21 mm (500-710-microns pore size, day 28) and 0.038 +/- 0.024 mm (150-300-microns, day 28), respectively. As much as 11% of the foam volume penetrated by bone tissue was filled with mineralized tissue. No significant trends over time were observed for any of the measured values (penetration depth, bone volume/surface area, or percent mineralized bone volume). These results suggest the feasibility of bone formation by osteoblast transplantation in an orthotopic site where not only bone formation from transplanted cells but also ingrowth from adjacent bone may occur.
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Materiales Biocompatibles , Ácido Láctico , Osteoblastos/trasplante , Osteogénesis , Ácido Poliglicólico , Polímeros , Animales , Células de la Médula Ósea , Masculino , Ensayo de Materiales , Mesenterio , Minerales/metabolismo , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas , Ratas Endogámicas F344RESUMEN
Three strains of male and female mice, CFLP, BALB/c and CBA/ca, were infected i.p. with the tetrathyridia of Mesocestoides corti and examined in groups of 5 at 0, 7, 14, 21, 28 and 42 days post-infection. At post-mortem the numbers of parasites both loose in the peritoneal cavity and in the liver tissue were counted, the livers weighted fresh and sections of liver stained to examine the inflammatory response, encapsulation of the tetrathyridia and for eosinophils, neutrophils, mast cells and plasma cells. BALB/c mice had significantly more parasites loose in the peritoneal cavity than CFLP and CBA/ca mice. Infected livers of all three strains were significantly heavier than control livers; the heaviest livers were those of the CFLP followed by the CBA/ca mice which also showed the greatest rate of weight increase. The tetrathyridia from the peritoneal cavities of the CBA/ca and CFLP mice were covered with a 'mucilage-like' substance. Tetrathyridia within the host capsules of the CBA/ca mice contained host cells. There was no strain difference with regard to numbers of tetrathyridia in the liver but male mice harboured significantly more parasites than females. Differences in the numbers of cell types within the liver were detected between the strains but no one strain showed any consistent pattern. There was an overall increase in total white blood counts as well as an increase in the number of eosinophils, monocytes, neutrophils and lymphocytes during the course of the infection.
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Sangre/parasitología , Infecciones por Cestodos/parasitología , Hígado/parasitología , Mesocestoides/crecimiento & desarrollo , Ratones Endogámicos/parasitología , Animales , Recuento de Células Sanguíneas , Carga Corporal (Radioterapia) , Movimiento Celular , Femenino , Masculino , Mesocestoides/anatomía & histología , Ratones , Tamaño de los Órganos , Cavidad Peritoneal/parasitología , Especificidad de la EspecieRESUMEN
Degeneration of the cochlear nerve before and after placement of the cochlear implant might influence the efficacy of the device. We examined histological characteristics, including the caliber of the cochlear nerve fibers of the central segment proximal to the porus acusticus, in three profoundly deaf patients. Two of them used a cochlear implant for many years longer in one ear than in the other, and one used an implant in one ear only. No qualitative or quantitative differences between the two sides were found. However, in all three cases we found that the cochlear nerves on both sides were substantially degenerated. These results indicated no noticeable effects of stimulation by the cochlear implant on the central portion of the cochlear nerve.