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1.
Biomaterials ; 34(12): 2947-59, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23375393

RESUMEN

Research in bone tissue engineering is focused on the development of alternatives to allogenic and autologous bone grafts that can stimulate bone healing. Here, we present scaffolds composed of the natural hydrophilic polysaccharides pullulan and dextran, supplemented or not with nanocrystalline hydroxyapatite particles (nHA). In vitro studies revealed that these matrices induced the formation of multicellular aggregates and expression of early and late bone specific markers with human bone marrow stromal cells in medium deprived of osteoinductive factors. In absence of any seeded cells, heterotopic implantation in mice and goat, revealed that only the composite macroporous scaffold (Matrix + nHA) (i) retained subcutaneously local growth factors, including Bone Morphogenetic Protein 2 (BMP2) and VEGF165, (ii) induced the deposition of a biological apatite layer, (iii) favored the formation of a dense mineralized tissue subcutaneously in mice, as well osteoid tissue after intramuscular implantation in goat. The composite scaffold was thereafter implanted in orthotopic preclinical models of critical size defects, in small and large animals, in three different bony sites, i.e. the femoral condyle of rat, a transversal mandibular defect and a tibial osteotomy in goat. The Matrix + nHA induced a highly mineralized tissue in the three models whatever the site of implantation, as well as osteoid tissue and bone tissue regeneration in direct contact to the matrix. We therefore propose this composite matrix as a material for stimulating bone cell differentiation of host mesenchymal stem cells and bone formation for orthopedic and maxillofacial surgical applications.


Asunto(s)
Materiales Biocompatibles , Huesos , Dextranos/química , Durapatita/química , Glucanos/química , Polisacáridos/química , Ingeniería de Tejidos , Secuencia de Bases , Células Cultivadas , Cartilla de ADN , Microscopía Electrónica de Rastreo , Reacción en Cadena en Tiempo Real de la Polimerasa , Difracción de Rayos X
2.
Biofabrication ; 2(1): 014101, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-20811116

RESUMEN

We present the first attempt to apply bioprinting technologies in the perspective of computer-assisted medical interventions. A workstation dedicated to high-throughput biological laser printing has been designed. Nano-hydroxyapatite (n-HA) was printed in the mouse calvaria defect model in vivo. Critical size bone defects were performed in OF-1 male mice calvaria with a 4 mm diameter trephine. Prior to laser printing experiments, the absence of inflammation due to laser irradiation onto mice dura mater was shown by means of magnetic resonance imaging. Procedures for in vivo bioprinting and results obtained using decalcified sections and x-ray microtomography are discussed. Although heterogeneous, these preliminary results demonstrate that in vivo bioprinting is possible. Bioprinting may prove to be helpful in the future for medical robotics and computer-assisted medical interventions.


Asunto(s)
Biotecnología/métodos , Robótica/métodos , Fracturas Craneales/terapia , Terapia Asistida por Computador/métodos , Ingeniería de Tejidos/métodos , Animales , Sustitutos de Huesos/uso terapéutico , Encéfalo/efectos de la radiación , Diseño Asistido por Computadora , Durapatita/uso terapéutico , Histocitoquímica , Masculino , Ratones , Nanopartículas/uso terapéutico , Cráneo/diagnóstico por imagen , Cráneo/lesiones , Microtomografía por Rayos X
3.
Transplantation ; 76(1): 77-83, 2003 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-12865790

RESUMEN

BACKGROUND: The deleterious effect of steatosis on transplanted livers is mainly related to a microcirculation impairment. We investigated the effect of preservation duration on the recovery of isolated perfused rat steatotic livers and tested the effect of pentoxifylline (PTX), known to have a beneficial effect on hepatic microcirculation. MATERIALS AND METHODS: Fatty rat livers were obtained using a diet able to induce an 80% to 100% microvesicular steatosis within 7 days. We studied the effect of the duration of preservation (12 hr, 18 hr, and 24 hr) on fatty and normal isolated perfused rat liver. PTX was added to University of Wisconsin solution during cold storage (30 mM/kg of weight) and at reperfusion (3 mM) (n=5 livers in each group). Lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, bile production, and vascular resistance were evaluated. The liver injury at the end of perfusion was assessed by optical and electron microscopy. RESULTS: For a 24-hr preservation period, fatty livers demonstrated increased enzymatic release (aspartate aminotransferase: 42+/-16 vs. 17+/-5 IU/L/g of liver, P<0.005; alanine aminotransferase: 32+/-13 vs. 13+/-3 IU/L/g of liver, P<0.005; lactate dehydrogenase: 1,207+/-497 vs. 291+/-195 IU/L/g of liver, P<0.001). Vascular resistance (0.32 vs. 0.15 cm H(2)O/min/mL, P<0.0005) and bile output (67+/-24 vs. 141+/-61 mg/g of liver, P<0.05) were decreased. Peliosis appeared after an 18-hr preservation period for fatty livers compared with a 24-hr preservation period for controls. All these negative effects were suppressed by PTX. CONCLUSION: Diffuse microvesicular steatosis became deleterious only after long preservation times (24 hr). PTX prevented this effect.


Asunto(s)
Hígado Graso/tratamiento farmacológico , Depuradores de Radicales Libres/uso terapéutico , Isquemia/fisiopatología , Hígado , Pentoxifilina/farmacología , Adenosina , Alopurinol , Animales , Hígado Graso/patología , Glutatión , Insulina , Hígado/efectos de los fármacos , Hígado/patología , Hígado/ultraestructura , Circulación Hepática , Pruebas de Función Hepática , Masculino , Preservación de Órganos/métodos , Soluciones Preservantes de Órganos , Rafinosa , Ratas , Ratas Wistar , Factores de Tiempo
4.
Transpl Int ; 15(2-3): 89-95, 2002 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-11935165

RESUMEN

The effect of donor nutritional status on hepatic function recovery after cold ischemia is still debated. We demonstrated previously that a 48-h fast diminished the survival rate of liver-transplanted rats and that the deleterious effect of fasting was prevented by infusion of alanine to the recipient at reperfusion. Whether the duration of fasting influenced the protective effect of alanine and whether this effect was metabolic were not known, and the elucidation of these questions is the aim of this study. The effect on hepatic function recovery of fasting periods of 24 h, 48 h and 72 h prior to cold ischemia were studied in a model of isolated, perfused rat liver. After a cold-ischemic time of 24 h in University of Wisconsin (UW) solution at 4 degrees C, livers were reperfused for 3 h. The combined effect of alanine (8 mM) infusion at liver reperfusion was evaluated for each prior fasting period. The addition of pyruvate (8 mM), a metabolic intermediary of alanine, was only tested in the 72-h fasting group. The evaluation criteria were: liver weight after reperfusion, release of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in the perfusate, bile production, vascular resistance and liver histology after reperfusion. The enzyme release at reperfusion was significantly higher when livers were harvested from rats submitted to a 48-h fast (ALT) or a 72-h fast (ALT, AST, LDH), as compared to those from fed rats. Vascular resistance was increased in 72-h fasted livers. An addition of alanine (8 mM) at reperfusion lowered the release of AST, ALT and LDH. This effect was more obvious when the fasting duration was increased. By contrast, the addition of pyruvate at reperfusion did not improve the recovery of livers submitted to a 72-h fasting period before preservation. A long fasting period is deleterious as compared to feeding; however, this effect can be compensated by infusion of alanine at reperfusion. The mechanism involved is not metabolic. In a clinical setting, the infusion of alanine to the recipient at reperfusion may be a convenient way to compensate for donor undernutrition, especially after a long stay in an intensive care unit.


Asunto(s)
Alanina/farmacología , Ayuno/fisiología , Isquemia , Trasplante de Hígado/fisiología , Hígado/irrigación sanguínea , Preservación de Órganos/métodos , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Bilis/metabolismo , Peso Corporal , Frío , L-Lactato Deshidrogenasa/sangre , L-Lactato Deshidrogenasa/metabolismo , Hígado/efectos de los fármacos , Hígado/fisiología , Pruebas de Función Hepática , Masculino , Estado Nutricional , Tamaño de los Órganos , Perfusión , Piruvatos/farmacología , Ratas , Ratas Wistar , Resistencia Vascular
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