RESUMEN

Diabetic ischemic ulcer is an intractable diabetic complication. Bone marrow mesenchymal stem cells (BMSCs) have great potential in variety of tissue repair. In fact, poor cell viability and tolerance limit their ability for tissue repair. In addition, it is difficult for stem cells to home and locate to the lesion. In this study, we explore whether over-expression of heme oxygenase-1 (HO-1) in BMSCs complexed with collagen play an important role in treatment of diabetic ischemic ulcers. In vitro, over-expression of HO-1 promoted the proliferation and paracrine activity of BMSCs and the conditioned medium of BMSCs accelerated HUVECs migration and proliferation. These processes were closely related to Akt signaling pathway and were not dependent on Erk signaling pathway. In vivo, in order to make BMSCs directly act on the wound, we choose a solid collagen as a carrier, BMSCs were planted into it, ischemic wounds of diabetic mice were covered with the complex of BMSCs and collagen. The results indicate that the complex of HO-1-overexpressing BMSCs and collagen biomaterials can significantly promote angiogenesis and wound healing. These preclinical findings open new perspectives for the treatment of diabetic foot ulcers.

Asunto(s)

Colágeno/farmacología , Diabetes Mellitus Experimental/terapia , Hemo-Oxigenasa 1/metabolismo , Isquemia/terapia , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Úlcera/terapia , Animales , Materiales Biocompatibles/farmacología , Vasos Sanguíneos/efectos de los fármacos , Capilares/efectos de los fármacos , Capilares/patología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Medios de Cultivo Condicionados/farmacología , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/patología , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Isquemia/complicaciones , Isquemia/patología , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Metaloporfirinas/farmacología , Ratones , Ratones Endogámicos C57BL , Neovascularización Fisiológica/efectos de los fármacos , Comunicación Paracrina/efectos de los fármacos , Protoporfirinas/farmacología , Úlcera/complicaciones , Úlcera/patología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Cicatrización de Heridas/efectos de los fármacos

RESUMEN

The patency rate of small-diameter tissue-engineered blood vessels is the determinant for their application in coronary artery bypass grafting. The coronary artery is innervated by vagus nerves. The origin of vagus nerves is rich in brain-derived neurotrophic factors (BDNF). We have investigated whether BDNF could improve the patency rate of small-diameter tissue-engineered blood vessels through promoting stem cell homing and paracrine activity. In vitro, we isolated early and late endothelial progenitor cells (EPCs) and found BDNF could promote single clone formation and paracrine function of EPCs, and could also induce the proliferation, migration and differentiation of late EPCs. BDNF could enhance the capturing of EPCs in parallel-plate flow chamber. Flow cytometric analysis and laser-scanning confocal microscope showed BDNF could enhance the mobilization and homing of C57BL/6 mouse EPCs after wire injury. Based on it, BDNF was coupled to the lumen surface of the blood vessel matrix material incubated with collagen through SPDP to construct BDNF-modified small-diameter tissue-engineered blood vessel. The blood vessel patency rate was significantly higher than that of control group 8 weeks after implantation in rats and the endothelialization level was superior to control. These results demonstrate that BDNF can effectively improve patency of small-diameter tissue-engineered blood vessels through stem cell homing and paracrine, and it is expected to play an important role in the construction of substitutes for coronary artery bypass grafting.

Asunto(s)

Vasos Sanguíneos/fisiología , Factor Neurotrófico Derivado del Encéfalo/farmacología , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/efectos de los fármacos , Ingeniería de Tejidos/métodos , Grado de Desobstrucción Vascular , Animales , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Colágeno/metabolismo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Humanos , Ratones , Ratones Endogámicos C57BL , Microscopía Confocal , Ratas , Ratas Wistar

RESUMEN

Endothelial progenitor cells (EPCs) mobilization and homing are critical to the development of an anti-thrombosis and anti-stenosis tissue-engineered blood vessel. The growth and activation of blood vessels are supported by nerves. We investigated whether nerve growth factors (NGF) can promote EPCs mobilization and endothelialization of tissue-engineered blood vessels. In vitro, NGF promoted EPCs to form more colonies, stimulated human EPCs to differentiate into endothelial cells, and significantly enhanced EPCs migration. Flow cytometric analysis revealed that NGF treatment increased the number of EPCs in the peripheral circulation of C57BL/6 mice. Furthermore, the treatment of human EPCs with NGF facilitated their homing into wire-injured carotid arteries after injection into mice. Decellularized rat blood vessel matrix was incubated with EDC cross-linked collagen and bound to NGF protein using the bifunctional coupling agent N-succinmidyl3-(2-pyridyldit-hio) propionate (SPDP). The NGF-bound tissue-engineered blood vessel was implanted into rat carotid artery for 1 week and 1 month. NGF-bound blood vessels possessed significantly higher levels of endothelialization and patency than controls did. These results demonstrated that NGF can markedly increase EPCs mobilization and homing to vascular grafts. Neurotrophic factors such as NGF have a therapeutic potential for the construction of tissue-engineered blood vessels in vivo.

Asunto(s)

Vasos Sanguíneos/citología , Vasos Sanguíneos/efectos de los fármacos , Células Endoteliales/citología , Factores de Crecimiento Nervioso/farmacología , Células Madre/citología , Células Madre/efectos de los fármacos , Ingeniería de Tejidos/métodos , Animales , Biomarcadores/metabolismo , Vasos Sanguíneos/trasplante , Arterias Carótidas/efectos de los fármacos , Arterias Carótidas/patología , Diferenciación Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ensayo de Unidades Formadoras de Colonias , Células Endoteliales/efectos de los fármacos , Células Endoteliales/trasplante , Movilización de Célula Madre Hematopoyética , Humanos , Ratones , Ratones Endogámicos C57BL , Neovascularización Fisiológica/efectos de los fármacos , Ratas

RESUMEN

OBJECTIVE: To determine if A20, a zinc finger protein that mediates the inflammatory response, affects monocyte-endothelial cell-cell interactions induced by low shear flow. METHODS: Primary cultured endothelial cells (EC) were transfected with an A20 expression vector, and the VCAM-1, ICAM-1 and IL-8 mRNA, and protein expression levels in A20-transfected EC lysates were checked by PCR array and ELISA, respectively. CD14-positive monocyte migration toward and adhesion to EC were measured using a parallel plate flow chamber. RESULTS: Low shear stress, defined as 0.2 Pa for 6 h, normally increases VCAM-1, ICAM-1 and IL-8 expression in EC and allows for binding of monocytes to EC. We found that overexpression of A20 in EC inhibits their normal expression of VCAM-1, ICAM-1 and IL-8 under low shear stress conditions. We also found that A20 inhibits IkappaBalpha degradation in EC following low shear stress exposure and that it attenuates the rolling and EC adhesive properties of shear-induced monocytes as compared with untransfected control EC. The results demonstrate that A20 overexpression in EC inhibits low shear flow-induced monocyte-EC interactions. These findings may be useful in the development of therapeutic agents aimed at treating chronic inflammatory diseases like atherosclerosis.

Asunto(s)

Células Endoteliales/citología , Péptidos y Proteínas de Señalización Intracelular/fisiología , Receptores de Lipopolisacáridos/inmunología , Monocitos/citología , Proteínas Nucleares/fisiología , Aorta , Adhesión Celular , Células Cultivadas , Proteínas de Unión al ADN , Células Endoteliales/metabolismo , Ensayo de Inmunoadsorción Enzimática , Expresión Génica , Humanos , Proteínas I-kappa B/metabolismo , Molécula 1 de Adhesión Intercelular/análisis , Molécula 1 de Adhesión Intercelular/genética , Interleucina-8/análisis , Interleucina-8/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Monocitos/inmunología , Proteínas Nucleares/genética , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estrés Mecánico , Transfección/métodos , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa , Molécula 1 de Adhesión Celular Vascular/análisis , Molécula 1 de Adhesión Celular Vascular/genética

RESUMEN

Endothelial progenitor cell (EPC)-seeded intravascular stents may reduce or prevent in-stent restenosis. A20 can play an important role for preventing vascular restenosis. Therefore, it is very important how to enhance the seeding efficiency of A20-modified EPCs on the stent for preventing in-stent restenosis. To approach this problem, we developed a novel transgenic EPC-seeded stent and evaluated its feasibility and efficiency. EPCs were isolated and purified from umbilical blood using immunomagnetic beads and then transfected with the A20 gene. One stent type (type 1) was coated with EDC cross-linked collagen, and another stent type (type 2) was coated with EDC cross-linked collagen and bound to the CD34 antibody using the bifunctional coupling agent N-succinmidyl3-(2-pyridyldithio) propionate (SPDP). Then, the stents were seeded with EPCs transfected with the A20 gene. The stents were implanted in biological artificial vessels, and cell adhesion was determined in a flow chamber. Cell growth was also measured. EPCs were transfected successfully with the A20 gene. The cells covered both types of stents with favorable biological function. After placement in a flow chamber, the number of cells attached to type 1 stents significantly dropped and their distribution was scattered. Type 2 stents were basically covered with cells and there were more cells on type 2 stents than on type 1 stents (p < 0.01). Collagen-coupled antibody effectively improves the seeding of transgenic EPCs, offering a new choice of stents to prevent restenosis caused by vascular disease after interventional treatment.

Asunto(s)

Células Endoteliales/citología , Péptidos y Proteínas de Señalización Intracelular/genética , Proteínas Nucleares/genética , Células Madre/citología , Stents , Antígenos CD34/química , Adhesión Celular , Proliferación Celular , Células Cultivadas , Colágeno/química , Proteínas de Unión al ADN , Humanos , Proteínas Inmovilizadas/química , Transfección , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa

RESUMEN

To investigate whether decellularized vascular tissues and A20-regulated endothelial progenitor cells can be used for constructing a transgenic tissue-engineered blood vessel with anti-atherosclerotic vascular stenotic properties. A20 gene-transfected endothelial progenitor cells differentiated endothelial cells and smooth muscle cells attached to and migrated into the decellularized porcine vascular scaffolding in a bioreactor. The histology of the conduits revealed viable and layered tissue. Scanning electron microscopy showed confluent, homogeneous tissue surfaces. The mechanical strength of the pulsed constructs was similar to that of the human artery. In vivo, the A20 gene-transfected tissue-engineered blood vessels were transplanted into the carotid artery of a rat for 6 months. Blood vessel xenotransplantation caused hyperacute rejection; all transplanted control blood vessels were completely rejected, but A20-transfected tissue-engineered blood vessels demonstrated good flow on implantation, and remained open for 6 months postoperatively, as assessed by Doppler. The HE stain demonstrated that the vessels were patent, without evidence of stenosis or dilatation after 6 months. These results demonstrate that transgenic tissue-engineered blood vessels have long-term patency and unique anti-stenotic properties.

Asunto(s)

Aterosclerosis/prevención & control , Vasos Sanguíneos/fisiología , Células Endoteliales/citología , Técnicas de Transferencia de Gen , Células Madre Hematopoyéticas/citología , Ingeniería de Tejidos/métodos , Grado de Desobstrucción Vascular , Animales , Anticuerpos Monoclonales/metabolismo , Arterias Carótidas/trasplante , Técnicas de Cultivo de Célula , Diferenciación Celular , Células Cultivadas , Células Endoteliales/fisiología , Células Endoteliales/ultraestructura , Endotelio Vascular/citología , Matriz Extracelular/química , Colágenos Fibrilares/química , Rechazo de Injerto , Células Madre Hematopoyéticas/fisiología , Humanos , Miocitos del Músculo Liso/citología , Miocitos del Músculo Liso/metabolismo , Ratas , Ratas Wistar , Sus scrofa , Transfección , Trasplante Heterólogo , Venas Umbilicales/citología

RESUMEN

OBJECTIVE: To study the effect of core-binding factor alpha1 (Cbfa1) on the mesenchymal stem cells (MSCs) osteoblastic differentiation. METHODS: The MSCs were isolated from Japan white rabbits and cultured in vitro. The 3rd generation MSCs were infected with Cbfa1 recombinant adenovirus. The expression of Cbfa1 was detected by immunofluorescence after being infected for 3 days and the proliferation was estimated by MTT method from the 1st day to the 7th day. Then the MSCs were divided into four groups: the commonly cultured group, the simply induced group, the control adenovirus treatment group, and the Cbfa1 adenovirus treatment group. The expressions of mRNA for a various of osteoblast gene markers such as alkaline phosphatase, osteocalcin, osteopontin and type I collagen were analyzed based on reverse transcriptase polymerase chain reaction (RT-PCR). The change of adipose and myoblastic differentiation gene marker PPAR gamma 2 and MyoD expression were detected by RT-PCR respectively. RESULTS: Positive staining of Cbfa1 was found in the MSCs infected with Cbfa1 adenovirus, and there was no significant difference in cell proliferation among the experimental groups (P>0.05). The RT-PCR indicated that all the osteoblast gene markers except type I collagen were up-regulated in the Cbfa1 adenovirus treatment group. In contrast, the expressions of PPAR gamma 2 and MyoD were restrained. CONCLUSION: Cbfa1 can directly promote the differentiation of MSCs into osteoblasts.

Asunto(s)

Células de la Médula Ósea/citología , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Células Madre Mesenquimatosas/citología , Osteoblastos/citología , Adenoviridae/genética , Animales , Células de la Médula Ósea/metabolismo , Diferenciación Celular , Células Cultivadas , Femenino , Masculino , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/metabolismo , Conejos , Transfección

RESUMEN

To explore the human smooth muscle cells seeding in blood vessel of minor pig after trypsin treatment and provide data for xenotransplantation and for using pig vessel in tissue engineering. HE and silver stain were used for checking the smooth muscle cells seeding in acellular blood vessel. The results showed that the smooth muscle cells seeding succeeded and the smooth muscle cells were in normal morphological distribution. These demonstrate that the pig aorta can be used for smooth muscle cells seeding, and hence for constructing new vascular grafts.

Asunto(s)

Bioprótesis , Prótesis Vascular , Trasplante de Células , Músculo Liso Vascular/citología , Ingeniería de Tejidos , Animales , Vasos Sanguíneos/citología , Separación Celular , Femenino , Humanos , Masculino , Porcinos , Porcinos Enanos , Ingeniería de Tejidos/métodos , Trasplante Heterólogo

RESUMEN

This study was aimed to examine the effectiveness of a gene transfer of human TGFbeta1 gene into endothelial cells and to determine whether TGFbeta1 increases ECM expression of endothelial cells. With the help of DOTAP, endothelial cells were transfected with pMAMneoTGFbeta1. The positive cell clones were selected with G418. The stable transfection and expression of TGFbeta1 in the endothelial cells were determined by immunofluorescence analysis. The expression levels of collagen I and fibronectin in the transfected and untransfected endothelial cells were determined by Western blot. The adhesion force between endothelial cells and matrix was determined by a micropipette technical system. The results showed abundant TGFbeta1 stable expression in the endothelial cells. TGFbeta1 gene was noted to increase collagen I and fibronectin expression and increase the adhesion between endothelial cells and matrix. These findings indicated that TGFbeta1 can be used in vascular tissue engineering for the enhancement of endothelial cells adhesion.

Asunto(s)

Endotelio Vascular/fisiología , Matriz Extracelular/metabolismo , Factor de Crecimiento Transformador beta/biosíntesis , Adhesión Celular , Células Cultivadas , Colágeno Tipo I/biosíntesis , Colágeno Tipo I/genética , Endotelio Vascular/citología , Matriz Extracelular/fisiología , Fibronectinas/biosíntesis , Fibronectinas/genética , Humanos , Ingeniería de Tejidos , Factor de Crecimiento Transformador beta/genética , Venas Umbilicales/citología

RESUMEN

OBJECTIVE: Endothelial angiogenesis in the intima of the arterial wall is one of key events in the pathogenesis of arteriosclerosis. The molecular mechanisms by which transforming growth factor beta 1 (TGFbeta1) and endothelial progenitor cells may be responsible for angiogenesis of arteriosclerosis lesions are poorly understood. MATERIALS AND METHODS: Primary culture smooth muscle cells were transfected with pMAMneoTGFbeta1. ELISA checked VEGF expression in smooth muscle cells. Human EPCs (CD34+ cells) were cultured in pMAMneoTGFbeta1 or pMAMneo transfected smooth muscle cells conditional medium. After 21 days, differentiated endothelial colonies were confirmed by immunofluorescence for von Willebrand factor (vWF) and vascular-endothelial (VE)-cadherin. The VEGFR-1 expression in differentiated endothelial colonies was detected by ELISA. Cells migration and adhesion toward pMAMneoTGFbeta1 and pMAMneo transfected smooth muscle cells were also measured in parallel flow chamber. RESULTS: Abundant TGFbeta1 stable expressed in smooth muscle cells. TGFbeta1 transfected smooth muscle cells expressed significantly higher level VEGF than pMAMneo group. As judged by positive staining for endothelial markers vWF and VE-cadherin, the combination of TGFbeta1 transfected smooth muscle cells conditional medium produced significantly more endothelial colonies (P<0.05) than did pMAMneo group. The adhesion force between endothelial progenitor cells and smooth muscle cells in TGFbeta1 group was higher than control. CONCLUSION: TGFbeta1 expressed smooth muscle cells can be helpful for increasing endothelial progenitor cells adhesion and differentiation. It may be responsible for angiogenesis of arteriosclerosis lesions and useful for blood vessel tissue engineering.

Asunto(s)

Células Endoteliales/metabolismo , Músculo Liso Vascular/metabolismo , Células Madre/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Antígenos CD , Arteriosclerosis/metabolismo , Cadherinas/metabolismo , Adhesión Celular , Técnicas de Cultivo de Célula , Diferenciación Celular , Ensayo de Inmunoadsorción Enzimática , Humanos , Músculo Liso Vascular/fisiopatología , Neovascularización Patológica/metabolismo , Factor de Crecimiento Transformador beta1 , Factor A de Crecimiento Endotelial Vascular/metabolismo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismo , Factor de von Willebrand/metabolismo

RESUMEN

In regions of a vessel that experience low shear stress and reversing flow patterns, early features in the pathogenesis of atherosclerosis include the accumulation of oxidized LDL (OxLDL) and adhesion of monocytes to endothelial cells (EC). Here we investigated the hypothesis that low shear stress (2 dyn/cm2) and OxLDL are synergistic for enhanced expression of vascular cell adhesion molecule (VCAM-1) and human aortic endothelial cell (HAEC)-monocyte adhesion. This study shows low shear stress can significantly reduce IkappaBalpha levels, activate NF-kappaB, increase the expression of VCAM-1 in HAEC and binding of monocytes. OxLDL itself cannot significantly increase the expression of VCAM-1 in HAEC and binding of monocytes, but through activation of NF-kappaB and degradation of IkappaBalpha induced by low shear stress it can significantly enhance VCAM-1 expression and monocyte adhesion, over that in unmodified LDL or control. These results suggest that low shear stress can regulate monocyte adhesion to oxidized lipid-induced endothelial cells via an IkappaBalpha-dependent pathway, and that low shear stress together with OxLDL may likely play an important role in atherogenesis.

Asunto(s)

Endotelio Vascular/fisiología , Proteínas I-kappa B/fisiología , Lipoproteínas LDL/farmacología , Monocitos/fisiología , Adhesión Celular/efectos de los fármacos , Células Cultivadas , Células Endoteliales/metabolismo , Células Endoteliales/fisiología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Humanos , Inhibidor NF-kappaB alfa , FN-kappa B/antagonistas & inhibidores , Transducción de Señal/fisiología , Estrés Mecánico , Molécula 1 de Adhesión Celular Vascular/metabolismo

RESUMEN

Burn injuries as well as skin damages are often associated with immune suppression and often cause multiple organ failures. The monolayer endothelium is vulnerable to injuries from circulating factors resulting from remote wounds. Endothelial cell activation and apoptosis can alter microvascular permeability and intensify organ damage. A20, as a physiological cytoprotective gene is essential for preventing spontaneous innate immune cell-mediated inflammation and tissue destruction. It is not known whether A20 has the function to protect endothelial cells from the effect of burns serum challenge on endothelial function in vitro. This study shows that A20 can express in endothelial cells after burns serum stimulation and inhibit endothelial cell activation and apoptosis induced by burns serum. These results suggest that A20 may be beneficial in limiting the response to burn injuries.

Asunto(s)

Quemaduras/metabolismo , Células Epiteliales/metabolismo , Proteínas/fisiología , Dedos de Zinc/fisiología , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Quemaduras/patología , Bovinos , Proteínas de Unión al ADN , Selectina E/metabolismo , Células Epiteliales/patología , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Péptidos y Proteínas de Señalización Intracelular , FN-kappa B/metabolismo , Proteínas Nucleares , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Venas Umbilicales/metabolismo

RESUMEN

This study sought to explore the change of the major histocompatibility complex (MHC) antigen expression and the endothelization of blood vessel in minor pig after trypsin treatment, and to provide data for xenotransplantation and pig vessel for use in tissue engineering. Western blot assays were conducted for detecting the expression of MHC xenoantigens. Scanning electron microscopy was used for checking the endothelization of decellularized blood vessel. The results showed that MHC antigen is not expressed after trypsin treatment. The endothelization is accomplished. The endothelial cells have normal morphological distribution. These demonstrate that the antigen of pig aorta is significantly decreased and it can be used for constructing new vascular grafts.

Asunto(s)

Bioprótesis , Prótesis Vascular , Complejo Mayor de Histocompatibilidad/fisiología , Animales , Células Cultivadas , Células Endoteliales/citología , Femenino , Masculino , Porcinos , Porcinos Enanos , Ingeniería de Tejidos/métodos , Andamios del Tejido

RESUMEN

This study was conducted to examine the effectiveness of a gene transfer of human TGF beta 1 gene into smooth muscle cells and whether the TGF beta 1 can increase elastin expression of smooth muscle cells. With the help of DOTAP, smooth muscle cells were transfected with pMAMneoTGF beta 1. The positive cell clones were selected with G418. The stable transfection and expression of TGF beta 1 in the smooth muscle cells were determined by immunofluorescence analysis. The expression of elastin in the transfected and untransfected cells were determined by in situ hybridization. The adhesion force between smooth muscle cells and matrix was detected by micropipette system. The results showed abundant TGF beta 1 stable expression in smooth muscle cells. TGF beta 1 gene can increase two-three times elastin expression and increase the adhesion between smooth muscle cells and matrix. TGF beta 1 can be used in vascular tissue engineering to increase smooth muscle cells adhesion.

Asunto(s)

Elastina/biosíntesis , Músculo Liso Vascular/citología , Factor de Crecimiento Transformador beta/fisiología , Adhesión Celular , Células Cultivadas , Humanos , Hibridación in Situ , Músculo Liso Vascular/metabolismo , Transfección , Factor de Crecimiento Transformador beta/biosíntesis , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta1

RESUMEN

To explore the changes of the antigen expression and the biomechanical characteristics of blood vessel in Banna little ear pig before and after trypsin treatment, and provide data for xenotransplantation and pig vessel using for tissue engineering. Geometric morphology and microstructure of pig cartoid artery were stuided quantitatively by histologic method and computer image analysis. The relationship between pressure and diameter was observed at different period of time before and after trypsin treatment. Affinity-immunohistochemistry assay was conducted to detect the expression of xenoantigens (alpha-Gal). The results showed that alpha-Gal antigen is only expressed in vascular endothelial cellsouly. There is no significant difference in blood vessel compliance. These demonstrate that the antigenicity of pig carotid artery is significantly reduced, however, the mechanical characteristics did not change significantly. We suppose that pig vessels treated by trypsin can be used as the substrate material for vascular tissue engineering.

Asunto(s)

Antígenos Heterófilos/biosíntesis , Vasos Sanguíneos/fisiología , Animales , Animales Endogámicos , Vasos Sanguíneos/efectos de los fármacos , Femenino , Masculino , Estrés Mecánico , Porcinos , Ingeniería de Tejidos , Tripsina/farmacología

RESUMEN

OBJECTIVE: To investigate the expression and distribution of xenoantigen in intervertebral disk of Chinese banna minipig inbred line, and to study the availability of xenograft transplantation of intervertebral disk. METHODS: Samples of intervertebral disk were collected from six Banna pigs of 8 to 11-month-old. The fixation, embedment and slice were performed. alpha-Gal specific binding lection (BSI-B4) were used as affinity reagents and affinity-immunohistochemistry assays (SABC methods and DAB stain) were conducted to detect the expression and distribution of xenoantigen (alpha-Gal). RESULTS: alpha-Gal was found in chondrocyte cell and chondrocyte-like cell in intervertebral disk which have the positive yellow-stained particulate aggradation. There was no stain in the matrix, elastic fiber and collagen fiber. CONCLUSIONS: The distribution of xenoantigen is locally in the tissue of intervertebral disk and its expression is weak. This suggests that the intervertebral disk of Banna pig may be alternative donor for xenotransplantation.

Asunto(s)

Antígenos Heterófilos/biosíntesis , Galactosa/inmunología , Disco Intervertebral/inmunología , Trasplante Heterólogo/inmunología , Animales , Animales Endogámicos , Condrocitos/inmunología , Galactosa/análisis , Inmunohistoquímica/métodos , Disco Intervertebral/trasplante , Masculino , Porcinos Enanos

RESUMEN

OBJECTIVE: To explore the kidney anatomic structure of banna minipig inbred-lines, and to provide data for kidney xenotransplantation. METHODS: The fresh and infused kidneys of banna minipig (including the vessel and the ureter) were checked by anatomic microscope and vernier caliper in original location and away body. The tissue structure was observed by HE stain. RESULTS: The structure of kidney of banna minipig inbred-lines (including the vessel and the ureter) are similar to that of human being. The fascia propria of kidney is divided into three layers including capsula fibrosa, capsula adipose and fascia renalis. The thickness of cortex renalis is (20.0 +/- 2.4) mm. The average diameter of renal artery is 5.1 mm and is similar to that of human being. All the kidneys of banna minipig inbred-lines have a single branch renal artery. The diameters of left and right ureters are 5.1 mm and 4.7 mm, respectively. CONCLUSION: The kidney of banna minipig inbred-lines is an ideal replacement of human kidney for xenotransplantation.

Asunto(s)

Riñón/anatomía & histología , Porcinos Enanos/anatomía & histología , Trasplante Heterólogo , Animales , China , Femenino , Trasplante de Riñón , Masculino , Arteria Renal/anatomía & histología , Venas Renales/anatomía & histología , Porcinos Enanos/genética , Uréter/anatomía & histología