ABSTRACT
BACKGROUND: Changes of elastic fibers in middle cerebral artery(MCA)is close related withthe aged cerebrovascular disease.OBJECTIVE: To observe the changes of elastic fibers of MCA in different aging rats.DESIGN: A descriptive and controlled study based on experimental animals.SETTING: Department of anatomy and central laboratory in a university.MATERIALS: Totally 36 healthy Wistar rats with either gender, weighing 200 - 280 g, were selected from the Animal Institute of the third medical military university of Chongqing[certification SCXX (army) 2002-007].INTERVENTIONS: Changes of elastic fibers of MCA of different aging rats were observed with light microscope, transmission electron microscope and image analysis system. MAIN OUTCOME MEASURES: ①) Major outcome: changes of elastic lamella in MCA of different aging rats; ②) Secondary outcome: ultramicrostructural changes of internal lamella under the transmission electron microscope. RESULTS: With the increase of age, the folded extent and quantity of internal elastic lamella were decreased, and the content of elastic fibers were also decreased significantly( P ＜ 0.01 ). However, the ratio of collagen fibers to elastic fibers was increased significantly( P ＜ 0.01 ) . In the aging group above 24 months, the internal elastic lamina thinned, delaminated and disrupted, and the lipid deposited in it. Endothelial cells and smooth muscle cells passed through the internal elastic lamina. CONCLUSION: Changes of elastic fibers may be related with the increased susceptibility to the cerebrovascular disease in aged people.
ABSTRACT
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.
Subject(s)
Animals , Female , Humans , Male , Bioprosthesis , Blood Vessel Prosthesis , Blood Vessels , Cell Biology , Cell Separation , Cell Transplantation , Muscle, Smooth, Vascular , Cell Biology , Swine , Swine, Miniature , Tissue Engineering , Methods , Transplantation, HeterologousABSTRACT
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.
Subject(s)
Humans , Cell Adhesion , Cells, Cultured , Collagen Type I , Genetics , Endothelium, Vascular , Cell Biology , Physiology , Extracellular Matrix , Metabolism , Physiology , Fibronectins , Genetics , Tissue Engineering , Transforming Growth Factor beta , Genetics , Umbilical Veins , Cell BiologyABSTRACT
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.
Subject(s)
Animals , Female , Male , Bioprosthesis , Blood Vessel Prosthesis , Cells, Cultured , Endothelial Cells , Cell Biology , Major Histocompatibility Complex , Physiology , Swine , Swine, Miniature , Tissue Engineering , Methods , Tissue ScaffoldsABSTRACT
Objective To investigate the effects of zinc finger protein gene A20 on the inhibition of lipopolysaccharide (LPS) induced interleukin 8 (IL 8) expression in endothelial cells. Methods Plasmid pcDNA3.1EHA20 was transfected into human umbilical vein endothelial cells (HUVECs) by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of A20 in HUVECs were determined by immunofluorescent analysis. IL 8 expression was detected by sandwich ELISA with double monoclonal antibodies. Results High expression of A20 gene in HUVECs transfected with pCDNA3.1EHA20 was confirmed by immunofluorescent analysis. IL 8 expression increased in LPS induced endothelial cells. A20 gene could inhibit more than 70% LPS induced IL 8 expression ( P
ABSTRACT
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.
Subject(s)
Humans , Cell Adhesion , Cells, Cultured , Elastin , In Situ Hybridization , Muscle, Smooth, Vascular , Cell Biology , Metabolism , Transfection , Transforming Growth Factor beta , Genetics , Physiology , Transforming Growth Factor beta1ABSTRACT
Objective Previous studies have demonstrated that LPS can induce endothelial cell activation and the expression of E-selectin. In this study, we examined whether A20 gene could inhibit the expression of E-selectin in endothelial cells induced by LPS. Methods With the help of DOTAP, endothelial cells were transfected with pCDNA3.1 EHA20. The postive cell clones were selected with G418.The stable transfection and expression of A20 in the endothelial cells were determined by immunofluorescence analysis. The E-selectin expression was checked by immunofluorescence, Western blot and in situ hybridization. Results Abundant A20 stable expression in endothelial cells transfected with pCDNA3.1 EHA20 was confirmed by immunofluorescence analysis. E-selectin expression increased in LPS-inducible endothelial cells. A20 gene inhibited 90% LPS-inducible E-selectin expression(P
ABSTRACT
Objective To isolate the bone marrow mesenchymal stem cells(MSCs) from the GFP-expressing mouse, and to study the osteoblastic differentation of the cells. Methods MSCs were isolated by density gradient centrifugation, then the clutrued cells were induced to osteoblastic differentiation using the conditional medium. We detectd the expression of GFP and MSCs differentiation into osteoblasts by histochemistry and immunochemistry. Results The MSCs maintained the expression of GFP during expanded and induced process. After induced for 10 days, lots of alkaline phosphatase and osteoclcin staining positive cells were observed.Conclusion The MSCs of GFP-expressing mouse were successfully isolated and differentiated into ostoblasts. It may be valuable for tracing the seeding cells in tissue engineering bone.
ABSTRACT
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.
Subject(s)
Animals , Female , Male , Animals, Inbred Strains , Antigens, Heterophile , Blood Vessels , Physiology , Stress, Mechanical , Swine , Tissue Engineering , Trypsin , PharmacologyABSTRACT
The sectional morphology of the human brachial artery, ulnar artery, radial artery, palmar digital artery of the thumb and the middle finger were observed under microscopy. Excepting the brachial artery, the intima of the other 4 arteries are thick, especially the ulnar artery. The histological variations must be noticed during the artery is anastomosed. The percentage of the adventitia of the whole wall thickness decreases and the elastic fibers become looser gradually in accordance with the size of the arteries. The relative contents of elastin, collagen and smooth muscle of the above 5 arteries were measured by microspectrophotometer. The content of elastin of these arteries decreases(P
ABSTRACT
A study on abdominal skin flaps of the fresh cadavers of young men under the operation, light and scanning electron microscope, and MAS image analysis system has been made. Methods include Chinese ink, ABS or methyl methacrylate injection and histological slides, transparent specimens as well as casts preparation. The course of the vessels in the human abdominal skin flap is in the "three steps form". Trunks and main branches of the cutaneous vessels run in the deep part of the superficial fascia, their branches and terminals form anastomoses in the middle of the reticular layer of dermis, and branches from the anastomoses form microvascular network in the subpapillary layer. The vascular networks in the skin flap are concentrated in five layers: the deep fascial, superficial fascial, profund dermal, subpapillary, and papillary layers. Between the subpapillary and the profund dermal vascular networks, there is a layer devoid of vascular network and crossed by vascular arteries only. Therefore, this layer may be called as "vascular network devoid area". The differences of the area fraction (Aa) or number of vessels are not significant among various areas of the abdominal skin flap. Design in detaching the abdominal skin flaps and skin grafts is discussed.