Objective To evaluate the feasibility of utilizing vascular cells combined with folded andframed culture model to develop completely autologous human tissue without using any scaffold material under the principles of ...Objective To evaluate the feasibility of utilizing vascular cells combined with folded andframed culture model to develop completely autologous human tissue without using any scaffold material under the principles of Tissue Engineering. Methods Human vascular cells cultured from ascending aorta (group A) and saphenous vein (group B) were seeded into 15cm-dishes (each n =12) and cultured to form cell sheets over a period of four weeks with Dulbecco's modified Eagle's medium supplemented with Immol/L L-ascorbic acid 2-phos-phate. Thereafter, cell sheets (6 samples of each group) were four-layer folded and cultured in a newly developed frame device for additional four weeks. Controls remained under standard culture conditions. Tissue development was evaluated by light and electron microscopy, biochemical assays. Results The formation of multi-layered cell sheets and production of extracellular matrix were observed in each group after the initial four weeks. Analysis of the folded and framed neo-tissue revealed a solid structure with increased matrix formation and tissue organization compared to the control groups after additional four weeks. DNA assay indicated significantly lower cell proliferation in folded and framed cell sheets than in that of unframed counterparts. Yet hydroxyproline assay demonstrated significant increase of collagen content in the framed aortic and venous derived tissues, which contained 82% and 42% that of human pericardium. Conclusion It is feasible to obtain completely autologous human cardiovascular tissue with the alternative new approach. Numerous issues including improvement of mechanical strength of neo-tissue remain to be investingated.展开更多
Background:The development of materials for cardiovascular surgery that would improve the effectiveness of surgical interventions remains an important task.Surgical intervention during the implantation of vascular pro...Background:The development of materials for cardiovascular surgery that would improve the effectiveness of surgical interventions remains an important task.Surgical intervention during the implantation of vascular prostheses and stents,and the body’s reaction to artificial materials,could lead to chronic inflammation,a local increase in the concentration of proinflammatory factors,and stimulation of unwanted tissue growth.The introduction of nonsteroidal anti-inflammatory drugs into implantable devices could be used to obtain vascular implants that do not induce inflammation and do not induce neointimal tissue outgrowth.Methods:The scaffolds were made by electrospinning from mixtures of polyurethane(PU)with diclofenac(DF).The kinetics of DF release from the scaffolds composed of 3%PU/10%HSA/3%DMSO/DF and 3%PU/DF were studied.The biocompatibility and anti-inflammatory effects of the obtained scaffolds on human gingival fibroblasts and umbilical vein endothelial cells were studied.Results:Both types of scaffolds are characterized by fast DF release.The viability of cells cultured on scaffolds is 2 times worse than that of cells cultured on plastic.The level of the proinflammatory cytokine IL-6 in the culture medium of cells cultured on DF-containing scaffolds was lower than that of cells cultured on scaffolds without DF.Conclusion:The introduction of DF into scaffolds minimizes the inflammation caused by cell reactions to an artificial material.展开更多
文摘Objective To evaluate the feasibility of utilizing vascular cells combined with folded andframed culture model to develop completely autologous human tissue without using any scaffold material under the principles of Tissue Engineering. Methods Human vascular cells cultured from ascending aorta (group A) and saphenous vein (group B) were seeded into 15cm-dishes (each n =12) and cultured to form cell sheets over a period of four weeks with Dulbecco's modified Eagle's medium supplemented with Immol/L L-ascorbic acid 2-phos-phate. Thereafter, cell sheets (6 samples of each group) were four-layer folded and cultured in a newly developed frame device for additional four weeks. Controls remained under standard culture conditions. Tissue development was evaluated by light and electron microscopy, biochemical assays. Results The formation of multi-layered cell sheets and production of extracellular matrix were observed in each group after the initial four weeks. Analysis of the folded and framed neo-tissue revealed a solid structure with increased matrix formation and tissue organization compared to the control groups after additional four weeks. DNA assay indicated significantly lower cell proliferation in folded and framed cell sheets than in that of unframed counterparts. Yet hydroxyproline assay demonstrated significant increase of collagen content in the framed aortic and venous derived tissues, which contained 82% and 42% that of human pericardium. Conclusion It is feasible to obtain completely autologous human cardiovascular tissue with the alternative new approach. Numerous issues including improvement of mechanical strength of neo-tissue remain to be investingated.
基金supported by the Russian state-funded project for ICBFM SB RAS(grant number 125012300656-5)。
文摘Background:The development of materials for cardiovascular surgery that would improve the effectiveness of surgical interventions remains an important task.Surgical intervention during the implantation of vascular prostheses and stents,and the body’s reaction to artificial materials,could lead to chronic inflammation,a local increase in the concentration of proinflammatory factors,and stimulation of unwanted tissue growth.The introduction of nonsteroidal anti-inflammatory drugs into implantable devices could be used to obtain vascular implants that do not induce inflammation and do not induce neointimal tissue outgrowth.Methods:The scaffolds were made by electrospinning from mixtures of polyurethane(PU)with diclofenac(DF).The kinetics of DF release from the scaffolds composed of 3%PU/10%HSA/3%DMSO/DF and 3%PU/DF were studied.The biocompatibility and anti-inflammatory effects of the obtained scaffolds on human gingival fibroblasts and umbilical vein endothelial cells were studied.Results:Both types of scaffolds are characterized by fast DF release.The viability of cells cultured on scaffolds is 2 times worse than that of cells cultured on plastic.The level of the proinflammatory cytokine IL-6 in the culture medium of cells cultured on DF-containing scaffolds was lower than that of cells cultured on scaffolds without DF.Conclusion:The introduction of DF into scaffolds minimizes the inflammation caused by cell reactions to an artificial material.
