Objective To study the feasibility of transplanting human saphanous vein endothelial cells to luminal surface of blood vessel prosthesis and to play a theoretical foundation for the clinical application of autologous endothelial cell transplantation. Methods Human saphanous vein endothelial cells were harvested with 0.1% collagenase and cultivated in vitro for 13.08±1.24 days. The cultures were confirmed as endothelial cells with the fourescent linked anti-Ⅷ antigen antibodies. The content of both 6-keto-PGF1α and Von Willebrand factor (vWF) in the supernatant were detected with ELISA and radioimmunoassay. The multiplied cells were lined in vitro onto the luminal surface of expanded polytetraflouroethylene (ePTFE) grafts precoated with fibrin glue and fibronectin, then cultivated again for 9 days. Results 11.46±2.69×１０６ of available endothelial cells could be regularly obtained, the number of endothelial cells increased 147.93±88.68 times when culture were terminated. All the cells diploid cells with a purity of 99%. The content of both 6-keto-PGF1α and vWF in the media showed no significant difference between the primary and subculture passages. The luminal surface of grafts was covered completely by a spindlelike endothelial monolayer and an even fibrin glue matrix could be seen underneath. Conclusion Endothelial cells derived from human saphanous veins might be feasible to be transplanted onto the luminal surface of ePTFE and present a potential clinical application.
Purpose To determine the effect of exogenous interleukin-1alpha; (IL-1alpha;) on the retina and its vasculature and VEGF expression in SD rats． Methods IL-1alpha;2．0 ng (20 mu;l) were injected into the vitreous of 8 left eyes of 8 SD rats while steriled PBS were injected into 8 right contralateral eyes of the same rats as control． All eyes were assessed by direct ophthalmoscopy every day and enucleated on the 7 thpostoperative day． Histological examination (hemato xylineosin staining) and immunohistochemical staining with antibody against VEGF antigen were performed， and sections were observed and photographed under light microscopy． Results ①All 8 IL-1alpha; inject ed eyes developed epiretinal membranes and extraretinal neovascularization on the 3 rd postoperative days while none of the 8 control eyes exhibited any a bnormal retinal vascular changes and they were confirmed by HE staining;②Immuno staining identified VEGF express mainly in the inner layer of vessel walls， the epiretinal membranes， the neuroganglional layer and the photoreceptor layer of retina， while the control eyes showed only weak positive staining in the photo receptor layer． Conclusions IL-1alpha; is capable of inducing vitreo retinal neovascularization，and increasing the expression of VEGF in the retina and epiretinal membranes． (Chin J Ocul Fundus Dis, 2001,17:135-137)
Objective To understand the value of pre-coating in artificial vessel endothelialization. Methods Literature concerning precoating in artificial vessel endothelialization was extensively reviewed. Results Pre-coating included chemical coatings(collagen, fibronectin, laminin, poly-l-lysin, gelatin andextracellular matrix), pre-clotting(plasma, blood, serum and fibrin glue), chemical bonding (heparin, RGD and lectins) and surface modification. Most of them could enhance the adhesion of the endothelial cells. Conclusion Pre-coating couldimprove endothelialization, but further research is needed to search for the appropriate concentration and incubation time.
OBJECTIVE: To prepare the compound biodegradable matrices, polyglycolic acid (PGA), polylactic acid (PLA) mesh and poly-beta-hydroxybutyrate(PHB) which precoated with collagen, and to observe the growth and differentiation of bovine vascular endothelial cells on these scaffolds. METHODS: By enzymatic digestion methods, bovine vascular endothelial cell (VEC) were isolated from calf thoracic aorta, then cultured and purified. PGA, PLA, PHB meshes were dipped into cross-linked type I collagen solution, dried under vacuum frozen condition. VEC were seeded into these scaffolds. The growth of VEC on scaffolds was analyzed by MTT method. RESULTS: The collagen, PGA/collagen, PLA/collagen scaffolds were elasticity and tenacity. VEC grew better on collagen, PGA/collagen, and PLA/collagen membranes than on the PHB/collagen one. CONCLUSION: The PGA/collagen scaffold has elasticity, plasticity and tenacity. VEC grow best on it. It is an ideal scaffold for tissue engineered vessel reconstruction for it integrating both advantages of biomaterials and degradable materials.
ObjectiveTo investigate the ability of autologous peripheral blood endothelial progenitor cells (EPCs) in promoting neovascularization of tissue engineered bone and osteogenesis of bone marrow mesenchymal stem cells (BMSCs). MethodThe peripheral blood EPCs and BMSCs from No. 1-9 New Zealand rabbits were isolated, cultured, and identified. According to the cell types, the third generation of cells were divided into 3 groups:EPCs (group A), BMSCs (group B), and co-cultured cells of EPCs and BMSCs (group C, EPCs:BMSCs=1:2) . Then cells were seeded on the partially deproteinised bone (PDPB) packaged with fibronectin to construct tissue engineered bone. After 4 days, autologous heterotopic transplantation of tissue engineered bone was performed in the rabbit's muscles bag of groups A, B, and C (the right arm, left arm, right lower limb respectively, 2 pieces each part). At 2, 4, and 8 weeks after transplantation, the growth of tissue engineered bone was observed, and the rate of bone ingrowth was calculated by HE staining; the expressions of CD34, CD105, and zonula occludens protein 1(ZO-1) were compared by immunohistochemical staining at each time point in tissue engineered bone among 3 groups. ResultsThe EPCs and BMSCs were isolated and identified successfully; immunofluorescent staining showed that EPCs were positive for CD34, CD133, and von Willebrand factor (vWF), and BMSCs were positive for CD29 and CD90 and were negative for CD34. The tissue engineered bone constructed in 3 groups was transplanted successfully. At 2, 4, and 8 weeks after autologous heterotopic transplantation, the general observations showed that the soft tissue around the tissue engineered bone increased and thickened gradually in each group with time passing; the boundary between bone and soft tissue was not clear; the pore space of tissue engineered bone gradually was filled, especially in group C, the circuitous vascular network could be seen in the tissue engineered bone. HE staining showed capillaries and collagen fibers increased gradually, tissue engineered bone ingrowth rate was significantly higher in group C than groups A and B at 4 and 8 weeks (P<0.05) , and group B was significantly higher than group A (P<0.05) . Immunohistochemical staining showed that the expressions of CD34, CD105, and ZO-1 in tissue engineered bone of 3 groups all increased with the extension of time, showing significant differences between groups at each time point (P<0.05) . At 2 weeks after transplantation, the expression of CD105 in group C was significantly higher than that in groups A and B (P<0.05) ; at 4 and 8 weeks, CD34, CD105, and ZO-1 expressions showed significant differences between 2 groups (P<0.05) ; the expression was the highest in group C, and was the lowest in group B. ConclusionsAutologous peripheral blood EPCs and BMSCs have synergistic effect, and can promote neovascularization and osteogenesis of tissue engineered bone in vivo.
【Abstract】Objective To introduce the possible effect of endogenous angiogenesis inhibitive factors in the therapy of hepatocarcinoma. Methods Recent relevant literatures were reviewed. ResultsEndogenous angiogenesis inhibitive factors can suppress the growth of tumor blood vessels, which might head off the development and metastasis of hepatocarcinoma effectively. This might provide a new approach to the therapy of hepatocarcinoma. ConclusionRecent studies on endogenous angiogenesis inhibitive factors will be helpful in the prevention and treatment of hepatocarcinoma.
Objective To study the biological behavior of osteoblast and vascular endothelial cell culture. Methods The osteoblasts and vascular endothelial cells were obtained from calvarial bone and renal cortox of 2-week rabbits respectively. The experiment were divided into group A (osteoblasts), group B (vascular endothelial cells) and group C(co-cultured osteoblasts and vascular endothelial cells). The cells were identified with cytoimmunochemical staining. The cellular biological behavior and compatibilitywere observed under inverted phase contrast microscope and with histological staining. The cells viability and alkaline phosphatase(ALP) activity were measured. Results The cytoimmunochemical staining showed that the cultured cells were osteoblasts and vascular endothelial cells .The cellular compatibility of osteoblasts and vascular endothelial cells was good. The ALP activity was higher in group C than in group A and group B(P＜0.01), and it was higher in group A than in group B(P＜0.05). In group C, the cellproliferation were increased slowly early, but fast later. Conclusion Thecellular compatibility of osteoblasts and vascular endothelial cells were good. The vascular endothelial cells can significantly increased the osteoblast viability and ALP activity,and the combined cultured cells have greater proliferation ability.