This study was aimed to compare the differences of adhesion properties of endothelial cells (EC) from arteries (AEC), veins (VEC) and capillaries (MVEC) under shear stress condition, and to explore whether they can get the same adhesive ability as graft in similar shear stress conditions. With mended parallel plate flow apparatus and peristalsis pump providing fluid shear stress used, endothelial culture models were established in vitro with the same environmental factors as steady culture. To compare the adhesion among three kinds of endothelial cells under dynamic condition and static condition, the dynamic change of cytoskeletal actin filaments and the effects of different adhesive proteins coated on the adhesion of EC to the glass were studied. The cultured endothelial cells under flow conditions were extended and arranged along the direction of flow. The adhesive ability from high to low under static condition were AEC, MVEC and VEC (VEC compared with AEC or MVEC, P < 0.05), sequentially. The adhesion of endothelial cells from variety sources under dynamic culture condition was significantly increased than that of the static groups. The ratio of cell retention was not significantly different between AEC and MVEC. But VEC was significantly different (P < 0.05) compared with AEC or MVEC. The actin filaments (F-actin) were bundled together and arranged along the direction of flow after fluid culture. Dense peripheral band (DPB) gradually disappeared and distinct stress fibers were formed, which even interconnected to form a whole in the MVEC. The adhesion of AEC, VEC and MVEC under shear stress conditions are more significantly increased than those under the static culture conditions, and the MVEC can achieve the same adhesion as AEC.
Arteries ; cytology ; Capillaries ; cytology ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cells, Cultured ; Cytoskeleton ; physiology ; Endothelial Cells ; cytology ; physiology ; Humans ; Shear Strength ; Stress, Mechanical ; Veins ; cytology

OBJECTIVETo investigate the influence of vacuum-assisted closure (VAC) technique on the growth of capillaries in the wound of the pig produced by explosion.

METHODSFour small white pigs were inflicted with 16 explosion wounds [(7.3 +/- 1.0) cm2 in area] on both sides of the buttocks, shoulders and hips by detonation of a specific type of explosive, and the wounds were randomly divided into 2 groups, i. e, control (C, with conventional treatment from 2 post-injury day (PID) on and treatment (T, with VAC treatment after debridement from 2 PID on) groups, with 8 wounds in each group. Wound tissues of 2mm x 2mm x 2mm in size were harvested for pathological examination before treatment and on 1 and 3 post-treatment day (PTU). The differentiation of adventitial cells were examined with light microscope, and the pixel value of desmin positive particles and the luminal area of newly formed capillaries were assessed with Image C software.

RESULTSMost of vessels in the wound of both groups were in elliptic shape when observed in longitudinal section. In C group, few newly formed capillaries vessels with lack of pericytes were observed before treatment and on 1, 3 PTD, then the number began to increase on 6 PTD. In T group, the number of newly formed capillaries with pericytes was increased on 1 PTD, and it continued to increase thereafter. The pixel values of desmin positive particles in C group on 1, 3, and 6 PTD were (91 +/- 54), (199 +/- 85), and (1552 +/- 298), respectively, which were obviously higher than those in T group [(2569 +/- 330), (3984 +/- 377), (9611 +/- 960), P < 0.01]. The area of vessel lumen in C group was (59 +/- 36), (250 +/- 70), and (938 +/- 287) microm2, respectively on 1, 3, and 6 PTD, which was also smaller than those in T group [(818 +/- 234), (4518 +/- 1080), and (9058 +/- 1656) microm2, P < 0.01].

CONCLUSIONCompared with conventional therapy, VAC can not only accelerate the formation of new capillaries, but also enhance the differentiation of pericytes and the process of enwrapping them around the vessels, and increase the luminal area of newly formed capillaries.

Animals ; Blast Injuries ; therapy ; Capillaries ; cytology ; growth & development ; Female ; Male ; Negative-Pressure Wound Therapy ; Neovascularization, Physiologic ; Swine ; Wound Healing

OBJECTIVETo investigate the mechanisms of baicalin on anti-cerebral ischemic through observing the effect of baicalin on human brain microvascular endothelial cell under the glucose deprivation combined with hypoxia condition.

METHODSHuman brain microvascular endothelial cells (HBMVECs) cultured in vitro were divided into the following groups: normal group, model group, baicalin high dose group, baicalin middle dose group, baicalin low dose group, nimodipine group. The kits were used to detect the cell viability, leakage rate of lactate dehydrogenase (LDH), Na(+)-K(+)-ATPase, Ca(2+)-ATPase, the concentration of Ca2+ in each group, and apoptosis rates of each group were analyzed by flow cytometry (FCM).

RESULTSCompared with the normal group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase in model group decreased significantly (P < 0.01, P < 0.05). But the leakage rate of LDH, Ca2+ in cells and apoptosis rates increased remarkably (P < 0.01). Compared with the model group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase increased obviously (P < 0.01, P < 0.05) in baicalin high dose group. But the leakage rate of LDH and Ca2+ in cells in baicalin high dose group decreased significantly comparing with that of model group (P < 0.05, P < 0.01). And the reduction of intracellular Ca2+ was superior to that of nimodipine group. Meanwhile, the apoptosis rates decreased significantly in both baicalin high and middle dose groups (P < 0.01).

CONCLUSIONBaicalin could improve the cell viability of HBMVECs under the glucose deprivation combined with hypoxia condition. And the mechanisms were related with improving the energy metabolism, inhibiting intracellular calcium overload and decreasing the apoptosis rate of cells further.

Apoptosis ; Brain ; blood supply ; Calcium ; metabolism ; Capillaries ; cytology ; Cell Hypoxia ; Cell Survival ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Flavonoids ; pharmacology ; Glucose ; chemistry ; Humans

OBJECTIVETo globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogenesis system with Affymetrix oligonucleotide array.

METHODSA microcarrier-based in vitro angiogenesis system was developed, in which endothelial cells (ECs) migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed network. The total RNA samples from the HMVECs at the selected time points (0.5 h, 24 h, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the software provided by the manufactory. The expression patterns of some genes were validated and confirmed by Semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines/chemokine receptors were specially examined and their functional implications were analyzed.

RESULTSAbout 1500 genes were found up- or down- regulated 2-folds or above detected by the arrays, and among them, about 400 genes regulated 3-folds or above. The regulated genes could be grouped into categories based on their molecular functions such as growth factor and receptor, cell proliferation, extracellular matrix, cell cycle and apoptosis, signaling molecule and transcription factor, and so on, using the Gene Ontology Mining Tool in The NetAffx Analysis Center. The regulated genes were also clustered into six groups based on their patterns of expression. As for chemokines, the CCL2/MCP-1, CCL5/RANTES and CX3CL1 were identified to be specially upregulated at 24 h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis.

CONCLUSIONSBased on our angiogenesis model, and by oligonucleotide arrays, the present study demonstrates global profiles of the gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be encouraged.

Capillaries ; cytology ; Cells, Cultured ; Chemokines ; genetics ; Endothelial Cells ; cytology ; Endothelium, Vascular ; cytology ; physiology ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; In Vitro Techniques ; Neovascularization, Physiologic ; genetics ; Oligonucleotide Array Sequence Analysis ; Receptors, Chemokine ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Adenocarcinoma ; blood supply ; physiopathology ; Capillaries ; metabolism ; ultrastructure ; Cell Separation ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; ultrastructure ; Humans ; Microcirculation ; Stomach Neoplasms ; blood supply ; physiopathology

OBJECTIVETo investigate the therapeutic effectiveness of intracoronary implantation of autologous bone marrow mononuclear cells (BM-MNC) in miniswine model of reperfused myocardial infarction.

METHODSSixteen miniswine myocardial ischemic reperfusion injury models made by ligation of the distal one third segment of left anterior descending artery for 90 minutes were randomized into 2 groups. In BM-MNC group (n = 9), (3.54 +/- 0.90) X 10(8) BM-MNC were intracoronary injected, and in the control group (n = 7), phosphate buffered saline was injected by the same way. Echocardiographic and hemodynamic results, vessel density, and myocardial infarction size were evaluated and compared before and 4 weeks after cell transplantation.

RESULTSIn BM-MNC group, there were no differences between before and 4 weeks after transplantation in aspects of left ventricular ejection fraction (LVEF), interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, or +dp/dtmax. In control group, LVEF, interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, and +dp/dtmax decreased significantly 4 weeks after transplantation (P < 0.05). Left ventricular end-diastolic pressure and -dp/dtmax, did not change significantly before and after cell transplantation in both groups. Capillary density in BM-MNC group was greater than that in control group [(13.39 +/- 6.96)/high power field vs. (3.50 +/- 1.90)/high power field, P < 0.05]. Infarction area assessed by tetrazolium red staining and the infarction percentage decreased in BM-MNC group compared with those in control group (P < 0.05).

CONCLUSIONSTransplantation of BM-MNC into myocardium with ischemic reperfusion injury increases capillary density and decreases infarction area. It has significantly beneficial effect on cardiac systolic function rather than on diastolic function.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Bone Marrow Transplantation ; Capillaries ; physiology ; Echocardiography ; Heart ; anatomy & histology ; physiology ; physiopathology ; Hemodynamics ; Myocardial Ischemia ; Random Allocation ; Swine ; Systole ; physiology ; Transplantation, Autologous ; physiology

In this work, blank polylactic acid (PLA) nanoparticles with unstained surface were prepared by the nano-deposition method. On the basis of the preparation, the effect of surface modification on brain microvascular endothelial cells (BMECs) targeting was examined by in vivo experiments and fluorescence microscopy. The results showed that PLA nanoparticles are less toxic than PACA nanoparticles but their BMECs targeting is similar to PACA nanoparticles. The experiments suggest that drugs can be loaded onto the particles and become more stable through adsorption on the surface of PLA nanoparticles with high surface activity. The surface of PLA nanoparticles was obviously modified and the hydrophilicity was increased as well in the presence of non-ionic surfactants on PLA nanoparticles. As a targeting moiety, polysobate 80 (T-80) can facilitate BMECs targeting of PLA nanoparticles.
Brain/*blood supply ; Brain/drug effects ; Capillaries/cytology ; Capillary Permeability/drug effects ; Drug Delivery Systems ; Endothelium, Vascular/*cytology ; Lactic Acid/*pharmacology ; Mice, Inbred Strains ; Microscopy, Fluorescence ; Nanoparticles ; Polymers/*pharmacology

OBJECTIVE: To determine the effects of naoyian (NYA) serum on the expression of vascular endothelial growth factor (VEGF) protein in cultured rat cerebral microvascular endothelial cell (RCMEC) with hypoxia. METHODS: NYA serum was separated from rat heart which had been filled stomach with NYA successively for 3 days. The rat cerebral microvascular endothelial cells were taken from the Sprageu-Dawley rat brain at postborn 7 days. The rat cerebral microvascular endothelial cells were incubated at anaerobic incubator to establish the hypoxia models. The vigo of RCMEC was determined by MTT. The level of expression of VEGF protein was measured by cell immunohistochemistry and Western blot. RESULTS: The OD value of NYA serum group was higher than the control groups after hypoxia for 18 hours. VEGF protein was increased by hypoxia in cerebral microvascular endothelial cells (P < 0.05). The content of VEGF protein in NYA serum containing medium was more significantly elevated than those cultured in other control media (P < 0.01). CONCLUSION VEGF protein was induced by hypoxia in rat cerebral microvascular endothelial cells, and NYA could upregulate the expression of VEGF protein, which may be one of the protection mechanisms for cerebral microvascular endothelial cells.
Animals ; Animals, Newborn ; Capillaries ; cytology ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; blood supply ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; cytology ; metabolism ; Female ; Male ; Rats ; Rats, Sprague-Dawley ; Serum ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVETo study the influence of conditioned medium of rat brain microvascular endothelial cells on mitochondrial function of cortical neurons and the protective effect of Tongluo Jiunao Injection (TJI) on it.

METHODSFour kinds of conditioned endothelial cell (EC) cultured medium were prepared, i.e. the N-CM medium prepared by EC cultured in the normal conditioned medium without any treatment; the NT-CM prepared by EC cultured in N-CM and treated with TJI 1 microl/ml for 10 h; the I-CM prepared by EC cultured in the non-glucose kreb medium under hypoxia condition; and the IT-CM by EC pre-treatce with TJI 1 microl/ml for 4 h and cultured as that of I-CM. The levels of neuronic mitochondrial activity, membrane potential (MMP) and cytochrome C (Cyt C) were determined before and after the glucose-oxygen deprived model neurons of brain cortex being cultured with different kinds of conditioned EC cultured medium for assessing the effects of these media on mitochondria of injured neuron.

RESULTSAs compared with those of the normal neuron, the mitochondrial activity and MMP of all injured neurons decreased and Cyt C level increased significantly. But comparison of these indexes among neurons cultured with different conditioned EC culture media showed that the greatest extent abnormality revealed in the N-CM cultured neurons, which even greater than that in the model neuron; while that was less in the N-CM cutured neuron than in model neuron; as for those cultured in the NT-CM and IT-CM, i.e. the TJI treated cuture medium, the abnormal changes were reduced significantly when compared with those cultured in medium untreated with TJI (N-CM and I-CM), respectively (all P < 0.05).

CONCLUSIONThe paracrine secretion of the brain microvascular endothelial cells has evident regulatory effect on survival of the injured neurons, which might possibly be related to its protective effect on neuron mitochondrial function, and TJI could enhance the protective effect.

Animals ; Capillaries ; cytology ; Cells, Cultured ; Cerebral Cortex ; blood supply ; cytology ; Culture Media, Conditioned ; pharmacology ; Cytochromes c ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; ultrastructure ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; metabolism ; physiology ; Neurons ; cytology ; drug effects ; ultrastructure ; Rats

OBJECTIVE: To evaluate the fixation strength and tissue reaction of the glue fixation and self-stabilizing leg fixation methods and to compare the results with those of the conventional tagging suture fixation method. MATER AND METHODS: Twelve healthy rabbits were selected and three different methods of implanting the port chamber were employed on the back of each rabbit. A total of thirty six port chambers were implanted with these three different methods, viz. the glue fixation method using tissue adhesive, the self-stabilizing leg method using a self-expandable stabilizing leg, and the suture fixation method. The fixation strength and the gross and histopathologic changes of each fixation method were evaluated at three days, one week, two weeks and four weeks after port implantation. RESULTS: The glue fixation method showed a good fixation strength, which was similar to that of the tagging suture method (p=0.3486). Five of the six ports (83%) implanted with the glue fixation method which were examined after two weeks showed cracks on the external surface, but this had no adverse effects on their function. A large amount of granulation tissue reaction was found at the bottom of the chamber (p=0.0025). The fixation with the self-stabilizing leg showed relatively lower fixation strength (p=0.0043), but no turning-over of the chamber occurred. The fixation strength improved with time after the first week, and minimal granulation tissue reaction was observed with this method. CONCLUSION: The glue fixation method exhibited equal fixation strength compared to the suture fixation, but showed cracking and a large amount of granulation tissue, whereas the fixation with a self-stabilizing leg showed weaker fixation strength.
Alloys ; Animals ; Capillaries/cytology/metabolism/pathology ; Cell Proliferation ; Device Removal ; Enbucrilate/therapeutic use ; *External Fixators ; Fibroblasts/metabolism/pathology ; Granulation Tissue/blood supply/metabolism/pathology ; *Implants, Experimental ; Models, Animal ; Rabbits ; Sutures/*utilization ; Time Factors ; Tissue Adhesives/*therapeutic use