Abstract: The mineral in human bone is not pure stoichiometric calcium phosphate (Ca/P) as it is partially substituted by in organic elements. In this study, the copper ions (Cu2+) substituted hydroxyapatite (CuHA) powder has been synthesized by the co-precipitation method. The CuHA powder has been used to fabricate CuHA fiber scaffolds by sol-gel process and the following sinter process. The resulted CuHA fibers have slightly different microstructure (i.e. porosity) compared to HA fiber scaffold, which is denser. The mechanical properties test was used to evaluate CuHA, and the results showed decreases in both compression strength and hardness tests. Moreover, the in vitro used endothelial cells to evaluate the angiogenesis of CuHA. The result illustrated that the viability of endothelial cell on CuHA fiber scaffold surfaces tends to antigenic behavior. The results obtained with CuHA scaffold give this material benefit in biological applications such as antimicrobial, antitumor, antigens, compacts, filling cavities of the tooth and for the deposition of metal implants anti-tumor, anti-cancer, bone filler, and scaffold.
Abstract: Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.
Abstract: 10 clinically healthy hemal nodes were collected from male bulls aged 2-3 years. Light microscopy revealed a capsule of connective tissue consisted mainly of collagen fiber surrounding hemal node, numerous erythrocytes were found in wide subcapsular sinus under the capsule. The parenchyma of the hemal node was divided into cortex and medulla. Diffused lymphocytes, and lymphoid follicles, having germinal centers were the main components of the cortex, while in the medulla there was wide medullary sinus, diffused lymphocytes and few lymphoid nodules. The area occupied with lymph nodules was larger than that occupied with non-nodular structure of lymphoid cords and blood sinusoids. Electron microscopy revealed the cellular components of hemal node including elements of circulating erythrocytes intermingled with lymphocytes, plasma cells, mast cells, reticular cells, macrophages, megakaryocytes and endothelial cells lining the blood sinuses. The lymphocytes were somewhat triangular in shape with cytoplasmic processes extending between adjacent erythrocytes. Nuclei were triangular to oval in shape, lightly stained with clear nuclear membrane indentation and clear nucleoli. The reticular cells were elongated in shape with cytoplasmic processes extending between adjacent lymphocytes, rough endoplasmic reticulum, ribosomes and few lysosomes were seen in their cytoplasm. Nucleus was elongated in shape with less condensed chromatin. Plasma cells were oval to irregular in shape with numerous dilated rough endoplasmic reticulum containing electron lucent material occupying the whole cytoplasm and few mitochondria were found. Nuclei were centrally located and oval in shape with heterochromatin emarginated and often clumped near the nuclear membrane. Occasionally megakaryocytes and mast cells were seen among lymphocytes. Megakaryocytes had multilobulated nucleus and free ribosomes often appearing as small aggregates in their cytoplasm, while mast cell had their characteristic electron dense granule in the cytoplasm, few electron lucent granules were found also, we conclude that, the main function of the hemal node of cattle is proliferation of lymphocytes. No role for plasma cell in erythrophagocytosis could be suggested.
Abstract: The future and the development of science is therefore
seen in interdisciplinary areas such as biomedical engineering. Selfassembled
structures, similar to stem cell niches would inhibit fast
division process and subsequently capture the stem cells from the
blood flow. By means of surface topography and the stiffness as well
as microstructure progenitor cells should be differentiated towards
the formation of endothelial cells monolayer which effectively will
inhibit activation of the coagulation cascade. The idea of the material
surface development met the interest of the clinical institutions,
which support the development of science in this area and are waiting
for scientific solutions that could contribute to the development of
heart assist systems. This would improve the efficiency of the
treatment of patients with myocardial failure, supported with artificial
heart assist systems. Innovative materials would enable the redesign,
in the post project activity, construction of ventricular heart assist.
Abstract: Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al2O3) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al2O3 at curing temperature 50˚C for 4h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.
Abstract: Heme oxygenase-1 (HO-1), an enzyme degrading heme to carbon monoxide, iron, and biliverdin, has been recognized as playing a crucial role in cellular defense against stressful conditions, not only related to heme release. In the present study, the effects of TNF-a on the expression of heme oxygenase-1 (HO-1) in human aortic endothelial cells (HAECs) as well as the related mechanisms were investigated. 10 ng/mL TNF-α treatment significantly increased HO-1 expression after 6h, then a further increase at 12h and declined at 24h. Treatment with 2 ng/mL of TNF-a after 12 h resulted in a significant increase in HO-1 expression, which peaked at 10 ng/mL, then declined at 20 ng/mL. TNF-α induced HO-1 expression and then HO-1 expression reduced vascular cell adhesion molecule-1 (VCAM-1) expression. Phosphorylation studies of ERK1/2, JNK, and p38, three subgroups of mitogen-activated protein kinases (MAPKs) demonstrated TNF-α-induced ERK1/2, JNK, and p38 phosphorylation. The increase in HO-1 expression in response to TNF-α treatment was affected by pretreatment with SP600125 (a JNK inhibitor) and SB203580 (a p38 inhibitor), not with PD98059 (an ERK1/2 inhibitor). The expression of HO-1 was stronger in aortas of TNF-α-treated apo-E deficient mice when compared with control mice. These results suggest that low dose of TNF-α treatment notably induced HO-1 expression was mediated through JNK/p38 phosphorylation and may have a protective potential in cardiovascular diseases and inflammatory response through the regulation of HO-1 expression.
Abstract: Hyperglycemia-mediated accumulation of advanced glycation end-products (AGEs) play a pivotal role in the development of diabetic complications by inducing inflammation. In the present study, we evaluated the possible effects of water/ethanol (1/1, v/v) extracts (WEE) and its fractions from Canarium album Raeusch. (Chinese olive) which is a fruit used on AGEs-stimulated oxidative stress and inflammation in monocytes and vascular endothelial cells. Co-incubation of EA.hy926 endothelial cells with WEE and its fractions for 24h resulted in a significant decrease of monocyte–endothelial cell adhesion, the expression of ICAM-1, generation of intracellular ROS and depletion of GSH induced by AGEs. Chinese olive fruit extracts also reduced the expression of pro-inflammatory mediates, such as TNF-α, IL-1β and IL-6 in THP-1 cells. These findings suggested that Chinese olive fruit was able to protect vascular endothelium from dysfunction induced by AGEs.
Abstract: Atherosclerosis was identified as a chronic inflammatory process resulting from interactions between plasma lipoproteins, cellular components (monocyte, macrophages, T lymphocytes, endothelial cells and smooth muscle cells) and the extracellular matrix of the arterial wall. Several types of genes were known to express during formation of atherosclerosis. This study is carried out to identify unknown differentially expressed gene (DEG) in atherogenesis. Rabbit’s aorta tissues were stained by H&E for histomorphology. GeneFishing™ PCR analysis was performed from total RNA extracted from the aorta tissues. The DNA fragment from DEG was cloned, sequenced and validated by Real-time PCR. Histomorphology showed intimal thickening in the aorta. DEG detected from ACP-41 was identified as cathepsin B gene and showed upregulation at week-8 and week-12 of atherogenesis. Therefore, ACP-based GeneFishing™ PCR facilitated identification of cathepsin B gene which was differentially expressed during development of atherosclerosis.
Abstract: Hyperglycaemia is a key factor that contributes to the
development of diabetes-related microvascular disease and a major
risk factor for endothelial dysfunction. In the current study, we have
explored glucose-induced abnormal intracellular calcium (Ca2+
i)
homeostasis in mouse microvessel endothelial cells (MMECs) in
high glucose (HG) (40mmol/L) versus control (low glucose, LG) (11
mmol/L). We demonstrated that the exposure of MMECs to HG for 3
days did not change basal Ca2+
i, however, there was a significant
increase of acetylcholine-induced Ca2+ entry. Western blots
illustrated that exposure to HG also increased STIM1 (Stromal
Interaction Molecule 1), but not Orai1 (the pore forming subunit),
protein expression levels. Although the link between HG-induced
changes in STIM1 expression, enhanced Ca2+ entry and endothelial
dysfunction requires further study, the current data are suggestive
that targeting these pathways may reduce the impact of HG on
endothelial function.
Abstract: Mechanical interaction between endothelial cells (ECs) and the extracellular matrix (or collagen gel) is known to influence the sprouting response of endothelial cells during angiogenesis. This influence is believed to impact on the capability of endothelial cells to sense soluble chemical cues. Quantitative analysis of endothelial-cell-mediated displacement of the collagen gel provides a means to explore this mechanical interaction. Existing analysis in this context is generally limited to 2D settings. In this paper, we investigate the mechanical interaction between endothelial cells and the extracellular matrix in terms of the endothelial-cellmediated displacement of the collagen gel in both 2D and 3D. Digital image correlation and Digital volume correlation are applied on confocal reflectance image stacks to analyze cell-mediated displacement of the gel. The skeleton of the sprout is extracted from phase contrast images and superimposed on the displacement field to further investigate the link between the development of the sprout and the displacement of the gel.
Abstract: Human immunodeficiency virus infection and
acquired immunodeficiency syndrome is a global pandemic with
cases reporting from virtually every country and continues to be a
common infection in developing country like India.
Microalbuminuria is a manifestation of human immunodeficiency
virus associated nephropathy. Therefore, microalbuminuria may be
an early marker of human immunodeficiency virus associated
nephropathy, and screening for its presence may be beneficial. A
strikingly high prevalence of microalbuminuria among human
immunodeficiency virus infected patients has been described in
various studies. Risk factors for clinically significant proteinuria
include African - American race, higher human immunodeficiency
virus ribonucleic acid level and lower CD4 lymphocyte count. The
cardiovascular risk factors of increased systolic blood pressure and
increase fasting blood sugar level are strongly associated with
microalbuminuria in human immunodeficiency virus patient. These
results suggest that microalbuminuria may be a sign of current
endothelial dysfunction and micro-vascular disease and there is
substantial risk of future cardiovascular disease events. Positive
contributing factors include early kidney disease such as human
immunodeficiency virus associated nephropathy, a marker of end
organ damage related to co morbidities of diabetes or hypertension,
or more diffuse endothelial cells dysfunction. Nevertheless after
adjustment for non human immunodeficiency virus factors, human
immunodeficiency virus itself is a major risk factor. The presence of
human immunodeficiency virus infection is independent risk to
develop microalbuminuria in human immunodeficiency virus patient.
Cardiovascular risk factors appeared to be stronger predictors of
microalbuminuria than markers of human immunodeficiency virus
severity person with human immunodeficiency virus infection and
microalbuminuria therefore appear to potentially bear the burden of
two separate damage related to known vascular end organ damage
related to know vascular risk factors, and human immunodeficiency
virus specific processes such as the direct viral infection of kidney
cells.The higher prevalence of microalbuminuria among the human
immunodeficiency virus infected could be harbinger of future
increased risks of both kidney and cardiovascular disease. Further
study defining the prognostic significance of microalbuminuria
among human immunodeficiency virus infected persons will be
essential. Microalbuminuria seems to be a predictor of cardiovascular
disease in diabetic and non diabetic subjects, hence it can also be
used for early detection of micro vascular disease in human
immunodeficiency virus positive patients, thus can help to diagnose
the disease at the earliest.
Abstract: To identify an endothelial cell-specific promoter suitable for vascular-specific targeting, we tested five promoters in vitro--Tie2SE, Tie2LE, ICAM2, Flt-1 and vWF--for promoter activity and specificity in endothelial cells, smooth muscle cells and non-vascular resident cells as well as tissues. These promoters, except for vWF, exhibited good endothelial activity and specificity in vitro. In a syngenic heart transplantation model, the ICAM2 promoter was variably functional in coronary endothelial cells of donor hearts. Thus, the ICAM2, Flt-1, Tie2SE and Tie2LE promoters hold promise for endothelial-specific targeting, but in vitro expression may not predict in vivo expression.