Tumor Necrosis Factor-α Regulates Heme Oxygenase-1 Expression in Endothelial Cells via the Phosphorylation of JNK/p38

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.

Human Elastin-derived Biomimetic Coating Surface to Support Cell Growth

A new sythetic gene coding for a Human Elastin-Like Polypeptide was constructed and expressed. The recombinant product was tested as coating agent to realize a surface suitable for cell growth. Coatings showed peculiar features and different human cell lines were seeded and cultured. All cell lines tested showed to adhere and proliferate on this substrate that has been shown also to exert a specific effect on cells, depending on cell type.

Alignment of MG-63 Osteoblasts on Fibronectin-Coated Phosphorous Doping Lattices in Silicon

A major challenge in biomaterials research is the regulation of protein adsorption which is a key factor for controlling the subsequent cell adhesion at implant surfaces. The aim of the present study was to control the adsorption of fibronectin (FN) and the attachment of MG-63 osteoblasts with an electronic nanostructure. Shallow doping line lattices with a period of 260 nm were produced for this purpose by implantation of phosphorous in silicon wafers. Protein coverage was determined after incubating the substrate with FN by means of an immunostaining procedure and the measurement of the fluorescence intensity with a TECAN analyzer. We observed an increased amount of adsorbed FN on the nanostructure compared to control substrates. MG-63 osteoblasts were cultivated for 24h on FN-incubated substrates and their morphology was assessed by SEM. Preferred orientation and elongation of the cells in direction of the doping lattice lines was observed on FN-coated nanostructures.

Evaluation of Antiglycation Effects of Extracts Obtained from Canarium album Raeusch Fruit and Beneficial Activity on Advanced Glycation Endproduct-Mediated Oxidative Stress and Inflammation in Monocytes and Vascular Endothelial Cells

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. 

Interactions between Cells and Nanoscale Surfaces of Oxidized Silicon Substrates

The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

Expression of Gen Extracellular Matrix and Cell Adhesion Molecule of Brain Embrio Mice at GD-10 By Real Time RT-PCR

research goal was to determine the expression levels cDNA of brain embrio at gestation days 10 (GD-10). The Electroforesis DNA results showed that GAPDH, Fibronectin1, Ncam1, Tenascin, Vimentin, Neurofilament heavy, Neurofilament medium and Neurofilament low were 447 bp, 462 bp, 293 bp. 416 bp, 327 bp, 301 bp, 398 bp and 289 bp. Result of real-time RT-PCR on brain Embryo at gestation days 10 showed that the expression of copy gen Fibronectin 36 copies, Ncam 21,708 copies; Tenascin 24,505 copies; Vimentin 538,554 copies; Neurofilament heavy 2,419 copies; Neurofilament medium 92,928 copies; Neurofilament low 125,809 copies. Vimentin expressed gene copies is very high compared with other gene copies. This condition are caused by Vimentin, that contribute to proliferate of brain development. The vimentin role to cell proliferation of brain.

Effect of Cold Plasma-Surface Modification on Surface Wettability and Initial Cell Attachment

A thin coating of hexamethyldisiloxane and subsequent O2-plasma treatment was performed on mirror-polished titanium in order to regulate the wide range of wettability including 106 and almost 0 degrees of contact angles. The adsorption behavior of fibronectin and albumin in both individual and competitive mode, and initial attachment of fibroblasts and osteoblasts were investigated. Individually, fibronectin adsorption showed a biphasic inclination, whereas albumin showed greater adsorption to hydrophobic surfaces. In competitive mode, in solution containing both fibronectin and albumin, fibronectin showed greater adsorption on hydrophilic surfaces, whereas Alb predominantly adsorbed on hydrophobic surfaces. Initial attachment of both cells increased with increase in surface wettability, in particular, on super-hydrophilic surface, which correlated well with fibronectin adsorption in competitive mode. These results suggest that a cold plasma-surface modification enabled to regulate the surface wettability, and fibronectin adsorption may be responsible for increasing cell adhesion on hydrophilic surfaces in a body fluid