Bovine Aorta Research Articles

An in vitro feasibility study of 355 nm laser atherectomy for the treatment of peripheral atherosclerotic lesions.

In this study, we utilized a novel 355 nm laser to ablate porcine aortas in the presence of physiological saline and contrast agent. Subsequently, we investigated the shape and depth of the resulting injuries. After ablating bovine tendons and aortas with the laser, we analyzed the size and quantity of particles postablation. Finally, we conducted ablation experiments using human ex vivo plaques. The analysis revealed minimal damage to porcine aortas within 2 s of exposure to the 355 nm laser. The degree of injury in the presence of contrast agent was higher than that in the presence of physiological saline but significantly lower than the damage caused by 308 nm laser. Regardless of whether it was bovine tendon or porcine aorta tissue, the proportion of particles <25 μm postlaser ablation exceeded 99%. Lastly, the 355 nm laser successfully opened three types of plaques: chronically occluded, stent restenosis, and stale thrombosis.

Transcriptomics of Besnoitia besnoiti-Infected Fibroblasts Reveals Hallmarks of Early Fibrosis and Cancer Progression.

Endothelial injury, inflammatory infiltrate and fibrosis are the predominant lesions in the testis of bulls with besnoitiosis that may result in sterility. Moreover, fibroblasts, which are key players in fibrosis, are parasite target cells in a Besnoitia besnoiti chronic infection. This study aimed to decipher the molecular basis that underlies a drift toward fibrosis during the disease progression. Transcriptomic analysis was developed at two times post-infection (p.i.), representative of invasion (12 h p.i.) and intracellular proliferation (32 h p.i.), in primary bovine aorta fibroblasts infected with B. besnoiti tachyzoites. Once the enriched host pathways were identified, we studied the expression of selected differentially expressed genes (DEGs) in the scrotal skin of sterile infected bulls. Functional enrichment analyses of DEGs revealed shared hallmarks of cancer and early fibrosis. Biomarkers of inflammation, angiogenesis, cancer, and MAPK signaling stood out at 12 h p.i. At 32 h p.i., again MAPK and cancer pathways were enriched together with the PI3K-AKT pathway related to cell proliferation. Some DEGs were also regulated in the skin samples of naturally infected bulls (PLAUR, TGFβ1, FOSB). We have identified potential biomarkers and host pathways regulated during fibrosis that may hold prognostic significance and could emerge as potential therapeutic targets.

An aorta ECM extracted hydrogel as a biomaterial in vascular tissue engineering application.

Biological scaffolds have been undergoing significant growth in tissue engineering applications over the last years. Biopolymers extracted from ECM with various protein factors and other biological agents have been active in restoring damaged tissue. In the present study, bioactive scaffold is prepared from bovine aorta extracted natural polymeric hydrogel with advantages of availability and cost-effectiveness. The biological scaffolds were prepared through freeze-drying method to make a 3D sponge with appropriate structure, well-defined architecture and interconnected pores for vascular tissue engineering, and studied the effect of aorta hydrogel concentrations (1, 2, 3, and 4% w/v) on the scaffolds. The prepared biological scaffolds were analyzed by mechanical tests, FTIR, SEM, porosity and PBS absorption. Moreover, the morphology and proliferation of human umbilical vein cord cells on the 3D sponges were investigated. Histological analysis including, Masson trichrome (MT), hematoxylin and eosin (H&E), Verhoeff/Van Gieson (VVG) and alcian blue (AB) revealed that during this process the main components of aorta extracellular matrix containing collagen, elastin, and glycosaminoglycan were well preserved. The obtained results revealed that the scaffolds porosity were more than 90%. The Aorta-ECM4% enabled HUVECs to survive, proliferate and migrate better than 2% and 3% aorta-ECM.

Decellularized bovine aorta as a promising 3D elastin scaffold for vascular tissue engineering applications.

Aim: To evaluate the suitability of using aorta elastin scaffold, in combination with human adipose-derived mesenchymal stem cells (hAd-MSCs), as an approach for cardiovascular tissue engineering. Materials & Methods: Human adipose-derived MSCs were seeded on elastin samples of decellularized bovine aorta. The samples were cultured in vitro to investigate the inductive effects of this scaffold on the cells. The results were evaluated using histological, and immunohistochemical methods, as well as MTT assay, DNA content, reverse transcription-PCR and scanning electron microscopy. Results: Histological staining and DNA content confirmed the efficacy of decellularization procedure (82% DNA removal). MTT assay showed the construct's ability to support cell viability and proliferation. Cell differentiation was confirmed by reverse transcription-PCR and positive immunohistochemistry for alfa smooth muscle actin and von Willebrand. Conclusion: The prepared aortic elastin samples act as a potential scaffold, in combination with MSCs, for applications in cardiovascular tissue engineering. Further experiments in animal models are required to confirm this.

Reversible electroadhesion of hydrogels to animal tissues for suture-less repair of cuts or tears

Electroadhesion, i.e., adhesion induced by an electric field, occurs between non-sticky cationic and anionic hydrogels. Here, we demonstrate electroadhesion between cationic gels and animal (bovine) tissues. When gel and tissue are placed under an electric field (DC, 10 V) for 20 s, the pair strongly adhere, and the adhesion persists indefinitely thereafter. Applying the DC field with reversed polarity eliminates the adhesion. Electroadhesion works with the aorta, cornea, lung, and cartilage. We demonstrate the use of electroadhesion to seal cuts or tears in tissues or model anionic gels. Electroadhered gel-patches provide a robust seal over openings in bovine aorta, and a gel sleeve is able to rejoin pieces of a severed gel tube. These studies raise the possibility of using electroadhesion in surgery while obviating the need for sutures. Advantages include the ability to achieve adhesion on-command, and moreover the ability to reverse this adhesion in case of error.

Studying the anticancer activity in vitro of doxorubicin-encapsulated pH-sensitive redox nanoparticles

Doxorubicin (DOX) is known as a chemotherapeutic agent for treating a variety of cancers. However, due to non-specific distribution, DOX causes severe side effects, leading to the limitation of its clinical application. In this study, pH-sensitive redox nanoparticles containing DOX were studied to form drug-coated nanoparticles with the size of about 40-60 nm that increases the concentration of drugs accumulated at the tumor site in response to low pH conditions. The optimal properties of the nanoparticles were investigated by analysing the antioxidant activity, sensitivity to pH, drug delivery in a low pH environment, and toxicity on cancer cells, thereby observing the possibility of inhibiting the migration of cancer cells. Results showed that the combination of DOX and the pH-sensitive redox nanoparticles not only increases anticancer in colorectal cancer (C-26), breast cancer (MCF-7) but also reduces bovine aorta endothelial cells (BAEC) and inhibits C-26 cell migration in vitro.Thepaper results reveal the application potential of the pH-sensitive redox nanoparticles in improving the effectiveness of cancer treatment.

Revisiting Ehrlichia ruminantium Replication Cycle Using Proteomics: The Host and the Bacterium Perspectives.

The Rickettsiales Ehrlichia ruminantium, the causal agent of the fatal tick-borne disease Heartwater, induces severe damage to the vascular endothelium in ruminants. Nevertheless, E. ruminantium-induced pathobiology remains largely unknown. Our work paves the way for understanding this phenomenon by using quantitative proteomic analyses (2D-DIGE-MS/MS, 1DE-nanoLC-MS/MS and biotin-nanoUPLC-MS/MS) of host bovine aorta endothelial cells (BAE) during the in vitro bacterium intracellular replication cycle. We detect 265 bacterial proteins (including virulence factors), at all time-points of the E. ruminantium replication cycle, highlighting a dynamic bacterium–host interaction. We show that E. ruminantium infection modulates the expression of 433 host proteins: 98 being over-expressed, 161 under-expressed, 140 detected only in infected BAE cells and 34 exclusively detected in non-infected cells. Cystoscape integrated data analysis shows that these proteins lead to major changes in host cell immune responses, host cell metabolism and vesicle trafficking, with a clear involvement of inflammation-related proteins in this process. Our findings led to the first model of E. ruminantium infection in host cells in vitro, and we highlight potential biomarkers of E. ruminantium infection in endothelial cells (such as ROCK1, TMEM16K, Albumin and PTPN1), which may be important to further combat Heartwater, namely by developing non-antibiotic-based strategies.

Abstract 14618: Bicuspid Aortic Valve Morphology and Risk Factors of Abnormal Hemodynamics: An Experimental Investigation of Velocity, Vorticity and Eccentricity of Systolic Jet Stream, Using a Magnetic Resonance Imaging Compatible Pulsatile Flow Circulation System

Introduction: Abnormal hemodynamics with bicuspid aortic valve (BAV) is influenced by the BAV morphology. However, due to the morphological variety, the relationship between BAV morphology and hemodynamics has not been well clarified. We experimentally investigated influences of BAV morphology on hemodynamics. Methods: An MRI compatible pulsatile flow circulation system incorporating an aortic valve model and morphologically relevant aortic arch model was developed. Two types of BAV models with cusp angles of 240-120 (asymmetric BAV) and 180-180 (symmetric BAV) were prepared using bovine aorta and pericardium. The vorticity and eccentricity of jet in the aortic arch model in systolic phase were assessed using 4D-flow MRI. Streamlines at peak systole were compared among 5 BAV morphologies defined based on the symmetry and the position of leaflets (Fig). Results: Eccentric supra-valvular jets directed to the aortic wall faced to the smaller leaflet were present in the 3 asymmetric BAVs. In the ABAV-1, a markedly eccentric jet impinging on the aortic outer curvature was present. In the ABAV-2, a left-posterior directed jet shifting to the outer curvature of proximal arch was present. The asymmetric BAVs induced larger flow vorticity in the ascending aorta than the symmetric BAVs (ABAV-1 vs -2 vs -3 vs SBAV-1 vs -2, 0.018 vs 0.019 vs 0.014 vs 0.010 vs 0.011 m 2 /s). The ABAV-2 had a larger vortex in the middle arch than the ABAV-1 (ABAV-2 vs -1, 0.010 vs 0.005 m 2 /s). Conclusion: Our study indicated that the angles and orientations of the BAV impacted on the locations of jets impinging on the aortic wall and magnitudes of vorticities in systole. In the asymmetric BAVs, the direction of jet was influenced by the position of smaller leaflet. Our data suggests that the ABAV-1 morphology may be a risk factor inducing asymmetric aneurysm bulged toward the aortic outer curvature, while the ABAV-2 morphology may be a risk factor of an aortic aneurysm involving the transvers arch.

Synthesis, characterization and biological evaluation of octyltrimethylammonium tetrathiotungstate.

Octyltrimethylammonium tetrathiotungstate salt (ATT-C8) was synthesized and its ability to chelate copper was evaluated. The biological and toxic aspects were evaluated by in vitro and in vivo assays, using bovine aorta endothelial cells (BAEC) and zebrafish (Danio rerio) embryos. The obtained results suggest that ATT-C8 has better biocompatibility, showing a significantly lower lethal concentration 50 (LC50) value in comparison to ammonium tetrathiotungstate (ATT). Zebrafish embryos assay results indicate that both tetrathiotungstate salts at the studied concentrations increase the hatching time. Even more, an in vivo assay showed that synthesized materials behave as copper antagonists and have the ability to inhibit its toxicological effects. Also, both materials were found to be active for the in vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The characterization of the materials was carried out using the following spectroscopic techniques: Ultraviolet-Visible (UV-Vis), Fourier Transform Infrared (FTIR) and proton nuclear magnetic resonance (1H-NRM).

Modulating electrospun polycaprolactone scaffold morphology and composition to alter endothelial cell proliferation and angiogenic gene response.

The aim of this study was to look at how the composition and morphology of polymer scaffolds could be altered to create an optimized environment for endothelial cells. Four polycaprolactone (PCL) scaffolds were electrospun with increasing fibre diameters ranging from 1.64 μm to 4.83 μm. The scaffolds were seeded with human umbilical vein endothelial cells (HUVEC) and cultured for 12 days. PCL scaffolds were then electrospun incorporating decellularized bovine aorta ECM and cultured in a hypoxic environment. We noted deeper cell infiltration on the largest fibre diameter compared to the other three scaffolds which resulted in an increase in the gene expression of CD31; a key angiogenic marker. Increased cell viability and cell proliferation were also noted on the largest fibre. Furthermore, we noted that the incorporation of extracellular matrix (ECM) had minimal effect on cell viability, both in normoxic and hypoxic culture conditions. Our results showed that these environments had limited influences on hypoxic gene expression. Interestingly, the major findings from this study was the production of excretory ECM components as shown in the scanning electron microscopy (SEM) images. The results from this study suggest that fibre diameter had a bigger impact on the seeded HUVECs than the incorporation of ECM or the culture conditions. The largest fibre dimeter (4.83 μm) is more suitable for seeding of HUVECs.

Selection of biological prosthesis for abdominal wall repair on the basis of in vitro biocompatibility determination.

Research on prostheses for repairing abdominal wall defects has progressed through past decades for developing an ideal prosthesis. The study was designed to compare different extracellular matrix (ECM) derived biological prostheses as alternate to conventional synthetic polymeric prostheses for the repair of full thickness abdominal wall defects. Five biological scaffolds derived from bovine diaphragm, bovine aorta, bovine gall bladder, porcine gall bladder, and rabbit skin were prepared and screened for their in vitro biocompatibility. Decellularized ECMs were subjected to various biocompatibility analyses, namely, water absorption potential, matrix degradation analysis, biomechanical testing, and cytocompatibility analysis. Though the rabbit skin displayed maximum biomechanical strength, due to its rapid degradation, it failed to fulfill the criteria of an ideal prosthesis. ECMs derived from bovine diaphragm and aorta were found to be superior than others based upon hydration and matrix degradation analysis, with best scores for bovine diaphragm followed by bovine aorta. The bovine diaphragm and aorta also displayed sufficient biomechanical strength, with diaphragm being the second highest (next to rabbit skin), in biomechanical strength followed by aorta. None of the biological prosthesis revealed any cytotoxicity. Thus, bovine diaphragm and aorta derived ECM fulfill the necessary criteria for their use as biological prosthesis. Because these prostheses are biocompatible, apart from their low cost, ease of availability, and simple preparation, they present a potential alternative to synthetic prosthesis for repair of abdominal wall defects, especially in veterinary patients.

Efficient isolation and identification of primary endothelial cells from bovine aorta by collagenase P

Introduction: Endothelial cells are widely used among researchers for investigation of cardiovascular diseases, particularly atherosclerosis. Since aortic endothelial cells are able to be cultured in high passages, these cells are suitable for physiological and pathological studies of blood vessels. Objectives: The aim of this study was employing a digestion method to isolate the endothelial cells from bovine aorta by utilizing collagenase P and, establishing and characterizing isolated primary endothelial cells (IPECs) cultures. Materials and Methods: IPECs were isolated from fresh bovine aorta via enzymatic digestion method using collagenase P. Cell morphology and its functions were assessed by measuring the gene expression of endothelial nitric-oxide synthase (eNOS) and endothelin-1. In order to validate them as IPECs, they were compared with bovine aortic endothelial cells (BAECs) and vascular smooth muscle cells (VSMCs). Effects of synthetic endothelin-1 (100 nm) were assessed on the phosphorylation of Smad2 transcription factor via western blotting in IPECs for periods of one to four hours. Results: In this study, the morphology of IPECs from bovine aorta was identical to that of the BAECs. The gene expressions of endothelin-1 and eNOS were higher than those of BAECs and VSMCs. In addition, synthetic endothelin-1 resulted in the time-dependent boost of phosphorylation of carboxy-terminal Smad2 in the IPECs for periods of two and four hours. Conclusion: The results of this study confirm the efficacy of the enzymatic digestion method in isolating a large number of endothelial cells with morphological and functional characteristics.

20-hydroxyeicosatetraenoic acid alters endothelial cell barrier integrity independent of oxidative stress and cell death

Unregulated inflammation during bovine mastitis is characterized by severe mammary tissue damage with systemic involvement. Vascular dysfunction underlies tissue pathology because of concurrent oxidative stress mediated by several inflammatory mediators. We recently demonstrated increased production of 20-hydroxyeicosatetraenoic acid (20-HETE), a cytochrome P450-derived (CYP) oxylipid that correlated with oxidative stress during severe bovine coliform mastitis. The hypothesis for this study was that 20-HETE-induced oxidative stress disrupts barrier function of endothelial cells. Primary endothelial cells from the bovine aorta were utilized to investigate the effects of 20-HETE on barrier integrity in an in-vitro model of oxidative stress. The effects of various antioxidants on modulating the 20-HETE barrier integrity effects also were investigated. Our results showed that 20-HETE decreased endothelial barrier integrity, which was associated with increased reactive metabolite production and decreased total glutathione. The antioxidant, vitamin E, partially delayed the loss of endothelial resistance upon exposure to 20-HETE but did not prevent complete loss of barrier integrity. The decrease in barrier resistance due to 20-HETE was neither associated with oxidative stress as assessed by oxidative protein or lipid damage nor endothelial cell apoptosis; however, selenium supplementation conferred resistance to loss of barrier integrity suggesting a role for shifts in redox status. Specific mechanisms by which 20-HETE alters vascular barrier integrity require further investigation to identify targets for therapy during inflammatory conditions with enhanced CYP450 activity.

Autophagy activation is required for homocysteine-induced apoptosis in bovine aorta endothelial cells

An elevated level of homocysteine (Hcy) in plasma is an independent risk factor for cardiovascular disease and central nervous system disease. Endothelial dysfunction as a result of apoptosis in endothelial cells is involved in the development and progression of these diseases. In this study, we aimed to investigate the effect of autophagy activation by amino acid starvation on Hcy-induced cytotoxicity in bovine aorta endothelial cells (BAECs). Hcy-induced lactate dehydrogenase (LDH) release was promoted by amino acid starvation. In addition, Hcy increased cleaved caspase-3 level, an indicator of apoptosis, by amino acid starvation. We revealed that oxidative stress is not involved in the Hcy-induced cytotoxicity promoted by amino acid starvation. Salazosulfapyridine (SASP), an SLC7A11 inhibitor, protected against the Hcy-induced LDH release promoted by amino acid starvation. SASP decreased the Hcy-induced cleaved caspase-3 level by amino acid starvation. We demonstrate for the first time that autophagy activation by amino acid starvation promotes Hcy-induced apoptosis in BAECs. Moreover, SLC7A11 inhibitor SASP, which is an amino acid transporter, protects against Hcy-induced apoptosis due to autophagy.

Role of Resveratrol on Indoxyl Sulfate-Induced Endothelial Hyperpermeability via Aryl Hydrocarbon Receptor (AHR)/Src-Dependent Pathway.

Resveratrol (RES), a dietary polyphenol compound, has been shown to possess health benefits due to its anti-inflammatory, antioxidative, and antiatherosclerosis properties. Tryptophan metabolite-derived indoxyl sulfate (IS) is identified as one of the uremic toxins and physiological endogenous ligand/activator of aryl hydrocarbon receptor (AHR), associated with atherosclerosis in chronic kidney disease (CKD) patients. Studies have shown that a high serum level of IS causes deleterious effects on health primarily by inducing oxidative stress and endothelial dysfunction. However, the precise mechanisms are still unclear. Here, we investigated the underlying mechanism of IS effect on endothelial permeability and the role of RES on IS-induced endothelial hyperpermeability via the AHR/Src-dependent pathway. Bovine aorta endothelial cells (BAECs) were cultured and incubated with IS in the presence or absence of RES, and transendothelial electrical resistance (TEER) and permeability of cells were measured. Alongside, AHR, Src kinase, and Vascular Endothelial Cadherin (VE-Cadherin) activation were examined. Our data showed that IS reduced TEER of cells resulting in increased permeability. VE-Cadherin, a vital regulator of endothelial permeability, was also significantly activated in response to IS, which appeared to be associated with changes of endothelial permeability and AHR/Src kinase. Interestingly, in this setting, RES reversed the effect of IS and inhibited the increased activation of Src induced by IS-activated AHR and modulated VE-Cadherin and permeability. CH223191, an inhibitor of AHR, significantly inhibits IS-induced endothelial hyperpermeability. Further analysis with treatment of PP2, an inhibitor of Src abolishing Src activation, suggests downstream factors. All our data indicated that IS upregulated the AHR/Src kinase pathway, and increased endothelial permeability and phosphorylation of VE-Cadherin may be represented and provide new strategies for addressing protective properties of RES against Src kinase involved in AHR-mediated endothelial hyperpermeability. The findings may be crucial for managing diseases in which endothelial permeability is compromised, and the dietary polyphenols are involved.

Fabrication of aortic bioprosthesis by decellularization, fibrin glue coating and re-endothelization: a cell scaffold approach

Aortic dysfunctions (aneurysm, aortitis) lead to the most serious conditions related to aortic wall with life-threatening complications. The most common modality of management for such conditions is replacement (diseased part) of aorta by a larger diameter stent (reconstructive vascular surgery) which in itself is a big trial. The most natural way is to use a re-endothelized scaffold. Developing a scaffold with biomimetic properties is an experimental aim for most of the scientists and surgeons. We aim to structure a strategy to overcome the well-known problems associated with aorta. In this study, we plan to remold a larger diameter blood vessel such as aorta from xenogeneic origin using different protocols to decellularize and comparing them with normal aorta. The chemicals and enzymes used for bovine aorta decellularization are 1% SDS (group II), 70% ethanol + 0.25% trypsin (group III), 70% ethanol (group IV), and 0.25% trypsin (group V). Group I served as control (without decellularization). Histology and SEM study were conducted for cellular presence/absence in all scaffolds. Later, the scaffolds were coated with the fibrin glue (FG) and endothelial cells were proliferated over them. 3D images were taken showing the remolding of the endothelial cells on FG-coated surfaces. The re-endothelization was confirmed by lectin and vWF+/+ expression. Graft elasticity and burst pressure were confirmed by biomechanical tensile testing. Further, the absence of host tissue DNA and presence of cellular DNA after re-endothelialization were confirmed by PicoGreen assay. The acceptability for metabolically active cellular proliferation on scaffolds and its non-toxicity were proved by cell viability assay. Current findings accomplish that larger diameter aorta extracellular matrix scaffold (group II) can be fabricated and re-endothelialized to develop non-thrombotic surfaces with improved graft patency with promising results compared to other fabricated scaffold groups.

Hybrid cardiovascular sourced extracellular matrix scaffolds as possible platforms for vascular tissue engineering.

The aim when designing a scaffold is to provide a supportive microenvironment for the native cells, which is generally achieved by structurally and biochemically imitating the native tissue. Decellularized extracellular matrix (ECM) possesses the mechanical and biochemical cues designed to promote native cell survival. However, when decellularized and reprocessed, the ECM loses its cell supporting mechanical integrity and architecture. Herein, we propose dissolving the ECM into a polymer/solvent solution and electrospinning it into a fibrous sheet, thus harnessing the biochemical cues from the ECM and the mechanical integrity of the polymer. Bovine aorta and myocardium were selected as ECM sources. Decellularization was achieved using sodium dodecyl sulfate (SDS), and the ECM was combined with polycaprolactone and hexafluoro‐2‐propanol for electrospinning. The scaffolds were seeded with human umbilical vein endothelial cells (HUVECs). The study found that the inclusion of aorta ECM increased the scaffold's wettability and subsequently lead to increased HUVEC adherence and proliferation. Interestingly, the inclusion of myocardium ECM had no effect on wettability or cell viability. Furthermore, gene expression and mechanical changes were noted with the addition of ECM. The results from this study show the vast potential of electrospun ECM/polymer bioscaffolds and their use in tissue engineering.

Curcumin Inhibited Endothelin mRNA Expression Induced by TGF-β in Bovine Endothelial Cell

Objectives: Curcumin is a plant polyphenol compound used as a traditional supplement in many countries. The potential therapeutic or preventive effects of curcumin may be related to its antioxidant and anti-inflammatory properties. The current in vitro study aimed to investigate the effect of curcumin on the expression of endothelin-1 (ET-1) stimulated by transforming growth factorbeta (TGF-β) in bovine aortic endothelial cells (BAECs). Methods: Bovine aortic endothelial cells derived from bovine aorta were maintained in low glucose DMEM (Dulbecco’s modified Eagle’s medium). The time-course and different concentrations of TGF-β (2 ng/mL and 10 ng/mL) were used to evaluate ET-1 mRNA expression. Also, the BAECs were treated with 10 µM of SB431542 (chemical inhibitor of TGF-β receptor) as positive control and different doses of curcumin (5 µM, 10 µM, and 15 µM). The expression of ET-1 mRNA was quantified by quantitative real-time polymerase chain reaction (qPCR). Statistical analysis was performed by one-way ANOVA with SPSS software. Results: ET-1 mRNA expression significantly increased in six hours in the TGF-β-treated group (2.79 ± 0.9). SB431542, as well as curcumin 10 µM and 15 µM significantly decreased the expression of ET-1 mRNA by 2.3±0.15, 1.5±0.16, and 1.02±0.01, respectively. Conclusions: Curcumin downregulated ET-1 mRNA; this result suggested a possible underlying molecular mechanism mediated through ET-1 to exert its anti-inflammatory and antioxidant properties.

Phenotypic Modulation of Cultured Primary Human Aortic Vascular Smooth Muscle Cells by Uremic Serum.

Patients with chronic kidney disease (CKD) have a markedly increased incidence of cardiovascular disease (CVD). The high concentration of circulating uremic toxins and alterations in mineral metabolism and hormone levels produce vascular wall remodeling and significant vascular damage. Medial calcification is an early vascular event in CKD patients and is associated to apoptosis or necrosis and trans-differentiation of vascular smooth muscle cells (VSMC) to an osteogenic phenotype. VSMC obtained from bovine or rat aorta and cultured in the presence of increased inorganic phosphate (Pi) have been extensively used to study these processes. In this study we used human aortic VSMC primary cultures to compare the effects of increased Pi to treatment with serum obtained from uremic patients. Uremic serum induced calcification, trans-differentiation and phenotypic remodeling even with normal Pi levels. In spite of similar calcification kinetics, there were fundamental differences in osteochondrogenic marker expression and alkaline phosphatase induction between Pi and uremic serum-treated cells. Moreover, high Pi induced a dramatic decrease in cell viability, while uremic serum preserved it. In summary, our data suggests that primary cultures of human VSMC treated with serum from uremic patients provides a more informative model for the study of vascular calcification secondary to CKD.

Evaluation of Laser Tissue Welding and Laser-Tissue Soldering for Mucosal and Vascular Repair.

Context:Laser tissue bonding (LTB) is believed to have certain advantages over conventional sutures such as fluid-tight closure and minimal scarring and fibrosis.Aim:The aim of the present study was to evaluate the bond strength of laser tissue welding and laser tissue soldering in mucosal and vascular repair.Materials and Methods:A total of 85 samples of bovine oral mucosa and 85 bovine aortas were bonded using a CO2 laser and different laser powers. Human serum albumin was used as solder. The breaking load for mucosal samples and the bursting pressure for aorta samples were evaluated. Few specimens were evaluated histologically for thermal damage and other microscopic changes.Statistical Methods:Two-way ANOVA was performed as the data were normally distributed and analyzed for significant differences between the groups. This was followed by Simple Main effects (Tuckey's post hoc test) to determine the individual variation between groups and also the significant differences within the groups.Results:Significantly higher values of breaking load (44.2 ± 3.03 g) and bursting pressure (70.8 ± 12.33 mmHg) were noted when 50% albumin was used. When reinforcing sutures were given the bond strength was further increased (68.0 ± 4.0 g for breaking load) (108.0 ± 12.56 mmHg for bursting pressure). Microscopically, a bridge of solder coagulum formed across the wound. Thermal damage was restricted to the top layers only although it did extend much more laterally adjacent to the wound edges. Few areas of vacuolization and carbonization were seen.Conclusion:LTB seems to be a promising new method of wound closure and warrants further evaluation in the form of in vivo and clinical studies.