PubMed Health⌕ Search

Biomedical subjects

Vincent Chan

Publications and source records attributed to Vincent Chan.

At least 19 recordsLinked to original sources
PubMed2006

Dynamics of smooth muscle cell deadhesion from thermosensitive hydroxybutyl chitosan.

Thermoresponsive polymer (TRP) enables the enzyme-free harvesting of cells through an acute increase in surface hydrophilicity of TRP across its lower critical solution temperature (LCST), rendering feasible the generation of polymer-free cell sheets for regenerative medicine applications. To date, the intricate mechanisms of cell deadhesion/detachment on TRP surface remain obscure. Elucidation of such biophysical responses would be valuable for the cell sheet technology. In this study, integrative biophysical techniques are applied to probe the thermal-induced deadhesion kinetics of smooth muscle cell (SMC) on thermoresponsive hydroxybutyl chitosan (HBC29) against different periods of pre-culture time at 37 degrees C. Atomic force microscopy demonstrates that both the surface topography and mechanical property of HBC29 film in water are acutely modulated across its LCST. Firstly, cells show negligible changes in adhesion contact area during low-temperature incubation on unmodified tissue culture polystyrene (TCPS). Secondly, the recession of adhesion contact and retraction of cell body for cells with different pre-culture times are triggered by HBC29 coating on TCPS. Interestingly, the initial rate of reduction in the normalized adhesion contact area of SMC is negatively correlated with the pre-culture time. Thirdly, the degree of cell deformation and average adhesion energy are reducing functions of time only for SMCs with the lowest pre-culture time. In contrast, adhesion energy per cell is a reducing function of time irrespective of the change of pre-culture time. Lastly, the temporal dynamics of cytoskeleton organization and beta-actin/smoothelin-B mRNA expression for SMCs is strongly dependent on the pre-culture time. Overall, this study demonstrates that the thermal-induced deadhesion of SMC on TRP is characterized by the evolution of its contractile phenotypes.

Animals↗
PubMed2006

The HBSP gene is expressed during HBV replication, and its coded BH3-containing spliced viral protein induces apoptosis in HepG2 cells.

The mechanisms of liver injury in hepatitis B virus (HBV) infection are defined to be due not to the direct cytopathic effects of viruses, but to the host immune response to viral proteins expressed by infected hepatocytes. We showed here that transfection of mammalian cells with a replicative HBV genome causes extensive cytopathic effects, leading to the death of infected cells. While either necrosis or apoptosis or both may contribute to the death of infected cells, results from flow cytometry suggest that apoptosis plays a major role in HBV-induced cell death. Data mining of the four HBV protein sequences reveals the presence of a Bcl-2 homology domain 3 (BH3) in HBSP, a spliced viral protein previously shown to be able to induce apoptosis and associated with HBV pathogenesis. HBSP is expressed at early stage of our cell-based HBV replication. When transfected into HepG2 cells, HBSP causes apoptosis in a caspase dependent manner. Taken together, our results suggested a direct involvement of HBV viral proteins in cellular apoptosis, which may contribute to liver pathogenesis.

Apoptosis↗
PubMed2006

Adhesion contact kinetics of HepG2 cells during Hepatitis B virus replication: Involvement of SH3-binding motif in HBX.

It has been shown that Hepatitis B virus (HBV) replication directly alters the expression of key cytoskeleton-associated proteins which play key roles in mechanochemical signal transduction. Nevertheless, little is known on the correlation between HBV replication and the subsequent adhesion mechanism of HBV-replicating cells. In this study, it is demonstrated that the lag time of adhesion contact evolution of HepG2 cells with HBV replication is significantly increased by two times compared to that of normal HepG2 cell on collagen coated substrate. During the initial 20 min of cell seeding, only diffuse forms of vinculin was detected in HBV replicating cells while vinculin-associated focal complexes were found in normal and control cells. Similar delay in cell adhesion in HBV-replicating cells was observed in cells transfected with HBX, the smallest HBV protein, suggesting its involvement in this cellular process. In addition, a proline rich region found in many SH3 binding proteins was identified in HBX. HBX was found to interact with the focal adhesion protein, vinexin-beta, through the SH3 binding. Furthermore, HepG2 cells with HBV replication showed evidence of cell rounding up, possibly resulting from cytoskeletal reorganizations associated with interaction between HBX and vinexin-beta. Taken together, our results suggest that HBX is involved in the cytoskeletal reorganization in response to HBV replication.

Adaptor Proteins, Signal Transducing↗
PubMed2006

Improved cardiovascular function with aminoguanidine in DOCA-salt hypertensive rats.

The ability of aminoguanidine (AG), an inhibitor of collagen crosslinking, to prevent changes in cardiac and vascular structure and function has been determined in the deoxycorticosterone acetate (DOCA)-salt hypertensive rat as a model of the cardiovascular remodelling observed in chronic human hypertension. Uninephrectomized rats (UNX) administered DOCA (25 mg every fourth day s.c.) and 1% NaCl in drinking water for 28 days developed cardiovascular remodelling shown as systolic hypertension, left ventricular hypertrophy, increased thoracic aortic and left ventricular wall thickness, increased left ventricular inflammatory cell infiltration together with increased interstitial collagen and increased passive diastolic stiffness, impaired contractility, prolongation of the action potential duration and vascular dysfunction. Treatment with AG (0.05-0.1% in drinking water; average 182+/-17 mg kg(-1) day(-1) in DOCA-salt rats) decreased blood pressure (DOCA-salt 176+/-4; +AG 144+/-5 mmHg; (*)P<0.05 vs DOCA-salt), decreased left ventricular wet weights (DOCA-salt 3.17+/-0.07; +AG 2.66+/-0.08 mg g(-1) body wt(*)), reduced diastolic stiffness constant (DOCA-salt 30.1+/-1.2; +AG 24.3+/-1.2(*) (dimensionless)), improved cardiac contractility (DOCA-salt 1610+/-130; +AG 2370+/-100 mmHg s(-1)(*)) and vascular reactivity (3.4-fold increase in maximal contractile response to noradrenaline, 3.2-fold increase in maximal relaxation response to acetylcholine, twofold increase in maximal relaxation response to sodium nitroprusside) and prolonged the action potential duration at 50% repolarization without altering collagen content or inflammatory cell infiltration.Thus, cardiovascular function in DOCA-salt hypertensive rats can be improved by AG independent of changes in collagen content. This suggests that collagen crosslinking is an important cause of cardiovascular dysfunction during cardiovascular remodelling in hypertension.

Action Potentials↗
PubMed2006

UV-embossed microchannel in biocompatible polymeric film: application to control of cell shape and orientation of muscle cells.

This article shows that ultra violet (UV) micro-embossing can be successfully used for fabricating biocompatible micropatterned films with microchannels separated by high aspect ratio microwalls. Eight series of micropatterns were investigated; the width of the microwall was either 10 or 25 microm and that of the microchannel either 40, 80, 120, or 160 microm. The material investigated was principally polyurethane diacrylate. The UV-embossed micropattern was extracted with methanol, converting the micropatterns from cytotoxic to biocompatible. The typical UV embossing method was modified by using a marginally adhesive polyester substrate, which facilitates demolding but is removable before methanol extraction to avoid fragmentation of the embossed micropatterns. The effect of the micropatterns on A7r5 smooth muscle cells and C2C12 skeletal muscle cells was investigated. The dimensions of both channel and wall have significant effects on the elongation of both muscle cells. In the narrower 40-microm channel, the C2C12 cells merged together to form myofibers. These results indicate that UV-embossed micropatterns may present a useful scaffold for in vitro cell shape and orientation control needed in vascular and muscle tissue engineering.

Animals↗
PubMed2006

Valve replacement surgery in end-stage renal failure: mechanical prostheses versus bioprostheses.

BACKGROUND: The 1998 American College of Cardiology/American Heart Association Guidelines recommend mechanical prostheses for valve replacement in patients with end-stage renal disease requiring dialysis. The aim of the study is to evaluate the combined experience at two academic centers. METHODS: Sixty-nine valve replacements (aortic 40; mitral 22; multiple 7; 47 bioprostheses, 22 mechanical prostheses) were performed. Total follow-up was 128.7 patient-years (bioprostheses, 68.4; mechanical prostheses, 60.4). RESULTS: Patient populations were homogeneous, except for age (bioprostheses greater than mechanical prostheses, p = 0.012), previous myocardial infarction (bioprostheses greater than mechanical prostheses, p = 0.040), and concomitant CABG (bioprostheses greater than mechanical prostheses, p = 0.019). A survival advantage was observed in favor of mechanical prostheses (p = 0.0299) at 5 years. Freedom from valve-related complications at 5 years was calculated for thromboembolism plus thrombosis plus hemorrhage (bioprostheses, 93.0% +/- 3.9%; mechanical prostheses, 76.4% +/- 12.7%), thromboembolism excluding thrombosis (bioprostheses, 93.0% +/- 3.9%; mechanical prostheses, 88.9% +/- 10.5%), and hemorrhage (bioprostheses, 100%; mechanical prostheses, 95.2% +/- 4.7%). One case of structural valve deterioration occurred in the bioprostheses group at 95 months after surgery. Five-year freedom from all valve-related complications was 82.8% +/- 8.1% for bioprostheses and 76.4% +/- 12.7% for mechanical prostheses. CONCLUSIONS: Overall survival was poor. Differences between populations were related to age at operation and coronary artery disease. Structural valve deterioration was not accentuated with bioprostheses. Considering lack of homogeneity between prostheses groups there was no superiority of mechanical prostheses over bioprostheses in terms of freedom from composites of complications. Bioprostheses should be considered in the management of valvular disease in end-state renal disease patients.

Adult↗
PubMed2006

Three-dimensional microchannels in biodegradable polymeric films for control orientation and phenotype of vascular smooth muscle cells.

The poor mechanical strength and vasoactivity of current small-diameter tissue engineered blood vessels (TEBVs) remain unsolved problems. Given the plasticity of smooth muscle cells (SMCs), 1 of the main limitations of current scaffolding techniques is the difficulty in controlling SMC phenotype shifts in vitro. A synthetic phenotype allows the cells to rapidly proliferate and produce extracellular matrix (ECM), whereas a shift to contractile phenotype with organized ECM ultimately provides a functional blood vessel. In this study, 3D deep (65 microm) and wide microchannels separated by high-aspect ratio (8) microwalls were successfully ultraviolet (UV) microembossed using a liquid UV polymerizable biodegradable macromer (poly(epsilon-caprolactone-r-L-lactide-r-glycolide) diacrylate) and the in vitro guidance effects of varying channel width (40-160 microm) on SMCs were verified. The results show that SMCs cultured in the wider microchannels (80-160 microm wide) switch from fibroblast morphology and random orientation to spindle-shaped morphology, and align along the direction of the microchannel nearing confluence achieved with similar cell density to unpatterned film. Further, an enhanced expression of smooth muscle alpha-actin of SMCs grown on micropatterns was found nearing confluence, which demonstrates a phenotype shift to a more contractile phenotype. These films are flexible and can be folded into tubular and lamellar structures for tissue engineering of small-diameter TEBVs as well as other organs such as esophagus or intestine. These results suggest that these micropatterned synthetic biodegradable scaffolds may be useful for guiding SMCs to grow into functional, small-diameter vascular grafts.

Animals↗
PubMed2006

Off-pump coronary surgery through mini-incisions: is it reasonable?

PURPOSE OF REVIEW: Coronary artery bypass grafting as currently performed is considered the gold standard of coronary revascularization. It also, however, is a potential source of complications, especially in patients with multiple co-morbid risk factors. To alleviate some of these downsides, cardiac surgeons designed minimally invasive procedures performed off-pump, without sternotomy, and through small incisions. The efficacy of these techniques is emerging in the literature. Our aim is to describe the new technique that we are using, and provide an objective review of the recent literature with regards to safety and related surgical outcomes. RECENT FINDINGS: Although no long-term follow-up data exist yet with respect to the specific technique described herein, current evidence suggests that both on-pump and off-pump coronary bypass grafting techniques, when done in experienced centers, provide similar rates of completeness of revascularization, long-term patency, and freedom from surgical reintervention. When applied to patients at high surgical risk, observational studies suggest a decrease in the incidence of stroke, bleeding, renal complications, and mortality with off-pump bypass grafting. SUMMARY: Cardiac surgery performed off-pump without sternotomy is a procedural extension of off-pump bypass grafting which has the potential to be as safe as conventional coronary bypass grafting. The technique could offer an advantage to the high-risk patient population, or to patients for whom return to physically demanding work must be expedient after surgery.

Coronary Artery Bypass, Off-Pump↗
PubMed2006

Primary malignant melanoma of the lung: a case report and review of the literature.

Melanomas of the respiratory tract are usually metastatic in origin, and finding a primary melanoma is very rare. We report the case of a 60-year-old man who presented with a 2-month history of cough with associated weight loss. The histopathologic findings were consistent with those of a melanoma. Before considering the diagnosis of primary melanoma of the lung, an extensive diagnostic workup was done to rule out metastasis from an occult primary tumor. In addition, previously reported cases of primary lung melanoma were reviewed, and the established criteria for determining the primary site were applied. Because the precise etiology has not yet been elucidated, the possible mechanism is discussed.

Antineoplastic Agents, Alkylating↗
PubMed2005

Adhesion contact dynamics of 3T3 fibroblasts on poly (lactide-co-glycolide acid) surface modified by photochemical immobilization of biomacromolecules.

A simple and effective method of biomacromolecule immobilization on biomaterial surface for direct tuning of biophysical parameters such as the initial cell deformation rate, degree of cell spreading and adhesion kinetics is important for tissue engineering. The photochemical immobilization of azide-chitosan (Az-CS) on poly (lactide-co-glycolide) acid (PLGA) is applied here. Chitosan immobilization on PLGA through the photoactive azide group further facilitates subsequent grafting of other biocompatible biomacromolecules like gelatin (Gel) through the active amine groups on CS. This study quantitatively compares the 3T3 fibroblast adhesion dynamics on three PLGA surfaces (Gel-CS-PLGA, CS-PLGA and unmodified PLGA surfaces) using Confocal-Reflectance Interference Contrast Microscopy (C-RICM) together with phase contrast imaging. CS-PLGA and Gel-CS-PLGA surfaces developed were confirmed by X-ray photoelectron spectroscopy, atomic force microscopy and water contact angle and cell adhesion contact dynamics measurements. The cell adhesion was strongest on the Gel-CS-PLGA surface and lowest on unmodified PLGA. The steady state adhesion energy attained by the cells on gelatin modified PLGA surface is determined as 4.0 x 10(-8) J/m(2), which is about 400 times higher than that on PLGA surface (1.1 x 10(-10) J/m(2)). Significantly increased cell adhesion with Gel-CS-PLGA is postulated to result in increased cell spreading. Our integrated biophysical method can quantify the transient contact dynamics and is sufficiently accurate to discriminate even between Gel and CS modified surfaces.

3T3 Cells↗
PubMed2005

Adhesion contact dynamics of fibroblasts on biomacromolecular surfaces.

Biomacromolecules like gelatin and chitosan have emerged as highly versatile biomimetic coatings for applications in tissue engineering. The elucidation of the interfacial kinetics of cell adhesion on biomacromolecular surfaces will pave the way for the rational design of chitosan/gelatin-based systems for cell regeneration. Biomacromolecular ultra-thin films, chemically immobilized on fused silica are ideal experimental models for determining the effect of surface properties on the biophysical cascades following cell seeding. In this study, confocal reflectance interference contrast microscopy (C-RICM), in conjunction with phase contrast microscopy and fluorescence confocal microscopy, was applied to detect the adhesion contact dynamics of 3T3 fibroblasts on chitosan and gelatin ultrathin films. X-ray photoelectron spectroscopy (XPS) confirmed the immobilization of chitosan or gelatin on the silanized glass surface. Both the initial cell deformation rate and the change of two-dimensional spread area of the 3T3 fibroblasts are higher on gelatin-modified surfaces than on chitosan surfaces. The steady-state adhesion energy of 3T3 fibroblasts on gelatin film is three times higher than that on chitosan film. Immuno-staining of actin further demonstrates the different organization of cytoskeleton, likely induced by the change in cell signaling mechanism on the two biomacromolecular surfaces. The better attachment of 3T3 fibroblast to gelatin is postulated to be caused by the presence of adhesive domains on gelatin.

3T3 Cells↗
PubMed2005

Coupling bending and shear effects on liposome deformation.

Cell membrane deformation induced by external mechanical stimuli has been studied extensively over the past three decades. The present study focuses on the coupling of in-plane shear H and out-of-plane bending B of liposome membrane and its influences on the deformation of a single vesicle subjected to (i) external compressive load via two parallel platens and (ii) contact forces caused by a rigid substrate. Our results show that the increase of membrane resultant stress in both loading configurations causes the liposome to become more rigid and the degree of vesicle deformation decreases when the in-plane shearing effect is dominant. A theoretical approach is developed to facilitate cell membrane characterization under different biomechanical stimuli.

Animals↗
PubMed2005

Aortic valve replacement and concomitant mitral valve regurgitation in the elderly: impact on survival and functional outcome.

BACKGROUND: The impact of mitral regurgitation (MR) on elderly patients (> or = 70 years) undergoing isolated aortic valve replacement (AVR) is not clearly defined. This study investigates the long-term effects of preoperative, moderate MR on survival and functional outcome in elderly AVR patients. METHODS AND RESULTS: A retrospective review identified 408 consecutive elderly patients who underwent isolated AVR from January 1983 to February 2004. The pathologic etiology of MR was determined on preoperative echocardiogram, and patients were stratified into no/mild MR (Group I; n = 338) versus moderate MR (Group II; n = 70). Follow-up was 95.1% complete. Functional outcome was evaluated using the Short Form-12 questionnaire. On univariate analysis, Groups I and II differed in incidence of previous myocardial infarction (13.9% versus 28.6%; P = 0.004), hyperlipidemia (18.7% versus 33.3%; P = 0.009), and congestive heart failure (50.0% versus 70.0%; P = 0.002). On multivariate analysis, moderate MR was an independent risk factor impacting long-term survival (P = 0.04). Actuarial survival at 1, 5, and 10 years for Group I was 93.8%, 73.3%, and 40.1% versus 92.3%, 58.2%, and 14.6% for Group II (P = 0.04). Available postoperative echocardiograms for Group II (n = 37) demonstrated improvement in MR in 81.8% of functional MR patients. However, MR persisted or worsened in 65.4% of patients with intrinsic mitral valve disease (myxomatous, calcific, or ischemic MR). Functional outcomes showed 77% of Group I versus 78.6% of Group II rated their health as good to excellent post-AVR. CONCLUSIONS: Moderate MR is an independent risk factor impacting long-term survival in elderly patients undergoing AVR. Therefore, patients with intrinsic mitral valve disease should be considered for concomitant MV surgery.

Age Factors↗
PubMed2005

pH responsive adhesion of phospholipid vesicle on poly(acrylic acid) cushion grafted to poly(ethylene terephthalate) surface.

Polymer-supported lipid bilayer is a key enabling technology for the design and fabrication of novel biomimetic devices. To date, the physical driving force underlying the formation of polymer-supported lipid bilayer remains to be determined. In this study, the interaction between dipalmitoylphosphocholine (DPPC) vesicle and poly(ethylene terephthalate) [PET] surface with or without grafted poly(acrylic acid) [PAA] layer is examined with several biophysical techniques. First, vesicle deformation analysis shows that the geometry of adherent vesicle on either plain PET or PAA-grafted PET surface is best described by a truncated sphere model. At neutral pH, the degree of deformation and adhesion energy are unaltered by the grafted polymerization of acrylic acid on PET surface. Interestingly, the average magnitude of adhesion energy is increased by 185% and -43% on PAA-grated PET and plain PET surface, respectively, towards an increase of pH at room temperature. Our results demonstrate the possibility of tuning the adhesive interaction between vesicle and polymer cushion through the control of polyelectrolyte ionization on the solid support.

Acrylic Resins↗
PubMed2005

Adhesion contact dynamics of primary hepatocytes on poly(ethylene terephthalate) surface.

The design of bioartificial liver assist device requires an effective attachment of primary hepatocytes on polymeric biomaterials. A better understanding of this cell-surface interaction would aid the optimal choice of biomaterials. In this study, the adhesion contact dynamics of primary hepatocytes on poly(ethylene terephthalate) (PET) surface with grafted poly(acrylic acid) (PAA) and coated collagen is probed with confocal reflectance interference contrast microscopy (C-RICM) in conjunction with phase contrast microscopy. An increase of acrylic acid density from 0 to 12 nmole/cm2 raises both the root-mean-square surface roughness and amount of adsorbed collagen of PET surface. C-RICM demonstrates that hepatocytes form tight adhesion contacts upon seeding on both plain PET and PAA-grafted PET (both with collagen coating) despite the insignificant two-dimensional cell spreading. At two hours after cell seeding, the normalized contact area and adhesion energy of hepatocytes on 12 nmole/cm2 PAA-grafted-PET (with collagen coating) is 27% and 114% higher, respectively, than that on collagen coated plain PET. Interestingly, the growth kinetics of adhesion patch for hepatocyte on PAA-grafted PET with collagen coating is best fitted by R proportional to t0.5 and is significantly different from that on collagen coated plain PET, which is best fitted by R proportional to t0.25. Overall, this study demonstrates the modulation of biophysical response of adherent hepatocytes through the control of the biomaterial surface properties.

Biocompatible Materials↗
PubMed2005

The influence of GFP-actin expression on the adhesion dynamics of HepG2 cells on a model extracellular matrix.

Integrins belong to a family of important cell surface receptors which mediate the adhesion of most anchorage-dependent cells to nature extracellular matrix (ECM) and biomaterials. It is known that the binding of integrin with ECM proteins triggers mechanochemical responses of cytoskeleton. To date, the intricate interplay between integrin-ECM interaction and cytoskeleton dynamics leading to the regulation of cell morphogenesis on biomaterials remains largely unknown. In this study, green fluorescence protein (GFP)-actins were expressed in HepG2 cells for the temporal visualization of cytoskeletal structure of adherent cells on naturally derived materials. By combining confocal reflectance contrast microscopy and fluorescence microscopy, the adhesion contact dynamics, cytoskeleton remodeling and two-dimensional spreading of intact and GFP-actin expressing HepG2 cells on collagen and fibronectin-coated substrates are simultaneously probed during the initial cell seeding. First of all, our results show that the evolution of adhesion contact of HepG2 cells upon integrin-collagen or integrin-fibronectin interaction is impaired by GFP-actin expression. Also, the initial rate of cell deformation is reduced by 70% and 43% on fibronectin and collagen, respectively, upon GFP-actin expression. Interestingly, the steady-state adhesion energy of HepG2 cells remains unchanged and increases on fibronectin- and collagen-coated substrate, respectively, upon GFP-actin expression. Our highly integrated biophysical approach demonstrates that GFP-actins diffusively concentrate in the cytoplasmic cortex during initial cell seeding while adhesion contact evolves and cell spreads. Kinetics analysis on the adhesion contact formation demonstrates the intricate interplay between cytoskeleton property and ECM proteins in cell adhesion.

Actins↗
PubMed2005

Interaction between O-carboxymethylchitosan and dipalmitoyl-sn-glycero-3-phosphocholine bilayer.

O-carboxylmethylchitosan (OCMCS), a chitosan derivative, has emerged as a strong polymeric biomembrane perturbant. In this study, the interaction between OCMCS and dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was examined with cross-polarization microscopy, differential scanning calorimetry (DSC) and the surface pressure-area isotherms techniques. Cross-polarized light images showed that OCMCS induced the fusion of small DPPC multilamellar vesicles (MLV) to form large lamellar structures. From DSC measurement, the highest degree of fusion was found at the optimum OCMCS concentrations between 0.0625 and 0.2 mg/ml which are orders of magnitude lower than those required for similar reductions with unmodified chitosan as perturbant. At these concentrations, the association of DPPC and OCMCS enhances the fusion of DPPC vesicles. Surface pressure-area isotherms of DPPC monolayer in the presence of OCMCS imply interactions between OCMCS and DPPC in neutral condition. In comparison with chitosan, OCMCS proved to be a more effective membrane perturbant not only in neutral but also in acidic and basic conditions. The physical driving forces for OCMCS-induced perturbation of DPPC bilayer in neutral conditions are mainly hydrogen bonding and hydrophobic interactions. In acidic or basic conditions, the physical driving forces are dominated by the electrostatic interactions. The strong OCMCS-DPPC interaction will potentially increase the effectiveness of OCMCS for gene or drug delivery.

1,2-Dipalmitoylphosphatidylcholine↗