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Find video protocols related to scientific articles indexed in Pubmed.
The effect of feet position on standing balance in patients with diabetes.
Proc Inst Mech Eng H
PUBLISHED: 09-11-2014
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Feet displacement is recognized to be an important element in standing and is also linked to postural instability in elderly people with diabetes. This study investigates standing balance in diabetic patients in four asymmetric feet displacements. Quiet standing balance was investigated using the Biodex Balance System in 18 diabetic patients and compared with 18 control elderly subjects. The four standing conditions, namely, comfortable feet position, preferred feet position with a stance width of 17?cm and 15° angle between the medial borders, feet side by side, and heel side by side with a 30° angle between medial edges of feet were evaluated (i.e. eyes opened, eyes closed). The overall stability was calculated by measuring anterior-posterior and medial-lateral indices in standing conditions. Differences among feet positions were compared using an analysis of variance and the independent t-test. The diabetic patients were unstable in the medial-lateral direction when standing with feet side by side versus heel side by side with a 30° angle between medial edges of feet (p?=?0.012 and 0.011, respectively), while in controls the anterior-posterior stability scores between standing in preferred foot position with stance width of 17?cm and 15° angle between the medial borders versus feet side by side, and heel side by side with a 30° angle between medial edges of feet versus preferred foot position with stance width of 17?cm and 15° angle between the medial borders had significant difference (p?
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A proteomic view to characterize the effect of chitosan nanoparticle to hepatic cells: is chitosan nanoparticle an enhancer of PI3K/AKT1/mTOR pathway?
Biomed Res Int
PUBLISHED: 02-10-2014
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Chitosan nanoparticle, a biocompatible material, was used as a potential drug delivery system widely. Our current investigation studies were the bioeffects of the chitosan nanoparticle uptake by liver cells. In this experiment, the characterizations of chitosan nanoparticles were measured by transmission electron microscopy and particle size analyzer. The average size of the chitosan nanoparticle was 224.6 ± 11.2?nm, and the average zeta potential was +14.08 ± 0.7?mV. Moreover, using proteomic approaches to analyze the differential protein expression patterns resulted from the chitosan nanoparticle uptaken by HepG2 and CCL-13 cells identified several proteins involved in the PI3K/AKT1/mTOR pathway. Our experimental results have demonstrated that the chitosan nanoparticle may involve in the liver cancer cell metastasis and proliferation.
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Characterization of silk fibroin modified surface: a proteomic view of cellular response proteins induced by biomaterials.
Biomed Res Int
PUBLISHED: 01-18-2014
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The purpose of this study was to develop the pathway of silk fibroin (SF) biopolymer surface induced cell membrane protein activation. Fibroblasts were used as an experimental model to evaluate the responses of cellular proteins induced by biopolymer material using a mass spectrometry-based profiling system. The surface was covered by multiwalled carbon nanotubes (CNTs) and SF to increase the surface area, enhance the adhesion of biopolymer, and promote the rate of cell proliferation. The amount of adhered fibroblasts on CNTs/SF electrodes of quartz crystal microbalance (QCM) greatly exceeded those on other surfaces. Moreover, analyzing differential protein expressions of adhered fibroblasts on the biopolymer surface by proteomic approaches indicated that CD44 may be a key protein. Through this study, utilization of mass spectrometry-based proteomics in evaluation of cell adhesion on biopolymer was proposed.
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Adenosine diphosphate-decorated chitosan nanoparticles shorten blood clotting times, influencing the structures and varying the mechanical properties of the clots.
Int J Nanomedicine
PUBLISHED: 01-01-2014
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Chitosan nanoparticles (NPs) decorated with adenosine diphosphate (ADP) (ANPs) or fibrinogen (FNPs) were used to fabricate hemostatic NPs that can shorten blood clotting time and prevent severe local hemorrhage. The structure and mechanical properties of the blood clot induced with ANP (clot/ANP) or FNP (clot/FNP) were also investigated. The NPs, ANPs, and FNPs, which had particle sizes of 245.1 ± 14.0, 251.0 ± 9.8, and 326.5 ± 14.5 nm and zeta potentials of 24.1 ± 0.5, 20.6 ± 1.9, and 15.3 ± 1.5 mV (n=4), respectively, were fabricated by ionic gelation and then decorated with ADP and fibrinogen. The zeta potentials and Fourier transform infrared (FTIR) spectroscopy of the NPs confirmed that their surfaces were successfully coated with ADP and fibrinogen. The scanning electron microscope (SEM) micrographs of the structure of the clot induced with "undecorated" chitosan NPs (clot/NP), clot/ANP, and clot/FNP (at 0.05 wt%) were different, after citrated bloods had been recalcified by a calcium chloride solution containing NPs, ANPs, or FNPs. This indicated that many NPs adhered on the membrane surfaces of red blood cells, that ANPs induced many platelet aggregates, and that FNPs were incorporated into the fibrin network in the clots. Measurements of the blood clotting times (Tc) of blood clot/NPs, clot/ANPs, and clot/FNPs, based on 90% of ultimate frequency shifts measured on a quartz crystal microbalance (QCM), were significantly (P<0.05) (n=4) shorter than that of a clot induced by a phosphate-buffered solution (PBS) (clot/PBS) (63.6% ± 3.1%, 48.3% ± 6.2%, and 63.2% ± 4.7%, respectively). The ?F2 values in the spectra of frequency shifts associated with the propagation of fibrin networks in the clot/ANPs and clot/FNPs were significantly lower than those of clot/PBS. Interestingly, texture profile analysis of the compressional properties showed significantly lower hardness and compressibility in clot/NPs and clot/ANPs (P<0.05 or better) (n=4) compared with clot/PBS and clot/FNPs. Accordingly, among the hemostatic NPs, ANP substantially reduced blood clotting times, ?F2 values, and compression flow properties of the clot. Hence, ANPs have potential applications for preventing severe local hemorrhage.
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Bilateral diabetic Charcot foot.
Aust Fam Physician
PUBLISHED: 03-27-2013
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Charcot neuro-osteoarthropathy (CNO) of the foot is a devastating neuropathic complication of diabetes. It is characterised by deformity of the foot architecture,which can be initiated by trauma to the neuropathic limb or occur spontaneously.The acute phase of the disease is often misdiagnosed and can rapidly lead to deformity and amputation. The aim of management is to halt further bone destruction through immobilisation of the affected limb.
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Poly (?-caprolactone) scaffolds functionalized by grafting NGF and GRGD promote growth and differentiation of PC12 cells.
J Biomed Mater Res A
PUBLISHED: 02-22-2013
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Poly(?-caprolactone) (PCL) scaffolds functionalized by grafting nerve growth factor (NGF) and Asp-Arg-Gly-Asp (GRGD)(PCL-NGF/GRGD) for neural tissue engineering. The influences of PCL-NGF/GRGD scaffolds on the growth and differentiation of PC12 cells were investigated. The successfully grafting NGF and GRGD into PCL-CS scaffold were verified by FTIR spectra. The densities of GRGD and NGF in the scaffolds were about 2.10×10(-1) ?mol/cm(2) and 1.51×10(-3) nmol/cm(2) . Growths of PC12 cells in PCL-GRGD and PCL/NGF-GRGD scaffolds via MTS measurements were significantly higher (p < 0.01, n = 4) than that in PCL-CS or PCL-NGF ones for three days of cultivation that was consistent with SEM observations. Moreover, the differentiation of PC12 cells, induced by NGF at 50 ng/mL for four days, in PCL-NGF/GRGD scaffolds were qualitatively more numbers and longer outgrowth of neurites than those in PCL-CS, PCL-GRGD, and PCL-NGF ones by SEM observations. The synergistic effects of grafting both NGF and GRGD ligands to PCL-CS scaffolds on the growth and differentiation of PC12 cells provide a new biomaterial for neural tissue engineering. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 315-323, 2014.
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Urinary protein profiling by liquid chromatography/tandem mass spectrometry: ADAM28 is overexpressed in bladder transitional cell carcinoma.
Rapid Commun. Mass Spectrom.
PUBLISHED: 09-14-2011
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Bladder cancer is the most common urological cancer with higher incidence rate in the endemic areas of Blackfoot disease (BFD) in southern Taiwan. The aim of this study was to utilize the proteomic approach to establish urinary protein patterns of bladder cancer. The experimental results showed that most patients with bladder cancer had proteinuria or albuminuria. The urine arsenic concentrations of bladder cancer patients in BFD areas were significantly higher than those patients from non-BFD areas. In the proteomic analysis, the urinary proteome was identified by nano-high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (nano-HPLC/ESI-MS/MS) followed by peptide fragmentation pattern analysis. We categorized 2782 unique proteins of which 89 proteins were identified with at least three unique matching peptide sequences. Among these 89 proteins, thirteen of them were not found in the control group and may represent proteins specific for bladder cancer. In this study, three proteins, SPINK5, ADAM28 and PTP1, were also confirmed by Western blotting and showed significant differential expression compared with the control group. ADAM28 may be used as a possible biomarker of bladder cancer.
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Characterization of ADAM28 as a biomarker of bladder transitional cell carcinomas by urinary proteome analysis.
Biochem. Biophys. Res. Commun.
PUBLISHED: 06-25-2011
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Human urine contains a large number of proteins and peptides (the urinary proteome). Global analysis of the human urinary proteome is important for understanding urinary tract diseases. Bladder cancer is the most common urological cancer with higher incidence rates in endemic areas of Blackfoot disease (BFD) in southern Taiwan. The aim of this study was to use the proteomic approach to establish urinary protein biomarkers of bladder cancer. ADAM28, identified by proteomic approaches and confirmed by Western blotting, showed significant differences compared with normal individuals, so it may be a biomarker of bladder cancer.
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Protein profiling of human nonpigmented ciliary epithelium cell secretome: the differentiation factors characterization for retinal ganglion cell line.
J. Biomed. Biotechnol.
PUBLISHED: 04-05-2011
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The purpose of this paper was to characterize proteins secreted from the human nonpigmented ciliary epithelial (HNPE) cells, which have differentiated a rat retinal ganglion cell line, RGC-5. Undifferentiated RGC-5 cells have been shown to express several marker proteins characteristic of retinal ganglion cells. However, RGC-5 cells do not respond to N-methyl-D aspartate (NMDA), or glutamate. HNPE cells have been shown to secrete numbers of neuropeptides or neuroproteins also found in the aqueous humor, many of which have the ability to influence the activity of neuronal cells. This paper details the profile of HNPE cell-secreted proteins by proteomic approaches. The experimental results revealed the identification of 132 unique proteins from the HNPE cell-conditioned SF-medium. The biological functions of a portion of these identified proteins are involved in cell differentiation. We hypothesized that a differentiation system of HNPE cell-conditioned SF-medium with RGC-5 cells can induce a differentiated phenotype in RGC-5 cells, with functional characteristics that more closely resemble primary cultures of rat retinal ganglion cells. These proteins may replace harsh chemicals, which are currently used to induce cell differentiation.
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Characterization of surface modification on self-assembled monolayer-based piezoelectric crystal immunosensor for the quantification of serum ?-fetoprotein.
J Mater Sci Mater Med
PUBLISHED: 03-30-2011
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Self-assembled monolayers (SAMs) on coinage metallic material can provide versatile modeling systems for studies of interfacial electron transfer, biological interactions, molecular recognition and other interfacial phenomena. Recently, a bio-sensing system has been produced by analysis of the attachment of antibody using alkanethiols, to form SAMs on the face of Au-quartz crystal microbalance (QCM) surfaces. In this study, the attachment of anti-?-fetoprotein monoclonal antibody to a SAMs surface of 11-mercaptoundecanoic acid was achieved using water-soluble N-ethyl-N-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide as coupling agents. Surface analyses were utilized by X-ray photoelectron spectroscopy and atomic force microscopy. The quantization of immobilized antibody was characterized by the frequency shift of QCM and the radioactivity change of ¹²?I labeled antibody. The limit of detection and linear range of the calibration curve of the QCM method were 15 ng/ml and 15-850 ng/ml. The correlation coefficients of ?-fetoprotein concentration between QCM and radioimmunoassay were 0.9903 and 0.9750 for the standards and serum samples, respectively. This report illustrates an investigation of SAMs for the preparation of covalently immobilized antibody biosensors.
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Promoting regeneration of peripheral nerves in-vivo using new PCL-NGF/Tirofiban nerve conduits.
Biomaterials
PUBLISHED: 03-08-2011
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Poly(?-caprolactone) (PCL) scaffolds were modified by grafting nerve growth factor (NGF) and Tirofiban (TF), a clinical anti-thrombosis drug, as a new biomaterial for producing nerve conduits to promote the regeneration of sciatic nerves. The successful grafting of NGF and TF onto PCL scaffolds was confirmed by FTIR and ESCA spectra. In-vitro growths of the PC12 cells in PCL-NGF and PCL-NGF/TF scaffolds, determined by MTS, were significantly higher (P < 0.05, n = 4) than those in the PCL scaffolds following three days of cultivation. Interestingly, this study evaluation of the PCL, PCL-NGF, and PCL-NGF/TF nerve conduits in a 12 mm long gap of the rat sciatic nerve defect model that the gastrocnemius muscle mass of the tested rats in the PCL-NGF/TF groups significantly exceeded those in the PCL-NGF and PCL group. In the rats that had been implanted with PCL-NGF/TF conduits, the generated nerves passed through those conduits, expressing beta-III tubulin (TB), growth association protein-43 (GAP-43) and myelin basic protein (MBP) along their longitudinal axis, and the proximal and distal nerve ends of the rats were successfully connected. Those that had been implanted with PCL and PCL-NGF conduits did not exhibit these effects, as revealed by an immunochemical study of the expressions of the proteins in the conduits. Moreover, counting within the dorsal horn of the spinal cord (C(5)) demonstrated that the numbers of CTB-HRP-labeled neurons in the rats that had been implanted with PCL-NGF/TF conduits were significantly higher than those in the other groups. In this study, in-vivo examinations of the use of newly designed PCL-NGF/TF conduits to promote the generation of nerves in a defective rat model significantly increased the gastrocnemius muscle mass, and led to the successful regeneration of nerves that bridged a 12 mm long defected gap of nerves in rats. However, more rats must be tested to confirm the efficacy the newly designed nerve conduits.
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Evaluation of transdifferentiation from mesenchymal stem cells to neuron-like cells using microfluidic patterned co-culture system.
Biomed Microdevices
PUBLISHED: 02-25-2011
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We design a microfluidic patterned co-culture system for mouse mesenchymal stem cells (mMSCs) and neural cells to demonstrate the paracrine effects produced by the neural cells in facilitating the transdifferentiation from mMSCs to neuron-like cells. Neural cells and mMSC are orderly patterned in the microfluidic co-culturing system without direct cell contact. This configuration provides us to calculate the percentage of neural marker transdifferentiated by mMSCs easily. We obtain higher transdifferentiated ratio of mMSC in the microfluidic co-culturing system (beta III tubulin: 67%; glial fibrillary acidic protein (GFAP): 86.2%) as compared with the traditional transwell co-culturing system (beta III tubulin: 59.8%; GFAP: 52.0%), which is similar to the spontaneous neural marker expression in the undifferentiated MSCs (beta III tubulin: 47.5%; GFAP: 60.1%). Furthermore, mMSCs expressing green fluorescent protein and neural cells expressing red fluorescent protein were also used in our co-culture system to demonstrate the rarely occurring or observed cell fusion phenomenon. The results show that the co-cultured neural cells increased the transdifferentiation efficiency of mMSCs from soluble factors secreted by neural cells.
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PCP copolymers grafted with RGD enhance the rates of RGD-PCP micelles internalized into cells.
J Microencapsul
PUBLISHED: 06-22-2010
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RGD-PCP copolymers were fabricated by grafting Arg-Gly-Asp (RGD) peptide to poly(epsilon-caprolactone)-b-chitooligosaccharide-b-poly(ethylene glycol) (PCP) copolymers and the rate of internalization of RGD-PCP micelles by PC 12 cells were examined. Increasing intensity of the absorbance of amine groups in FT-IR spectra of RGD-PCP copolymers compared with those of PCP copolymers indicated the presence of RGD in new copolymers. Moreover, the grafting efficiency and molar ratio of RGD peptides to PCP copolymers were 88.2% and 0.45, respectively, analysed with HPLC. The RGD-PCP copolymers self-assemble to micelles at the critical micelle concentration (CMC) of 0.018 wt% (178 mg L(-1)) and with a mean diameter of 90 nm using a dynamic light-scattering (DLS) analyser. Interestingly, the internalization of DPH-loaded RGD-PCP micelles into PC 12 cells is much faster (e.g. within 5 min) than that of PCP micelles. The new RGD-PCP micelles may potentially be used in cellular drug delivery.
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Cerebral infarction in acute anemia.
J. Neurol.
PUBLISHED: 03-01-2010
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There are few previous studies on the relationship between cerebral infarction and acute anemia. This study presents patients with cerebral infarction in acute anemia due to marked blood loss and aims to clarify the stroke nature and possible mechanism. Patients with acute cerebral infarction and anemia following marked blood loss without systemic hypotension were recruited from 2001 to 2009. Clinical characteristics, particularly hemoglobin level, and neuroimaging findings were reviewed in detail to analyze the stroke nature and verify the possible pathogenesis. Twelve patients (males 8; mean age 74.9 years) were included. Eleven patients had cerebral infarction after acute massive gastrointestinal bleeding, and one had cerebral infarction following postoperative extensive hematoma during hospitalization. In all patients, borderzone infarction was the most characteristic finding: six had unilateral and six had bilateral borderzone infarction. Mean hemoglobin at infarction after acute blood loss was 5.8 g/dl, with 46% reduction from baseline. Of nine patients receiving detailed extracranial and intracranial vascular studies, none had severe carotid stenosis and six had intracranial stenosis. The arterial borderzones are the most vulnerable regions to a fall in cerebral perfusion. Acute anemia may produce cerebral blood flow insufficiency, reduce oxygen-carrying capacity, and result in distal-field tissue ischemic injury when hemoglobin level decreases below a critical level, especially in patients with intracranial stenosis.
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Silk fibroin/chitosan-hyaluronic acid versus silk fibroin scaffolds for tissue engineering: promoting cell proliferations in vitro.
J Mater Sci Mater Med
PUBLISHED: 02-05-2010
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The feasibility of silk fibroin protein (SF) scaffolds for tissue engineering applications to promote cell proliferation has been demonstrated, as well as the ability to mimic natural extra-cellular matrix (ECM), SF/chitosan (CS), a polysaccharide, scaffolds for tissue engineering. However, the response of cells to SF/CS-hyaluronic acid (SF/CS-HA) scaffolds has not been examined, which this study attempts to do and then compares those results with those of SF scaffolds. SF/CS-HA microparticles were fabricated to produce scaffolds in order to examine the proliferations of human dermal fibroblasts (HDF) in the scaffolds. Positive zeta potentials and ATR-FTIR spectra confirmed the co-existence of SF and CS-HA in SF/CS-HA microparticles. HDF proliferated well and migrated into SF/CS-HA scaffolds for around 160 mum in depth, as well as those in SF scaffolds after 7 days of cultivation, as observed using confocal microscopy. Interestingly, HDF grown in SF/CS-HA scaffolds had a markedly higher cell density than that in SF ones. Additionally, MTT assay revealed that the growth rates of HDF in SF/CS-HA scaffolds significantly exceeded (P < 0.01, n = 5) those in scaffolds of SF and SF/CS. The daily glucose consumptions and lactate formations, metabolic parameters, of HDF grown in SF/CS-HA and SF/CS scaffolds were significantly higher (P < 0.01, n = 3) than those in SF ones in most culturing days. Results of this study suggest that SF/CS-HA scaffolds have better cell responses for tissue engineering applications than SF ones.
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The influence of rat mesenchymal stem cell CD44 surface markers on cell growth, fibronectin expression, and cardiomyogenic differentiation on silk fibroin - Hyaluronic acid cardiac patches.
Biomaterials
PUBLISHED: 07-17-2009
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Since MSCs contain an abundant of CD44 surface markers, it is of interesting to investigate whether CD44 on rat MSC (rMSCs) influenced cell growth, fibronectin expression and cardiomyogenic differentiation on new SF/HA cardiac patches. For this investigation, we examined the influences of rMSCs with or without a CD44-blockage treatment on the aforementioned issues after they were cultivated, and further induced by 5-aza on SF and SF/HA patches. The results showed that the relative growth rates of rMSCs cultured on cultural wells, SF/HA patches without or with a CD44-blockage treatment were 100%, 208.9+/-7.1 (%) or 48.4+/-6.0 (%) (n=3, for all), respectively, after five days of cultivations. Moreover, rMSCs cultivated on SF/HA patches highly promoted fibronectin expressions (e.g., 1.8x10(5)/cell, in fluorescent intensity) while cells with a CD44-blockage treatment markedly diminished the expressions (e.g., 1.1x10(4)/cell, in fluorescent intensity) on same patches. For investigating possible influences of CD44 surface markers of rMSCs on their cardiomyogenic differentiation, the expressions of specific cardiac genes of cells were examined by using real-time PCR analysis. The results indicated that 5-aza inducing rMSCs significantly promoted the expressions of Gata4, Nkx2.5, Tnnt2 and Actc1 genes (all, P<0.01 or better, n=3) on SF/HA patches compared with those expressions on SF patches and for cells with a CD44-blockage treatment on SF/HA patches. Furthermore, the intensity of the expressions of cardiotin and connexin 43 of 5-aza inducing rMSCs were markedly higher than those of cells with a CD44-blockage treatment after they were cultured on SF/HA patches. Through this study, we reported that CD44 surface markers of rMSCs highly influenced the proliferations, fibronectin expressions and cardiomyogenic differentiation of rMSCs cultivated on cardiac SF/HA patches.
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Sustained release of 5-FU from Poloxamer gels interpenetrated by crosslinking chitosan network.
Int J Pharm
PUBLISHED: 05-07-2009
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This study investigates in vitro the drug delivery characteristics of new thermo-sensitive gels, P-CS/GA gels, in which a chitosan (CS) network is crosslinked with various concentrations of glutaraldehyde (GA) that interpenetrates Poloxamer (P) gels. The results indicate that the swelling ratios of all P-CS/GA gels are markedly superior to those of non-swelling P and P-CS gels. For example, P-CS/GA (0.1 wt.%) gels have swelling ratios of 13.2+/-1.0, which are maintained for approximately 18 h in water at 37 degrees C. In vitro releases of 5-FU from P-CS/GA (0.1 wt.%) gels had significantly lower initial burst release (P<0.01) and lasted much longer than those from gels without a CS network. For example, the duration of release of 5-FU was in a significantly sustained manner for up to 52 h, which was about 10 times or longer than the period of delivery using P or P-CS gels. The release of drugs from gels with an interpenetrating CS network could be modeled by Fickian diffusion; the characteristic constant k of drug-gel systems decreased as increasing GA concentrations in the P-CS/GA gels, and increasing the viscosities of the P, P-CS and P-CS/GA solutions.
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Mucoadhesive liposomes for intranasal immunization with an avian influenza virus vaccine in chickens.
Biomaterials
PUBLISHED: 04-15-2009
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The aim of this study was to characterize a nasally delivered bioadhesive liposome using an inactivated H5N3 virus as a model antigen. Bioadhesive liposomes were developed using tremella (T) or xanthan gum (XG) as the bioadhesive polysaccharide. Using chickens as the target animal, we evaluated whether delivery of a bioadhesive liposomal influenza vaccine via a mucosal site of infection could improve vaccine effectiveness. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cytotoxicity assays demonstrated that T, XG and liposomes were non toxic to chicken spleen macrophages. Enzyme-linked immunosorbent assay (ELISA) was used to determine the adjuvant effect of the bioadhesive liposomal-vaccines. Chickens immunized with a low dose (200 microL) of bioadhesive liposomal influenza vaccine had significantly higher mucosal and serum antibody levels (P<0.05). In addition, liposomes mixed with a low-viscosity bioadhesive gel used for nasal delivery resulted in superior antibody responses compared with liposomes mixed with a high-viscosity gel (P<0.05). This suggest that a low-viscosity gel mixed with liposomes is more suitable for nasal delivery, and that chickens elicit higher mucosal secretory immunoglobulin A (s-IgA) and serum IgG after two vaccinations.
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Enhancing growth and proliferation of human gingival fibroblasts on chitosan grafted poly (epsilon-caprolactone) films is influenced by nano-roughness chitosan surfaces.
J Mater Sci Mater Med
PUBLISHED: 04-03-2009
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The bioactivity of poly (epsilon-caprolactone) (PCL) films is improved by grafting chitosan (CS) surfaces with various values of nano-roughness on PCL surfaces. To examine the effects of the design, growing human gingival fibroblasts (HGFs) on the films was conducted. Various values of nano-rough CS surfaces were cast using nano-rough PCL molds that had been fabricated using a solvent-etched technique. The features of nano-CS/PCL surfaces were characterized using an atomic force microscope (AFM) to observe the topography and to determine the value of centerline average roughness of a surface, R(a). The R(a) values of the nano-CS/PCL films were 36.8 +/- 1.6, 100.0 +/- 3.0, and 148 +/- 7.0 nm, while that of the smooth CS/PCL film was 12.5 +/- 1.6 nm. The growth and proliferation of HGFs on the films are elucidated by fluorescent staining and analyzed by MTT viability assay following three and 7 days of culture. The viability assay of the cells reveals that the growth rates of HGFs on both CS/PCL and nano-CS/PCL films significantly exceed (95% or more; P < 0.001) those of PCL on both days, demonstrating the improvement of the bioactivity of PCL films by grafting CS. Additionally, the growth rates and proliferations of HGFs on nano-CS/PCL films of roughness 100 and 148 nm markedly exceed (15% or more; P < 0.001) those on 36.8 nm nano-CS/PCL and CS/PCL films, after both periods of culturing, indicating that the high nano-roughness CS surfaces further enhance the growth rate of HGFs. In conclusion, markedly improving the bioactivity of PCL films by grafting CS is demonstrated. Moreover, high nano-roughness of nano-CS/PCL films can further accelerate the growth and proliferation of HGFs compared with those of CS/PCL films. This work presents a new concept for designing biomaterials in tissue engineering.
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Effect of lipopolysaccharide on intranasal administration of liposomal Newcastle disease virus vaccine to SPF chickens.
Vet. Immunol. Immunopathol.
PUBLISHED: 03-31-2009
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In order to potentiate the low immunogenicity of the inactivated Newcastle disease virus immunized into chickens by mucosal route, liposomes as a drug delivery system and LPS (lipopolysaccharide) as an immuno-stimulator were evaluated. Here, we report a new nasal delivery system of inactivated Newcastle disease virus (NDV) vaccine. The intranasal vaccine was based on different lipids to form MLV (multi-lamellar vehicles) liposomes. The liposomes had combined carrier and adjuvant activities, which induced strong systemic (serum) and local (lung and nasal) humoral responses in SPF (specific-pathogen-free) chickens, and provided protective immunity. PC-Lip (phosphatidylcholine-liposome) elicited significant mucosal secretary immunoglobulin A (s-IgA) levels (p<0.05) in tracheal lavage fluid and serum IgG levels (p<0.05). In response to virulent viral challenge, birds treated with PBS (phosphate buffered saline) as control group died, whereas 80% of chickens which received PC-Lip, PC-Lip-LPS, PS-Lip (phosphatidylserine-liposome), and PS-Lip-LPS survived. HAI titers were 1:2560 in the PS-Lip-LPS group and 1:1280 in the PC-Lip, PC-Lip-LPS, and PS-Lip groups after two vaccinations. The results suggest that PC-Lip or PS-Lip might thus be suitable as a potential adjuvant for mucosal vaccination against NDV in chickens.
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The cardiomyogenic differentiation of rat mesenchymal stem cells on silk fibroin-polysaccharide cardiac patches in vitro.
Biomaterials
PUBLISHED: 02-09-2009
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Polysaccharides and proteins profoundly impact the development and growth of tissues in the natural extra-cellular matrix (ECM). To mimic a natural ECM, polysaccharides were incorporated to/or co-sprayed with silk fibroin (SF) to produce SF/chitosan (CS) or SF/CS-hyaluronic acid (SF/CS-HA) microparticles that were further processed by mechanical pressing and genipin cross-linking to produce hybrid cardiac patches. The ATR-FTIR spectra confirm the co-existence of CS or CS-HA and SF in microparticles and patches. For evaluating the cellular responses of rMSCs to the SF/CS and SF/CS-HA cardiac patches, the growth of rMSCs and cardiomyogenic differentiation of 5-aza inducing rMSCs cultured on patches was examined. First, the isolated rMSCs were identified with various positive and negative surface markers such as CD 44 and CD 31 by a flow cytometric technique, respectively. For examining the growth of rMSCs on the patches, MTT viability assay was performed, and the results demonstrated that the growth of rMSCs on SF and SF-hybrid patches significantly exceeded (P<0.001) that on culture wells after seven days of cultivation. Additionally, the relative growth rates of rMSCs on SF/CS and SF/CS-HA hybrid patches were significantly better (P<0.01) than that on SF patches that were also observed by using vimentin stain to the cells. For instance, the relative cell growth rates (%) in cell culture wells, SF, SF/CS and SF/CS-HA patches were 100%, 282.9+/-6.5%, 337.0+/-8.0% and 332.6+/-6.6% (n=6, for all), respectively. For investigating the effects of the hybrid patches on cardiomyogenic differentiation of 5-aza inducing rMSCs, the expressions of specific cardiac genes of cells such as Gata4 and Nkx2.5 were examined by real-time quantitative polymerase chain reaction (real-time PCR) analysis. The results of cardiomyogenic differentiation of induced rMSCs on SF/CS and SF/CS-HA hybrid patches significantly improved the expressions of cardiac genes of Gata4, Nkx2.5, Tnnt2 and Actc1 genes (all, P<0.01 or better, n=3) than those on SF patches and culture wells. Interestingly, the results of cardiac gene expressions of the cells on the SF/CS-HA hybrid patches were the most pronounced in promoting cardiomyogenic differentiations in this investigation. Furthermore, immunofluorescence staining of cardiac proteins such as cardiotin and connexin 43 for induced rMSCs cultured on SF/CS and SF/CS-HA hybrid patches were much pronounced compared with SF patches, indicating the improvements of cardiomyogenic differentiation on the hybrid patches. The results of this study demonstrate that the SF/CS and SF/CS-HA hybrid patches may be promising biomaterials for regenerating infarcted cardiac tissues.
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The t-PA-encapsulated PLGA nanoparticles shelled with CS or CS-GRGD alter both permeation through and dissolving patterns of blood clots compared with t-PA solution: an in vitro thrombolysis study.
J Biomed Mater Res A
PUBLISHED: 02-03-2009
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Accelerated thrombolysis by pressure-driven permeation has been demonstrated in in vitro and in vivo animal models by using plasminogen activators (PAs) encapsulated liposomes or PEG microparticles. Recent reports have also described acceleration of thrombolysis using tissue type PA (t-PA) encapsulated in PLGA nanoparticles (NPs) coated with chitosan (CS) or CS-GRGD by interactions between the NPs and blood clots. However, the permeation through and dissolving patterns in thrombolysis with the aforementioned microparticles or NPs, which may be clinically relevant to the recovery status of the posttreatments, have not been reported. Therefore, this work studied such phenomena in thrombolysis with t-PA encapsulated in NPs. The t-PA solution and the NPs exhibited distinctly different permeation patterns of dissolved clots. Plasma permeates through clots showed a stream flow or burst flow phenomena when lyzed with NPs shelled with CS or CS-GRGD, respectively, whereas a diffusion pattern was observed in those lyzed with t-PA solution. At the outlet position of clots, the clots dissolved with PLGA/CS and PLGA/CS-GRGD NPs revealed extremely rough surfaces to a depth of 100 mum, indicating that a cross-permeation direction of clot lysis occurred, while those dissolved with t-PA solution showed slightly rough surfaces to a depth of 12 mum. Permeation through and clot dissolution patterns of thrombolysis with t-PA encapsulated in NPs shelled with CS or CS-GRGD distinctly differed from those dissolved with t-PA solutions in this in vitro thrombolysis model, These findings may be relevant to posttreatment of patients with conventional PA thrombolysis.
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Cardiac repair using chitosan-hyaluronan/silk fibroin patches in a rat heart model with myocardial infarction.
Carbohydr Polym
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The cardiac repair of myocardial infarction (MI) hearts of rats using chitosan-hyaluronan/silk fibroin (chitosan-HYA/SF) cardiac patches was examined after eight weeks of implantation. Rats with implantations of chitosan-HYA/SF patches (CHS group) significantly (P<0.05) reduced the dilation of the inner diameter of left ventricle (LV) (4.27 ± 0.29 mm), increased wall thickness of LV (1.5 ± 0.13 mm) and improved the fractional shortening of LV of hearts (LVFS) (42.8 ± 2.4%) compared with those values of LVs of rats without implants (MI group) (e.g., 5.92 ± 0.39 mm, 1.2 ± 0.06 mm and 31.5±1.4%, respectively). Moreover, blood vessel-like structures in MI regions of LVs in the CHS group were widely distributed while none was found in the MI group. The CHS group significantly improved the secretion of paracrine factors, such as VEGF in the MI regions of LVs (P<0.05, n=4), relative to that in the MI group. In conclusion, chitosan-HYA/SF cardiac patches are promising biomaterials for the cardiac repair of MI rat hearts.
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Assessing human urinary proteome using a mass spectrometry-based profiling system combined with magnetic nanoparticles.
Clin. Chim. Acta
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Samples originating from body fluids often contain a complex mixture of inorganic salts, buffers, chaotropic agents, surfactant/detergents, preservatives, and other solubilizing agents. The presence of those contaminants often precludes direct analysis by mass spectrometry. Urine, a blood filtrate produced by the urinary system, is an ideal bio-sample and a rich source of biomarkers for diagnostic information.
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Utilizing isotope dilution-matrix-assisted laser desorption ionization-time of flight mass spectrometry as a reference procedure for the radioimmunoassay of serum thyroxine.
Clin. Chim. Acta
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The thyroid hormone, thyroxine (T4), is a tyrosine-based hormone produced by the thyroid gland, which is essential in regulating a number of biological processes, including growth, neurodevelopment, carbohydrate metabolism, oxygen consumption and protein synthesis. Data on human thyroid hormone metabolism were gathered since the middle of the 1970s mainly by the use of radioactive iodinated ((125)I or (131)I) hormones.
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Determining early adhesion of cells on polysaccharides/PCL surfaces by a quartz crystal microbalance.
J Mater Sci Mater Med
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The early adhesions of cells to various biopolymers are important to their growths and proliferations. Here, the adhesion of cells (e.g., fibroblasts) on the electrode of a quartz crystal microbalance (QCM) that was coated by PCL or PEG/PCL and further adsorbed by chitosan (CS) or CS/hyaluronic acid (HA) layers, was examined by cell-counting technique, QCM method and MTS assay under a serum-free condition for 3 h. The surfaces on electrodes of the QCM were confirmed to have been modified by measuring their contact angles, FT-IR spectra and the weights of biopolymers affected the frequency shifts of the QCM. Among tested surfaces on electrodes, the adhesion of fibroblasts on a HA/CS/PCL surface was the most (e.g., 3.08 × 10(5) cells/cm(2)) while that on a PEG/PCL surface was the least (e.g., 0.7 × 10(5) cells/cm(2)), as determined by cell-counting technique. The frequency shift and the mass of adhering fibroblasts on HA/CS/PCL electrodes were -3,537 ± 770 Hz and 3.78 ± 0.22 ?g (n = 3), respectively, that were significantly exceeded those on other electrodes (-393 ± 58 Hz and 0.32 ± 0.06 ?g, n = 3, respectively, for PEG/PCL electrodes). These results were consistent with cell-counting technique. Although MTS assay yielded similar results, it was less sensitive than the two aforementioned methods. In conclusion, modified electrodes of a QCM provide a convenient and sensitive method for examining the early adhesion of cells (e.g., 3 h) to biopolymer surfaces.
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Assessing the responses of cellular proteins induced by hyaluronic acid-modified surfaces utilizing a mass spectrometry-based profiling system: over-expression of CD36, CD44, CDK9, and PP2A.
Analyst
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The cell responses to biopolymer surface at the early adhesion stages can be critical for cell survival. The purpose of this research was to assess formation of hyaluronic acid (HA) biopolymer surface, the fibroblasts were used as an experimental model to evaluate the responses of cellular proteins induced by biopolymer materials using a mass spectrometry-based profiling system. Surfaces were covered by multi-walled carbon nanotubes (CNT), chitosan (CS), and HA to increase the surface area, enhance the adhesion of biopolymer and promote the rate of cell proliferation. The amount of adhered fibroblasts on CNT/CS/HA electrodes of quartz crystal microbalance (QCM) were greatly exceeded those on other surfaces that were consistent with cell-count technique. Moreover, analyzing differential protein expressions of adhered fibroblasts on those biopolymer surfaces by proteomic approaches identified CD36, CD44, PP2A, and CDK9 as key proteins. To validate the influences of those four proteins on adhesions of fibroblasts on biopolymers, the cells were blocked by antibodies of the proteins and the adhesions of cells on the tested biopolymer surfaces were examined using a QCM technique, flow cytometric analysis and morphological observations. The results of significantly decreasing the weights and densities of the blocked fibroblasts adhering to CNT/CS/HA surfaces were obtained, and validate those proteins found by proteomic approaches. Utilizing mass spectrometry-based proteomics to evaluate cell adhesions on biopolymers is proposed.
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Cardiac repair achieved by bone marrow mesenchymal stem cells/silk fibroin/hyaluronic acid patches in a rat of myocardial infarction model.
Biomaterials
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Bone marrow mesenchymal stem cells/silk fibroin/hyaluronic acid (BMSC/SH) patches were implanted into myocardial infarction (MI) rat hearts to investigate the efficacies of them on enhancing left ventricular (LV) remodeling and cardiac repair. 45 rats were divided into four groups: Sham, MI (MI hearts, induced by a cryo-injury technique), SH and BMSC/SH (MI hearts with implantations of SH and BMSC/SH patches, respectively). After eight weeks of post-implantation, the patches for the SH and BMSC/SH groups were intact and well adhered on the MI zones with no and minor immunological responses, respectively, examined by a CD68 marker, while severe inflammation on the zones was observed for the MI group. The SH group showed the efficacy of cardiac repair on MI zones. Moreover, BMSC/SH group significantly improved the wall thickness of LV, assessed by echocardiography, and had high viability of delivery BMSC, largely reduced apoptosis, significantly promoted neo-vascularization and stimulated the secretions of various paracrine factors such as VEGF, examined by real-time PCR, in MI zones compared with those of the SH and MI groups. In conclusion, the therapeutic efficacies of using BMSC/SH patches for repairing MI hearts were demonstrated by showing the advantages of both bioactive SH patches and BMSC-based therapy.
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Activity-dependent neuroprotector homeobox protein: A candidate protein identified in serum as diagnostic biomarker for Alzheimers disease.
J Proteomics
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Alzheimers disease (AD) is the most common cause of dementia of late life. To enhance our understanding of AD proteome, the serum proteins were analyzed using two-dimensional gel electrophoresis (2DE) combined with nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (nano-HPLC-ESI-MS/MS) followed by peptide fragmentation patterning. In this study, six protein spots with differential expression were identified. Five up-regulated proteins were identified as actin, apolipoprotein A-IV (Apo A-IV), inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), alpha-1-antitrypsin (AAT), and antithrombin-III (AT-III); one protein, activity-dependent neuroprotector homeobox protein (ADNP) was down-regulated in AD patients. These proteins with differential expression in the serum may serve as potential indicators of AD. Our results suggested that ADNP may play an important role in slowing the progression of clinical symptoms of AD.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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