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ABSTRACT: In this study, the four commonly used cytotoxicity assays and the mechanical properties as evaluated by atomic force microscopy (AFM) were compared in a cellular system. A cytotoxicity assay is the first and most essential test to evaluate biocompatibility of various toxic substances. Many of the cytotoxicity methods require complicated and labor-intensive process, as well as introduce experimental error. In addition, these methods cannot provide instantaneous and quantitative cell viability information. AFM has become an exciting analytical tool in medical, biological, and biophysical research due to its unique abilities. AFM-based force-distance curve measurements precisely measure the changes in the biophysical properties of the cell. Therefore, we observed the morphological changes and mechanical property changes in L929 cells following sodium lauryl sulfate (SLS) treatment utilizing AFM. AFM imaging showed that the toxic effects of SLS changed not only the spindle-like shape of L929 cells into a round shape, but also made a rough cell surface. As the concentration of SLS was increased, the surface roughness of L929 cell was increased, and stiffness decreased. We confirmed that inhibition of proliferation clearly increased with increases in SLS concentration based on results from MTT, WST, neutral red uptake, and LIVE/DEAD viability/cytotoxicity assays. The estimated IC50 value by AFM analysis was similar to those of other conventional assays and was included within the 95% confidence interval range. We suggest that an AFM quantitative analysis of the morphological and biophysical changes in cells can be utilized as a new method for evaluating cytotoxicity.
Micron 03/2013; · 1.53 Impact Factor
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ABSTRACT: The aim of this study was to quantitatively investigate the effects of extracorporeal shockwave therapy (ESWT) on the nanostructure and adhesion force of collagen fibrils in a rat model of collagenase-induced Achilles tendinitis (CIAT) using histology and atomic force microscopy. A total of 45 rats were divided into experimental groups of three rats each: a control group, 27 CIAT rats with nine time points, and 15 ESWT rats with five time points. Progressive changes in nanostructure including the fibrillary diameter and D-periodicity, and biomechanical properties including the fibrillary adhesion forces in each healing phase were investigated over a 5-week period after collagenase injection. On postoperative day 3, CIAT rats showed granulomatous tissue associated with subacute inflammation, and a deterioration in nanostructure and mechanical properties compared to controls. On postoperative day 12, the ESWT group showed increased vascularity, fibroblastic activity, lymphocyte and plasma cell infiltration, dense histocytes, and disorganization of the fibers compared to the CIAT group. The ESWT group showed and improvement in nanostructure and mechanical properties compared to controls, while the CIAT group showed a deterioration in nanostructure and mechanical properties compared to controls. On postoperative day 26, the ESWT group showed 30% inflamed tissue and 70% fibrotic tissue, while the CIAT group showed chronic inflammation. By the end of the experiments, in both groups the changes had reversed and the tissues were similar in appearance to those in the control group. Following ESWT the deformed and irregular collagen network returned to a well-aligned normal collagen network nanostructure. These results suggest that ESWT may promote the healing response in Achilles tendinitis.
Lasers in Medical Science 11/2012; 27(6):1195-204. · 2.00 Impact Factor
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ABSTRACT: Because oxygen plays a critical role in the pathophysiology of myocardial injury during subsequent reperfusion, as well as ischemia, the accurate measurement of myocardial oxygen tension is crucial for the assessment of myocardial viability by ischemia-reperfusion (IR) injury. Therefore, we utilized a sol-gel derived electrochemical oxygen microsensor to monitor changes in oxygen tension during myocardial ischemia-reperfusion. We also analyzed differences in oxygen tension recovery in post-ischemic myocardium depending on ischemic time to investigate the correlation between recovery parameters for oxygen tension and the severity of IR injury. An oxygen sensor was built using a xerogel-modified platinum microsensor and a coiled Ag/AgCl reference electrode. Rat hearts were randomly divided into 5 groups: control (0 min ischemia), I-10 (10 min ischemia), I-20 (20 min ischemia), I-30 (30 min ischemia), and I-40 (40 min ischemia) groups (n = 3 per group, respectively). After the induction of ischemia, reperfusion was performed for 60 min. As soon as the ischemia was initiated, oxygen tension rapidly declined to near zero levels. When reperfusion was initiated, the changes in oxygen tension depended on ischemic time. The normalized peak level of oxygen tension during the reperfusion episode was 188 ± 27 in group I-10, 120 ± 24 in group I-20, 12.5 ± 10.6 in group I-30, and 1.24 ± 1.09 in group I-40 (p < 0.001, n = 3, respectively). After 60 min of reperfusion, the normalized restoration level was 129 ± 30 in group I-10, 88 ± 4 in group I-20, 3.40 ± 4.82 in group I-30, and 0.99 ± 0.94 in group I-40 (p < 0.001, n = 3, respectively). The maximum and restoration values of oxygen tension in groups I-30 and I-40 after reperfusion were lower than pre-ischemic values. In particular, oxygen tension in the I-40 group was not recovered at all. These results were also demonstrated by TTC staining. We suggest that these recovery parameters could be utilized as an index of tissue injury and severity of ischemia. Therefore, quantitative measurements of oxygen tension dynamics in the myocardium would be helpful for evaluation of the cardioprotective effects of therapeutic treatments such as drug administration.
The Analyst 09/2012; 137(22):5312-9. · 4.23 Impact Factor
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ABSTRACT: Background/Aims: Recent studies have shown that angiotensin II (Ang II) type 1 receptor blockers (ARB) may provide renal protection independent of their blood pressure-lowering effect. However, evidence for this comes from indirect methods, such as genetic or protein expression studies. In this study, we hypothesized that telmisartan, a specific ARB, applied to Ang II-stimulated mesangial cell (MC) would exert a renoprotective effect via modulation of MCs' mechanical properties. Methods: We investigated the effect of telmisartan on Ang II-induced changes in MCs utilizing real-time atomic force microscopy (AFM) imaging and force-distance curve measurements. Results: Real-time AFM images of live MCs demonstrated that cells contracted towards the center after Ang II exposure, and telmisartan treatment abolished this change. Cellular spring constants showed that telmisartan prevented Ang II-induced MC stiffening (Ang II: 0.109 ± 0.019 N/m, Ang II + telmisartan: 0.051 ± 0.016 N/m, p < 0.005). Telmisartan-treated MCs had a significantly lower adhesion force than those of the control group (control: 0.49 ± 0.22 nN, telmisartan: 0.22 ± 0.06 nN, Ang II: 0.40 ± 0.25nN, Ang II + telmisartan: 0.27 ± 0.14 nN, p < 0.005). These results demonstrate that the dynamic contraction and mechanical properties of Ang II-stimulated MCs are restored by telmisartan. Conclusions: We report for the first time the use of AFM force-distance curves on live MCs to directly monitor changes in surface adhesion and stiffness of cells after treatment with telmisartan in real time.
Kidney and Blood Pressure Research 08/2012; 35(6):573-582. · 1.46 Impact Factor
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ABSTRACT: Mitochondrial dysfunction plays a central role in mediating both the necrotic and apoptotic components of reperfusion injury. Because mitochondrial swelling is one of the most important indicators of the beginning of mitochondrial permeability transition, quantification of morphological changes in mitochondria would be useful in evaluating the degree of IR injury, as well as the protective effects of various therapies. In this study, we characterized the morphological changes in heart mitochondria caused by the duration and severity of ischemia utilizing particle shape analysis on atomic force microscopy (AFM) topographic images. We also simultaneously investigated the nano-mechanical changes in rat heart mitochondria by injury using force-distance curve measurements. Rats were randomly divided into 3 groups: control group (n=3), myocardial ischemia without reperfusion (PI group, n=3), and myocardial ischemia with reperfusion (IR group, n=4). Normal mitochondria appeared ellipsoidal with a mean area of 3551±1559nm(2) and mean perimeter of 217.54±52.09nm (n=60). The mean area and perimeter of mitochondria in the IR groups increased to 28,181±21,248nm(2) and 595.74±234.29nm (n=40, p<0.0001 vs. control group, respectively), maintaining oval in shape. But, in the PI group, all parameters showed significant differences compared to parameters of the control group (n=35, p<0.0001). In particular, the mean axial ratio and roundness were significantly different from those in the IR group. Mitochondria in the PI group looked more spherical than those of control and IR groups. Adhesion force is the force before the last event on the retraction half of force-distance curve measurements, corresponding to the point where the tip and the surface loose contact. The adhesion forces of heart mitochondria in the IR and PI groups significantly decreased to 19.56±1.08nN (n=30, p<0.0001) and 18.65±3.18nN (n=30, p<0.0001), compared to normal mitochondria which had an adhesion force of 27.64±0.88nN (n=30). Adhesion force is governed by the attractive portion of the interacting forces between the surface atoms of the contacts. From the morphological and nano-mechanical changes in heart mitochondria, we suggested that the outer membranes of mitochondria were broken by myocardial ischemic injury before they became swollen, and the swelling might be correlated with the ischemic injury. We inferred that the breakage of membranes leads to uptake of water and matrix swelling. As a result, shape measurement parameters for the quantitative analysis of mitochondrial swelling could be very effective for evaluating the myocardial injury.
Micron 06/2012; · 1.53 Impact Factor
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ABSTRACT: V-AKT murine thymoma viral oncogene homolog 1 (AKT1) is an important downstream target of the insulin-signaling pathway and
may be an important regulator of pancreatic beta cell growth. This study investigated the association of theAKT1 gene with susceptibility to type 2 diabetes mellitus and its related traits. By sequencing theAKT1 gene in 24 unrelated individuals, we iden-tified 32 genetic variations including 30 single nucleotide polymorphisms and 2
deletions. For the association analysis, we selected seven single nucleotide polymorphisms (rs10138227, −726G>A; rs3730358,
+12574C>T; rs2494737, +12656T>A; rs2498796, +15761T>C; rs2498799, +19087 A>G; rs2494732, +19789G>A; rs3803304, +19835G>C)
based on minor allele frequency (>0.05) and linkage disequilibrium status. The study included 483 type 2 diabetes patients
(206 men and 277 women with mean age 64±2.8 years and mean age at onset 56 ± 8.1 years) and 1,138 non-diabetic control subjects
(516 men and 622 women with mean age 64 ±2.9 years). Two single nucleotide polymorphisms (rs2498796, +15761T>C and rs2494732,
+19789G>A) were found to be associated with risk of type 2 diabetes mellitus, and showed an increased risk of type 2 diabetes
mellitus in a recessive model (OR=1.343, 95% CI 1.021–1.765,p=0.035 and OR=1.534, 95% CI 1.058–2.225,p=0.024, respectively). These SNPs were also associated with diabetes-related traits such as levels of fasting blood glucose
and hemoglobin A1c. In addition, type 2 diabetes mellitus patients who also have dyslipidemia or high blood pressure showed
significant association with single nucleotide polymorphisms in AKT1 when compared with healthy controls. These results indicate
that genetic variation in AKT1 influences the development of type 2 diabetes mellitus in the Korean population.
Key wordsT2DM-AKT1-insulin-signaling pathway-single nucleotide polymorphism-dyslipidemia-high blood pressure
Genes & genomics 04/2012; 31(1):73-83. · 0.44 Impact Factor
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ABSTRACT: The aim of this study was to quantitatively investigate the effects of extracorporeal shockwave therapy (ESWT) on the nanostructure and adhesion force of collagen fibrils in a rat model of collagenase-induced Achilles tendinitis (CIAT) using histology and atomic force microscopy. A total of 45 rats were divided into experimental groups of three rats each: a control group, 27 CIAT rats with nine time points, and 15 ESWT rats with five time points. Progressive changes in nanostructure including the fibrillary diameter and D-periodicity, and biomechanical properties including the fibrillary adhesion forces in each healing phase were investigated over a 5-week period after collagenase injection. On postoperative day 3, CIAT rats showed granulomatous tissue associated with subacute inflammation, and a deterioration in nanostructure and mechanical properties compared to controls. On postoperative day 12, the ESWT group showed increased vascularity, fibroblastic activity, lymphocyte and plasma cell infiltration, dense histocytes, and disorganization of the fibers compared to the CIAT group. The ESWT group showed and improvement in nanostructure and mechanical properties compared to controls, while the CIAT group showed a deterioration in nanostructure and mechanical properties compared to controls. On postoperative day 26, the ESWT group showed 30% inflamed tissue and 70% fibrotic tissue, while the CIAT group showed chronic inflammation. By the end of the experiments, in both groups the changes had reversed and the tissues were similar in appearance to those in the control group. Following ESWT the deformed and irregular collagen network returned to a well-aligned normal collagen network nanostructure. These results suggest that ESWT may promote the healing response in Achilles tendinitis.
Lasers in Medical Science 01/2012; · 2.00 Impact Factor
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ABSTRACT: The aim of this study is to quantitatively investigate the short-term effects of RF tissue-tightening treatment in in vivo rabbit dermal collagen fibrils. These effects were measured at different energy levels and at varying pass procedures on the nanostructural response level using histology and AFM analysis. Each rabbit was divided into one of seven experimental groups, which included the following: control group, and six RF group according to RF energy (20 W and 40 W) and three RF pass procedures. The progressive changes in the diameter and D-periodicity of rabbit dermal collagen fibrils were investigated in detail over a 7-day post-treatment period. The dermal tissues treated with the RF tissue-tightening device showed more prominent inflammatory responses with inflammatory cell ingrowth compared to the control. This effect showed more prominent with the passage of day after treatment. Although an increase in the diameter and D-periodicity of dermal collagen fibrils was identified immediately after the RF treatment, a decrease in the morphology of dermal collagen fibrils continued until post-operative day 7. Furthermore, RF treatment led to the loss of distinct borders. Increases in RF energy with the same pass procedure, as well as an increase in the number of RF passes, increased the occurrence of irreversible collagen fibril injury. A multiple-pass treatment at low energy rather than a single-pass treatment at high energy showed a large amount of collagen fibrils contraction at the nanostructural level.
Lasers in Medical Science 10/2011; 27(5):923-33. · 2.00 Impact Factor
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ABSTRACT: This study presents a novel algorithm to measure joint space widths (JSWs) in patients with rheumatoid arthritis (RA) using radiographic hand images. Radiographic images were first preprocessed, and then phalangeal regions corresponding to the bone structures of each finger were extracted using step-wedge functions. Phalangeal branch paths were also extracted. Each of the five extracted phalangeal branch paths matched the bone structures of each finger exactly and ran through the center of each finger. The algorithm automatically detected 14 joints, which were identified as sharp changes in gray scale intensity along phalangeal branch paths through the profile plot. The regions of interest corresponding to the 14 joints were subsequently extracted. A total of 35 radiographic images from three groups were tested. The performance of our algorithm was evaluated by measuring joint location percentage errors and mean JSWs for three joints in the phalanges. The algorithm correctly detected 94.69% of total joints and had a low detection rate in RA patients with severe deformities or ankylosis. The mean JSW in the control group was significantly greater than that in the RA group (p<0.05). In contrast, the standard deviation of JSW in the control group was lower than that in the RA groups (p<0.005). Control and seropositive RA groups showed significant symmetry in JSW values.
Computers in biology and medicine 09/2011; 41(10):987-98. · 1.27 Impact Factor
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ABSTRACT: The exact mechanism of rheumatoid arthritis (RA) is unclear, but a combination of genetic, environmental and hormonal factors is thought to be involved. This study examined the progressive arthritic reaction of murine type II collagen-induced arthritis (CIA), a representative animal model of RA. Arthritic reactions, including inflammation and bone erosion were examined using an objective non-invasive method. Two scoring systems were used to evaluate changes in cutaneous inflammation and bone erosion during RA progression. The severity of inflammation was evaluated by visual scoring of erythema and edema, while the degree of bone erosion was quantified by macroradiographical erosion analysis of specific bones. A significant difference was observed in both visual (P = 0.0001, n = 7) and radiographic (P < 0.0001, n = 7) examinations for the RA group as compared to the control. The relationship between inflammatory change and erosive change in bone showed a significant positive correlation, r = 0.9550 (P < 0.0001, n = 7). The overall rate of asymmetry was 25.23% in both fore- and hindpaws. The results generated from these experiments show that murine CIA is a promising model for elucidating the mechanism of RA. In addition, the results of this study may be used for monitoring RA progression as well as screening therapy efficacy in the joint pathology.
Rheumatology International 05/2011; 31(5):595-603. · 1.88 Impact Factor
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ABSTRACT: The Achilles tendon consists mainly of type I collagen fibers that contain collagen fibrils. When the Achilles tendon is injured, it is inflamed. The collagenase-induced model has been widely used to study tendinitis. The major advantages of atomic force microscopy (AFM) over conventional optical and electron microscopy for bio-imaging include its non-requirement of a special coating and vacuum, and its capability to perform imaging in all environments. AFM force-distance measurements have become a fundamental tool in the fields of surface chemistry, biochemistry and materials science. Therefore, the changes in the ultrastructure and adhesion force of the collagen fibrils on the Achilles tendons of rats with Achilles tendinitis were observed using AFM. The changes in the structure of the Achilles tendons were evaluated based on the diameter and D-banding of the collagen fibrils. Collagenase-induced Achilles tendinitis was induced with the injection of 30 microl crude collagenase into 7-week-old male Sprague-Dawley rats. The animals were each sacrificed on the first, second, third, fifth and seventh day after the collagenase injection. The normal and injured Achilles tendons were fixed in 4% buffered formalin and dehydrated with increasing concentrations of ethanol. AFM was performed using the non-contact mode at the resolution of 512 x 512 pixels, with a scan speed of 0.8 line/sec. The adhesion force was measured via the force-distance curve that resulted from the interactions between the AFM tip and the collagen fibril sample using the contact mode. The diameter of the collagen fibrils in the Achilles tendons significantly decreased (p < 0.05) after the collagenase injection, and the pattern of the D-banding of the collagen fibrils was similar to that of the diameter changes. The adhesion force decreased until the fifth day after the collagenase injection, but increased on the seventh day after the collagenase injection (p < 0.0001).
Journal of Nanoscience and Nanotechnology 01/2011; 11(1):773-7. · 1.56 Impact Factor
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ABSTRACT: This study examined the neuroprotective effects of magnesium-sulfate (MgSO(4)) on the cerebral blood flow (CBF) and extracellular glutamate concentration in an eleven vessel occlusion (11VO) rat model. Twenty-one male Sprague-Dawley rats (250-350g) were used for the 11VO ischemic model, which was induced by a 10-min transient occlusion. The animals were divided into 3 groups, including ischemic-induced animals (ischemia group), ischemic-induced and MgSO(4) treated animals (MgSO(4) group), and sham animals for comparison. The real-time extracellular glutamate concentration was measured using a microdialysis biosensor, and the CBF was monitored by laser Doppler flowmetry. Neuronal cell death in the hippocampal region was observed 72h after ischemia by several stains (Nissl, DAPI, NeuN, and cleaved caspase3). A significant decrease in %CBF was observed in both the ischemia and MgSO(4) groups, such as ~10% during the ischemic period. However, the MgSO(4) group showed a significant decrease in the initial reperfusion %CBF compared to the ischemia group. A significantly lower level of glutamate release was observed in the MgSO(4) group than in the ischemia group during the ischemic and reperfusion episode. Our staining results revealed a significant decrease in neuronal cell death in the hippocampus in the MgSO(4) group compared to the ischemia group. These results suggest that MgSO(4) is responsible for the protection of neuronal cells by suppressing the release of extracellular glutamate under ischemic conditions and the CBF response during the initial reperfusion period.
Brain research 01/2011; 1371:121-8. · 2.46 Impact Factor
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ABSTRACT: Glutamate is a key excitatory neurotransmitter in the brain, and its excessive release plays a key role in the development of neuronal injury. In order to define the effect of nimodipine on glutamate release, we monitored extracellular glutamate release in real-time in a global ischemia rat model with eleven vessel occlusion.
TWELVE RATS WERE RANDOMLY DIVIDED INTO TWO GROUPS: the ischemia group and the nimodipine treatment group. The changes of extracellular glutamate level were measured using microdialysis amperometric biosensor, in coincident with cerebral blood flow (CBF) and electroencephalogram. Nimodipine (0.025 µg/100 gm/min) was infused into lateral to the CBF probe, during the ischemic period. Also, we performed Nissl staining method to assess the neuroprotective effect of nimodipine.
During the ischemic period, the mean maximum change in glutamate concentration was 133.22±2.57 µM in the ischemia group and 75.42±4.22 µM (p<0.001) in the group treated with nimodipine. The total amount of glutamate released was significantly different (p<0.001) between groups during the ischemic period. The %cell viability in hippocampus was 47.50±5.64 (p<0.005) in ischemia group, compared with sham group. But, the %cell viability in nimodipine treatment group was 95.46±6.60 in hippocampus (p<0.005).
From the real-time monitoring and Nissl staining results, we suggest that the nimodipine treatment is responsible for the protection of the neuronal cell death through the suppression of extracellular glutamate release in the 11-VO global ischemia model of rat.
Journal of Korean Neurosurgical Society 01/2011; 49(1):1-7. · 0.60 Impact Factor
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ABSTRACT: Glutamate is the principal excitatory neurotransmitter in the brain, and its excessive release plays a key role in neuronal death associated with a wide range of neural disorders. Real-time monitoring of extracellular glutamate levels would be very helpful in understanding the excitotoxic process of neurotransmitters on brain injury. Toward the detection of L: -glutamate, we describe in this chapter the preparation of carbon nanotube (CNT) network probes with immobilized L: -glutamate oxidase (GLOD) by using a non-covalent functionalized method. Such GOLD-CNT network probes are evaluated with real-time electronic responses corresponding to standard glutamate solutions in vitro and a 11-vessel occlusion (11 VO) rat model in vivo. The ultrahigh sensitivity, selectivity, and fast response time of GLOD-CNT network probes are greatly promising for the real-time electronic detection of extracellular glutamate levels in brain.
Methods in molecular biology (Clifton, N.J.) 01/2011; 743:65-75.
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ABSTRACT: In orthodontics, the surface roughnesses of orthodontic archwire and brackets affect the effectiveness of arch-guided tooth movement, corrosion behavior, and the aesthetics of orthodontic components. Atomic force microscopy (AFM) measurements were used to provide quantitative information on the surface roughness of the orthodontic material. In this study, the changes in surface roughness of various orthodontic bracket slots before and after sliding movement of archwire in vitro and in vivo were observed through the utilization of AFM. Firstly, we characterized the surface of four types of brackets slots as follows: conventional stainless steel (Succes), conventional ceramic (Perfect), self-ligating stainless steel (Damon) and self-ligating ceramic (Clippy-C) brackets. Succes) and Damon brackets showed relatively smooth surfaces, while Perfect had the roughest surface among the four types of brackets used. Secondly, after in vitro sliding test with beta titanium wire in two conventional brackets (Succes and Perfect), there were significant increases in only stainless steel bracket, Succes. Thirdly, after clinical orthodontic treatment for a maximum of 2 years, the self-ligating stainless steel bracket, Damon, showed a significant increase in surface roughness. But self-ligating ceramic brackets, Clippy-C, represented less significant changes in roughness parameters than self-ligating stainless steel ones. Based on the results of the AFM measurements, it is suggested that the self-ligating ceramic bracket has great possibility to exhibit less friction and better biocompatibility than the other tested brackets. This implies that these bracket slots will aid in the effectiveness of arch-guided tooth movement.
Micron 10/2010; 41(7):775-82. · 1.53 Impact Factor
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ABSTRACT: Mitochondria play critical roles in both the life and the death of cardiac myocytes. Various factors, such as the loss of ATP synthesis and increase of ATP hydrolysis, impairment in ionic homeostasis, formation of reactive oxygen species (ROS), and release of proapoptotic proteins are related to the generation of irreversible damage. It has been proposed that the release of cytochrome c is caused by a swelling of the mitochondrial matrix triggered by the apoptotic stimuli. However, there is a controversy about whether or not the mitochondria, indeed, swell during apoptosis. The major advantages of atomic force microscopy (AFM) over conventional optical and electron microscopes for bio-imaging include the fact that no special coating and vacuum are required and imaging can be done in all environments--air, vacuum or aqueous conditions. In addition, AFM force-distance curve measurements have become a fundamental tool in the fields of surface chemistry, biochemistry, and material science. In this study, we used AFM to observe the morphological and property changes in heart mitochondria that were isolated from a rat myocardial infarction model. From the shape parameters of the mitochondria in the AFM topographic image, it seemed that myocardial infarction caused the mitochondrial swelling. Also, the results of force-distance measurements showed that the adhesion force of heart mitochondria was significantly decreased by myocardial in infarction. Therefore, we suggested that myocardial infarction might be the cause of mitochondrial swelling and the changes in outer membrane of heart mitochondria.
Micron 09/2010; 42(3):299-304. · 1.53 Impact Factor
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ABSTRACT: A thorough characterization of the morphological structure and physical properties is essential for an understanding of human hair. A number of techniques such as scanning electron microscopy, transmission electron microscopy and confocal microscopy have been used to study hair surfaces. Recently, atomic force microscopy (AFM) has emerged as an ideal method for the non-invasive examination of hair surfaces.
To investigate the effects of aging on normal Korean hair diameter and surface features using AFM.
We enrolled 60 Korean volunteers of various ages who had no hair diseases. We analyzed hair diameter, AFM images of the hair surface, cuticular descriptors and micro-scale mechanical properties for their associations with aging. Results: Hair diameter was found to increase for the first 20-30 years of life, after which it began to decrease. AFM images of most of the younger subjects showed typical step-like topographic properties with clear scale edges. The AFM images of most of the older subjects revealed dilapidated structures, poorly demarcated scale edges and undulated surfaces. Among the cuticular descriptors, surface roughness increased significantly with age. Force to distance analysis demonstrated a dependence on age.
These results suggest that aging causes changes in hair diameter and surface structure.
Skin Research and Technology 07/2010; 17(1):63-8. · 1.71 Impact Factor
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ABSTRACT: The aim of this study was to define the effects of glutamate release on cell death in an eleven vessel rat occlusion model. Male Sprague-Dawley rats (250-350g) were used for the 11 vessel occlusion ischemic model, which was induced by a 5- and 10-min transient occlusion. During the surgical procedure, the extracellular glutamate concentration was measured in real-time using a microdialysis amperometirc biosensor with cerebral blood flow. In order to confirm neuronal cell death, brains were removed 72h after ischemia for the detection of the neuron-specific nuclear protein and cleaved caspase-3 levels, using double-immunofluorescence. A significant decrease in % cerebral blood flow was observed in both the 5- and 10-min 11 vessel occlusion models, while an increase in glutamate release was detected after the onset of ischemia that continued to rise during the ischemic period. However, a significantly higher level of glutamate release was observed in the 10-min ischemia group compared to the 5-min group. Unlike the small amount of brain damage in the 5-min group, the increased glutamate levels in the 10-min group resulted in ischemic cell death in the hippocampal region with the activation of cleaved caspase-3 and the inhibition of neuron-specific nuclear protein expression. This study suggests that the increased level of glutamate release induces apoptotic cell death in the 11 vessel occlusion ischemic model.
Brain research 06/2010; 1342:160-6. · 2.46 Impact Factor
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ABSTRACT: Acupuncture (ACU) is known to be effective in ischemia treatment, and glutamate (GLU) excitotoxicity is an important factor in neuronal cell death. We observed the effect of ACU on cerebral blood flow (%CBF) and DeltaGLU (the changes in GLU release) in the ischemic stroke rat model of diabetic mellitus (DM). A global ischemia was induced using the eleven-vessel occlusion (11-VO) method in 14 Sprague-Dawley rats (DM), which were randomly divided into two groups: the control group and the ACU-treatment group. Extracellular DeltaGLU was assessed using an intra-cerebral biosensor system measuring 256 samples per second, simultaneously with %CBF and electroencephalogram. ACU stimulation was applied to ACU points GB34 and GB39 during the ischemic period. Twenty-three diagnostic parameters were proposed first for a detailed analysis of changes in %CBF and GLU release during ischemia/reperfusion. ACU rats showed a significant decrease in ischemic (p < 0.05) and reperfusion %CBF (p < 0.0001) than control rats, and a significantly larger decrease in ischemic DeltaGLU (p < 0.05) and peak level of reperfusion DeltaGLU (p < 0.005) than control rats. From these results, we suggest that ACU stimulation is responsible for the potential protection of neurons through suppression of %CBF response in the increased plasma osmolality and extracellular DeltaGLU in diabetic rats under ischemic conditions.
Physiological Measurement 03/2010; 31(5):633-47. · 1.68 Impact Factor
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ABSTRACT: An increase in excitotoxic amino acid glutamate (GLU) concentration associated with neuronal damage might be the cause of the ischemic damage observed in stroke patients suffering from hyperglycemia. However, the effect has never been investigated by real-time in vivo monitoring. Therefore, this study examined the effects of the functional responses of ischemia-evoked electroencephalography (EEG), cerebral blood flow (%CBF) and DeltaGLU in hyperglycemia through real-time in vivo monitoring. Five Sprague-Dawley rats were treated with streptozocin (hyperglycemia) and five normal rats were used as the controls. Global ischemia was induced using an 11-vessel occlusion model. The experimental protocols consisting of 10 min pre-ischemic, 10 min ischemic and 40 min reperfusion periods were applied to both groups. Under these conditions, the responses of the ischemia-evoked EEG, %CBF and DeltaGLU were monitored in real time. The EEG showed flat patterns during ischemia followed by poor recovery during reperfusion. The peak reperfusion %CBF was decreased significantly in the hyperglycemia group compared to the control group (p < 0.05, n = 5). The extracellular DeltaGLU releases increased significantly during ischemia (p < 0.0001, n = 5) and reperfusion (p < 0.001, n = 5) in the hyperglycemia group compared to the control group. The decrease in reperfusion %CBF during short-term hyperglycemia might be related to the increased plasma osmolality, decreased adenosine levels and swollen endothelial cells with decreased vascular luminal diameters under hyperglycemic conditions. And, the increase in DeltaGLU during short-term hyperglycemia might be related to the neurotoxic effects of the high extracellular concentrations of DeltaGLU and the inhibition of GLU uptake.
Physiological Measurement 02/2010; 31(3):439-50. · 1.68 Impact Factor
