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Purpose: While initial fixation using a press-fit of the acetabular cup is critical for the durability of the component, restoration of the hip center is regarded as an attributable factor for implant survival and successful outcome. In protrusio acetabuli (PA), obtaining both restoration of the hip center and the press-fit of the acetabular cup simultaneously might be difficult during total hip arthroplasty (THA). We tested the hypothesis that use of a medialized cup, if press-fitted, will not result in compromise of the implant stability and outcome after cementless THA of PA. Materials and Methods: A total of 26 cementless THAs of 22 patients with PA were reviewed. During THA, press-fit of the cup was prioritized rather than hip center restoration. A press-fit was obtained in 24 hips. A press-fit could not be obtained in the two remaining hips; therefore, reinforcement acetabular components were used. Restoration of the hip center was achieved in 17 cups; 15 primary cups and two reinforcement components; it was medialized in nine cups. Implant stability and modified Harris hip score (mHHS) between the two groups were compared at a mean follow-up of 5.1 years (range, 2-16 years). Results: Twenty-six cups; 17 restored cups and nine medialized press-fitted cups, remained stable at the latest follow-up. A similar final mHHS was observed between the restored group and the medialized group (83.6±12.1 vs 83.8±10.4, P=0.786). Conclusion: Implant stability and favorable results were obtained by press-fitted cups, irrespective of hip center restoration. THA in PA patients showed promising clinical and radiological results.
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PURPOSE: The purposes of this study are (1) to measure the ischiofemoral distance (IFD) in nondiseased hips and (2) to reveal patient demographic and anatomical factors associated with IFD. METHODS: In this retrospective study, we identified patients who had unilateral osteonecrosis of the femoral head on computed tomography (CT) scan from November 2005 to July 2018 and assessed the unaffected contralateral hips of the patients. Among the contralateral hips, we excluded hips with hip pain, incomplete or poor-quality CT image, incomplete medical record, degenerative arthritis of the hip, or previous hip surgery. IFD was measured on the axial CT image, and correlated demographic factors (age, sex, height, weight, and body mass index) and anatomical parameters (neck-shaft angle of the femur and femoral anteversion) with IFD were evaluated. RESULTS: Five hundred seventeen patients (517 hips) were evaluated. There were 302 men and 215 women, and their mean age was 51.7 years (range 15-83 years). The mean IFD was 33.2 (±9.2) mm in men and 24.3 (±8.9) mm in women (P < .001). Interobserver and intraobserver reliability (intraclass correlation coefficients) were 0.99 (95% confidence interval 0.98-1.0) and 0.98 (95% confidence interval 0.97-0.99), respectively. The mean neck-shaft angle was 129.31° ± 5.04° in males and 129.93° ± 6.29° in females. The mean femoral anteversion was 9.72° ± 7.95° in males and 12.61° ± 8.91° in females. IFD was positively correlated with height (correlation coefficient [r] = 0.464, P < .001) and weight (0.286, P < .001), whereas it was negatively correlated with age (-0.198, P < .001), neck-shaft angle (-0.123, P = .005), and femoral anteversion (-0.346, P < .001). There was no correlation between body mass index and IFD (P = .522). In multivariate regression analysis, IFD was positively associated with height (ß = .632), and negatively associated with neck-shaft angle of the femur and femoral anteversion (ß = -0.155 and -0.328. respectively). CONCLUSIONS: In asymptomatic hips, the mean IFD was 33.2 ± 9.2 mm in males and 24.3 ± 8.9 mm in females. The IFD was positively correlated with height and negatively with neck-shaft angle of the femur and femoral anteversion. LEVEL OF EVIDENCE: Level III, retrospective comparative study.
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Pinzamiento Femoroacetabular/diagnóstico , Fémur/diagnóstico por imagen , Isquion/diagnóstico por imagen , Tomografía Computarizada por Rayos X/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Enfermedades Asintomáticas , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Curva ROC , Reproducibilidad de los Resultados , Estudios Retrospectivos , Adulto JovenRESUMEN
The osseous regeneration of large bone defects is still a major clinical challenge in maxillofacial and orthopedic surgery. Previous studies demonstrated that biphasic electrical stimulation (ES) stimulates bone formation; however, polyimide electrode should be removed after regeneration. This study presents an implantable electrical stimulation bioreactor with electrodes based on liquid crystal polymer (LCP), which can be permanently implanted due to excellent biocompatibility to bone tissue. The bioreactor was implanted into a critical-sized bone defect and subjected to ES for one week, where bone regeneration was evaluated four weeks after surgery using micro-CT. The effect of ES via the bioreactor was compared with a sham control group and a positive control group that received recombinant human bone morphogenetic protein (rhBMP)-2 (20 µg). New bone volume per tissue volume (BV/TV) in the ES and rhBMP-2 groups increased to 132% (p < 0.05) and 174% (p < 0.01), respectively, compared to that in the sham control group. In the histological evaluation, there was no inflammation within the bone defects and adjacent to LCP in all the groups. This study showed that the ES bioreactor with LCP electrodes could enhance bone regeneration at large bone defects, where LCP can act as a mechanically resistant outer box without inflammation. Graphical abstract To enhance bone regeneration, a bioreactor comprising collagen sponge and liquid crystal polymer-based electrode was implanted in the bone defect. Within the defect, electrical current pulses having biphasic waveform were applied from the implanted bioreactor.
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Reactores Biológicos , Regeneración Ósea/fisiología , Mandíbula/patología , Mandíbula/fisiopatología , Polímeros/química , Animales , Hueso Esponjoso/diagnóstico por imagen , Hueso Esponjoso/patología , Hueso Esponjoso/fisiopatología , Estimulación Eléctrica , Electrodos , Masculino , Mandíbula/diagnóstico por imagen , Osteogénesis , Conejos , Microtomografía por Rayos XRESUMEN
BACKGROUND: Hip fracture surgery (HFS) is often associated with perioperative blood loss, and it frequently necessitates transfusion. However, the hemoglobin (Hb) threshold for transfusion remains controversial in hip fracture patients. We evaluated the usefulness of the restrictive strategy and preoperative intravenous iron supplementation in HFS. METHODS: We retrospectively reviewed the medical records of 1,634 patients (> 60 years of age) who underwent HFS between May 2003 and June 2014 and were followed up for 1 year or more after surgery. We used the liberal transfusion strategy until May 2009 to determine the transfusion threshold; afterwards, we switched to the restrictive transfusion strategy. Patients with the restrictive transfusion strategy (restrictive group) received intravenous iron supplementation before surgery. We compared the transfusion rate, morbidity, and mortality of the restrictive group with those of the patients with the liberal transfusion strategy (liberal group). RESULTS: Preoperative intravenous iron supplementation was not associated with any adverse reactions. The transfusion rate was 65.3% (506/775) in the liberal group and 48.2% (414/859) in the restrictive group (p < 0.001). The mean hospital stay was shorter in the restrictive group (21.5 vs. 28.8 days, p < 0.001). There was no significant difference in the postoperative medical complications including myocardial infarction and cerebrovascular event. Mortality at postoperative 30, 60, and 90 days was similar between the two groups. CONCLUSIONS: Our blood management protocol involving restrictive strategy combined with preoperative intravenous iron supplementation appears to be effective and safe in HFS of elderly patients.
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Anemia/terapia , Transfusión Sanguínea/normas , Fracturas de Cadera/cirugía , Compuestos de Hierro/administración & dosificación , Administración Intravenosa , Anciano , Anciano de 80 o más Años , Anemia/etiología , Pérdida de Sangre Quirúrgica , Suplementos Dietéticos , Femenino , Hemoglobinas/análisis , Fracturas de Cadera/sangre , Humanos , Masculino , Procedimientos Ortopédicos , Hemorragia Posoperatoria/terapia , Cuidados Preoperatorios , Estudios RetrospectivosRESUMEN
PURPOSE: Hip fractures in hemodialysis patients are accompanied by high rates of complications and morbidities. Previous studies have mainly reported on nonunion and avascular necrosis of femoral neck fractures in this patient group. In this study the complication and clinical results of hemodialysis patients with intertrochanteric fractures treated with proximal femoral intramedullary nailing have been investigated through comparison with patients with normal kidney function. MATERIALS AND METHODS: Forty-seven patients were included; the hemodialysis group (n=17) and the control group with normal kidney function (n=30). The medical history and clinical findings including preoperative and postoperative blood examinations, radiological examinations and ambulatory status (measured using the Koval score). The rate of complications and morbidities were also investigated and compared. RESULTS: Preoperative hemoglobin/hematocrit was lower but a significant increase in partial thromboplastin time was observed in the hemodialysis group. The amount of bleeding/transfusions were higher and operative time was longer in the hemodialysis group. Upon radiologic examination, there was no significant difference in rate of unstable fracture and nonunion between the two groups. However the postoperative Koval score was significantly worse and the odds ratio of inability to walk after surgery was 13.5 times higher in the hemodialysis group. CONCLUSION: There was no significant difference in radiological results, but the risk of inability to walk after surgery was 13.5 times higher in the hemodialysis group. Hemodialysis patients have more morbidities and are hemodynamically unstable therefore require special attention. Accurate reduction and firm fixation is required and attentive postoperative rehabilitation is needed.
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Existence of the genomically and epigenomically diverse subclones in a tumor severely limits the therapeutic efficacy of targeted agents. To overcome such a limitation, we prepared a novel targeted prodrug, EMC-DEVD-S-DOX, which comprises two important features: radiation-induced apoptosis targeting and albumin-binding properties. In particular, the prodrug binds circulating albumin after intravenous administration and then activated by caspase-3, which is upregulated from apoptotic cells that responded to radiotherapy. The prodrug was designed to bind circulating albumin to extend half-life and facilitate tumor accumulation in order to increase the possibility of contacting caspase-3, which is only transiently upregulated during apoptosis. Our results showed that EMC-DEVD-S-DOX had a prolonged half-life with enhanced tumor accumulation, which clearly benefited the therapeutic effect of the prodrug. Also, agreeing with the in vitro studies that showed ignorable cytotoxic effect in the absence of caspase-3, the prodrug was effective only when combined with radiotherapy without any noticeable systemic toxicity in vivo. Due to the highly selective action of EMC-DEVD-S-DOX independent to the complex genomic profiles of tumor, the prodrug would overcome the limitation of current targeted therapy and potentiate radiotherapy in the clinical oncology.
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Albúminas/metabolismo , Caspasa 3/metabolismo , Neoplasias/tratamiento farmacológico , Neoplasias/radioterapia , Profármacos/uso terapéutico , Animales , Caproatos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Doxorrubicina/uso terapéutico , Humanos , Masculino , Ratones , Neoplasias/patología , Péptidos/química , Profármacos/farmacocinética , Profármacos/farmacología , Regulación hacia ArribaRESUMEN
BACKGROUND: To evaluate the effect of spondylolisthesis on lumbar lordosis on the OSI (Jackson; Orthopaedic Systems Inc.) frame. Restoration of lumbar lordosis is important for maintaining sagittal balance. Physiologic lumbar lordosis has to be gained by intraoperative prone positioning with a hip extension and posterior instrumentation technique. There are some debates about changing lumbar lordosis on the OSI frame after an intraoperative prone position. We evaluated the effect of spondylolisthesis on lumbar lordosis after an intraoperative prone position. METHODS: Sixty-seven patients, who underwent spinal fusion at the Department of Orthopaedic Surgery of Gwangmyeong Sungae Hospital between May 2007 and February 2012, were included in this study. The study compared lumbar lordosis on preoperative upright, intraoperative prone and postoperative upright lateral X-rays between the simple stenosis (SS) group and spondylolisthesis group. The average age of patients was 67.86 years old. The average preoperative lordosis was 43.5° (± 14.9°), average intraoperative lordosis was 48.8° (± 13.2°), average postoperative lordosis was 46.5° (± 16.1°) and the average change on the frame was 5.3° (± 10.6°). RESULTS: Among all patients, 24 patients were diagnosed with simple spinal stenosis, 43 patients with spondylolisthesis (29 degenerative spondylolisthesis and 14 isthmic spondylolisthesis). Between the SS group and spondylolisthesis group, preoperative lordosis, intraoperative lordosis and postoperative lordosis were significantly larger in the spondylolisthesis group. The ratio of patients with increased lordosis on the OSI frame compared to preoperative lordosis was significantly higher in the spondylolisthesis group. The risk of increased lordosis on frame was significantly higher in the spondylolisthesis group (odds ratio, 3.325; 95% confidence interval, 1.101 to 10.039; p = 0.033). CONCLUSIONS: Intraoperative lumbar lordosis on the OSI frame with a prone position was larger in the SS patients than the spondylolisthesis patients, which also produced a larger postoperative lordosis angle after posterior spinal fusion surgery. An increase in lumbar lordosis on the OSI frame should be considered during posterior spinal fusion surgery, especially in spondylolisthesis patients.
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Cuidados Intraoperatorios/métodos , Vértebras Lumbares/cirugía , Complicaciones Posoperatorias/prevención & control , Posición Prona/fisiología , Estenosis Espinal/cirugía , Espondilolistesis/cirugía , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Postura/fisiología , Estudios RetrospectivosRESUMEN
BACKGROUND: Both accurate measurement of HbA1c and minimal reagent lot-to-lot variability are essential for point-of-care HbA1c assays. The accuracy of three different cartridge lots of the Samsung LABGEO PT HbA1c Test was investigated to determine whether the results can be used for follow-up and screening of patients with diabetes. METHODS: The LABGEO PT10 device and three different lots of the LABGEO PT HbA1c Test cartridge were used. Seven levels of reference materials were measured using each cartridge in a duplicate manner for 3 days. The bias, within-laboratory precision, and total error were calculated. The medical decision point analysis was performed. RESULTS: The mean absolute bias, within-laboratory precision, and total error of each cartridge were 3.3%, 2.5%, and 8.1% for Lot1; 1.9%, 2.6%, and 7.1% for Lot2; and 2.7%, 2.8%, and 8.1% for Lot3. The predicted value (95% confidence interval) of each cartridge at an HbA1c of 6.5% was 6.74% (6.66, 6.83) for Lot1, 6.60 (6.51, 6.70) for Lot2, and 6.51 (6.39, 6.63) for Lot3. CONCLUSIONS: Our data suggest that the LABGEO PT HbA1c Test can be used to monitor patients with diabetes and perform diabetes screening when false-positive results are obtained in the doctor's office.
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Hemoglobina Glucada/análisis , Juego de Reactivos para Diagnóstico/normas , Toma de Decisiones Clínicas , Humanos , Estándares de ReferenciaRESUMEN
While recombinant human bone morphogenetic protein (rhBMP)-2-based bone therapy presents potential osteoinductivity, it also leads concern due to transient osteoclast activation during early healing periods, ultimately limiting its clinical use. Therefore, we investigated in vivo and in vitro rhBMP-2 signaling which mediates early bone resorbing effect, depending on the dose, and attempted to inhibit this resorption phenomenon using NFAT inhibitor as a target molecule. High-dose of rhBMP-2 (20 µg/defect) enhanced osteoclast activation and the expression of bone resorption markers, compared to low dose (5 µg/defect) at one week after surgery in collagen sponge-delivered rat calvarial defect models. Interestingly, this trend was also observed in the expression of bone formation markers. In particular, rhBMP-2 upregulated RANKL expression, while it downregulated osteoprotegerin (OPG) expression, resulting in a dose-dependent increase in the ratio of RANKL to OPG. NFAT inhibitor (150 µm) treatment in vivo suppressed the high-dose effect of rhBMP-2 on both resorption and formation. In vitro results of rhBMP-2 signaling and NFAT inhibitor effects in rat mesenchymal stem cells showed similar trends as in vivo results. Microcomputer tomography-based evaluation at 4 weeks showed that combined treatment of NFAT inhibitor with 20 µg rhBMP-2 in vivo increased bone volume (BV) more than 20 µg rhBMP-2 alone, which showed little difference in BV compared to 5 µg of rhBMP-2. These results demonstrated that rhBMP-2 implantation concurrently signalized into enhanced osteoclastogenesis and osteoblastogenesis in vivo, dose-dependently. Ratio of RANKL/OPG might be an index for early bone resorbing activity of implanted rhBMP-2. A local cocktail treatment of NFAT inhibitor and high-dose rhBMP-2 might be an alternative to overcome early bone resorbing effects, thereby accelerating bone formation.
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Proteína Morfogenética Ósea 2/farmacología , Resorción Ósea/metabolismo , Osteoclastos/citología , Osteoclastos/metabolismo , Factor de Crecimiento Transformador beta/farmacología , Animales , Proteína Morfogenética Ósea 2/uso terapéutico , Colágeno/metabolismo , Osteoclastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/farmacología , Proteínas Recombinantes/uso terapéutico , Factor de Crecimiento Transformador beta/uso terapéutico , Cicatrización de Heridas/efectos de los fármacosRESUMEN
We report a fully integrated device that can perform both multiple biochemical analysis and sandwich type immunoassay simultaneously on a disc. The whole blood is applied directly to the disposable "lab-on-a-disc" containing different kinds of freeze-dried reagents for the blood chemistry analysis as well as reagents required for the immunoassay. The concentrations of different kinds of analytes are reported within 22 min by simply inserting a disc to a portable device. Using the innovative laser irradiated ferrowax microvalves together with the centrifugal microfluidics, the total process of plasma separation, metering, mixing, incubation, washing, and detection is fully automated. The analyzer is equipped with an optical detection module to measure absorbances at 10 different wavelengths to accommodate the various kinds of reaction protocols. Compared to the conventional blood analysis done in clinical laboratories, it is advantageous for point-of-care applications because it requires a smaller amount of blood (350 µL vs. 3 mL), takes less time (22 min vs. several days), does not require specially trained operators or expensive instruments to run biochemical analysis and immunoassay separately.
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Análisis Químico de la Sangre/instrumentación , Inmunoensayo/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Análisis Químico de la Sangre/economía , Análisis Químico de la Sangre/métodos , Diseño de Equipo , Humanos , Inmunoensayo/economía , Inmunoensayo/métodos , Técnicas Analíticas Microfluídicas/economía , Técnicas Analíticas Microfluídicas/métodosRESUMEN
BACKGROUNDS/AIMS: Choledochal cyst of the bile duct is characterized by cystic dilatation of the intra- or extrahepatic bile ducts. It is a relatively uncommon disease and there is still much controversy regarding its etiology as being congenital or acquired. METHODS: The medical records of 60 patients who underwent surgical treatments for choledochal cyst between April 1995 and April 2009 at the Gachon University Gil Hospital were reviewed retrospectively. To compare the clinical characteristics, patients under 19 years of age were grouped into children and the others were grouped into adults. RESULTS: Of the overall 60 patients, 24 were grouped into children and 36 were grouped into adults. Female predominance was common in both groups (M : F=1 : 6.5). The most common clinical symptom was abdominal pain (73.3%) in both groups. Children had remarkable jaundice (33.3% vs. 0%) and gastrointestinal symptoms including nausea and vomiting. Fever and chills were more common in children because of the associated complications of cholangitis. According to the Todani classification, type I was the most common form of choledochal cyst in both groups, and type IVa was significantly more common in children than adults (45.3% vs. 16.7%). Thirty patients (50%) had anomalous pancreaticobiliary ductal union which was confirmed by preoperative imaging studies or intraoperative cholangiography. All patients except for one child underwent cyst excision with hepaticojejunostomy. CONCLUSION: There was no significant difference in the clinical characteristics of choledochal cysts between children and adults. However, combined diseases especially bile duct malignancy were common in aged patients, early detection and more aggressive surgery is necessary for patients with choledochal cysts.
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The centrifugal microfluidic platform has been a focus of academic and industrial research efforts for almost 40 years. Primarily targeting biomedical applications, a range of assays have been adapted on the system; however, the platform has found limited commercial success as a research or clinical tool. Nonetheless, new developments in centrifugal microfluidic technologies have the potential to establish wide-spread utilization of the platform. This paper presents an in-depth review of the centrifugal microfluidic platform, while highlighting recent progress in the field and outlining the potential for future applications. An overview of centrifugal microfluidic technologies is presented, including descriptions of advantages of the platform as a microfluidic handling system and the principles behind centrifugal fluidic manipulation. The paper also discusses a history of significant centrifugal microfluidic platform developments with an explanation of the evolution of the platform as it pertains to academia and industry. Lastly, we review the few centrifugal microfluidic-based sample-to-answer analysis systems shown to date and examine the challenges to be tackled before the centrifugal platform can be more broadly accepted as a new diagnostic platform. In particular, fully integrated, easy to operate, inexpensive and accurate microfluidic tools in the area of in vitro nucleic acid diagnostics are discussed.
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Centrifugación/instrumentación , Centrifugación/tendencias , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/tendencias , Diseño de EquipoRESUMEN
Dispensing uniform pico-to-nanoliter droplets has become one of essential components in various application fields from high-throughput bio-analysis to printing. In this study, a new method is suggested and demonstrated for dispensing a droplet on the top plate with an inverted geometry by using electric field. The process of dispensing droplets consists of two stages: (i) formation of liquid bridge by moving up the charged fluid mass using the electrostatic force between the charges on the fluid mass and the induced charges on the substrate and (ii) its break-up by the motion of the top plate. Different from conventional electrohydrodynamic methods, electric induction enables the droplets to be dispensed on various surfaces including non-conducting substrate. The use of capillarity with an inverted geometry removes the need of external pumps or elaborates control for constant flow feed. The droplet diameter has been characterized as a function of the nozzle-to-plate distance and the plate moving velocity. The robustness of the present method is shown in terms of nozzle length and applied voltage. Finally, its practical applicability is confirmed by rendering a 19 by 24 array of highly uniform droplets with only 1.8% size variation without use of any active feedback control.
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Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Campos Electromagnéticos , Tamaño de la PartículaRESUMEN
A portable, disc-based, and fully automated enzyme-linked immuno-sorbent assay (ELISA) system is developed to test infectious diseases from whole blood. The innovative laser irradiated ferrowax microvalves and centrifugal microfluidics were utilized for the full integration of microbead-based suspension ELISA assays on a disc starting from whole blood. The concentrations of the antigen and the antibody of Hepatitis B virus (HBV), HBsAg and Anti-HBs respectively, were measured using the lab-on-a-disc (LOD). All the necessary reagents are preloaded on the disc and the total process of the plasma separation, incubation with target specific antigen or antibody coated microbeads, multiple steps of washing, enzyme reaction with substrates, and the absorbance detection could be finished within 30 minutes. Compared to the conventional ELISA, the operation time was dramatically reduced from over 2 hours to less than 30 minutes while the limit of detection was kept similar; e.g. the limit of detection of Anti-HBs tests were 8.6 mIU mL(-1) and 10 mIU mL(-1) for the disc-based and the conventional ELISA respectively.
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Ensayo de Inmunoadsorción Enzimática/instrumentación , Anticuerpos contra la Hepatitis B/sangre , Antígenos de la Hepatitis B/sangre , Técnicas Analíticas Microfluídicas/instrumentación , Automatización , Centrifugación , Simulación por Computador , Ensayo de Inmunoadsorción Enzimática/métodos , Diseño de Equipo , Antígenos de Superficie de la Hepatitis B/sangre , Humanos , Técnicas Analíticas Microfluídicas/métodos , Microesferas , Sensibilidad y EspecificidadRESUMEN
Valving is critical in microfluidic systems. Among many innovative microvalves used in lab-on-a-chip applications, phase change based microvalves using paraffin wax are particularly attractive for disposable biochip applications because they are simple to implement, cost-effective and biocompatible. However, previously reported paraffin-based valves require embedded microheaters and therefore multi-step operation of many microvalves was a difficult problem. Besides, the operation time was relatively long, 2-10 s. In this paper, we report a unique phase change based microvalve for rapid and versatile operation of multiple microvalves using a single laser diode. The valve is made of nanocomposite materials in which 10 nm-sized iron oxide nanoparticles are dispersed in paraffin wax and used as nanoheaters when excited by laser irradiation. Laser light of relatively weak intensity was able to melt the paraffin wax with the embedded iron oxide nanoparticles, whereas even a very intense laser beam does not melt wax alone. The microvalves are leak-free up to 403.0 +/- 7.6 kPa and the response times to operate both normally closed and normally opened microvalves are less than 0.5 s. Furthermore, a sequential operation of multiple microvalves on a centrifugal microfluidic device using a single laser diode was demonstrated. It showed that the optical control of multiple microvalves is fast, robust, simple to operate, and requires minimal chip space and thus is well suited for fully integrated lab-on-a-chip applications.
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Centrifugación/instrumentación , Centrifugación/métodos , Iluminación , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Nanoestructuras/química , Rayos LáserRESUMEN
We report a fully integrated, pathogen-specific DNA extraction device utilizing centrifugal microfluidics on a polymer based CD platform. By use of the innovative laser irradiated Ferrowax microvalve (LIFM) together with the rapid cell lysis method using laser irradiation on magnetic particles, we could, for the first time, demonstrate a fully integrated pathogen specific DNA extraction from whole blood on a CD. As a model study, DNA extraction experiments from whole blood spiked with Hepatitis B virus (HBV) and E.coli were conducted. The total process of the plasma separation, mixing with magnetic beads conjugated with target specific antibodies, removal of plasma residual, washing and DNA extraction was finished within 12 min with only one manual step, the loading of 100 microL of whole blood. Real-time PCR results showed that the concentration of DNA prepared on a CD using a portable sample preparation device was as good as those by conventional bench top protocol. It demonstrates that our novel centrifugal microfluidics platform enables a full integration of complex biological reactions that require multi-step fluidic control.
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Centrifugación/instrumentación , Centrifugación/métodos , ADN Bacteriano/sangre , ADN Bacteriano/aislamiento & purificación , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Escherichia coli/genética , Escherichia coli/patogenicidad , Humanos , MagnetismoRESUMEN
Experimental results are presented for the study of drop formation mechanism in a newly proposed electrohydrodynamic (EHD) method of drop generation in an AC electric field. In the method, a small drop is generated in two stages. A pendant drop is elongated with large oscillation by an electric force in the first stage. Then, it undergoes formation and breakup of a liquid bridge between the upper nozzle and the insulator-coated lower flat plate in the second stage. It is found that there exists a resonant frequency for maximum oscillation, which leads to an efficient drop formation in the latter stage. It is also found that breakup of liquid bridge is accelerated by the electrowetting tension acting on the drop perimeter contacting the insulator-coated flat plate. Thus the whole procedure of drop formation depends heavily on the frequency of AC field and the properties of the insulator such as hydrophilicity, thickness, and the dielectric constant. It is demonstrated that a wide range of drop size, from picoliter to nanoliter, can be obtained by controlling such key parameters without changing the nozzle diameter.
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Campos Electromagnéticos , ADN/química , Electrodos , Análisis de Secuencia por Matrices de Oligonucleótidos/instrumentación , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Oscilometría , Tamaño de la Partícula , Propiedades de Superficie , Agua/químicaRESUMEN
In the present paper, we first demonstrated the possibility of electrohydrodynamic (EHD) dispensing method for preparing nanoliter probe DNA droplets on surfaces in DNA microarrays. To study the effect of an electric field on the dynamic behavior of pendent DNA droplet, visualization experiments with three kinds of electrode shapes are performed. In the early stage of droplet dispensing, it is shown that applied electric field assists a gravitational force exerted on DNA droplet. The pendent droplet is elongated in the parallel direction of applied electric field. However, after making fluid bridge between electrodes, it is shown that the electric force accelerates the capillary breaking of droplet by assisting a surface tension force exerted on droplet surface. Specifically, nanoliter dispensing volume (2 nL) is obtained in the needle-type electrode configuration. In addition, for the case of hydrophobic electrode surface, it is shown that the dispensing volume and spot size are remarkably decreased. Under the high relative humidity condition, it is observed that spot size is rapidly increased because of reduction in evaporation rate on droplet surface during the dispensing procedure. On the other hand, it is obtained that the spot size is not changed significantly in the wide range of DNA concentration from 1 to 10,000 nM. To monitor the influence of high electric voltage on DNA stability, we prepared a silicon-based chip with five capture probes for pathogens related with respiratory infectious diseases by EHD dispensing method. From the examination, it is clearly confirmed that pathogens are detected and the effective signal levels of pathogenic bacteria after hybridization are retained. Consequently, it is found that EHD dispensing method can be used to make cost-effective DNA microarrays with no thermal and electrical influences on DNA properties.