Controlled cellular orientation on PLGA microfibers with defined diameters.

In this study, we investigated the effects of the diameter of microfibers on the orientation (angle between cells' major axis and the substrate fiber long axis) of adhered cells. For this purpose, mouse fibroblast L929 cells were cultured on the surface of PLGA fibers of defined diameters ranging from 10 to 242 mum, and their adhesion and alignment was quantitatively analyzed. It was found that the mean orientation of cells and the spatial variation of cell alignment angle directly related to the microfiber diameter. Cells that were cultured on microfibrous scaffolds oriented along the long axis of the microfiber and the orientation increased as the fiber diameter decreased. For the fiber diameter of 10 microm, the mean orientation was 3.0 +/- 0.2 degrees (mean +/- SE), whereas for a diameter of 242 microm, it decreased to 37.7 +/- 2.1 degrees . Using these studies we demonstrate that fibroblasts have a characteristic alignment on microscale fibers and that the microscale fiber diameter plays a critical role in cellular orientation. The ability to control cellular alignment on engineered tissue scaffold can be a potentially powerful approach to recreate the microscale architecture of engineered tissues. This may be important for engineering a variety of human tissues such as tendon, muscle and nerves as well as applications in 3D tissue culture and drug screening.

Development of a new pulsatile extracorporeal life support device incorporating a dual pulsatile blood pump.

A new pulsatile extracorporeal life support device (ECLS) has been developed, designed to sustain pulsatile blood flow during emergency cardiopulmonary resuscitations and cardiopulmonary operations. This device features two identical pulsatile pumps that operate alternately and can therefore provide blood inflow in a more uniform manner than similar systems featuring a single-pump configuration. In order to confirm the presumed benefits of this newly-developed dual pulsatile pump configuration, we have conducted a series of in vitro experiments designed to compare the properties of the new system with a single pump system, specifically with regard to pump delivery rate and active filling efficiency. Our results reveal that the dual pump configuration can, indeed, deliver a higher flow than can the single-pump system, and exhibits an active filling efficiency superior to that of the single-pump configuration. We performed a series of animal experiments to measure the pulsatility of the dual-pump configuration in terms of equivalent energy pressure (EEP). In order to measure EEP, we measured femoral arterial pressure and pump outflow. The results of our animal experiments revealed that the newly-developed pulstile ECLS exhibits sufficient pulsatility in terms of the EEP considerations.

The effects of pulsatile versus non-pulsatile extracorporeal circulation on the pattern of coronary artery blood flow during cardiac arrest.

BACKGROUND: In sudden cardiac arrest, the effective maintenance of coronary artery blood flow is of paramount importance for myocardial preservation as well as cardiac recovery and patient survival. The purpose of this study was to directly compare the effects of pulsatile versus non-pulsatile circulation to coronary artery flow and myocardial preservation in a cardiac arrest condition. METHODS: A cardiopulmonary bypass circuit was constructed in a ventricular fibrillation model using fourteen Yorkshire swine weighing 25-35 kg each. The animals were randomly assigned to group I (n=7, non-pulsatile centrifugal pump) or group II (n=7, pulsatile T-PLS pump). Extracorporeal circulation was maintained for two hours at a pump flow of 2 L/min. The left anterior descending coronary artery flow was measured with an ultrasonic coronary artery flow measurement system at baseline (before bypass) and at every 20 minutes after bypass. Serologic parameters were collected simultaneously at baseline, 1 hour, and 2 hours after bypass in the systemic arterial and coronary sinus venous blood. The Mann-Whitney U test of STATISTICA 6.0 was used to determine intergroup significances using a p value of <0.05. RESULTS: The resistance index of the coronary artery was lower in group II and the difference was significant at 40 min, 80 min, 100 min and 120 min (p<0.05). The mean velocity of the coronary artery was higher in group II throughout the study, and the difference was significant from 20 min after starting the pump (p<0.05). The coronary artery blood flow was higher in group II throughout the study, and the difference was significant from 40 min to 120 min (p<0.05) except at 80 min. Serologic parameters showed no differences between the groups at 1 hour and 2 hours after bypass in the systemic and coronary sinus blood (p=NS). CONCLUSION: In the cardiac arrest condition, pulsatile extracorporeal circulation provides more blood flow, higher flow velocity and less resistance to coronary artery than non-pulsatile circulation.

Biliary ascariasis: MR cholangiography findings in two cases.

We describe the imaging features of two cases of biliary ascariasis. Ultrasonography and CT showed no specific abnormal findings, but MR cholangiography clearly demonstrated an intraductal linear filling defect that led to the correct diagnosis. MR cholangiography is thus a useful technique for the diagnosis of biliary ascariasis.

In vivo evaluation of lumbrokinase, a fibrinolytic enzyme extracted from Lumbricus rubellus, in a prosthetic vascular graft.

BACKGROUND: Lumbrokinase (LK) is a fibrinolytic enzyme purified from the earthworm Lumbricus rubellus. To investigate the fibrinolytic and antithrombotic effects of lumbrokinase, a series of animal experiments were performed. METHODS: The Dacron graft (3 mm in diameter, 3 cm in length) were treated with LK via two different methods, simple dipping and covalent bonding METHODS: Covalent bonding was performed by UV reaction to polyacrylic acid. The grafts were interposed into the inferior vena cava of the rabbits and harvested for 5 hours, 1, 2 and 4 weeks after the implantation. RESULTS: The LK non-treated graft (n=4) were totally occluded with thrombus 5 hours after the implantation. Both types of LK treated graft (n=8) were patent 1 week after the implantation. The grafts treated with the simple dipping method (n=4) were occluded with thrombus 2 weeks after the implantation. The grafts treated with covalent bonding (n=4) were patent 4 weeks after the implantation. Ultrastructural analysis of the luminal surface of the patent grafts by scanning electron microscopy revealed the thin plasma protein layer to be about 5 micro in thickness with platelet adhesions. CONCLUSIONS: Lumbrokinase has potential antithrombotic effects in a small diameter vascular prosthesis. The covalent bonding method proved to be more effective than the simple dipping method.

Tissue-engineered heart valve leaflets: an effective method for seeding autologous cells on scaffolds.

BACKGROUND: A precondition for the successful formation of tissue-engineered heart valves is the generation of a proper matrix on biodegradable scaffolds over a limited period of time. The aim of this study was to find an effective method of seeding autologous cells on these scaffolds to create a new matrix for heart valves. METHODS: Myofibroblasts and endothelial cells were isolated and cultured from an ovine artery. A synthetic biodegradable scaffold consisting of polyglycolic and polylactic acids was seeded first with the myofibroblasts, then coated with endothelial cells. Three different methods of myofibroblast seeding were compared: I) daily seeding of myofibroblasts (1x10(6)) for ten days and culture for four days; II) seeding of myofibroblasts (1x10(7)) and culture for 14 days with the use of a simple medium; III) seeding of myofibroblasts (1x10(7)) with the use of a medium containing collagen and culture for 14 days. Light and electron microscopic analyses were performed. RESULTS: The group that used the medium containing collagen showed the best results in terms of seeding efficiency. CONCLUSION: Seeding autologous cells with a medium containing collagen onto the scaffold showed the largest cell population and might generate the best matrix on the scaffold.

Application of the moving-actuator type pump as a ventricular assist device: in vitro and in vivo studies.

A moving actuator type pump has been developed as a multifunctional Korean artificial heart (AnyHeart). The pump consists of a moving actuator as an energy converter, right and left sacs, polymer (or mechanical) valves, and a rigid polyurethane housing. The actuator containing a brushless DC motor moves back and forth on an epicyclical gear train to produce a pendular motion, which compresses both sacs alternately. Of its versatile functions of ventricular assist device and total artificial heart use, we have evaluated the system performance as a single or biventricular assist device through in vitro and in vivo experiments. Pump performance and anatomical feasibility were tested using various animals of different sizes. In the case of single ventricular assist device (VAD) use, one of the sacs remained empty and a mini-compliance chamber was attached to either an outflow or inflow port of the unused sac. The in vitro and in vivo studies show acceptable performance and pump behavior. Further extensive study is required to proceed to human application.

Applications of the pulsatile flow versatile ECLS: in vivo studies.

INTRODUCTION: T-PLS (Twin-Pulse Life Support) is the first commercial pulsatile ECLS (Extra Corporeal Life Support) device (1). The dual sac structure of T-PLS can effectively reduce high membrane oxygenator inlet pressure and hemolysis. To verify both the use of T-PLS for ECLS and the advantages of T-PLS, we tested various models. METHOD AND RESULTS: In the partial CPB (cardio pulmonary bypass) model (swine), T-PLS (N = 6), and Biopump (N = 2), a single pulsatile pump (N = 2), were compared. In the case of single pulsatile flow, during pump systole, pressure increased to 700 - 800 mmHg at the inlet port of the membrane oxygenator. fHb, a hemolysis measurement value, was about 80 mg/dL at 3 hours. On the contrary, because of T-PLS's dual sac system, the pressure of T-PLS had a maximum value of about 250 mmHg and fHb was similar to that of the commercial centrifugal pumps. In the total CPB model (bovine, N = 6), the heart was stopped via cardioplegia (Kcl). T-PLS flow was maintained at 3.0-4.5 L/min. T-PLS functioned like a natural heart, having a pulse pressure of 26-43 mmHg and a pulse rate of 40-60 bpm (beats per minute). In the emergency case model (canine, N = 6), T-PLS was started 10 minutes after cardiac arrest from electronic shock. In spite of cardiac arrest for a period of 40 minutes, the heart was recovered after defibrillation. In the ARDS (Acute Respiratory Distress Syndrome) model (canine, N = 6), minimal ventilator parameters were set: tidal volume 130 ml, respiration rate = bpm, FiO2 = 10%. Three hours after starting T-PLS, PO2 of the carotid artery blood (after 2 hours: 195 +/- 89.4; after 3 hours: 258 +/- 99.3 mmHg) was above half the value of the femoral artery but was within normal range. CONCLUSION: It is suggested that a portable pulsatile ECLS like T-PLS may be used as a CPB device and as an alternative CPR (cardiopulmonary resuscitation) device in the case of cardiac arrest. Due to the pulsatile flow, oxygenated blood is delivered to the patient without overloading the ARDS patients heart.

Prenatal diagnosis of Hb Bart's hydrops fetalis in West Malaysia: the identification of the alpha thal 1 defect by PCR based strategies.

Haemoglobin Bart's hydrops fetalis is the result of complete absence of functional alpha-globin genes where the fetus is homozygous for the alpha 0-thal gene. Prenatal diagnosis can be made by analysis of fetal DNA from chorionic villus, amniotic cells and fetal blood. Earlier studies for analysing genomic DNA needed digestion with restriction enzymes and hybridisation to radiolabelled probes which took 2 weeks. We have used the polymerase chain reaction (PCR) and non-radioactive primers to identify specific target sequences with results available within 1-3 days for the diagnosis of haemoglobin Bart's syndrome. With fetal blood samples, complete absence of alpha-chain synthesis is confirmed by globin chain electrophoresis on cellulose acetate pH 6.0.

Assessments of injectable alginate particle-embedded fibrin hydrogels for soft tissue reconstruction.

Soft tissue reconstruction is often needed after massive traumatic damage or cancer removal. In this study, we developed a novel hybrid hydrogel system consisting of alginate particles and a fibrin matrix that could maintain tissue volume long term. Alginate particles were fabricated by mixing 5% alginate with a 20 mM calcium solution. Cells and these alginate particles were then embedded in fibrin (alginate-fibrin) hydrogels using a dual syringe mixer. Cell-hydrogel constructs were evaluated in terms of cell survival and proliferation in the constructs in vitro. The results indicated that cellular viability, spreading and proliferation in the alginate-fibrin hydrogels were significantly higher compared to constructs fabricated with fibrin or alginate only. In vivo explants showed that cells contained within fibrin-only hydrogels did not contribute to neo-tissue formation, and the fibrin was fully degraded within a 12 week period. In the alginate-fibrin system, higher cellularity and vascular ingrowth were observed in vivo. This resulted in neo-tissue formation in the alginate-fibrin hydrogels. These results demonstrate that fibrin may enhance cell proliferation and accelerate the formation of extracellular matrix proteins in the alginate-fibrin system, while the alginate particles could contribute to volume retention. This injectable hybrid system composed of degradable and non-degradable hydrogels may be a preferable approach to the repair of soft tissue defects.

Study of undifferentiated spondyloarthropathy among first-degree relatives of ankylosing spondylitis probands.

OBJECTIVE: To estimate in a Chinese population the prevalence of undifferentiated spondyloarthropathy (USpA) among first-degree relatives (FDRs) of ankylosing spondylitis (AS) probands, and to compare the clinical features of familial USpA with those of sporadic USpA. METHODS: The FDRs of two separate cohorts of consecutive AS probands were evaluated for the prevalence of USpA, using the Modified New York criteria and the European Spondylitis Study Group criteria for AS and SpA, respectively. Sporadic USpA and FDRs of non-SpA rheumatic patient probands served as separate controls. RESULTS: Among the 301 FDRs of 102 AS probands, 7.0% were USpA. This was 1000 times higher than the 147 FDRs of 40 non-SpA probands (P = 0.00230). Within the AS families, USpA was less male-dominated than AS (33.3 vs 72.5%) (P = 0.006). The only feature distinguishing familial from sporadic USpA was that the percentages of HLA B27 were 100 and 50%, respectively (P<0.001). CONCLUSION: USpA and AS coexist in the same Chinese families, both being predisposed by HLA B27. In these families, a female gender favours the development of USpA rather than AS. A significant subset of sporadic USpA (HLA B27-negative group) has a different genetic predisposition compared with familial USpA.

Changes in the clinical and laboratory features of lupus patients after the big earthquake in Taiwan.

In order to understand the impact of the massive earthquake on disease activity, 60 patients with systemic lupus erythematosus (SLE) living in the earthquake area (group 1) and 31 patients living away from the area (group 2) were enrolled in this study. A questionnaire was used to record demographic data and clinical manifestations before and 6 months after the earthquake. Laboratory tests, including serum C3, C4, creatinine, anti-DNA antibody, CBC and 24 h urine protein were taken. The results showed that nine patients in group 1 had obvious disease flare-up as evidenced by fever, arthralgia, malar rash and photosensitivity, increased protein loss or edema, and other symptoms. However, there were no significant changes in either group 1 or 2 patients. In contrast, the comparison of laboratory parameters in group 1 patients before and after the earthquake showed a significant decrease of C4 from 17.3 +/- 15.3 to 14.1 +/- 7.33 and a significant increase of creatinine from 1.20 +/- 1.54 to 1.27 +/- 1.70 (P < 0.05), respectively. No significant difference for either C4 or creatinine was found in group 2 patients. In conclusion, there was no striking change in the clinical aspects for lupus patients. However, some laboratory results became abnormal in group 1 patients after the earthquake, indicating that their lupus status may have worsened.

Recent progress of moving-actuator type mechanical circulatory support systems: AnyHeart and T-PLS.

The mechanical circulatory support system using moving-actuator mechanisms were developed by Seoul National University and Korea University. AnyHeart is a fully implanted pulsatile bi-ventricular assist device, and T-PLS is a pulsatile flow versatile extracorporeal life support system. Through lots of in-vitro and in-vivo experiments, the developed mechanical systems are faced to produce on commercial scale. This paper describes the recent progress of two mechanical circulatory support systems, AnyHeart and T-PLS.