OBJECTIVE: To investigate the repairing ability of nano-hydroxyapatite/recombinant human bone morphogenetic protein-2 (Nano-HA/rhBMP-2) composite artificial bone for bone defect and to provide evidence for its applicatio...OBJECTIVE: To investigate the repairing ability of nano-hydroxyapatite/recombinant human bone morphogenetic protein-2 (Nano-HA/rhBMP-2) composite artificial bone for bone defect and to provide evidence for its application in clinical repair of bone defect. METHODS: The animal model of bone defect was made on unilateral radius of 90 New Zealand white rabbits, which were randomly divided into experimental group (group A, bone defect was repaired with Nano-HA/rhBMP-2 composite artificial bone), control group (group B, bone defect was repaired with Nano-HA artificial bone), and blank group (group C, bone defect was unrepaired). The repairing ability for bone defect was evaluated by gross observation, X-ray examination, scanning electron microscope, radionuclide bone imaging and biomechanical analysis at 4(th), 8(th), and 12th week postoperatively. RESULTS: Both Nano-HA/rhBMP-2 composite artificial bone and Nano-HA artificial bone could stimulate new bone formation, but the former could stimulate more new bone formation and had better repairing ability for bone defect than that of the latter, with statistically significant difference (P <0.05). CONCLUSION: Nano-HA/rhBMP-2 composite artificial bone has good repairing ability for bone defect and it is hopeful to become an ideal repairing material for bone defect.
An amperometric biosensor based on zucchini (Cucurbita pepo) tissue homogenate is presented. The zucchini tissue homogenate was crosslinked with gelatine using glutaraldehyde and fixed on a pretreated teflon membrane. Th...An amperometric biosensor based on zucchini (Cucurbita pepo) tissue homogenate is presented. The zucchini tissue homogenate was crosslinked with gelatine using glutaraldehyde and fixed on a pretreated teflon membrane. The zucchini tissue contained the enzyme ascorbate oxidase and this enzyme catalyzed the oxidation of ascorbic acid in the presence of dissolved oxygen. The principle of the measurements was based on the determination of the decrease in the dissolved oxygen level. Determinations were carried out by standard curves, which were obtained by the measurement of the decrease in the oxygen level related to ascorbic acid concentration. Optimization and characterization studies of the biosensor were carried out in detail. First of all, the amounts of zucchini tissue homogenate, gelatin, and glutaraldehyde percentage were optimized. Experimental parameters such as buffer system, pH, buffer concentration, and temperature were also optimized carefully. Thermal stability, storage stability, and repeatability of the biosensor were investigated. A linear response was observed from 5x10(-6) M to 1.2x10(-3) M ascorbic acid. Finally, the results of some plant and drug samples analyzed with the presented biosensor compared with the spectrophotometric method (Tillman reagent) used as a reference.
Amperometric biosensors using laccase from Trametes versicolor as a bioelement were developed for 2,4-dichloro phenoxy acetic acid (2,4-D). Laccase enzyme was immobilized by gelatin and glutaraldehyde onto a Clark oxygen...Amperometric biosensors using laccase from Trametes versicolor as a bioelement were developed for 2,4-dichloro phenoxy acetic acid (2,4-D). Laccase enzyme was immobilized by gelatin and glutaraldehyde onto a Clark oxygen probe and screen printed electrodes (SPEs). Amperometric and chronoamperometric measurements were carried out with the biosensors. First, the effect of laccase activity on the biosensor performances was investigated for both biosensors, and then optimum pH and temperature and also thermal stability of the biosensors were tested. In addition, the detection ranges of some phenolic compounds were obtained by the help of calibration graphs of them. In repeatability studies, variation coefficients and standard deviations for both biosensors were also calculated by the studies done for this purposes. Finally, the biosensors were applied to the determination of 2,4-D in a real herbicide sample.
In this study the biosensor was constructed by immobilizing tissue homogenate of banana peel onto a glassy carbon electrode surface. Effects of immobilization materials amounts, effects of pH, buffer concentration and te...In this study the biosensor was constructed by immobilizing tissue homogenate of banana peel onto a glassy carbon electrode surface. Effects of immobilization materials amounts, effects of pH, buffer concentration and temperature on biosensor response were studied. In addition, the detection ranges of 13 phenolic compounds were obtained with the help of the calibration graphs. Storage stability, repeatability of the biosensor, inhibitory effect and sample applications were also investigated. A typical calibration curve for the sensor revealed a linear range of 10-80 microM catechol. In reproducibility studies, variation coefficient and standard deviation were calculated as 2.69%, 1.44 x 10(-3) microM, respectively.
The whole blood immuno-adsorption (WBIA) system, using an adsorbent to remove pathogenic antibodies of Myasthenia Gravis (MG), was studied. Cellulose-tryptophan adsorbent was synthesized and purified in our lab. Experime...The whole blood immuno-adsorption (WBIA) system, using an adsorbent to remove pathogenic antibodies of Myasthenia Gravis (MG), was studied. Cellulose-tryptophan adsorbent was synthesized and purified in our lab. Experimental autoimmune myasthenia gravis (EAMG) rabbits were passively transferred with immunoglobulin from patients with myasthenia gravis. The rabbits underwent extracorporeal whole blood adsorption for 2 hours. Results showed no significant damage to blood cells and no changes in the concentrations of electrolytes. Total protein decreased by 12.6% (P<0.05) and globulin protein decreased 21.9% (P<0.05). The overall removal of antibodies against nicotinic acetylcholine receptor (nAChR) was 49.85%. The percentage of decrement of compound muscle action potential in 3, 5, 10 Hz of EAMG rabbits all dropped down after the treatment. The quantity of neuromuscular junctions per unit area (25 mm(2)) increased significantly after treatment (P<0.05). In conclusion, the adsorbent was biocompatible, safe for whole blood immuno-adsorption. Whole blood immuno-adsorption improved clinical manifestation and neuromuscular function of the passively transferred EAMG rabbits.
Numerous manufacturing techniques for autogenous fibrin glue used as scaffold material have been described. As there is no consensus regarding the influence of chemical additives on cell biology, it was the aim of this s...Numerous manufacturing techniques for autogenous fibrin glue used as scaffold material have been described. As there is no consensus regarding the influence of chemical additives on cell biology, it was the aim of this study to establish a method for manufacturing autologous fibrin glue without any additives. The serum part was separated from whole blood. After fibrinogen precipitation, centrifugation was performed to obtain the fibrinogen pellet. Various experimental series were run to examine influences of various temperatures or substituting centrifugation for sedimentation. The method as described here is effective, simple, and performed without any additives, which could potentially influence cell biology.
OBJECTIVE: To investigate the effects of recombinant Murine RANK on the osteoclast activity. METHODS: Osteoclast was observed with soluble RANK. Female Wistar rats were bilaterally ovariectomized, and intra-abdominally i...OBJECTIVE: To investigate the effects of recombinant Murine RANK on the osteoclast activity. METHODS: Osteoclast was observed with soluble RANK. Female Wistar rats were bilaterally ovariectomized, and intra-abdominally injected with 5 mg/Kg soluble RANK. The bone metabolism, bone density, and bone histomorphology were observed. RESULTS: Compared with the control group, the quantity of TRAP-positive osteoclasts and bone resorption pit counting in the rest groups were significantly reduced. The bone density of the dosed groups was significantly increased and TRAP-stained osteoclasts in bone tissue sections were almost inhibited. CONCLUSION: rh-Murine RANK was able to inhibit osteoclast differentiation and prevent ovariectomy-induced bone loss.
A highly sensitive, amperometric uric acid biosensor possessing unique physical, electro-conductive properties of carbon nanotubes is described for determination of uric acid level in serum. A uric acid biosensor was con...A highly sensitive, amperometric uric acid biosensor possessing unique physical, electro-conductive properties of carbon nanotubes is described for determination of uric acid level in serum. A uric acid biosensor was constructed after immobilization of uricase onto a polyaniline/CNT matrix by carbonamide linkage. The scanning electron micrographs confirmed the immobilization of globular enzyme onto the electroploymerized polyaniline/CNT composite. Employment of carbon nanotubes composite in the present electrode leads to a very quick response time of 8 sec with the minimum detection limit of 5 microM along with an increase in shelf-life of electrode system to >180 days with slight loss of enzyme activity. Efficiency of the present amperometric uric acid biosensor was validated by quantitative estimation of uric acid level in biological serum of healthy individuals and the persons suffering from hyperuricemia and gout.
The fiberoptical spatial filter anemometry (SFA) is a common technique based on an optical grid to measure the velocity of corpuscular components in a multiphase flow, e.g. in the microvessels. The technical innovation i...The fiberoptical spatial filter anemometry (SFA) is a common technique based on an optical grid to measure the velocity of corpuscular components in a multiphase flow, e.g. in the microvessels. The technical innovation is the analysis of flow velocities using an optical grid sensor and frequency analysis by Fast Fourier Transformation (FFT). This study describes a non-invasive, on-line technique to measure RBCV in the microcirculation. The sensor's validity was proven by in vitro measurements using a rotation disk of an exactly defined velocity with a correlation coefficient of 0.99967. For validation of RBCV measurements in the microcirculation in vivo, the setup was adapted to an intravital microscope. RBCV was measured in arterioles, capillaries, and postcapillary venules ranging from 8-140 microm diameter. As reference method for velocity measurements a computer assisted imaging system was used to measure the RBC-velocity in the identical vessels by frame to frame analysis. Both methods revealed a high significant correlation using transillumination technique for capillaries (r=0.986, p<0.001) and venules (r=0.952, p<0.001) as well as epiillumination technique (capillaries r=0.939, venules r=0.975, p<0.001).
This study was designed to investigate whether polymerized human placenta hemoglobin (PolyPHb) pretreatment provided protection to the heart after ischemia/reperfusion (I/R) injury. After 10-min basal perfusion, isolated...This study was designed to investigate whether polymerized human placenta hemoglobin (PolyPHb) pretreatment provided protection to the heart after ischemia/reperfusion (I/R) injury. After 10-min basal perfusion, isolated Sprague-Dawley rat hearts were pretreated with 0.1 gHb/dL PolyPHb and subjected to I/R injury. PolyPHb pretreatment greatly reduced the decreases in left ventricular developed pressure (LVDP), maximum LVDP increase and decrease rate (+/-dp/dt), and the increase in left ventricular end-diastolic pressure (LVEDP) as compared to the control group. Moreover, the myocardial infarction and troponin-I release were significantly reduced in the pre-HBOCs group. Therefore, PolyPHb pretreatment was protective to the isolated I/R heart.
Up to 32 Landrace ear marginal tissue samples were used for establishing a fibroblast cell bank by the means of primary explantation and cryopreservation. Biological analysis suggested that the Population Doubling Time (...Up to 32 Landrace ear marginal tissue samples were used for establishing a fibroblast cell bank by the means of primary explantation and cryopreservation. Biological analysis suggested that the Population Doubling Time (PDT) of the cell line was approximately 24h. The diploid accounted for 97.2% of the whole population; isozyme analysis of Lactic Dehydrogenase (LDH) and Malic Dehydrogenase(MDH) disproved cross-contamination from other cell lines. The results of bacterium, fungus, virus and mycoplasma tests were all negative. The transfection rates of three fluorescent proteins were high, indicating that the exogenous genes could be effectively expressed in the cells. It had not only preserved the precious germplasm resource of the Landrace on the cell level but also provided valuable material for the researches of genomics, postgenomics, somatic cloning and so on.
A novel encapsulated oxidative biocatalyst comprising glucose oxidase (GOD) coencapsulated with oxygen carriers within polyelectrolyte complex capsules was developed for the production of D-gluconic acid and delta-glucon...A novel encapsulated oxidative biocatalyst comprising glucose oxidase (GOD) coencapsulated with oxygen carriers within polyelectrolyte complex capsules was developed for the production of D-gluconic acid and delta-gluconolactone. The capsules containing immobilized GOD were produced by polyelectrolyte complexation with sodium alginate (SA) and cellulose sulfate (CS) as polyanions, poly(methylene-co-guanidine) (PMCG) as the polycation, CaCl(2) as the gelling agent and NaCl as the antigelling agent (GOD-SA-CS/PMCG capsules). Poly(dimethylsiloxane) (PDMS) and an emulsion of n-dodecane (DOD) or perfluorodecaline (PFD) with PDMS were used as the oxygen carriers and MnO(2) was used as a hydrogen peroxide decomposition catalyst. Water-soluble PDMS was found to act as both an oxygen carrier and an emulsifier of water-insoluble DOD and PFD. Stable microcapsules could be produced with concentrations of up to 4% (w/w) of PDMS, 10% (w/w) of DOD and PFD, and 25% (w/w) of MnO(2) in the polyanion solution of SA and CS. Roughly a two-fold increase in the GOD activity from 21.0+/-1.1 to 38.4+/-2.0 U*g(-1) and product space-time yields (STY) from 44.3+/-2.0 to 83.4+/-3.4 g*H*day(-1) could be achieved utilizing coencapsulated oxygen carriers compared to GOD encapsulated in the absence of oxygen carriers. This enhanced production does not significantly depend on the selected oxygen carrier under the conditions used in this study.
Artif Cells Blood Substit Immobil Biotechnol
· 2010 Apr · PMID 20205612
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Crosslinking of ultrapure hemoglobin, crystalline catalase, and superoxide dismutase resulted in a soluble nanodimensional complex of polyhemoglobin-catalase-superoxide dismutase. A less expensive and more convenient way...Crosslinking of ultrapure hemoglobin, crystalline catalase, and superoxide dismutase resulted in a soluble nanodimensional complex of polyhemoglobin-catalase-superoxide dismutase. A less expensive and more convenient way is to crosslink bovine stroma-free hemolysate (stroma-free hemolysate) that already contains hemoglobin, catalase, and superoxide dismutase into polyhemoglobin with catalase and superoxide dismutase activities (stroma-free polyhemolysate) [21]. The objective of the present study is to evaluate the immunological properties of this stroma-free polyhemolysate. Each of three groups of rats received weekly subcutaneous injections of one of the stroma-free polyhemolysate, stroma-free hemolysate, and saline for four weeks. One week after the four cycles of weekly immunization, serum and plasma were collected for C3a complement activation tests and Ouchterlony antibody-antigen precipitation tests, respectively. Results show that stroma-free polyhemolysate retained significant antioxidant enzyme activity. The C3a complement activation test and Ouchterlony test show that four weekly subcutaneous injections of bovine stroma-free polyhemolysate did not result in any immunological reaction in rats when tested this way.
Chitin conduits to bridge nerve injuries with a small gap (2 mm) give us a biological window in which the peripheral nerve regeneration process can be observed. In this study, the regeneration process was observed on dif...Chitin conduits to bridge nerve injuries with a small gap (2 mm) give us a biological window in which the peripheral nerve regeneration process can be observed. In this study, the regeneration process was observed on different intervals postoperatively. Histological analysis revealed that the early regeneration process occurred in three phase: degeneration and matrix phase, axonal and Schwann migration phase, myelination and maturation phase. Schwann cells grew into the lumen from both the proximal and distal nerve segments. Axonal regrowth progressed at an average rate of 1.4 mm/d. Regenerating axonal fibers were myelinated by Schwann cells from both sides of the conduit.
Artif Cells Blood Substit Immobil Biotechnol
· 2010 Apr · PMID 20196685
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Liposome-encapsulated hemoglobin (LEH) based on a novel, synthetic, non-phospholipid was developed, and evaluated for cerebral energy metabolism in a 40% hemorrhage rat model. The markers of tissue energetics were monito...Liposome-encapsulated hemoglobin (LEH) based on a novel, synthetic, non-phospholipid was developed, and evaluated for cerebral energy metabolism in a 40% hemorrhage rat model. The markers of tissue energetics were monitored by (1)H- and (31)P-magnetic resonance spectroscopy (MRS). After hemorrhage, (1)H-MRS showed an increase in the levels of lactate and pyruvate. These markers returned to baseline values following LEH resuscitation. Both LEH and saline were able to exert a neuron-protective effect as indicated by the recovery of N-acetylaspartate. (31)P MRS showed a fall in phosphocreatine after hemorrhage, which upon LEH or saline resuscitation returned to the baseline values. Similarly, inorganic phosphate increased after bleeding, but returned to normal after resuscitation. LEH resuscitation also recovered beta-ATP levels, but saline resuscitation provided only a modest recovery. The results indicate the utility of MRS to monitor cerebral metabolism in hemorrhage/resuscitation. The data is also supportive of the new LEH formulation as an oxygen carrier.
To investigate the diagnosis and microsurgical treatment of cavernous sinus hemangioma, the clinical data, including pathology, epidemiology, medical imaging, operation procedure, and post-operational complication of 12...To investigate the diagnosis and microsurgical treatment of cavernous sinus hemangioma, the clinical data, including pathology, epidemiology, medical imaging, operation procedure, and post-operational complication of 12 cavernous sinus hemangioma patients undergoing operations in Affiliated Hospital of Medical College of Qingdao University from 1999 to 2008, were analyzed. There were 2 males and 10 females. The patients were aged from 28 to 61 years. Headaches and deficits of the cranial nerves coursing through the cavernous sinus were the principal symptoms at presentation. The common clinical manifestations were visual loss, diplopia, facial numbness, and extraocular muscle palsy. The radiological features in all patients were similar with a characteristic pattern of extension and encasement of carotid artery. CT showed the lesion as hypodense to isodense with marked enhancement after contrast administration. T1-weighted MR imaging showed the lesions as hypointense with marked enhancement after contrast administration. T2-weighted MR imaging showed the lesions as hyperintense. The maximum size of the lesion was 9 to 57 mm (mean 45 mm). Basal pterional craniotomies were used for eight patients. Orbitozygomatic osteotomies were used for two patients. Pterional approach was used for two patients. The lesions were removed through incising the lateral wall of the cavernous sinus. The tumor was totally removed in five cases, subtotally removed in four cases, and partially removed in two cases. The main post-operational complications included oculomotor nerve paralysis (four cases) and trigeminal nerve lesions (three cases). No postoperative death occurred. Operation is the best choice for cavernous sinous hemangioma. It was helpful to control bleeding through intradura and incising the lateral wall of the cavernous sinus.
A zero-linked polymeric hemoglobin (OxyVita Hb) has been developed for application as an acellular therapeutic hemoglobin-based-oxygen-carrier (HBOC). For effective and safe oxygen binding, transport and delivery, an HBO...A zero-linked polymeric hemoglobin (OxyVita Hb) has been developed for application as an acellular therapeutic hemoglobin-based-oxygen-carrier (HBOC). For effective and safe oxygen binding, transport and delivery, an HBOC must meet essential molecular requirements related to its structural integrity and redox stability. OxyVita is a super polymer possessing an average M.wt. of 17 x 10(6) Da. Structural integrity was determined by unfolding studies of OxyVita in the presence of increasing concentrations of urea. The unfolding midpoints (D(1/2)) of different preparations of OxyVita (solution and powder forms) were compared to Lumbricus Hb (LtHb) and Arenicola Hb (ArHb), natural acellular polymeric hemoglobins, which are serving as models for an effective and safe acellular HBOC. Reduction studies of OxyVita Hb using endogenous reducing agents were also investigated. Results from these studies indicate that: 1) OxyVita Hb exhibits greater resistance to conformational change than either LtHb or ArHb in the reduced (oxyHb) state; and 2) the reduction of met OxyVita Hb to oxyHb occurs slowly in the presence of either ascorbic acid (70% reduction in 560 min.) or beta-NADH (40% reduction in 90 min.). These studies provide consistent evidence that OxyVita Hb possesses physiochemical properties that exhibit structural integrity and redox behavior necessary for functioning as an effective and safe HBOC within clinical applications. These results are in agreement with observations made by other investigators as to the reduction in heme-loss of OxyVita Hb, essential for the reversible binding/release of molecular oxygen within the circulatory system.
RATIONALE: Earlier work has shown that experimental conditions calling for improved tissue oxygenation could be assisted by i.v. infusion of a dodecafluoropentane emulsion (DDFPe) forming oxygen-transporting microbubbles...RATIONALE: Earlier work has shown that experimental conditions calling for improved tissue oxygenation could be assisted by i.v. infusion of a dodecafluoropentane emulsion (DDFPe) forming oxygen-transporting microbubbles. OBJECTIVES: The present work investigated the effect of DDFPe on hypoxia due to experimental shunts in the pig lung. METHODS: Nineteen O(2) breathing, anesthetized pigs had glass beads administered into the trachea so as to significantly depress arterial oxygen tension (PaO(2)). PaO(2) was recorded for up to 12 hrs while 0.1 ml/kg DDFPe was administered 1-3 times. MAIN RESULTS: The animals were divided into two groups based on arterial oxygen saturation (SaO(2)) after shunt induction, combined with oxygen breathing: the "SaO(2) >90% group" (n=6) and the "SaO(2) <90% group" (n=13). In the "SaO(2) <90% group," the PaO(2) increased stepwise with each infusion from 56.6+/-2.9 to 88.6+/-14.6 mmHG (P<or=0.001); improvements lasted about 2 hrs after each infusion. Mixed venous oxygenation also increased with the infusions, e.g. (1(st) infusion) from a PvO(2) of 41.4+/-2.3 to 49.9+/-4.2 mmHg (P<or=0.05) and SvO(2) 58.0+/-2.9% (P<or=0.01), the venous changes supporting arterial oxygenation. At the same time, arterial CO(2) levels fell. Arterial O(2) and CO(2) levels were paralleled by similar changes in muscle tissue. Pulmonary arterial pressures did not indicate any pulmonary embolization by bubbles. Toxic effects of the treatment were not observed. CONCLUSION: These results suggest that, on condition of successful toxicity testing, intravascular administration of a DDFPe and oxygen breathing may be beneficial in severe right-to-left shunting in humans.
BACKGROUND: The binding of anti-acetylcholine receptor antibodies (AChRAb) to the main immunogenic region (MIR) of AChR alpha-subunit in the neuromuscular junction is the major pathogenesis of myasthenia gravis (MG). MET...BACKGROUND: The binding of anti-acetylcholine receptor antibodies (AChRAb) to the main immunogenic region (MIR) of AChR alpha-subunit in the neuromuscular junction is the major pathogenesis of myasthenia gravis (MG). METHODS: A synthetic peptide of 10 amino acids corresponding to the MIR of human AChR was coupled with cellulose beads to make an antigen-specific immunoadsorbent (hMIR10-CB). RESULTS: The hMIR10-CB could remove AChRAb in MG sera by 40.3+/-2.3%, compared to a tryptophan nonspecific adsorbent Trp-CB by only 22.4+/-1.5% as determined in ELISA, and also showed good blood compatibility for blood cells, plasma ions and plasma proteins as checked in whole blood perfusion in rabbits. CONCLUSIONS: The antigen-specific immunoadsorbent hMIR10-CB can serve as a potential candidate in the immunoadsorption treatment of MG.