Artificial cells, blood substitutes, and immobilization biotechnology最新文献

Pectinesterase isolated from Malatya apricot pulp was noncovalently and covalently immobilized onto bentonite and glutaraldehyde-containing amino group functionalized porous glass beads surface at pH 8.0 and pH 9.0, respectively. The effect of various parameters such as pH, temperature, activation energy, heat and storage stability on immobilized enzyme were investigated. The optimum temperature of covalently and noncovalently immobilized PE was 50°C. This value was 60°C for free PE. Although optimum pH of covalently-immobilized PE was 8.0, this parameter was 9.0 for free and covalently-immobilized PE. The noncovalently immobilized enzyme exhibited better thermostability than the free and covalently immobilized PE.

Objective: Currently used decellularization procedures have negative effects on extracellular matrix (ECM) integrity. The objective of this study is to evaluate four decellularization methods and their effect on the collagen ultrastructure, mechanical behavior and antigenicity of porcine aortic valves.

Methods: Aortic valves were placed in a trypsin, osmotic, trypsin-osmotic or detergent-osmotic solution. Leaflets were processed for histology and mechanical testing. Matrices were implanted subdermally in rats to evaluate immune reaction and calcification.

Results: Trypsin-osmotic methodology effected near-complete decellularization. Trypsin treatment resulted in cell removal only in the spongiosa layer. Osmotic and detergent-osmotic treatments did not remove any cells from the cusps. Mechanical strength was significantly inferior in the trypsin (p50,03) and trypsin-osmotic treated group (p50,04). Trypsin and trypsin-osmotic decellularized matrices evoked a strong CD31 inflammatory cell infiltration.

Conclusion: Enzymatic-osmotic decellularization appears to be the only effective method to remove cellular components. However, the near cell free scaffolds still evokes a strong CD31 T-cell inflammatory reaction.

To prepare an NCECS/nHA composite for tissue-engineered trachea and investigate its biomechanical and biocompatibile properties. Biomechanical tests were performed on dry and wet NCECS/nHA composite specimens prepared in vitro. The cell adhesion rate on each composite surface after 2, 6, and 12 hours of culture was calculated, and cell proliferation activity was measured using an MTT assay. NCECS/nHA composites exhibited satisfactory tensile strength and Young's modulus values. The adhesion rate of rabbit tracheal chondrocytes on NCECS/nHA surfaces reached 88.4% after 12 hours of culture. NCECS/nHA composites are promising scaffold materials for tissue-engineered trachea owing to satisfactory biocompatible and biomechanical properties.

In a previous dose escalation study our group found that combining 90μg/kg rFVIIa with HBOC-201 reduced blood loss and improved physiologic parameters compared to HBOC alone. In this follow-up study in a swine liver injury model, we found that while there were no adverse hematology effects and trends observed in the previous study were confirmed, statistical significance could not be reached. Additional pre-clinical studies are indicated to identify optimal components of a multifunctional blood substitute for clinical use in trauma.

A method is described for construction of an amperometric polyphenol biosensor employing nitrocellulose membrane-bound laccase purified from cell-free extract of Ganoderma lucidum onto a Pt electrode. The biosensor showed optimum response within 10s, at 0.4 V in 0.1M acetate buffer, pH 6.0, and 35°C. Detection limit of the biosensor was 3.0 × 10(-8)M. Analytical recovery of added guaiacol was 97.00%. Within batch and between batch coefficients of variation were <0.97% and <1.26%, respectively. The sensor measured total phenolic content in fruit juices and alcoholic beverages. The enzyme electrode was used 100 times over 4 months, when stored at 4°C.

Nanotechnology has potential in the development of novel and effective delivery of drugs within lungs. Different strategies have been utilized for pulmonary delivery of drugs, including the use of lipid-based delivery systems (liposomes, ISCOMs, SLNs), use of polymeric matrix (PLGA, poly caprolactone, cynoacrylates, gelatin), development of polysaccharide particulates (chitosan, alginates, Carbopol, etc.), biocompatible metallic inorganic particles (iron, gold, zinc), etc. This paper reviews various nanopaticulate approaches in the form of lipids, polymers, metals, polysaccharides, or emulsions based for pulmonary drug delivery that could provide an increased biological efficacy and better local and systemic action.

We report the application of Vitamin D3 (VD(3)) in nanoparticles of oleoyl alginate ester (OAE)(OAE-VD(3)). The internalization of fluorescent OAE-VD(3) by Caco-2 cells was visualized by confocal laser scanning microscopy. In vivo pharmacokinetic studies showed that incorporation into OAE nanoparticles resulted in increased absorption of VD(3). Its application in the treatment of rickets was assayed using a model of nutritionally induced vitamin D-deficiency rickets. The results showed that the encapsulated VD(3) had better efficacy than that of the free drug in vivo. Our studies provide evidence that OAE nanoparticles are valuable as nutraceutical delivery vehicles to enhance the absorption of VD(3).

Background: Primary osteoarthritis (OA) is a polygenic disease. To investigate the gene expression profile of cartilage and synovium from osteoarthritis and healthy rats using cDNA microarray is beneficial to recognize the pathogenesis of osteoarthritis and provide evidence for gene therapy of osteoarthritis.

Objective: The present study aimed to investigate the gene expression profile of the cartilage and synovium of chronic arthritis and healthy rats through cDNA microarray assay, and identify the differentially expressed genes. This study may be helpful for understanding the role of differentially expressed genes in osteoarthritis and the gene polymorphism of osteoarthritis.

Methods: A total of 24 male Wistar rats were randomly divided into control group and osteoarthritis group (n = 12 per group). The synovial and cartilage were obtained and total RNA was extracted. cDNA microarray assay was performed to identify the differentially expressed genes, and cluster analysis was conducted.

Results and conclusion: A total of 82 differentially expressed genes were identified, among which 27 were up-regulated and 55 down-regulated. Gene microarray assay is effective to identify differentially expressed genes and may find out novel osteoarthritis associated genes. Multiple genes are involved in the pathogenesis of osteoarthritis. The differentially expressed genes provide important information for further studies on the pathogenesis of osteoarthritis and gene therapy of osteoarthritis.

The temperature effects on enzyme activity at the conditions with and without inhibition, operation, and storage were investigated for free esterase and immobilized chicken liver esterase with three methods (adsorption, adsorption-microcapsule, and adsorption-crosslinking) used in a calorimetric biosensor. The results indicated that the temperature had significant effects on the enzyme activity by means of catalytic reaction, irreversible denaturation, mass transfer, and structure deformation of resin. Among all the esterase, the adsorption-crosslinking immobilized esterase had the best performances of enzyme activity retention ratio (EARR), above 96% in operation and 90% in storage on condition of maintaining its response to dichlorvos.

Poly(methyl methacrylate) (PMMA) microfibers were used as a template for development of a capillary-like network in agarose hydrogel. Microfibers with diameter 10-20 μm, which is comparable to the diameter of native capillary vessels, were fabricated using a wet spinning technique. The microfibers were embedded in agarose gel and dissolved by immersing the gel in dichloromethane. The resultant microchannels in the gel had the same diameter as the microfibers, and allowed an aqueous solution to be perfused through the gel. The methodology is promising for fabricating a capillary-like network in tissue engineering scaffolds of various water-soluble polymers.