We have assessed the biocompatibility of a new composite bone graft consisting of calcium phosphate cement (CPC) and poly(1actide-co-glycolide) (PLGA) microspheres (approximate diameter of 0.184.36 mm) using cell culture techniques. CPC powder is mixed with PLGA microspheres and water to yield a workable paste that could be sculpted to fit the contours of a wound. The cement then hardens into a matrix of hydroxyapatite microcrystals containing PLGA microspheres. The rationale for this design is that the microspheres will initially stabilize the graft but can then degrade to leave behind macropores for colonization by osteoblasts. The CPC matrix could then be resorbed and replaced with new bone. In the present study, osteoblast-like cells (MC3T3-El cells) were seeded onto graft specimens and evaluated with fluorescence microscopy, environmental scanning electron microscopy and the Wst-1 assay (an enzymatic assay for mitochondria1 dehydrogenase activity). Cells were able to adhere, attain a normal morphology, proliferate and remain viable when cultured on the new composite graft (CPC-PLGA) or on a control graft (CPC alone). These results suggest that our new cement consisting of CPC and PLGA microspheres is biocompatible. This is the first time that a 'polymer-in-mineral' (PLGA microspheres dispersed in a CPC matrix) cement has been formulated that is moldable, resorbable and that can form macropores after the cement has set.
The chemical, physical, and morphological complexity of atmospheric aerosol elemental carbon (EC) presents major problems in assuring measurement accuracy. Since EC and black carbon are often considered equivalent, methods based on thermal-optical analysis (TOA) are widely used for EC in ambient air samples because no prior knowledge of the aerosol's absorption coefficient is required. • C for 60 s, 850• C for 60 s, and 900• C for 90 s to 120 s, respectively.
A method for fabricating biomimetic surfaces from intact cell membranes is described. A monolayer of alkanethiol on gold is covered by a second layer derived from the components of erythrocyte membranes either by self-assembly or by Langmuir-Blodgett methods. The resulting asymmetric hybrid layer was characterized by ellipsometry, surface plasmon resonance (SPR), contact angle, capacitance, voltammetry, and electron and atomic force microscopy. The erythrocyte membrane layer was measured to be approximately 30-40 A in thickness. Using SPR, the presence of erythrocyte components on the surface was demonstrated by their selective removal by enzymatic action. The uniform deposition of membranous material on the substrate was shown by electron and atomic force microscopy. Demonstration of acetylcholinesterase (AChase) activity, a membrane-anchored enzyme, on the surface for at least 8 days, suggests that the outer leaflet of the erythrocyte membrane is present in its native form. Cyclic voltammetry demonstrates that enhanced electron transport from a solution redox species accompanies formation of the erythrocyte layer at the surface. This enhanced electron transport is blocked by 4,4'-diisothiocyanate stilbene-2,2'-disulfonic acid, a well known blocker of anion transport, suggesting that an erythrocyte anion transporter protein is incorporated into the surface layer in an active conformation.
Purpose
Nanoparticle size is important in drug delivery. Clearance of nanoparticles by cells of the reticuloendothelial system has been reported to increase with increase in particle size. Further, nanoparticles should be small enough to avoid lung or spleen filtering effects. Endocytosis and accumulation in tumor tissue by the enhanced permeability and retention effect are also processes that are influenced by particle size. We present the results of studies designed to optimize crosslinked biodegradable stealth polymeric nanoparticles fabricated by dispersion polymerization.
Methods
Nanoparticles were fabricated using different amounts of macromonomer, initiators, crosslinking agent and stabilizer in a dioxane/DMSO/water solvent system. Confirmation of nanoparticle formation was by scanning electron microscopy (SEM). Particle size was measured by dynamic light scattering (DLS). D-optimal mixture statistical experimental design was used for the experimental runs, followed by model generation (Scheffe polynomial) and optimization with the aid of a computer software. Model verification was done by comparing particle size data of some suggested solutions to the predicted particle sizes.
Results and Conclusion
Data showed that average particle sizes follow the same trend as predicted by the model. Negative terms in the model corresponding to the crosslinking agent and stabilizer indicate the important factors for minimizing particle size.
Decanethiol [CH3(CH2)9SH] self-assembled monolayer films on silver substrates have been irradiated in selected areas by focused ion or electron bombardment. Subsequent immersion of the irradiated sample in a solution of a fluoromercaptan [CF3(CF2)2(CH2)2SH] results in attachment of this molecule to the silver surface in the ion or electron-exposed regions, producing a micrometer spatial-scale pattern of two chemically distinct alkanethiol monolayers. The coverage of the fluoromercaptan on the bombarded areas was found to reach maximum levels of 70% at ion doses of 6×1013 ions/cm2 and 50% at electron doses of 2×1017 electrons/cm2 as determined by secondary ion mass spectrometry. These methods of maskless patterning may be useful for semiconductor or biosensor device fabrication.
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