Aim. The usefulness of photodynamic therapy (PDT) for treating sentinel lymph node (SLN) metastasis was evaluated. Materials and Methods. Verteporfin, a hydrophobic photosensitizer, forms a soluble ...aggregate with poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) (PMB). The concentrations of verteporfin were determined by measuring the fluorescence emitted at 700 nm. Seven days after the inoculation of A431 cells at the forearm of BALB/c nude mice, PMB-verteporfin was injected at dorsum manus and 75 J of light energy was delivered for 1 minute. Fifty-three mice were randomly assigned to the combination of PMB-verteporfin injection and light exposure, light exposure alone, PMB-verteporfin injection alone, and no treatment groups. Ten days after PDT, brachial lymph nodes, which were considered as SLNs, were harvested and evaluated. Results. The concentration of verteporfin in SLN was significantly higher than other organs. The combination of PMB-verteporfin injection and light exposure group significantly reduced the SLN metastasis (13%) comparing with no treatment group (52%), light exposure alone group (57%), and PMB-verteporfin injection alone group (46%). Conclusions. These data suggested that PDT using PMB as a nanotransporter of verteporfin could be a minimally invasive treatment of SLN metastasis in breast cancer and represent a potential alternative procedure to SLNB.
The optimum spikelet number in Oryza sativa L. “Datemasayume” cultivated in Miyagi Prefecture is 30,000–34,000 grains m−2. This number may be obtained (within the target range) by controlling ...nitrogen uptake during the panicle formation and full heading stages. Thus, to control nitrogen uptake and obtain the optimum spikelet number, we designed a fertilization method wherein soil nitrogen fertility is considered. Nitrogen uptake from the transplantation to panicle formation stages could be predicted using a multiple regression equation in which the amount of nitrogen in basic fertilizer and effective accumulative temperature were explanatory variables. Furthermore, nitrogen uptake from the panicle formation to full heading stages could be predicted using a multiple regression equation in which the amount of nitrogen in topdressing fertilizer and the amount of nitrogen applied to the surface soil were explanatory variables. These multiple regression equations were adapted to calculate the optimum amounts of basic and topdressing fertilizer nitrogen. According to verification analysis conducted in a local producer’s field, the more that the amount of applied fertilizer deviated from the calculated amounts of basic and topdressing fertilizer estimated using our formula, the more that the nitrogen uptake from the transplantation to the panicle formation stage and the spikelet number deviated from the target ranges. Thus, the optimum spikelet number of Datemasayume can be obtained if the optimum nitrogen richness, calculated according to our formula, is provided using basic and topdressing fertilizers.
BACKGROUND:Peritendinous adhesions are serious complications after surgical repair of tendons. As an anti-adhesion material, we focused on 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, our ...original biocompatible polymer, and prepared an aqueous solution of MPC-containing polymer called poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate-co-p-vinylphenylboronic acid) (PMBV), which can be formed into hydrogel properties by mixture with another aqueous polymer, poly(vinyl alcohol) (PVA). The objective of the present study was to examine the possible application of the MPC hydrogel for the reduction of peritendinous adhesions.
METHODS:The effects of the hydrogel on peritendinous adhesions and tendon healing were examined by means of histological and mechanical analyses in a rat Achilles tendon model and a rabbit flexor digitorum profundus tendon model. Cell migration and viability were examined with use of fibroblastic NIH3T3 cells cultured in a double chamber dish.
RESULTS:Among the concentrations examined, 2.5% and 5.0% PMBV formed hydrogel properties immediately after mixing with 2.5% PVA and maintained a honeycomb microstructure with nanometer-scaled pores for three weeks after implantation. In animal models, the hydrogel formed from 5.0% PMBV remained at the sutured site during the critical period up to three weeks and disappeared by six weeks. The MPC hydrogel reduced the peritendinous adhesions histologically and mechanically by >25% at three weeks, without impairing tendon healing as determined with mechanical analyses. In the cell culture, cell migration was reduced by the MPC hydrogel, although cell viability was unaffected, indicating physical prevention, rather than cytotoxicity, to be the anti-adhesion mechanism.
CONCLUSIONS:The MPC hydrogel that was formed by a local injection and mixture of two aqueous solutions, 5.0% PMBV and 2.5% PVA, reduced peritendinous adhesions without impairing tendon healing. This effect may be due to its excellent biocompatibility without a foreign-body reaction and the formation of a microstructure that physically prevents passage of cells but allows cytokines and growth factors to pass for healing.
CLINICAL RELEVANCE:This nanotechnology could potentially improve the quality of surgical repair of tendon, especially the zone-II area of the digital flexor tendon.
A carbon nanotube (CNT)‐modified electrode was fabricated by dropping a dispersion of multi‐walled CNTs in water‐soluble and amphiphilic phospholipid polymer with both dispersing ability and ...anti‐biofouling property onto a Au electrode. A poly(2‐methacryloyloxyethyl phosphorylcholine‐co‐n‐butyl methacrylate) (PMB) composed from 50 mol% of 2‐methacryloxylethyl phosphorylcholine and 50 mol% of n‐butyl methacrylate (PMB50) was used as dispersing reagent for CNTs. The dispersion of water‐insoluble material by PMB50 and its antifouling effects in electrochemical analysis were investigated. The CNT‐modified electrode showed an anodic peak potential that was shifted negatively and an increase in the current value for the electrolytic oxidation of nicotinamide adenine dinucleotide. In addition, the charge on PMB50 did not inhibit the electrochemical reaction of the redox compounds K3Fe(CN)6, Ru(NH3)6Cl3, and hydroxymethylferrocene. Cyclic voltammetry of K3Fe(CN)6 in 4 % bovine serum albumin (BSA) using a bare Au electrode, the anodic peak current was reduced to 47 % of that without BSA. In contrast, the antifouling effect of the PMB50‐coated electrode meant that the current was only reduced to 70 % of that without BSA.
Abstract The biological performances of a cell-containing phospholipid polymer hydrogel in bulk and miniaturized formats without an additional culture medium support were investigated and compared. ...The cell-containing hydrogel was formed spontaneously when solutions of commercial polyvinyl alcohol (PVA) and the phospholipid polymer poly2-methacryloyloxyethyl phosphorylcholine (MPC)- co - n -butyl methacrylate (BMA)- co - p -vinylphenylboronic acid (VPBA) (PMBV) suspended with cells in a cell culture medium are mixed together. Bulk and miniaturized hydrogels, with approximate thicknesses of 3.1 mm and 400 μm, respectively, were prepared in a 96-well microplate and a glass microchip, respectively. In both cases, the hydrogels were homogeneous, and cells were spatially encapsulated. The long-term observation (4 and 8 days) of cell morphology suggested that cells were passively attached to the interface of the hydrogel but were unable to spread and flatten, which inhibited cell growth in both hydrogels. Viability evaluations revealed that cells in both hydrogel formats maintained the same high viability levels after long-term encapsulation. Cytotoxicity assays indicated that the cells in the miniaturized hydrogel maintained a high degree of correlation in cytotoxic sensitivity with the cells in the bulk hydrogel and a routine medium culture. The PMBV/PVA hydrogel not only provides a beneficial cytocompatible microenvironment for long-term cell survival without an additional culture medium support but also creates a static condition for cell sustainment in a microchip similar to that in bulk. The uniform long-term performances of PMBV/PVA hydrogels in bulk and miniaturized formats make them ideal for the development of long-term, flexible, three-dimensional, living cell-based tools for routine cell-based assays and applications on bulk to microscale levels.
A photo-reactive polymer having a phospholipid polar group was prepared, and the polymer was photo-immobilized on polymeric surfaces, where its interactions with biocomponents were investigated. By ...using a photo-immobilization method, the polymer was used for surface modification of polyethylene and polypropylene, polymers whose surfaces were not treated in our previous development of the phosphorylcholine-derived polymer. The photo-reactive polymer was synthesized by a coupling reaction involving copolymer consisting of 2-methacryloyloxyethyl phosphorylcholine and methacrylic acid with 4-azidoaniline. When the polymer was unpattern immobilized on the surface, X-ray photo-electron spectroscopic analysis and static contact angle measurements were performed. It was shown that the surface was covered with phospholipid polar groups. Micropattern immobilization was carried out using a micropatterned photo-mask. Measurements using atomic force microscopy showed that the swelled micropatterned polymer was five times as thick as the dried one. Protein adsorption and platelet adhesion were reduced on the polymer-immobilized regions. Mammalian cells did not adhere, and formed aggregates on the immobilized regions. In conclusion, the photo-reactive phospholipid polymer was covalently immobilized on the conventional polymer surfaces and it tended to reduce interactions with proteins and cells.
This report describes a direct approach for cell micropatterning in a closed glass microchannel. To control the cell adhesiveness inside the microchannel, the application of an external stimulus such ...as ultraviolet (UV) was indispensible. This technique focused on the use of a modified 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which is known to be a non-biofouling compound that is a photocleavable linker (PL), to localize cells
via
connection to an amino-terminated silanized surface. Using UV light illumination, the MPC polymer was selectively eliminated by photochemical reaction that controlled the cell attachment inside the microchannel. For suitable cell micropatterning in a microchannel, the optimal UV illumination time and concentration for cell suspension were investigated. After selective removal of the MPC polymer through the photomask, MC-3T3 E1 cells and vascular endothelial cells (ECs) were localized only to the UV-exposed area. In addition, the stability of patterned ECs was also confirmed by culturing for 2 weeks in a microchannel under flow conditions. Furthermore, we employed two different types of cells inside the same microchannel through multiple removal of the MPC polymer. ECs and Piccells were localized in both the upper and down streams of the microchannel, respectively. When the ECs were stimulated by adenosine triphosphate (ATP), NO was secreted from the ECs and could be detected by fluorescence resonance energy transfer (FRET) in Piccells, which is a cell-based NO indicator. This technique can be a powerful tool for analyzing cell interaction research.
A simple and direct approach for cell patterning inside the microchannel using photochemical reaction.
The phosphorylcholine group functional methacrylate monomer, 2-methacryloyloxyethyl phosphorylcholine (MPC), was graft polymerized from the polydimethylsiloxane (PDMS) substrate using ultraviolet ...irradiation and using benzophenone as a photoinitiator. The varying monomer concentrations and irradiation times were investigated in order to verify the relationships between graft density and protein resistance under specific biological conditions. The ellipsometry analysis revealed that the layer thickness of the grafted polymer depended on the monomer concentrations after the irradiation for 1
min, however, it stabilized thereafter in all the specified conditions. The curve fitting of the C1
s spectrum obtained by X-ray photoelectron spectroscopy analysis showed that the amount of grafted polymer increased with an increase in both monomer concentration and irradiation time. Atomic force microscopic images revealed that the terminations among the graft chains became dominant due to magnified chain mobility followed by growth of their length.
In vitro albumin and fibrinogen adsorption results indicated that the resistance to protein adsorption was easily tuned by the specified conditions due to the controlled graft density. Lubrication was dramatically enhanced by the grafting and it was further promoted by an increase in the graft density in good solvents, indicating that the interactions between the graft chains and the solvents resulted in the lubrication system. These basic findings regarding the grafted PDMS surface are important for versatile applications, including its use as a biomaterial and microfluidic device.
The aim of this study is to improve the biocompatibility of titanium alloy (Ti) implants by immobilization of multilayered phospholipid polymer hydrogel able to reduce protein adsorption and cell ...adhesion. We fabricated and characterized a multilayered hydrogel on Ti substrate via a layer-by-layer self-assembly deposition method using a phospholipid polymer bearing a phenylboronic acid moiety and poly(vinyl alcohol) (PVA). The water-soluble phospholipid polymer (PMBV) was synthesized from 2-methacrylocyloxyethyl phosphorylcholine,
n-butyl methacrylate, and 4-vinylphenylboronic acid (VPBA). The PMBV reacted with PVA and formed a hydrogel due to covalent linkage between the VPBA units and hydroxyl groups of PVA. The hydrogel layer growth on the Ti surface was initialized by the deposition of one layer of photoreactive PVA bonded by UV irradiation to the Ti surface, which was modified with an alkylsilane compound. The multilayered hydrogel was built up by alternating the deposition of the PMBV and PVA; this was monitored by several methods: static contact angle measurement, X-ray photoelectron spectroscopy, and attenuated Fourier-transform infrared spectroscopy. The results revealed clearly the progressive construction of the multilayered hydrogel on the Ti substrate. The PMBV/PVA multilayer prepared on the Ti substrate reduced the adhesion of L929 cells compared with that on an untreated Ti substrate. Thus, we concluded that the formation of the multilayered hydrogel is effective to improve the biocompatibility on Ti-based medical devices.
Abstract Osteolysis caused by wear particles from polyethylene in artificial hip joints is of great concern. Various bearing couple combinations, bearing material improvements, and surface ...modifications have been attempted to reduce such wear particles. With the aim of reducing the wear and developing a novel artificial hip-joint system, we created a highly lubricious metal-bearing material: A 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer was grafted onto the surface of the cobalt–chromium–molybdenum (Co–Cr–Mo) alloy. For ensuring the long-term retention of poly(MPC) on the Co–Cr–Mo alloy, we used a 4-methacryloxyethyl trimellitate anhydride (4-META) intermediate layer and photo-induced graft polymerization technique to create a strong bonding between the Co–Cr–Mo substrate and the poly(MPC) chain via the 4-META layer. The Co–Cr–Mo alloy was pretreated with nitric acid and O2 plasma to facilitate efficient interaction between the 4-META carboxyl group and the surface hydroxyl group on the Cr oxide passive layer of the Co–Cr–Mo alloy. After MPC grafting, the MPC unit peaks were clearly observed in the Fourier-transform infrared spectroscopy with attenuated total reflection (FT-IR/ATR) and X-ray photoelectron spectroscopy (XPS) spectra of the Co–Cr–Mo surface. Tribological studies with a pin-on-plate machine revealed that surface MPC grafting markedly lowered the friction coefficient. We concluded that the grafted poly(MPC) layer successfully provided high lubricity to the Co–Cr–Mo surface.
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