Effect of epigallocatechin-3-o-gallate and quercetin on the cryopreservation of cartilage tissue.

The effects of epigallocatechin-3-o-gallate (EGCG) and quercetin on the contents of extracellular matrix (ECM) in porcine cartilage at 4 °C were investigated. The addition of quercetin at 0.01 mM for the incubation of porcine cartilage disks at 4 °C for 2 week could suppress the decrease in ECM and the compliance of the disks, markedly greater than those of EGCG (1.

Early tissue formation on whole-area osteochondral defect of rabbit patella by covering with fibroin sponge.

Large osteochondral defects have been difficult to repair via tissue engineering treatments due to the lack of a sufficient number of source cells for repairing the defect and to the severe mechanical stresses affecting the replacement tissue. In the present study, whole-area osteochondral defects of rabbit patella were covered and wrapped with a fibroin sponge containing chondrocytes, with or without Green Fluorescent Protein (GFP) transgenic marking, on the surface facing the osteochondral defect. Five of eight osteochondral defects that were covered with the chondrocyte-seeded fibroin sponges showed hyaline cartilage-like repair containing no fibroin fragments at 6 weeks after surgery.

Development of a FRET-based recombinant tension sensor to visualize cell-material interactions.

A Förster/fluorescence resonance energy transfer (FRET)-based molecular tension sensor was originally reported by the fusion of intracellular molecules, which has contributed to the elucidation of cell mechanotransduction. However, it is still unclear whether recombinant tension sensors can detect forces in the extracellular environment. Here, we developed a recombinant FRET-based tension sensor (rFRET-TS) and immobilized it to a glass surface.

Silk fibroin sponges with cell growth-promoting activity induced by genetically fused basic fibroblast growth factor.

Transgenic silkworm technology has enabled the biological properties of silk fibroin protein to be altered by fusion to recombinant bioactive proteins. However, few studies have reported the fabrication of genetically modified fibroin proteins into three-dimensional spongy structures to serve as scaffolds for tissue engineering. We generated a transgenic silkworm strain that produces fibroin fused to basic fibroblast growth factor (bFGF) and processed the fibroin into a spongy structure using a simple freeze/thaw method.

Static magnetic field effects on impaired peripheral vasomotion in conscious rats.

We investigated the SMF effects on hemodynamics in the caudal artery-ligated rat as an in vivo ischemia model using noninvasive near-infrared spectroscopy (NIRS) combined with power spectral analysis by fast Fourier transform. Male Wistar rats in the growth stage (10 weeks old) were randomly assigned into four groups: (i) intact and nonoperated cage control (n = 20); (ii) ligated alone (n = 20); (iii) ligated and implanted with a nonmagnetized rod (sham magnet; n = 22); and (vi) ligated and implanted with a magnetized rod (n = 22). After caudal artery ligation, a magnetized or unmagnetized rod (maximum magnetic flux density of 160 mT) was implanted transcortically into the middle diaphysis of the fifth caudal vertebra.

Quantification of cell co-migration occurrences during cell aggregation on fibroin substrates.

A quantitative analytical method was proposed for measuring cell co-migration, which was defined as two or more cells migrating together. To accurately identify and quantify this behavior, cell migration on fibroin substrates was analyzed with respect to intercellular distance. Specifically, cell size was characterized by major diameter, and then, based on these measurements and cell center data, a specific threshold distance for defining co-migration was determined after analyzing cell motion using the Voronoi diagram method.

How do chondrocytes aggregate on fibroin substrate.

The effects of substrate material on the spatio-temporal behavior of cells is an important issue. Although cell aggregation has been observed on various fibroin substrates, the mechanisms of this aggregation have yet to be fully clarified. In this study, cell aggregation behavior on fibroin substrates were evaluated, focusing on the distance between each cell and the direction of individual cell migration.

Evaluation of the clinical performance of ultrahigh molecular weight polyethylene fiber cable using a dog osteosynthesis model.

The purpose of this study was to assess the removability and biological reactivity of an ultrahigh molecular weight polyethylene (UHMWPE) fiber cable as a new biomaterial for osteosynthesis. We used a pull-out test and an implantation test to analyze the performance of the UHMWPE fiber cable using a dog model, and compared its characteristics with those of a wire cable and a soft wire. In the pull-out test, the UHMWPE fiber cable was as easy to remove as the soft wire, and both the UHMWPE fiber cable and the soft wire were significantly easier to remove than the wire cable.

Helical conformation endows poly-l-lactic acid fibers with a piezoelectric charge under tensile stress.

Poly-l-lactic acid (PLLA) has been clinically used as a bioabsorbable material and attains a piezoelectric charge upon molecular orientation by the application of a shear force to the C-axis of the crystal line region. Previous studies showed that implanted drawn PLLA films or rods accelerate the ossification due to piezoelectric effect. In this study, we originally designed helically-twisted PLLA fiber to produce piezoelectricity in bioabsorbable suture upon tensile stress.

Quantitative evaluation of fibroblast migration on a silk fibroin surface and TGFBI gene expression.

Cell migration plays important roles in natural processes involving embryonic development, inflammation, wound healing, cancer metastasis and angiogenesis. Cell migration on various biomaterials is also believed to improve the rate of wound healing and implant therapies in the tissue-engineering field. This study measured the distance traversed, or mileage, of mouse fibroblasts on a silk fibroin surface.

The antioxidant and non-antioxidant contributions of vitamin E in vitamin E blended ultra-high molecular weight polyethylene for total knee replacement.

Vitamin E (VE) blended ultra-high molecular weight polyethylene (UHMWPE) has been developed in Japan as a material for use in total knee replacement (TKR). Various results have demonstrated that VE blended UHMWPE reduces the incidence of delamination failure and lowers the amount of wear produced during knee simulator testing. It was also found that wear particles from VE blended UHMWPE elicited a reduced biological response compared to conventional UHMWPE.

Observation and quantification of chondrocyte aggregation behavior on fibroin surfaces using Voronoi partition.

Cell migration is one of the fundamental processes in histogenesis, and it is necessary to investigate such multicellular behavior quantitatively in cell regeneration studies. In this study, Voronoi diagram analysis was first confirmed in simulation testing, and then used to evaluate the multicellular behavior of chondrocytes on three different substrates: (1) wild-type fibroin (FIB); (2) L-RGDSx2 transgenic fibroin; (3) and collagen. The indices for the round factor average, round factor homogeneity, and area disorder (AD), calculated from Voronoi diagram analysis, were used to characterize the difference in spatiotemporal changes for the different chondrocyte populations, and a regression analysis of the AD index was used to measure the speed of cell aggregation.

A feasibility study for evaluation of mechanical properties of articular cartilage with a two-electrode electrical impedance method.

Background: Since articular cartilage has important mechanical properties such as load-bearing, shock absorption and lubrication for activities in daily life, it is important to evaluate the mechanical properties of repaired cartilage in terms of whether those properties are the same as those of natural cartilage. The purpose of this study was to investigate the effectiveness of an electrical impedance method for quantitatively measuring the mechanical properties of cartilage.

Methods: Cartilage specimens were harvested from porcine knee joint, and two kinds of enzyme-treated cartilages were prepared to investigate the correlation between mechanical and electrical properties resulting from changes in the structure of the extracellular matrix.

Adhesive force behavior of single ATDC5 cells in chondrogenic culture.

Cellular mechanical properties are implicated in numerous cell behaviors, but their involvement in cell differentiation process has remained unclear. Since mechanical interactions between chondrogenic cells and their surrounding environment heavily affect the maintenance of their differentiation phenotype, here, using a chondrogenic cell strain ATDC5, we evaluated cell mechanical properties (e.g.

Chondrocyte distribution and cartilage regeneration in silk fibroin sponge.

Chondrocytes distribution and cartilage formation in three types of fibroin sponges with different average pore sizes (40-80, 80-120 and 100-140 μm) was measured. The image processing was performed combining two methods to identify cells automatically: extraction of local maximum luminance and multi-threshold analysis. The results showed that initial accumulation of chondrocytes localized at surface area at 3 h in the small and medium-pore groups, however, the difference in the cell distributions become equivalent until 24 h after seeding.

Effect of RGDS-expressing fibroin dose on initial adhesive force of a single chondrocyte.

Initial chondrocyte-material interactions are important for cell behaviors such as proliferation, phenotypic expression and matrix synthesis. Previously, we showed that chondrocytes cultured in/on silk fibroin scaffolds proliferate without dedifferentiating into fibroblast-like cells and that RGDS sequences genetically interfused in the fibroin light chain protein enhance cartilage tissue formation. In the present study, the adhesive force of chondrocytes was measured on fibroin substrates containing RGDS-expressing fibroin molecules produced by transgenic silkworms at the different densities of 0, 0.

Recovery Effects of a 180 mT Static Magnetic Field on Bone Mineral Density of Osteoporotic Lumbar Vertebrae in Ovariectomized Rats.

The effects of a moderate-intensity static magnetic field (SMF) on osteoporosis of the lumbar vertebrae were studied in ovariectomized rats. A small disc magnet (maximum magnetic flux density 180 mT) was implanted to the right side of spinous process of the third lumbar vertebra. Female rats in the growth stage (10 weeks old) were randomly divided into 4 groups: (i) ovariectomized and implanted with a disc magnet (SMF); (ii) ovariectomized and implanted with a nonmagnetized disc (sham); (iii) ovariectomized alone (OVX) and (vi) intact, nonoperated cage control (CTL).

Effects of RGDS sequence genetically interfused in the silk fibroin light chain protein on chondrocyte adhesion and cartilage synthesis.

Initial chondrocyte-silk fibroin interactions are implicated in chondrogenesis when using fibroin as a scaffold for chondrocytes. Here, we focused on integrin-mediated cell-scaffold adhesion and prepared cell adhesive fibroin in which a tandem repeat of the Arg-Gly-Asp-Ser (RGDS) sequence was genetically interfused in the fibroin light chain (L-chain) (L-RGDSx2 fibroin). We investigated the effects of the sequence on chondrocyte adhesion and cartilage synthesis, in comparison to the effects of fibronectin.

Observation of chondrocyte aggregate formation and internal structure on micropatterned fibroin-coated surface.

Condensation/aggregation process of rabbit-derived chondrocytes on a fibroin-coated patterned substrate was observed to estimate initial aggregation process in fibroin sponge. Chondrocytes were seeded on array of 160 microm diameter pits in three densities: 5 cells/pit (2.5 x 10(4) cells/cm(2), LOW), 15 cells/pit (7.

Strain-induced crystallization and orientation of vitamin E-blended ultrahigh molecular weight polyethylene.

The effects of vitamin E addition on the strain-induced crystallization and molecular chain orientation of ultrahigh molecular weight polyethylene (UHMWPE) were examined in order to clarify the wear mechanism of vitamin E-blended UHMWPE in knee prostheses. The structure changes of vitamin E-blended UHMWPE before and after tensile strain were analyzed by X-ray diffraction, Raman spectroscopy and image correlation method. The vitamin E-blended UHMWPE exhibited lower strain-induced crystallization than virgin UHMWPE but a higher Ic value in Raman spectroscopic analysis.

The Influence of a gradient static magnetic field on an unstirred Belousov-Zhabotinsky reaction.

It is believed that static magnetic fields (SMF) cannot affect the pattern formation of the Belousov-Zhabotinsky (BZ) reaction, which has been frequently studied as a simplified experimental model of a nonequilibrium open system, because SMF produces no induced current and the magnetic force of SMF far below 1 T is too low to expect the effects on electrons in the BZ reaction. In the present study, we examined whether the velocity of chemical waves in the unstirred BZ reaction can be affected by a moderate-intensity SMF exposure depending on the spatial magnetic gradient. The SMF was generated by a parallel pair of attracting rectangular NdFeB magnets positioned opposite each other.

Spatial gradient effects of 120 mT static magnetic field on endothelial tubular formation in vitro.

This study investigated the spatial magnetic gradient effects of static magnetic fields (SMF) on endothelial tubular formation by applying the maximum spatial gradient to a target site of culture wells for cell growth. The respective maximum values of magnetic flux density (B(max)), magnetic flux gradient (G(max)) and the magnetic force product of the magnetic flux density and its gradient (a parameter of magnetic force) were 120 mT, 28 mT/mm and 1428 mT(2)/mm. The effects of gradient SMF on tubular formation were compared with those of uniform SMF that has no spatial gradients on the entire bottom area of culture wells.

Reduction of wear volume from ultrahigh molecular weight polyethylene knee components by the addition of vitamin E.

Wear performance and debris-size distribution of vitamin E (DL-alpha tocopherol, VE)-added ultrahigh molecular weight polyethylene (UHMWPE) was evaluated using a knee-simulator test. VE was mixed with GUR 1050 UHMWPE powder at 0.3 wt%, and the tibial components of the knee joint were made by direct compression molding.

In vivo mechanical condition plays an important role for appearance of cartilage tissue in ES cell transplanted joint.

The objective of this study was to evaluate the effects of the mechanical environment on the formation of cartilage tissue in transplanted embryonic stem (ES) cells. Full-thickness osteochondral defects were created on the patella groove of SD rats, and ES cells (CCE ES cells obtained from 129/Sv/Ev mice and Green ES FM260 ES cells obtained from 129SV [D3] - Tg [NCAG-EGFP] CZ-001-FM260Osb mice) were transplanted into the defects embedded in collagen gel. The animals were randomly divided into either the joint-free group (JF group) or the joint-immobilized group (JI group) for 3 weeks after a week postoperatively.