Biomedical subjects

Tetsuya Tateishi

Publications and source records attributed to Tetsuya Tateishi.

26 records · Page 2Linked to original sources

PubMed2002

Hydrostatic fluid pressure enhances matrix synthesis and accumulation by bovine chondrocytes in three-dimensional culture.

Monolayer cell cultures and cartilage tissue fragments have been used to examine the effects of hydrostatic fluid pressure (HFP) on the anabolic and catabolic functions of chondrocytes. In this study, bovine articular chondrocytes (bACs) were grown in porous three-dimensional (3-D) collagen sponges, to which constant or cyclic (0.015 Hz) HFP was applied at 2.8 MPa for up to 15 days. The effects of HFP were evaluated histologically, immunohistochemically, and by quantitative biochemical measures. Metachromatic matrix accumulated around the cells within the collagen sponges during the culture period. There was intense intracellular, pericellular, and extracellular immunoreactivity for collagen type II throughout the sponges in all groups. The incorporation of [(35)S]-sulfate into glycosaminoglycans (GAGs) was 1.3-fold greater with constant HFP and 1.4-fold greater with cyclic HFP than in the control at day 5 (P < 0.05). At day 15, the accumulation of sulfated-GAG was 3.1-fold greater with constant HFP and 2.7-fold with cyclic HFP than the control (0.01). Quantitative immunochemical analysis of the matrix showed significantly greater accumulation of chondroitin 4-sulfate proteoglycan (C 4-S PG), keratan sulfate proteoglycan (KS PG), and chondroitin proteoglycan (chondroitin PG) than the control (P < 0.01). With this novel HFP culture system, 2.8 MPa HFP stimulated synthesis of cartilage-specific matrix components in chondrocytes cultured in porous 3-D collagen sponges.

Animals↗

PubMed2002

Promotion of bone formation using highly pure porous beta-TCP combined with bone marrow-derived osteoprogenitor cells.

Beta-tricalcium phosphate (TCP) exhibits rapid degradation and weak mechanical properties, which has limited its application as bone graft substitutes, though it has good biocompatibility and osteoconductivity. We hypothesized that a composite of highly pure porous beta-TCP and bone marrow-derived osteoprogenitor cells (BMO) could improve bone formation, and slow down the degradation of beta-TCP. A highly pure porous beta-TCP with 75% porosity was fabricated. The pores averaged 200-400 microm in diameter, with interconnecting paths 100-200 microm. Blocks of beta-TCP 5 mm3 were combined with BMO, and incubated 2 weeks with (+) or without (-) osteogenic medium. They were then implanted into subcutaneous sites of syngeneic rats for 24 weeks. These composites were harvested at different time points. The alkaline phosphatase activity and bone osteocalcin content of the composites (+) were much higher than corresponding values in the composites (-) of the control group (p<0.01). Light microscopy revealed mature bone and lots of blood vessels only in the TCP/BMO composite (+). The amount of newly formed bone increased until week 24. Slow resorptive activity could be found. The mechanical parameters of the composites were much improved over those of dry beta-TCP blocks. These results showed that tissue engineering treatment on incubating the composites of beta-TCP and BMO cells in osteogenic medium results in a good osteogenic activity.

Alkaline Phosphatase↗

PubMed2002

Effects of different exposures of hyperbaric oxygen on ligament healing in rats.

Hyperbaric oxygen (HBO) is a method of augmenting, intermittently, oxygen availability to tissues. We examined the effect of three different HBO exposures on the healing of experimentally induced ligament lacerations in the right hind limb of 44 male Wistar rats. Animals were divided into four groups after ligament injury: (a) control group, animals breathed room air at 1 ATA (atmosphere absolute) in a hyperbaric chamber for 60 min; (b) HBO treatment at 1.5 ATA for 30 min once a day, (c) HBO treatment at 2 ATA for 30 min once a day, (d) 2 ATA for 60 min once a day. At 14 days post-ligament injury, we compared the ligaments of the four treatment groups for gross appearance, histology and expression of pro-alpha(I) mRNA by northern hybridization. Our results indicate that HBO was effective in promoting ligament healing compared to control (p < 0.01). Of these three exposures, HBO at 2 ATA for 60 min was the most effective, resulting in enhanced extra-cellular matrix deposition as measured by collagen synthesis.

Animals↗

PubMed2002

Long-term durability of porous hydroxyapatite with low-pressure system to support osteogenesis of mesenchymal stem cells.

In this study, we hypothesize that loading more marrow-derived mesenchymal stem cells (MSCs) into porous material by using a low-pressure system during subculture, creating a composite which combines MSCs and a novel mechanical reinforced porous hydroxyapatite, can result in more bone tissue formation in vivo. Within 26 weeks postimplantation, we examined in vivo bone formation of the experimental group with 100 mmHg pressure applied to porous HA blocks loaded with MSCs. For in vivo testing, the 2-week subcultured HA/MSC composites were implanted into subcutaneous sites of syngeneic rats. These implants were harvested at 13 and 26 weeks after implantation. SEM showed that the pore surface is covered by osteoblasts as well as collagenous extracellular matrix at 13 weeks. Light microscopy revealed the quantity of bone at 26 weeks was greater than at 13 weeks. These results showed that the novel mechanical reinforced porous HA combined with MSC has more potential for bone formation at 100 mmHg, making this method very efficient for bone reconstruction.

Animals↗

PubMed2002

Hybrid of gel-cultured smooth muscle cells with PLLA sponge as a scaffold towards blood vessel regeneration.

Although rapid formation of a smooth inner surface is important in constructing an artificial vascular graft, a conventional model that uses a biodegradable polymer such as poly-glycolic acid needs long-term culture to form it. In another model, which uses collagen gel, it is reported that prompt formation of the smooth inner surface was achieved. But the mechanical properties were not suitable, resulting in rupture under high pressure at the arterial level. Therefore, we propose a new artificial vascular graft model made of biodegradable polymer, gel, and cells. At first we manufactured an artificial vascular graft (i.d. 5 mm, o.d.7 mm) consisting of poly-L-lactic acid (PLLA) with open pore structures by using gas-forming methods. After mixing human normal aortic smooth muscle cells (SMCs) with type I collagen solution, pores of the PLLA scaffold were filled with the mixture. The collagen mixture was made into gel in the pores of the PLLA scaffold, incubating at 37 degrees C. WET-SEM analysis showed that the prompt formation of a smooth inner surface was achieved in the new model. The ratio of incorporation of SMCs into the artificial vascular graft became approximately 100% by using the cell-collagen mixture, whereas only 40% of SMCs were trapped in the conventional model where SMCs were inoculated as a cell-medium suspension. Therefore, it was suggested that the new artificial vascular graft model was superior in smooth inner surface formation and cell inoculation, compared with conventional models using either biodegradable polymer or gel.

Aorta↗

PubMed2002

Three-dimensional seeding of chondrocytes encapsulated in collagen gel into PLLA scaffolds.

Tissue engineering approaches have been clinically tried to repair damaged articular cartilages. It is an essential step to uniformly seed chondrocytes into 3D scaffolds in order to reconstruct tissue-engineered cartilages in vitro, but the tissue engineering could not have been provided with efficient cell seeding methods. Type I collagen is clinically used and known as a cytocompatible material, having recognition sites for integrins. Collagen gel encapsulating chondrocytes has been tried for making regenerated cartilages, but it is found difficult to have the gel keep its original shape after long-term culture, because of shrinking. On the other hand, 3D scaffolds, either of a nonwoven structure or a sponge-like structure, involve difficulty in that chondrocytes could not be uniformly seeded, although they have adequate initial mechanical properties. In this study, by combining collagen gelation with a nonwoven PLLA scaffold, we achieved uniform cell seeding into the 3D scaffold. Bovine articular chondrocytes were mixed with type I collagen solution, and the solution was poured into the nonwoven PLLA scaffold (1.5 mm thick, diameter 15 mm). The collagen-chondrocyte mixture was made into gel at 37 degrees C for 1 h. The 0.39% collagen mixture was viscous enough to prevent cells from precipitating during gelation. Almost all chondrocytes were able to be incorporated into the PLLA scaffolds by mixing with collagen solution and subsequently making into gel, while 30-40% of the chondrocytes seeded as a cell suspension were not trapped into the PLLA scaffolds. The method presented, where chondrocytes were mixed with collagen solution, and the mixture was incorporated into a 3D scaffold, then made into gel in the scaffold, could serve as an alternative for in vitro cartilage regeneration, also simultaneously having the advantages of both materials.

Animals↗

PubMedDate not supplied

L-Phe end-capped poly(L-lactide) as macroinitiator for the synthesis of poly(L-lactide)-B-poly(L-lysine) block copolymer.

A poly(L-lactide)-b-poly(Nepsilon-(Z)-L-lysine) (PLLA-b-PZLys) block copolymer was synthesized through the ring-opening polymerization of Nepsilon-(Z)-lysine-N-carboxyanhydride using L-Phe-terminated PLLA as a macroinitiator. The L-Phe-terminated PLLA was prepared through a novel three-step process. First, the hydroxyl-terminated PLLA was synthesized through the ring-opening polymerization of L-lactide initiated by n-butanol under the existence of tin(II) ethylhexanoate. Subsequently, the complete capping of the hydroxyl end group of PLLA with BOC-L-Phe was achieved by using a mixed anhydride of BOC-L-Phe under the catalysis of 4-(1-pyrrolidinyl) pyridine. Finally, the free amino end group was obtained by removal of the t-butoxycarbonyl group through trifluoroacetic acid treatment under anhydrous condition. All these treatments were conducted under mild conditions, thus avoiding the breakdown of the PLLA backbone. Poly(L-lactide)-b-poly(L-lysine) block copolymer was produced after deprotection treatment of PLLA-b-PZLys. The structure of the block copolymer was confirmed by 1H NMR, IR, and GPC. Adjustment of the ratio of the NCA monomer to the macroinitiator could control the chain length of the PLys block.

Biocompatible Materials↗

PubMedDate not supplied

Tissue engineering of cartilage using a hybrid scaffold of synthetic polymer and collagen.

A biodegradable hybrid scaffold of synthetic polymer, poly (DL-lactic-co-glycolic acid) (PLGA), and naturally derived polymer, collagen, was prepared by forming collagen microsponges in the pores of PLGA sponge. This was then used as the three-dimensional scaffold for tissue engineering of bovine articular cartilage, both in vitro and in vivo. In vitro studies show that hybridization with collagen facilitated cell seeding in the sponge and raised seeding efficiency. Chondrocytes adhered to the collagen microsponges, where they proliferated and secreted extracellular matrices with time, filling the space within the sponge. Hematoxylin and eosin staining revealed that most of the chondrocytes after 4 weeks of culture, and almost all cell types after 6 weeks of culture, maintained their phenotypically rounded morphology. While new tissue formed, the scaffold degraded and lost almost 36.9% of its original weight after 10 weeks. Subcutaneous implantation studies in nude mice demonstrated more homogeneous tissue formation in hybrid sponge than in PLGA sponge. The new tissue formed maintained the original shape of the hybrid sponge. The synthetic PLGA sponge, serving as a skeleton, facilitated easy formation into desired shapes and provided appropriate mechanical strength to define the ultimate shape of engineered tissue. Incorporation of collagen microsponges facilitated cell seeding and homogeneous cell distribution and created a favorable environment for cellular differentiation. The hybrid sponge could therefore represent a promising candidate as a three-dimensional scaffold for articular cartilage tissue engineering.

Animals↗