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Query: EC:3.1.3.1 (
alkaline phosphatase
)
47,916
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
The present study investigated the ectopic osteoinduction and early degradation of recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded porous beta-tricalcium phosphate (beta-TCP) in mice. The porous beta-
TCP
with 50 microg of rhBMP-2 (n = 25) and porous beta-
TCP
(control group, n = 25) were implanted into muscle pouches in the right and left thigh of 28-day-old mice (n = 25), respectively. At every time point (3, 7, 14, 21 and 28 days after implantation), five mice were euthanized and the histological examinations of implantation sites were performed. In addition, the
alkaline phosphatase
(
ALP
) activity was also quantitatively analyzed. For the rhBMP-2-loaded group, blood vessel formation and immature cartilage was observed within the porous beta-
TCP
3 days after implantation. Mature cartilage was observed 7 days after implantation of rhBMP-2-loaded porous beta-
TCP
. Newly formed woven bone, lamellar bone as well as marrow were observed 14 and 21 days after implantation of the rhBMP-2-loaded porous beta-
TCP
. Lamellar bone and marrow were observed 28 days after implantation of the rhBMP-2-loaded porous beta-
TCP
. For the control group, no bone or cartilage was observed at all time points. However, multinucleated giant cells and fibrous tissues were observed in the control group at 7 and 28 days after implantation, respectively. At 21 and 28 days after implantation, porous beta-
TCP
was observed to fragment indicating early degradation of the porous beta-
TCP
in both groups. In addition,
ALP
was observed to be significantly higher in the rhBMP-2-loaded beta-
TCP
as compared to the control beta-
TCP
. It was concluded from this study that the rhBMP-2-loaded porous beta-
TCP
induced blood vessel and ectopic bone formation.
...
PMID:Ectopic osteoinduction and early degradation of recombinant human bone morphogenetic protein-2-loaded porous beta-tricalcium phosphate in mice. 1568 50
Biodegradable gelatin sponges at different contents of beta-tricalcium phosphate (beta-TCP) were fabricated to allow bone morphogenetic protein (BMP)-2 to incorporate into them. The in vivo osteoinduction activity of the sponges incorporating BMP-2 was investigated, while their in vivo profile of BMP-2 release was evaluated. The sponges prepared had an interconnected pore structure with an average pore size of 200 microm, irrespective of the beta-
TCP
content. The in vivo release test revealed that BMP-2 was released in vivo at a similar time profile, irrespective of the beta-
TCP
content. The in vivo time period of BMP-2 retention was longer than 28 days. When the osteoinduction activity of gelatin or gelatin-beta-
TCP
sponges incorporating BMP-2 was studied following the implantation into the back subcutis of rats in terms of histological and biochemical examinations, homogeneous bone formation was histologically observed throughout the sponges, although the extent of bone formation was higher in the sponges with the lower contents of beta-
TCP
. On the other hand, the level of
alkaline phosphatase
activity and osteocalcin content at the implanted sites of sponges decreased with an increase in the content of beta-
TCP
. The gelatin sponge exhibited significantly higher osteoinduction activity than that of any gelatin-beta-
TCP
sponge, although every sponge with or without beta-
TCP
showed a similar in vivo profile of BMP-2 release. In addition, the in vitro collagenase digestion experiments revealed that the gelatin-beta-
TCP
sponge collapsed easier than the gelatin sponge without beta-
TCP
incorporation. These results suggest that the maintenance of the intrasponge space necessary for the osteoinduction is one factor contributing to the osteoinduction extent of BMP-2-incorporating sponges.
...
PMID:Enhanced osteoinduction by controlled release of bone morphogenetic protein-2 from biodegradable sponge composed of gelatin and beta-tricalcium phosphate. 1576 65
There are three key factors in tissue engineering: seeding cells, scaffold and their interaction. Although mesenchymal stem cells (MSCs) are potential seeding cells, the problem of what phase MSCs should be used is not yet solved. On the other hand, degradable porous scaffolds which have good mechanics and good biocompatibility are preferred. To choose the optimum seeding cells and the suitable ratio of beta-
TCP
/PLLA porous scaffold, we observed the phenotype of the male SD rat's osteoblastic MSCs and detected the amount of
alkaline phosphatase
, osteocalcin and type I collagen secreted by the osteoblastic rMSCs in different phase. About 10, 14 and 20 days after induction, the induced cells came into proliferative phase, matrix synthesis phase and mineralization phase, respectively. Then we chose the suitable cells and seeded them on beta-
TCP
/PLLA composite scaffolds with different ratios (beta-
TCP
/PLLA = 1:1; beta-
TCP
/PLLA = 1:2; and beta-
TCP
/PLLA = 2:1). Fluorescence microscope, scanning electron microscope and MTT assay were used to observe and to detect the biocompatibility of the scaffolds. The results indicated that all of these materials have biocompatibility to some extent. Cells can grow well on all of the scaffolds. However, scaffold beta-
TCP
/PLLA = 2:1 seems to be a more suitable tissue engineering scaffold on account of its minimal influence on cell growth and differentiation.
...
PMID:[Research of osteoblast-induced rat mesenchymal stem cells cocultured with beta-TCP/PLLA composite of different ratio]. 1588 27
Beta-tricalcium phosphate-reinforced high-density polyethylene (beta-
TCP
/HDPE) is a new biomaterial which was made as a copy of bone composition with the aim of replacement of bony tissues. The composite samples were prepared using medical grade
TCP
powder and granular polyethylene. The raw materials were first compounded and the resulting composite preforms were compression molded into desired shape. The biocompatibility of composite samples with different volume fractions of
TCP
(20, 30, and 40 vol %) was assessed by proliferation,
alkaline phosphatase
(
ALP
), and cell adhesion assays using G-292 osteoblast cells. Cell-material interaction on the surface of the composites was observed by scanning electron microscopy (SEM). The effect of beta-
TCP
/HDPE on the behavior of G-292 cells was compared with those of a composite and a negative control samples. Results showed the composite samples had a higher proliferation rate of G-292 cells in the presence of composite samples as compared to the composite control sample after 3, 7, and 14 days of incubation period.
ALP
production after incubation in the presence of composite samples was seen to peak on the day 7. The number of adhered cells on the composite samples was higher than the numbers adhered on composite and negative control samples after the above incubation periods. Morphology investigation of adhered cells by SEM indicated a normal morphology and also many of the cells were in the process of cell division. The above results indicate that beta-
TCP
/HDPE samples are biocompatible, nontoxic, and in some cases show an increase in the proliferation rate of the cells,
ALP
production, and cell adhesion as compared to the control counterparts.
...
PMID:In vitro evaluation of biocompatibility of beta-tricalcium phosphate-reinforced high-density polyethylene; an orthopedic composite. 1609 12
Current research in the field of tissue engineering utilizes biomaterial scaffolds, cells, and growth factors for the creation of a functional, biologically active tissue. This study examined the effect of two commercially available, three-dimensional scaffolds, ultraporous beta-tricalcium phosphate ceramics (beta-
TCP
, Vitoss) and open-celled poly(lactic acid) foams (OPLA, Drilac), on the osteogenic differentiation potential of human dermal fibroblasts. Serum-free, chemically-defined medium containing the metabolic factor 1alpha,25-dihydroxyvitamin D3 was used to promote an osteogenic phenotype in these cells. Osteoblast differentiation was assessed using PCR and immunohistochemical methods to detect gene and protein expression for the osteoblast markers
alkaline phosphatase
, osteopontin, and osteocalcin. Dermal fibroblasts cultured on beta-
TCP
scaffolds in chemically-defined medium with vitamin D3 exhibited up-regulated gene and protein expression compared to cells cultured on OPLA scaffolds. These results suggest that Vitoss (beta-
TCP
) scaffolds seeded with dermal fibroblasts and maintained in chemically-defined medium with vitamin D3 are better suited for bone tissue engineering applications than Drilac (OPLA) foams.
...
PMID:Influence of three-dimensional scaffold on the expression of osteogenic differentiation markers by human dermal fibroblasts. 1610 17
Trabecular bone is a material of choice for reconstruction after trauma and tumor resection and for correction of congenital defects. Autologous bone grafts are available in limited shapes and sizes; significant donor site morbidity is another major disadvantage to this approach. To overcome these limitations, we used a tissue engineering approach to create bone replacements in vitro, combining bone-marrow-derived differentiated mesenchymal stem cells (MSCs) suspended in hydrogels and 3-dimensionally printed (3DP) porous scaffolds made of beta-tricalcium-phosphate (beta-TCP). The scaffolds provided support for the formation of bone tissue in collagen I, fibrin, alginate, and pluronic F127 hydrogels during culturing in oscillating and rotating dynamic conditions. Histological evaluation including toluidine blue,
alkaline phosphatase
, and von Kossa staining was done at 1, 2, 4, and 6 weeks. Radiographic evaluation and high-resolution volumetric CT (VCT) scanning, expression of bone-specific genes and biomechanical compression testing were performed at 6 weeks. Both culture conditions resulted in similar bone tissue formation. Histologically collagen I and fibrin hydrogels specimens had superior bone tissue, although radiopacities were detected only in collagen I samples. VCT scan revealed density values in all but the Pluronic F127 samples, with Houndsfield unit values comparable to native bone in collagen I and fibrin glue samples. Expression of bone-specific genes was significantly higher in the collagen I samples. Pluronic F127 hydrogel did not support formation of bone tissue. All samples cultured in dynamic oscillating conditions had slightly higher mechanical strength than under rotating conditions. Bone tissue can be successfully formed in vitro using constructs comprised of collagen I hydrogel, MSCs, and porous beta-
TCP
scaffolds.
...
PMID:Hydrogel-beta-TCP scaffolds and stem cells for tissue engineering bone. 1637 62
To observe living cell morphology on ceramics by light microscopy, we fabricated a new material-transparent beta - tricalcium phosphate (t-beta
TCP
) ceramic-for the purpose of serving as a tissue culture substrate. Bone marrow stromal cells (BMSCs) were obtained from rat femora and cultured on both t-beta
TCP
ceramic disks and culture grade polystyrene (PS) dishes in an osteogenic medium. After 1 day of culture, cell attachment and spreading on both the t-beta
TCP
and PS substrata were equally and clearly detected by ordinary light microscopy. After 14 days of culture, extensive cell growth,
alkaline phosphatase
(
ALP
) staining, and bone mineral deposition could be detected on both substrata. In addition, quantitative biochemical analyses revealed high DNA content,
ALP
activity, and osteocalcin content of these cultures. This experiment is significant in that all of the results were similarly observed on both the t-beta
TCP
and PS substrata, indicating the excellent properties of beta
TCP
ceramics for BMSCs culture towards osteogenic differentiation.
...
PMID:Observation and quantitative analysis of rat bone marrow stromal cells cultured in vitro on newly formed transparent beta-tricalcium phosphate. 1638 70
The aim of this investigation was to test the biocompatibility of three-dimensional bioresorbable foams made of poly(L-lactic acid) (PLA), alone or filled with hydroxyapatite (HA) or beta-tricalcium phosphate (beta-TCP), with human primary osteoblasts, using a direct contact method. Porous constructs were processed by supercritical gas foaming, after a melt-extrusion of ceramic/polymer mixture. Three neat polymer foams, with pore sizes of 170, 310, and 600 microm, and two composite foams, PLA/5 wt% HA and PLA/5 wt% beta-
TCP
, were examined over a 4-week culture period. The targeted application is the bone tissue-engineering field. For this purpose, human fetal and adult bone cells were chosen because of their highly osteogenic potential. The association of fetal bone cells and composite scaffold should lead to in vitro bone formation. The polymer and composite foams supported adhesion and intense proliferation of seeded cells, as revealed by scanning electron microscopy. Cell differentiation toward osteoblasts was demonstrated by
alkaline phosphatase
(
ALP
) enzymatic activity, gamma-carboxylated Gla-osteocalcin production, and the onset of mineralization. The addition of HA or beta-
TCP
resulted in higher
ALP
enzymatic activity for fetal bone cells and a stronger production of Gla-osteocalcin for adult bone cells.
...
PMID:Biocompatibility of bioresorbable poly(L-lactic acid) composite scaffolds obtained by supercritical gas foaming with human fetal bone cells. 1641 9
To obtain more extensive bone formation in composites of porous ceramics and bone marrow stromal cells (BMSCs), we hypothesized that a low-pressure system would serve to facilitate the perfusion of larger number of BMSCs into the porous scaffold, enhancing bone formation within the composites. After culturing BMSCs in osteogenic medium, porous blocks of beta-tricalcium phosphate (beta-TCP) were soaked in the cell suspension. Composites of the block and BMSCs were put immediately into a vacuum desiccator. Low pressure was applied to the low pressure group, while controls were left at atmospheric pressure. Composites were incubated in vitro or subcutaneously implanted into syngeneic rats, then analyzed biologically and histologically. In the in vitro group, cell suspension volume, cell seeding efficiency,
alkaline phosphatase
(
ALP
) activity, and DNA content in the beta-
TCP
blocks were significantly higher in low pressure group than in the controls. Scanning electron microscopy (SEM) demonstrated that a greater number of cells covered the central parts of the composites in the low pressure group.
ALP
activity in the composites was increased at 3 and 6 weeks after implantation into rats. Histomorphometric analysis revealed more uniform and extensive bone formation in the low pressure group than in the controls. The application of low pressure during the seeding of BMSCs in perfusing medium into a porous scaffold is useful for tissue-engineered bone formation.
...
PMID:Enhancement of tissue engineered bone formation by a low pressure system improving cell seeding and medium perfusion into a porous scaffold. 1655 48
Beta-tricalcium phosphate-reinforced high-density polyethylene (beta-
TCP
/HDPE) is a new biomaterial, which was made to simulate bone composition and study its capacity to act like bony tissues. This material was produced by replacing mineral component and collagen soft tissue of bone with beta-
TCP
and HDPE, respectively. The biocompatibility of composite samples with different volume fractions of
TCP
(20, 30, and 40 vol %) and two different particle sizes (80-100 and 120-140 mesh size) was examined in vitro using the osteoblast cell line G-292 by proliferation,
alkaline phosphatase
(
ALP
) production, and cell adhesion assays. Cell-material interaction on the surface of the composites was observed by scanning electron microscopy (SEM). The effect of beta-
TCP
particle size on behavior of the osteoblast cell line was compared between two groups of the composite samples containing smaller and larger reinforcement particle sizes as well as with those of a negative control. In general, results showed that the composite samples containing larger particles supported a higher rate of proliferation and
ALP
production by osteoblast cells after 3, 7, and 14 days of incubation compared to the composite samples with smaller particle size and control. Furthermore, more cells were attached to the surface of composite samples containing larger particle size when compared to the smaller particle size composites (p<0.05). This number was nearly equal with numbers adhered on negative control [tissue culture polystyrene (TPS)] and significantly higher in comparison with composite control [polyethylene (PE)] (p<0.05). Adhered cells presented a normal morphology by SEM and many of the cells were seen to be undergoing cell division. These findings indicate that beta-
TCP
/HDPE composites are biocompatible, nontoxic, and in some cases, act to stimulate proliferation of the cells,
ALP
production, and cell adhesion when compared to the control counterparts. Furthermore, beta-
TCP
/HDPE samples with larger reinforcement particle size were shown to possess better biological properties.
...
PMID:Effect of reinforcement particle size on in vitro behavior of beta-tricalcium phosphate-reinforced high-density polyethylene: a novel orthopedic composite. 1661 17
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