Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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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)

To investigate the effects of the microstructure of collagenous carriers on the in vivo function of bone sialoprotein (BSP) in mineralization and osteoblast differentiation, we examined the ultrastructure of reconstituted type I collagen (collagen) and heat-denatured collagen (gelatin) and the in vivo responses to purified bone-derived BSP that was implanted with collagen or gelatin into surgically created 8-mm rat calvarial bone defects. Scanning and transmission electron microscopies revealed that the collagen displayed a fine fibrillar structure with interconnecting spaces between the fibrils/fibers, while the gelatin completely lost this unique three-dimensional structure after denaturation. The rates of in vivo release of BSP from the collagen scaffold were significantly lower than those from the gelatin. Collagen-BSP, but not gelatin-BSP, induced early mineral deposition in the matrix of proliferating repair cells in the calvarial defects at approximately 4-7 days after implantation. Expression levels of osteoblast markers, alkaline phosphatase activity and amounts of new bone synthesized in the collagen-BSP treated defects were significantly greater than that in the gelatin-BSP treated defects (p<0.001). The data suggest that the fibrillar microstructure of reconstituted collagen is essential for retaining BSP at a higher concentration within the defects, which enhances BSP-mediated matrix mineralization and osteoblast differentiation during the repair of rat calvarial defects.
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PMID:Role of fibrillar structure of collagenous carrier in bone sialoprotein-mediated matrix mineralization and osteoblast differentiation. 1704 34

This study investigates the cellular response of fetal osteoblasts to bioactive resorbable composite films consisting of a poly-D,L-lactide (PDLLA) matrix and bioactive glass 45S5 Bioglass (BG) particles at three different concentrations (0% (PDLLA), 5% (P/BG5), and 40% (P/BG40)). Using scanning electron microscopy (SEM) we observed that cells were less spread and elongated on PDLLA and P/BG5, whereas cells on P/BG40 were elongated but with multiple protrusions spreading over the BG particles. Vinculin immunostaining revealed similar distribution of focal adhesion contacts on all cells independent of substratum, indicating that all materials permitted cell adhesion. However, when differentiation and maturation of fetal osteoblasts was examined, incorporation of 45S5 BG within the PDLLA matrix was found to significantly (p < 0.05) enhance alkaline phosphatase enzymatic activity and osteocalcin protein synthesis compared to tissue culture polystyrene controls and PDLLA alone. Alizarin red staining indicated extracellular matrix mineralization on both P/BG5 and P/BG40, with significantly more bone nodules formed than on PDLLA. Real time RT-PCR revealed that expression of bone sialoprotein was also affected by the BG containing films compared to controls, whereas expression of Collagen Type I was not influenced. By performing these investigations in the absence of osteogenic factors it appears that the incorporation of BG stimulates osteoblast differentiation and mineralization of the extracellular matrix, demonstrating the osteoinductive capacity of the composite.
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PMID:Enhanced differentiation and mineralization of human fetal osteoblasts on PDLLA containing Bioglass composite films in the absence of osteogenic supplements. 1707 51

Adipose-derived stem cells (ASCs) are considered to be multipotent mesenchymal stem cells that are easily induced to differentiate into functional osteoblasts both in vitro and in vivo. Osterix (Osx) is a zinc finger-containing transcription factor of Sp gene family, which plays important roles in bone development and mineralization. In this study, we hypothesized that overexpression of Osx in murine ASCs would promote their osteogenic differentiation in vitro. A plasmid expressing Osx (pcDNA3.1-Osx) was constructed and applied to transfect monolayers of murine ASCs. Then expression of bone-related genes, nodule formation, proliferation rate, and alkaline phosphatase activity were examined to evaluate the osteogenic potential of ASCs with pcDNA3.1-Osx transfection. Results of RT-PCR and immunohistochemistry showed that pcDNA3.1-Osx transfection enhanced the expression of bone matrix proteins, such as bone sialoprotein, osteocalcin, osteopontin, and Collagen type I in ASCs. At the same time, overexpression of Osx in ASCs enhanced alkaline phosphatase activity and capability to form mineralized nodules, while not inhibited their proliferation rate. These results indicated that pcDNA3.1-Osx transfection promoted the osteogenic differentiation of ASCs, while not affecting their proliferative ability. Since they can be easily isolated and genetically modified, ASCs are hopeful cell sources in the further application of hard tissue engineering.
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PMID:Osteogenic differentiation of adipose derived stem cells promoted by overexpression of osterix. 1720 79

Microparticulate systems have attracted a great deal of attention over the past few years as a carrier for the delivery of cells and proteins in the treatment of defective tissues. The composition of microparticulates is regarded as being of utmost importance for the successful recruitment of the cells involved in the tissue regeneration process. Collagen-apatite nanocomposites mimicking the extracellular bone matrix are thus considered to be a potential vector for bone regeneration, either directly or through the delivery of osteogenic cells. In this study, we developed microspheres constituted of collagen and apatite for the treatment of skeletal defects. The apatite-precipitated collagen solution (30% apatite) was formed into microspheres under a water-in-oil emulsion condition. Spherical particles with diameters of tens to hundreds of micrometers (average of approximately 166 microm) were successfully produced. The internal structure of the microspheres featured a typical nanocomposite wherein apatite nanocrystalline precipitates were organized evenly within the reconstituted collagen matrix. The nanocomposite microspheres were observed to recruit favorable adhesion and growth of rat bone marrow derived stem cells. The cells supported on the nanocomposite microspheres stimulated the expression of a series of bone-associated genes. The osteogenic marker, alkaline phosphatase, was secreted to a significantly higher level on the nanocomposite microspheres than on the pure collagen counterpart. The present finding suggests that the collagen-apatite nanocomposite microspheres have high osteogenic potential and are useful for tissue-engineering applications.
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PMID:Microspheres of collagen-apatite nanocomposites with osteogenic potential for tissue engineering. 1742 98

Tendon to bone integration after rotator cuff repair is not a reproducible process. During repair, bioabsorbable and nonabsorbable suture material is universally used to facilitate the procedure. Improving the biological architecture of inert suture might aid in overall tendon to bone healing. The objective of our study is to enhance the bone to tendon union by absorbing type I collagen onto high strength nonabsorbable polyester/polyethylene suture commonly used in rotator cuff surgery. Our purpose was to evaluate the tendon and bone cellular response to this novel coated suture compared to uncoated suture. Primary human osteoblasts (HOBs) and tenocytes were plated onto polyester/polyethylene suture that was either uncoated or coated with type I bovine collagen. Cell adhesion to the sutures was assayed at 24 hours. Proliferation was determined at 48 hours by measuring [3H]- Thymidine incorporation in cells attached to the sutures. At 24 and 48 hours, respectively, cells grown on the collagen-coated suture showed a significantly greater response measured by adhesion and proliferation than cells grown on uncoated suture. At five days of culture, alkaline phosphatase activity and protein synthesis was significantly greater on the collagen-coated suture compared to uncoated. Collagen-coated polyester/polyethylene suture appears to stimulate adhesion, proliferation alkaline phosphatase, and protein synthesis more than uncoated sutures, and therefore may aid in the tendon to bone incorporation process critical to rotator cuff repair.
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PMID:Tendon and bone responses to a collagen-coated suture material. 1744 99

The objective of this study was to create a novel approach to promote bone induction through sustained release of growth factor from a 3-dimensional (3D) hybrid scaffold. Peptide-amphiphile (PA) was synthesized by standard solid-phase chemistry that ends with the alkylation of the NH2 terminus of the peptide. Collagen sponge was reinforced by incorporation of poly(glycolic acid) (PGA) fiber. A 3D network of nanofibers was formed by mixing basic fibroblast growth factor (bFGF) suspensions with dilute aqueous solutions of PA. A hybrid scaffold was fabricated by combination of self-assembled PA nanofibers and collagen sponge reinforced with incorporation of PGA fibers. The in vitro release profile of bFGF from hybrid scaffold was investigated, and ectopic bone formation induced by the released bFGF was assessed after subcutaneous implantation of hybrid scaffold into the backs of rats. Homogeneous bone formation was histologically observed throughout the hybrid scaffolds, in marked contrast to collagen sponge-incorporated bFGF. The level of alkaline phosphatase activity and osteocalcin content at the implanted sites of hybrid scaffolds were significantly high compared with collagen sponge incorporated with bFGF. The combination of bFGF incorporated in a collagen sponge self-assembled PA nanofiber hybrid scaffold is a promising procedure to improve bone regeneration.
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PMID:Bone regeneration on a collagen sponge self-assembled peptide-amphiphile nanofiber hybrid scaffold. 2655 49

The scaffold surface composition can be altered by the use of surface coatings. The use of thin coatings will give special surface properties, while the bulk properties of the scaffold are preserved. Collagen type I is known to play an important role during cell adhesion as well as osteoblast differentiation. A common way to coat surfaces is the adsorption method. An alternative way is the use of a protein immobilization method like p-nitrophenyl chloroformate. In this study, we investigated the effect of a collagen type I coating and p-nitrophenyl chloroformate as a protein immobilization method on osteoblast adhesion, proliferation, and differentiation. Titanium fiber meshes were treated with sodium hydroxide (NaOH), followed by p-nitrophenyl chloroformate, and coated with collagen type I. Osteoblast-like cells were seeded into the meshes and cultured for 24 days. The cell attachment, proliferation, and differentiation were measured by using Live and Dead assay, cell counting, DNA analysis, alkaline phosphatase activity assay, calcium content measurement, Real Time PCR (QPCR), and scanning electron microscopy (SEM). Results demonstrated that initially less cells were attached to the covalently bounded collagen meshes (NPC-Col) compared with titanium as control (Ti) and adsorbed collagen meshes (ABS-Col). Further, a decreased growth curve of cells cultured on the NPC-Col meshes was observed in comparison with Ti and ABS-Col meshes. The calcium measurements and SEM pictures revealed that all three surfaces showed differentiation of osteoblast-like cells after 8-24 days. On the basis of our results, we conclude that initially less cells were attached to the NPC-Col meshes and that they had a decreased proliferation rate. Further, we conclude that an adsorbed collagen type I coating stimulated the osteoblastic differentiation of rat bone marrow cells.
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PMID:The response of osteoblast-like cells towards collagen type I coating immobilized by p-nitrophenylchloroformate to titanium. 1755 25

In this study, we enhanced the expression of a plasmid DNA in mesenchymal stem cells (MSC) by the combination of three-dimensional (3D) tissue-engineered scaffold and nonviral gene carrier. To function as an enhanced delivery of plasmid DNA, acetic anhydride was reacted with polyethylenimine (PEI) to acetylate 80% of the primary and 20% of the secondary amines (PEI-Ac(80)). This acetylated PEI has been demonstrated to show enhanced gene-delivery efficiency over unmodified PEI. Collagen sponges reinforced by incorporating of poly(glycolic acid) (PGA) fibers were used as the scaffold material. DNA nanoparticles formed through simple mixing of plasmid DNA encoding bone morphogenetic protein-2 (BMP-2) and PEI-Ac(80) solutions were encapsulated within these scaffolds. MSC were seeded into each scaffold and cultured for several weeks. Within these scaffolds, the level of BMP-2 expression by transfected MSC was significantly enhanced compared to MSC transfected by DNA nanoparticles in solution (in 2D tissue culture plates). Homogeneous bone formation was histologically observed throughout the sponges seeded with transfected MSC by using DNA nanoparticles after subcutaneous implantation into the back of rats. The level of alkaline phosphatase activity and osteocalcin content at the implanted sites of sponges seeded with transfected MSC by using DNA nanoparticles were significantly higher when compared with those seeded with other agents.
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PMID:DNA nanoparticles encapsulated in 3D tissue-engineered scaffolds enhance osteogenic differentiation of mesenchymal stem cells. 1768 52

Collagen/hydroxyapatite (HA) nanocomposite thin films containing 10, 20, and 30 wt.% HA were prepared on commercially pure titanium substrates by the spin coating of their homogeneous sols. All of the nanocomposite coatings having a thickness of approximately 7.5 microm exhibited a uniform and dense surface, without any obvious aggregation of the HA particles. A minimum contact angle of 36.5 degrees was obtained at 20 wt.% HA, suggesting that these coatings would exhibit the best hydrophilicity. The in vitro cellular assays revealed that the coating treatment of the Ti substrates favored the adhesion of osteoblast-like cells and significantly enhanced the cell proliferation rate. The cells on the nanocomposite coatings expressed much higher alkaline phosphatase (ALP) levels than those on the uncoated Ti substrates. Increasing the amount of HA resulted in a gradual improvement in the ALP activity. The nanocomposite coatings on Ti substrates also exhibited much better cell proliferation behaviors and osteogenic potentials than the conventional composite coatings with equivalent compositions, demonstrating the greater potential of the former as implant materials for hard tissue engineering.
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PMID:Bioactive nanocomposite coatings of collagen/hydroxyapatite on titanium substrates. 1821 56

Surface properties affect the biological properties of cells modulating the expression of different factors. Osteoblasts contribute both to new bone formation and controlling haematopoiesis through cytokines and growth factors. We analyzed the effect of bone (calcium-phosphate bone slides), cartilaginous (hyaluronan-based scaffold), and plastic substrate culture on human osteoblast proliferation, bone matrix molecule, and inflammatory factor expression. Osteoblast proliferation increased to a greater extent when the cells were grown for 14 days on plastic and bone slides, whereas hyaluronan-based scaffold (HA-scaffold) induced only a minimal increase. Collagen type I, osteonectin, alkaline phosphatase and osteocalcin were expressed on osteoblasts grown on plastic and bone slides and down-modulated at mRNA and protein level by HA-scaffold. Bone slides showed the ability to increase osteopontin mRNA expression. The expression of CXCR4 and CXCL13 was upregulated by bone slides and HA-scaffold, while CXCL12 and CXCR5 expression was down-modulated. These data suggest a substrate-dependent modulation of human osteoblast proliferation, bone matrix and inflammatory factor expression, which might help to understand the active role played by osteoblasts in bone microenvironment by coupling bone extracellular matrix, chemokines and the haematopoietic system.
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PMID:Surface-dependent modulation of proliferation, bone matrix molecules, and inflammatory factors in human osteoblasts. 1844 23


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