Biological Activities Of Osteoblasts Research Articles

The Effect of Nano-Silica Gel on Biological Activity of Osteoblasts and Expression of Insulin-Like and Growth Factor-2.

This study assessed the effect nano-silica gel material on bioactivity of osteoblasts and expression of IGF-2. Methods: Silica gel nanoparticles (Nanjing Kike Company) were divided according to their concentrations as follows; 0 μg/mL a control group with cells without nanoparticle treatment, 25 μg/mL as group 1, 50 μg/mL as group 2, and 100 μg/mL as group 3. The transmission electron microscope was used to measure morphology, while particle size analyzer was used to measure particle size, and potential analyzer measured Zeta potential, and MTT measured proliferation.Moreover, ALP kit was used to measure ALP activity, and Alizarin red staining measured formation of wild flower nodules, while RT-PCR was used to measure expression of IGF-2. Results: The shape of silica gel nanoparticles was spherical, with uniform particle size distribution, and particle size was between 50-800 nm. The average particle size was 383 nm, and Zeta potential was -12.3. The growth rate of control group and group 1 was relatively close (t = 0.95, P = 0.37), and growth rate of groups 2 and 3 was higher than control (group t2 = 5.63, P < 0.05, group t3 = 10.38, P < 0.05). The value-added rate for group 3 was higher than group 2 (t = 4.41, P < 0.05). Group 1 had higher activity than control group (t = 10.29, P < 0.05) and lower activity than group 3 (t = 9.85, P < 0.05) which had higher activity than group 2 (t = 4.16, P < 0.05). Groups 1, 2, and 3 induced the growth of osteoblasts, promoted calcium salt deposition, and produced red mineralized nodules where the cells converged. The formation of mineralized nodules obviously depended on concentration of silica nanoparticles. Group 1 had higher IGF-2 expression than control (t = 19.99, P < 0.05) and lower level than group 2 (t = 16.69, P < 0.05). Silica gel nanoparticles promoted MC3T3-E1 cell proliferation and differentiation. The mechanism of action may be that, silica gel nanoparticles accelerate the growth of ALP activity and osteoblast extracellular matrix mineralization by promoting the level of IGF-2. The production of chemical nodules accelerates the proliferation and differentiation of osteoblasts.

A bone substitute with high affinity for vitamin D‐binding protein―relationship with niche of osteoclasts

The biological activity of osteoblasts and osteoclasts is regulated not only by hormones but also by local growth factors, which are expressed in neighbouring cells or included in bone matrix. Previously, we developed hydroxyapatite (HA) composed of rod-shaped particles using applied hydrothermal methods (HHA), and it revealed mild biodegradability and potent osteoclast homing activity. Here, we compared serum proteins adsorbed to HHA with those adsorbed to conventional HA composed of globular-shaped particles (CHA). The two ceramics adsorbed serum albumin and γ-globulin to similar extents, but affinity for γ-globulin was much greater than that to serum albumin. The chemotactic activity for macrophages of serum proteins adsorbed to HHA was significantly higher than that of serum proteins adsorbed to CHA. Quantitative proteomic analysis of adsorbed serum proteins revealed preferential binding of vitamin D-binding protein (DBP) and complements C3 and C4B with HHA. When implanted with the femur of 8-week-old rats, HHA contained significantly larger amount of DBP than CHA. The biological activity of DBP was analysed and it was found that the chemotactic activity for macrophages was weak. However, DBP-macrophage activating factor, which is generated by the digestion of sugar chains of DBP, stimulated osteoclastogenesis. These results confirm that the microstructure of hydroxyapatite largely affects the affinity for serum proteins, and suggest that DBP preferentially adsorbed to HA composed of rod-shaped particles influences its potent osteoclast homing activity and local bone metabolism.

Effects of extracts of oxyntic mucosa in rat on the biological activity of osteoblasts

We explored the relationship between extracts of oxyntic mucosa (EOM) and the biological activity of osteoblasts in rats. We found that EOM could enhance the activity of bone formation in osteoblast. Our results suggest that EOM likely play a role in the cases of osteopenia induced by gastrectomy. Surgical removal of the stomach (gastrectomy) leads to osteopenia in animals and in humans. It was demonstrated that EOM could induce transient hypocalcaemia and stimulate an uptake of Ca(2+) into bone in rats. The main aim of this study has been to clarify whether this procedure was performed through osteoblast, which is responsible for bone formation. Osteoblasts were isolated, cultured, and identified in vitro. Preparing the rats' EOM and diluting into low, middle, and high concentrations, respectively. After osteoblasts were treated by different concentration EOMs or saline (for control), the intracytoplasm [Ca(2+)]i was measured by laser scanning confocal microscopy; the proliferation of osteoblast cells were detected with cell counting kit 8 (CCK-8); and the expressions of collagen type I and osteocalcin were assayed by reverse transcriptase polymerase chain reaction and Western blot. EOMs were found to induce a dose-related rapid increase of intracytoplasm [Ca(2+)]i in osteoblasts and could stimulate osteoblasts to enhance proliferation and upregulate the expressions of collagen type I and osteocalcin significantly (p < 0.05) compared with the control group. It was confirmed that EOM could stimulate osteoblasts to elevate the cytoplasm [Ca(2+)]i and promote the multiplication and the activity of bone formation in osteoblasts.

Biological activities of osteoblasts on poly(methyl methacrylate)/silica hybrid containing calcium salt

The biological activity of osteoblasts on the newly developed bioactive poly(methyl methacrylate) (PMMA)/silica hybrid containing calcium salt was investigated. The attachment, proliferation, and differentiation of primary cultured mouse calvarial osteoblasts were evaluated by hexosaminase, MTT, and alkaline phosphatase activity assays, respectively. The PMMA/silica hybrid showed higher biological activities than those of pure PMMA with regard to all three parameters. Besides, the calcium phosphate layer, determined by scanning electron microscopy with energy dispersive spectroscopy, occurred only on the PMMA/silica hybrid. Better biological activities on the PMMA/silica hybrid than those on the PMMA were explained by the role of calcium phosphate layer formed on the PMMA/silica hybrid and the released calcium and silicon ions from it during the cell culture. These results suggest that the PMMA/silica hybrid might be useful as a bone substitute or filler.

Changes in biological activity of bone cells in ovariectomized rats revealed by in situ hybridization.

Twelve-week-old female rats were ovariectomized (OVX) and compared with sham-operated control rats at 3, 5, 7, 10, 14, 30, and 60 days postoperation with respect to the expression of type I collagen and osteopontin mRNAs, as well as bone structure and the number of osteoclasts. The trabecular number and separation were significantly decreased and increased, respectively, in the metaphyseal trabecular bones of OVX rat femurs. The number of osteoclasts was significantly increased in the same region of OVX rats at 3 and 5 days postoperation. Type I collagen mRNA was expressed in osteoblasts, and osteopontin mRNA was expressed in some osteoclasts, in mononuclear cells on the bone resorption surface, and in osteocytes near the resorption surface. In the metaphyseal trabecular bone, type I collagen and osteopontin mRNA expression levels in individual cells was initially increased in OVX rats from 7 to 10 days postoperation, and this was sustained for 60 days. The number of osteopontin mRNA-expressing osteocytes was also significantly increased at 10 days postoperation, which lasted until 60 days. In the epiphysis, an increase in type I collagen mRNA expression was initially observed in OVX rats at 14 days postoperation, which lasted until 60 days. The number of osteopontin mRNA-expressing osteocytes was virtually identical until 30 days postoperation in the epiphysis. These findings indicated that the biological activities of osteoblasts and osteocytes are stimulated in bones of the OVX rat and that the response for OVX differs between the metaphysis and epiphysis. Furthermore, the number of osteopontin mRNA-expressing osteocytes was increased only in bones that tended to be resorbed after OVX. This indicates that some osteocytes were stimulated to express osteopontin mRNA by estrogen deficiency and suggests that these osteopontin mRNA-expressing osteocytes may be involved in regulation of bone metabolism.

The effect of glucocorticoid on the synthesis of biglycan and decorin in human osteoblasts and bone marrow stromal cells.

We have previously demonstrated that glucocorticoids induce differentiation of human bone marrow stromal cells (BMSC) into cells expressing mature osteoblast phenotype. As glucocorticoids have marked effects on extracellular matrix protein synthesis in bone, and because proteoglycans are important components of bone matrix and may condition the differentiation and biological activities of osteoblasts, we studied the effects of dexamethasone (Dex) on the synthesis of small proteoglycans [decorin (DCN) and biglycan (BGN)] in adult human BMSC and human osteoblasts (HOB). First passaged HOB and BMSC were treated with either ethanol or 10(-7) M Dex for 7 days. After treatment, the cells were metabolically labeled with either [35S]SO4 alone or [35S]SO4 and [3H]leucine together for 24 h. Conditioned media were collected, and cell layers were extracted with 4 M guanidine HCl. The extracts and conditioned media were subjected to gel filtration and ion exchange chromatography. Fractions containing radiolabeled proteoglycans were analyzed either directly or after immunoprecipitation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Dex treatment resulted in a dramatic increase in DCN and an associated decrease in BGN in both the conditioned medium and the cell layer of HOB cultures. In Dex-treated BMSC cultures, BGN was decreased in both the conditioned medium and cell layer, whereas DCN was stimulated in the majority of cultures. Northern blot analysis indicated that steady state messenger RNA (mRNA) concentration of DCN was increased by Dex in all of the HOB cultures and in seven of eight BMSC cultures analyzed. The steady state mRNA level of BGN was decreased by Dex in both HOB and BMSC cultures. The regulation of DCN and BGN mRNA by Dex in both HOB and BMSC (when responsive) was dose dependent. Time-course analysis indicated that as little as 1 day of treatment with Dex was sufficient to decrease BGN mRNA and increase DCN mRNA (when observed) levels in BMSC; the regulation spanned a 4-week interval, during which the extracellular matrix of BMSC was mineralized. The effect of Dex on the steady state mRNA levels of DCN and BGN in HOB was also apparent after 1 day of treatment. These accumulated results suggest that Dex modulates the synthesis of small proteoglycans in both human bone marrow stromal osteoprogenitor cells and mature osteoblasts.

A review of bone cell and substratum interactions: An illustration of the role of scanning electron microscopy

AbstractThe first experimental studies concerning observations of changes in bone cell functional morphology were made using the SEM, and SEM has remained paramount in this field. Bone forming and resorbing cells only exist on surfaces – which are available for study after removal of adjacent tissue layers: The underlying matrix surface can then be studied after removal of the cells, and the mineral front examined after removing the matrix (with an appropriate solvent or by plasma ashing).In this review, we analyse the main findings which we have made in this laboratory concerning the biological activities of osteoblasts (bone forming cells) and osteoclasts (bone resorbing cells). The technical problems of specimen preparation of cells which shrink more than the substrate to which they are attached have been convered previously (Boyde et al. 1977). Such problems obviously affect the lateral, cell to cell inter‐relationships more than the cell to substrate effects which we cover here.At present, we can conclude that SEM has made a major contribution to bone biology by permitting observation of normal cells and natural and surrogate substrates. We confidently predict that it will continue to play a pivotal role in the closer observation of cell‐cell‐substrate interactions particularly in respect of local hormonal effects, as well as in bone pathology and implantology.