The optimal dose of recombinant human osteogenic protein-1 enhances differentiation of mouse osteoblast-like cells: An in vitro study

Zhang, Feng; Ren, Lingfei; Lin, Hongyu; Yin, Mei-nv; Tong, Yongqing; Shi, Guangming

doi:10.1016/j.archoralbio.2011.10.008

Cited by 24 publications

(40 citation statements)

References 40 publications

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“…On the other hand, qPCR experiments showed an absence of early gene expression regulation, which may be explained by the relatively low levels of hBMP‐7 secreted in cell culture supernatants of transfected cells. Exogenous human BMP 7 added to MC3T3‐E1 cultures, has been previously shown to induce the expressions of Alp , Runx2 , and ColIa1 markers in a dose‐ and time‐dependent manner . However, the range of concentrations in which the induction potential of hBMP‐7 was investigated (10–400 ng mL −1 ) in a previously published study, was much higher than the concentration measured in the transfected cell culture supernatants in our experiments (200 pg mL −1 ).…”

Section: Discussioncontrasting

confidence: 67%

“…Exogenous human BMP 7 added to MC3T3‐E1 cultures, has been previously shown to induce the expressions of Alp , Runx2 , and ColIa1 markers in a dose‐ and time‐dependent manner . However, the range of concentrations in which the induction potential of hBMP‐7 was investigated (10–400 ng mL −1 ) in a previously published study, was much higher than the concentration measured in the transfected cell culture supernatants in our experiments (200 pg mL −1 ). This difference may explain why we did not observe an increased gene expression in cells transfected with CaP/PEI/phBMP‐7/SiO 2 ‐SH nanoparticles.…”

Section: Discussioncontrasting

confidence: 67%

“…In addition, in this study, ascorbic acid or β‐glycerophosphate were not included in nanoparticle treated cell cultures in order to capture solely the effect of hBMP‐7 produced after transfection. In contrast, osteogenic conditions were used in previous studies alongside exogenous addition of BMP‐7 . The use of osteogenic medium (that is, primary medium supplemented with ascorbic acid and β‐glycerophosphate) may have enhanced the observed effect, since a synergistic effect between ascorbic acid and BMP‐7 was reported to stimulate osteoblast‐gene expression, specifically osteocalcin ( Ocn ) and Bsp mRNA levels …”

Section: Discussionmentioning

confidence: 99%

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Calcium phosphate nanoparticles carrying BMP‐7 plasmid DNA induce an osteogenic response in MC3T3‐E1 pre‐osteoblasts

Hadjicharalambous

Kozlova

Sokolova

et al. 2015

J Biomedical Materials Res

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Functionalized calcium phosphate nanoparticles with osteogenic activity were prepared. Polyethyleneimine-stabilized calcium phosphate nanoparticles were coated with a shell of silica and covalently functionalized by silanization with thiol groups. Between the calcium phosphate surface and the outer silica shell, plasmid DNA which encoded either for bone morphogenetic protein 7 (BMP-7) or for enhanced green fluorescent protein was incorporated as cargo. The plasmid DNA-loaded calcium phosphate nanoparticles were used for the transfection of the pre-osteoblastic MC3T3-E1 cells. The cationic nanoparticles showed high transfection efficiency together with a low cytotoxicity. Their potential to induce an osteogenic response by transfection was demonstrated by measuring the alkaline phosphatase (ALP) activity and calcium deposition with alizarin red staining. The expression of the osteogenic markers Alp, Runx2, ColIa1 and Bsp was investigated by means of real-time quantitative polymerase chain reaction. It was shown that phBMP-7-loaded nanoparticles can provide a means of transient transfection and localized production of BMP-7 in MC3T3-E1 cells, with a subsequent increase of two osteogenic markers, specifically ALP activity and calcium accumulation in the extracellular matrix. Future strategies to stimulate bone regeneration focus into enhancing transfection efficiency and achieving higher levels of BMP-7 produced by the transfected cells.

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Section: Discussioncontrasting

confidence: 67%

Section: Discussioncontrasting

confidence: 67%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Calcium phosphate nanoparticles carrying BMP‐7 plasmid DNA induce an osteogenic response in MC3T3‐E1 pre‐osteoblasts

Hadjicharalambous

Kozlova

Sokolova

et al. 2015

J Biomedical Materials Res

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“…3A), no more than 6% and 41% of the loaded BMP-7 would have been released from the trehalose and trehalose-free formulations, respectively, after 7 days (i.e., 28 and 238 ng BMP-7/mg of particles). Since 2.5 mg of particles and 1 mL solution were used in the cell assay, there was a theoretical cumulative amount of 70 and 595 ng/mL available, respectively, which according to 32 should have led to significant differences in the differentiation of MC3T3-E1 cells into osteoblasts, as measured by an ALP assay, but which were not observed here (both formulations were effective and nonstatistically different). Hence, it could be concluded that there were clearly different amounts of BMP-7 released and likely higher than predicted by the release assay.…”

Section: In Vitro Microparticle Direct Culturementioning

confidence: 77%

Growth Factor-Loaded Microparticles for Tissue Engineering: The Discrepancies of In Vitro Characterization Assays

Bock

Dargaville

Kirby

et al. 2016

Tissue Engineering Part C: Methods

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“…ALP staining was used as a marker of osteoblasts during maturation, and the specific activity of ALP was measured to evaluate cell growth and differentiation. In addition, we measured the expression levels of the Runx2 and OP-1 genes, which encode essential transcription factors involved in the regulation of osteoblast differentiation [23]. Treatment with 10 µM GC resulted in decreased Runx2, OP-1, and ALP expression (p < 0.05), as well as decreased ALP activity (▶ Fig.…”

Section: Resultsmentioning

confidence: 99%

Beta-Ecdysone Protects Mouse Osteoblasts from Glucocorticoid-Induced Apoptosis In Vitro

et al. 2017

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Glucocorticoid-induced osteoporosis is a common form of secondary osteoporosis. Glucocorticoids affect both bone formation and resorption, and prolonged glucocorticoid exposure can suppress osteoblast activities. beta-Ecdysone, found in many plants, is involved in protein synthesis, carbohydrate and lipid metabolism, and immunologic modulation. Here, we evaluated the effects of beta-ecdysone on osteoblast viability by assessing apoptosis following treatment with excess glucocorticoids. Mouse bone marrow stromal cells were induced to differentiate and grow into osteoblasts, and then treated with 10 µM glucocorticoid and 10, 1, or 0.1 µM beta-ecdysone. The expression levels of osteoblast growth and differentiation factors (runt-related transcription factor 2, osteogenic protein-1, and alkaline phosphatase), apoptosis-related genes (transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8), and Akt1 and phospho-Akt (Thr308) were then assessed via alkaline phosphatase staining, acridine orange-propidium iodide staining, annexin V/PI apoptosis assay, real-time RT-PCR, and Western blot analyses. Notably, treatment with 10 µM glucocorticoid resulted in reduced osteoblast viability and the specific activity of alkaline phosphatase as well as reduced runt-related transcription factor 2, osteogenic protein-1, and alkaline phosphatase mRNA expression , indicating that glucocorticoid inhibited osteogenic differentiation. Moreover, glucocorticoid treatment yielded increased transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8 expression and decreased Akt1 and phospho-Akt levels, indicating glucocorticoid-induced apoptosis. Meanwhile, beta-ecdysone inhibited glucocorticoid function, preserving the expression of Akt1 and phospho-Akt and reducing the expression of transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8. Thus, beta-ecdysone prevented glucocorticoid-induced osteoblast apoptosis. These data highlight the potential for beta-ecdysone as a treatment for preventing the effects of glucocorticoid on bone growth.

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The optimal dose of recombinant human osteogenic protein-1 enhances differentiation of mouse osteoblast-like cells: An in vitro study

Cited by 24 publications

References 40 publications

Calcium phosphate nanoparticles carrying BMP‐7 plasmid DNA induce an osteogenic response in MC3T3‐E1 pre‐osteoblasts

Calcium phosphate nanoparticles carrying BMP‐7 plasmid DNA induce an osteogenic response in MC3T3‐E1 pre‐osteoblasts

Growth Factor-Loaded Microparticles for Tissue Engineering: The Discrepancies of In Vitro Characterization Assays

Beta-Ecdysone Protects Mouse Osteoblasts from Glucocorticoid-Induced Apoptosis In Vitro

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