Preoperative Diagnosis of Infection in Patients with Nonunions

Stucken, Charlton; Olszewski, Dana; Creevy, William R.; Murakami, Akira M.; Tornetta, Paul

doi:10.2106/jbjs.l.01034

Cited by 77 publications

(56 citation statements)

References 12 publications

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“…The iMEFs were established and characterized as described [30]. Cell lines were maintained in in complete DMEM supplemented with 10% fetal bovine serum (FBS),100U/ml penicillin and 100mg/ml streptomycin in a humidified atmosphere of 5% CO 2 at 37°C as described [14,16,20,31]. Trichostatin A (TSA) was obtained from EMD Chemicals (Gibbstown, NJ).…”

Section: Methodsmentioning

confidence: 99%

“…Recombinant adenoviruses were generated using AdEasy technology as described [14,15,32,33,34]. The coding region of human BMP9 was PCR amplified and cloned into an adenoviral shuttle vector and subsequently used to generate recombinant adenovirus in HEK293 cells.…”

Section: Methodsmentioning

confidence: 99%

“…Quantitative ALP activity was assessed by a modified Great Escape SEAP Chemiluminescence assay (BD Clontech, Mountain View, CA) and/or histochemical staining assay (using a mixture of 0.1 mg/ml napthol AS-MX phosphate and 0.6 mg/ml Fast Blue BB salt) as described [14,15,17,18,19,20,31,33,37,42,43]. For the chemilluminescence assays, each assay condition was performed in triplicate and the results were repeated in at least three independent experiments.…”

Section: Methodsmentioning

confidence: 99%

“…Infected cells were cultured in the presence of ascorbic acid (50μg/mL) and β-glycerophosphate (10 mM). At 14 days after infection, mineralized matrix nodules were stained for calcium precipitation by means of Alizarin Red S staining, as described previously [14,15,17,18,19,20,31,33,37]. Cells were fixed with 0.05% (v/v) glutaraldehyde at room temperature for 10min.…”

Section: Methodsmentioning

confidence: 99%

“…We have demonstrated that BMP9 is one of the most potent BMPs among the 14 types of BMPs in inducing osteogenic differentiation of MSCs by regulating several important downstream targets [5,9,14,15,16,17,18,19,20,21,22]. BMP9 (also known as growth differentiation factor 2, or GDF-2) was originally identified in the developing mouse liver [23].…”

Section: Introductionmentioning

confidence: 99%

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Inhibition of Histone Deacetylases Potentiates BMP9-Induced Osteogenic Signaling in Mouse Mesenchymal Stem Cells

Wang

Liu

et al. 2013

Cell Physiol Biochem

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Background/Aims: We have demonstrated that bone morphogenetic protein 9 (BMP9) is one of the most potent BMPs in regulating osteoblast differentiation of mesenchymal stem cells (MSCs) although the molecular mechanism underlying BMP9-induced osteogenesis remains to be fully elucidated. It is known that epigenetic regulations play an important role in regulating the stem cell potency and lineage commitment. Here, we investigate if the inhibition of histone deacetylases (Hdacs) affects BMP9-induced osteogenic differentiation of MSCs. Methods: Using the Hdac inhibitor trichostatin A (TSA), we assess that TSA enhances BMP9-mediated osteogenic markers and matrix mineralization in MSCs, and bone formation in mouse embryonic limb explants. Results: We find that the endogenous expression of most of the 11 Hdacs is readily detectable in MSCs. BMP9 is shown to induce most Hdacs in MSCs. We demonstrate that TSA potentiates BMP9-induced early osteogenic marker alkaline phosphatase (ALP) activity in MSCs, as well as late osteogenic markers osteopontin (OPN) and osteocalcin (OCN) and matrix mineralization. Fetal limb explant culture studies reveal that TSA potentiates BMP9-induced endochondral bone formation, possibly by expanding hypertrophic chondrocyte zone of growth plate. Conclusion: Our findings strongly suggest histone deacetylases may play an important role in fine-tuning BMP9-mediated osteogenic signaling through a negative feedback network in MSCs. Thus, Hdac inhibitors may be used as novel therapeutics for bone fracture healing.

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