ABSTRACT
Sanguinarine, a benzophenanthridine alkaloid, has been previously demonstrated to exert antimicrobial, anti-inflammatory, and anti-tumor activities. A previous study has identified Sanguinarine as a potential drug candidate for osteoporosis treatment by computational bioinformatics analysis. This study further evaluated the effects of Sanguinarine on the differentiation of murine preosteoblast MC3T3-E1 cells and its anti-osteoporosis activity in an ovarietomized rat model. Sanguinarine treatment (0.25, 0.5, 1, and 2 µm) of MC3T3-E1 cells significantly increased alkaline phosphatase (ALP) activity and the phoshporalyation of AMP-activated protein kinase α subunit (AMPKα), but did not affect cell proliferation. The induction effects of Sanguinarine treatment (2 µm) on ALP activity, AMPKα phosphorylation, Smad1 phosphorylation, and the expression of three osteoblast differentiation-regulators (bone morphogenetic protein 2 [BMP2], osterix [OSX], and osteoprotegerin [OPG]) were partially reversed by Compound C treatment. More importantly, Sanguinarine treatment promoted bone tissue growth in an ovariectomized (OVX) osteoporosis rat model as evaluated by histological examination, micro-CT analysis, and serum parameter detection. In conclusion, these results indicate that Sanguinarine induces the differentiation of MC3T3-E1 cells through the activation of the AMPK/Smad1 signaling pathway. Sanguinarine can stimulate bone growth in vivo and may be an effective drug for osteoporosis treatment.
Subject(s)
Benzophenanthridines/pharmacology , Bone Density Conservation Agents/pharmacology , Bone Remodeling/drug effects , Cell Differentiation/drug effects , Isoquinolines/pharmacology , Osteoblasts/drug effects , Osteogenesis/drug effects , Osteoporosis, Postmenopausal/drug therapy , Ovariectomy , 3T3 Cells , AMP-Activated Protein Kinases/metabolism , Animals , Bone Density/drug effects , Disease Models, Animal , Dose-Response Relationship, Drug , Female , Humans , Mice , Osteoblasts/metabolism , Osteoblasts/pathology , Osteoporosis, Postmenopausal/metabolism , Osteoporosis, Postmenopausal/pathology , Osteoporosis, Postmenopausal/physiopathology , Phosphorylation , Rats, Sprague-Dawley , Signal Transduction/drug effects , Smad1 Protein/metabolismABSTRACT
This study aimed to prepare and characterize a silk fibroin/calcium sulfate (SF/CS) bone cement. SF solutions of the following concentrations 3, 4.5, 6, 7.5, and 9 g/L were used to prepare bone cement samples with SF-to-CS ratios of 0.35, 0.40, 0.45, 0.50, and 0.55 mL of SF solutions (or water as control) per g of CS. Compressive strength, setting time, degradation when immersed in phosphate-buffered saline, X-ray diffraction, Fourier infrared spectroscopy, and scanning electron microscopy were used to characterize the SF/CS bone cement. Biocompatibility was determined using rat bone mesenchymal stem cells (rBMSCs) and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The optimal bone cement was obtained with 0.4 mL of 6 g/L SF solution per g of α-hemihydrate CS powder. Compared with CS bone cement, compressive strength of SF/CS bone cement was 19.7% higher (p = 0.003). Samples prepared with SF had less degradation in phosphate-buffered saline than the ones prepared with deionized water. Using SF solution as the solidifying liquid increases the initial setting and final setting time of CS compared with deionized water. Using extracts from SF/CS bone cement, all cell relative proliferation rates were >100%, showing no cytotoxicity for any sample. In conclusion, using the 6 g/L SF solution at 0.40 mL/g of CS increased the cement compressive strength. SF solution added to CS achieved a bone cement with increased durability compared with CS bone cement. The SF/CS cement had no cytotoxicity on rBMSCs. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 512-519, 2018.
