ABSTRACT
Piper betle leaves possess several ethnomedicinal properties and are immensely used in traditional medicinal practices in regions of Asian and African subcontinents. However, their effects in treating skeletal complications are least known. In this study, we evaluated cellular and molecular effects of betel leaf extract (BLE) and its major phytoconstituent, hydroxychavicol (HCV) in promoting osteogenesis in vitro and alleviating glucocorticoid induced osteoporosis (GIO) in vivo. Both BLE and HCV markedly stimulated osteoblast differentiation of C3H10T1/2 cells with increased expression of RUNX2 and osteopontin through the GSK-3ß/ß-catenin-signaling pathway. Also, oral administration of BLE and HCV in GIO rats resulted in restoration of bone mass and tissue microarchitecture. Thus, with our findings we conclude that BLE and HCV promote osteogenesis of C3H10T1/2 cells via the GSK-3ß/ß-catenin pathway and alleviate GIO in rats.
Subject(s)
Eugenol/analogs & derivatives , Osteogenesis/drug effects , Osteoporosis/drug therapy , Piper betle/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Animals , Bone Density/drug effects , Cell Differentiation/drug effects , Cell Line , Core Binding Factor Alpha 1 Subunit/metabolism , Eugenol/pharmacology , Female , Glucocorticoids/adverse effects , Glycogen Synthase Kinase 3 beta/metabolism , Mice , Osteoblasts/drug effects , Osteoporosis/metabolism , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , beta Catenin/metabolismABSTRACT
Bone repair using BMP-2 is a promising therapeutic approach in clinical practices, however, high dosages required to be effective pose issues of cost and safety. The present study explores the potential of low dose BMP-2 treatment via tissue engineering approach, which amalgamates 3-D macro/microporous-nanofibrous bacterial cellulose (mNBC) scaffolds and low dose BMP-2 primed murine mesenchymal stem cells (C3H10T1/2 cells). Initial studies on cell-scaffold interaction using unprimed C3H10T1/2 cells confirmed that scaffolds provided a propitious environment for cell adhesion, growth, and infiltration, owing to its ECM-mimicking nano-micro-macro architecture. Osteogenic studies were conducted by preconditioning the cells with 50 ng/mL BMP-2 for 15 min, followed by culturing on mNBC scaffolds for up to three weeks. The results showed an early onset and significantly enhanced bone matrix secretion and maturation in the scaffolds seeded with BMP-2 primed cells compared to the unprimed ones. Moreover, mNBC scaffolds alone were able to facilitate the mineralization of cells to some extent. These findings suggest that, with the aid of 'osteoinduction' from low dose BMP-2 priming of stem cells and 'osteoconduction' from nano-macro/micro topography of mNBC scaffolds, a cost-effective bone tissue engineering strategy can be designed for quick and excellent in vivo osseointegration.
Subject(s)
Bone Morphogenetic Protein 2/pharmacology , Cellulose/chemistry , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Nanofibers/chemistry , Polysaccharides, Bacterial/chemistry , Tissue Engineering , Tissue Scaffolds , Transforming Growth Factor beta/pharmacology , Animals , Bone and Bones , Calcification, Physiologic , Cell Differentiation , Cell Proliferation/drug effects , Cell Survival/drug effects , Chemical Phenomena , Mice , Nanofibers/ultrastructure , Osteogenesis/drug effects , Recombinant Proteins/pharmacology , Thermogravimetry , X-Ray DiffractionABSTRACT
Boerhavia diffusa is a perennial herb belonging to the Nyctaginaceae family. This plant has been widely used in Indian traditional medicinal system to cure several human ailments. However, traditional use of this plant in the treatment and management of wounds has not been validated by any comprehensive scientific study. The present study was aimed to explore the in vitro and in vivo wound healing potential of methanol extract (ME) and chloroform extract (CE) from B. diffusa leaf and subsequent identification of the bioactive metabolites, which might be responsible for enhancement of wound healing property of the extracts. The study included in vitro cell viability and wound scratch assays as well as in vivo excision wound assays in rat models. Both ME and CE were analysed for their antioxidant properties and phenolics content. The gas chromatography-mass spectrometry analyses were performed for identification of bioactive metabolites present in the ME and CE. ME of B. diffusa leaf significantly enhanced viability and migration of human keratinocyte cells (HaCaT) as compared to the untreated and CE-treated groups. The topical application of ME of B. diffusa leaf in excision wound model significantly decreased the wound area by the 14th day (91%) as compared to control (22%) (pâ¯<â¯0.05). The GC-MS analysis revealed the presence of caffeic acid, ferulic acid and D-pinitol as the major bioactive metabolites in ME. These results suggest that ME of B. diffusa possesses significant wound healing potential, where D-pinitol and caffeic acid served as the lead constituent metabolites responsible for the healing.