Abstract
BackgroundSpinach (Spinacia oleracea) is a rich source of flavonoids and therefore widely used therapeutically as an antioxidant agent in traditional medicine. The present study was undertaken to study the bone regenerating property of dried Spinacia oleracea extract (DSE) and self-emulsifying formulation of the extract (FDSE) on drill-hole model of fracture repair in rats.Methods0.8 mm hole was drilled in the diaphyseal region of femur in adult SD rats. DSE and formulated extract (FDSE) was administered orally and fractured femur was collected after treatment regimen. Micro-CT, transcriptional analysis and measurement of calcein intensity of callus formed at the injured site was performed to study the efficacy of the extract and formulation on bone regeneration. Further, compounds from extract were assessed for in-vitro osteoblast activity.ResultsMicro-architecture of the regenerated bone at injured site exhibited 26% (p < 0.001) and 35% (p < 0.01) increased BV/TV (bone volume /tissue volume) and Tb.N. (trabecular number) for DSE (500 mg.kg− 1). Further, FDSE exhibited similar augmentation in BV/TV (p < 0.01) and Tb. N (p < 0.01) parameters at dose of 250 mg.kg− 1. Analogous results were obtained from transcriptional analysis and calcein intensity at the fractured site. 3-O-Methylpatuletin, one of the compound isolated from the extract stimulated the differentiation and mineralization of primary osteoblast and depicted concentration dependent antagonizing effect of H2O2 in osteoblast apparently, minimizing ROS generation thus affectivity in fracture repair.ConclusionsThe present study showed that bone regenerating property of spinach was augmented by formulating extract to deliverable form and can be further studied to develop as therapeutic agent for fracture repair.Graphical abstract
Highlights
S. oleracea L. commonly known as Spinach, is an edible flowering plant in the family of Amaranthaceae
Preparation of self-micro emulsifying drug delivery system (SEDDS) Different blank self emulsifying drug delivery system (SEDDS) formulations were prepared by varying the ratio of oil to co-surfactant to increase the loading and decrease the precipitation of extract after dilution
bone volume/tissue volume (BV/TV) was increased by ~ 26% (p < 0.01) at 500 mg.kg1dy1 of dried Spinacia oleracea extract (DSE) compared to control group whereas; no significant effect was observed at the lower dose
Summary
S. oleracea L. commonly known as Spinach, is an edible flowering plant in the family of Amaranthaceae. It is an erect herb about 30–60 cm height, native to South-West Asia and cultivated throughout world as vegetables. Spinach is recommended as the best source of iron, vitamins and minerals It is a well-known source of bioactive phytochemicals and nutrients such as ascorbate, carotenoids, tocopherols, phenolics, flavonoids, folate and minerals [1]. Our previous studies on ethanolic extract of Spinacia oleracea exhibits bone restoration in ovariectomy induced bone loss model of osteoporosis in rats and the effect was comparable to estrogen [5]. Bioactive constituents characterized from the extract exhibit significant chondrocyte protective effects in articular chondrocytes apparently preventing cartilage damage in MIA induced osteoarthritis [6]. The present study was undertaken to study the bone regenerating property of dried Spinacia oleracea extract (DSE) and self-emulsifying formulation of the extract (FDSE) on drill-hole model of fracture repair in rats
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