Ethyl acetate and n‐butanol fraction of Cissus quadrangularis promotes the mineralization potential of murine pre‐osteoblast cell line MC3T3‐E1 (sub‐clone 4)

Abstract

In a sequel to investigate osteogenic potential of ethanolic extract of Cissus quadrangularis (CQ), the present study reports the osteoblast differentiation and mineralization potential of ethyl acetate (CQ‐EA) and butanol (CQ‐B) extracts of CQ on mouse pre‐osteoblast cell line MC3T3‐E1 (sub‐clone 4) with an objective to isolate an antiosteoporotic compound. Growth curve, proliferation, and viability assays showed that both the extracts were nontoxic to the cells even at high concentration (100 µg/ml). The cell proliferation was enhanced at low concentrations (0.1 µg/ml and 1 µg/ml) of both the extracts. They also upregulated the osteoblast differentiation and mineralization processes in MC3T3‐E1 cells as reflected by expression profile of osteoblast marker genes such as RUNX2, Osterix, Collagen (COL1A1), Alkaline Phosphatase (ALP), Integrin‐related Bone Sialoprotein (IBSP), Osteopontin (OPN), and Osteocalcin (OCN). CQ‐EA treatment resulted in early differentiation and mineralization as compared with the CQ‐B treatment. These findings suggest that low concentrations of CQ‐EA and CQ‐B have proliferative and osteogenic properties. CQ‐EA, however, is more potent osteogenic than CQ‐B.

Osteogenic potential of hexane and dichloromethane fraction of Cissus quadrangularis on murine preosteoblast cell line MC3T3‐E1 (subclone 4)

“…Previously we have reported mitogenic concentrations of CQ‐E (0.1 and 1 µg/ml; Tasadduq et al, ) and two solvent fractions of CQ, CQ‐EA, and CQ‐B, (0.1, 1, 10, and 100 µg/ml; Toor et al, ). However, in the present study, we found that CQ‐D and CQ‐H are efficacious at lower concentrations (0.1, 1, 10 and 100 ng/ml) as suggested by analysis of growth parameters, cell viability, metabolic activity, and proliferation assays.…”

“…However, mineralization was significantly enhanced in CQ–H‐treated cells compared to CQ–D‐treated cells (Table ). Treatment with other solvent fractions of CQ, CQ‐EA, and CQ‐B (Toor et al, ) resulted in enhanced mineralization of MC3T3‐E1 cell after 14 days of osteogenic induction. Though enhanced, the mineral deposition was less in CQ‐EA‐treated cultures compared to CQ‐H treatment.…”

“…Treatment with other solvent fractions of CQ, CQ-EA, and CQ-B (Toor et al, 2018) upregulation of ALP activity when rat mesenchymal stem cells were differentiated into osteoblasts in response to petroleum ether extract of CQ.…”

“…Expression of genes ( SP7, IBSP , and BGLAP ) was upregulated in CQ–H‐ and CQ–D‐treated cultures, whereas expression of RUNX2, COL1A1 , and SPP1 was unaffected compared to positive control cultures. CQ‐EA and CQ‐B treatment, however, did not affect the expression of SP7 and IBSP , but an upregulated expression of RUNX2, COL1A1, ALP, SPP1 , and BGLAP (Toor et al, ). RUNX2 is the master regulator of osteogenesis and regulates the expression of osteoblast marker genes including osterix, collagen I, alkaline phosphatase, bone sialoproteins, osteopontin, and osteocalcin (Hayrapetyan et al, ).…”

A review on efficacy of Cissus quadrangularis in pharmacological mechanisms

“…Osteogenic potential of Cissus was studied by Toor, et al [41] and ethanolic extract of the plant accelerated fracture healing as well as early remodeling of fracture callus. The authors also studied the effect of hexane and dichloromethane fraction on the differentiation and mineralization of mouse pre-osteoblast cell line [42].…”