Counteractions of a Novel Hydroalcoholic Extract from Lens Culinaria against the Dexamethasone-Induced Osteoblast Loss of Native Murine Cells

Abstract

The cytoprotective effects of a novel hydroalcoholic extract (0.01–5 mg/mL) from Lens culinaria (Terre di Altamura Srl) were investigated within murine native skeletal muscle fibers, bone marrow cells, and osteoblasts, and in cell lines treated with the apoptotic agent staurosporine (2.14 × 10−6 M), the alkylating drug cisplatin (10−4 M), the topoisomerase I inhibitor irinotecan (10−4 M), the antimitotic pro-oxidant doxorubicin (10−6 M), and the immunosuppressant dexamethasone (2 × 10−6 M). An amount of 10g of plant material was used to obtain a 70% ethanol/water product, following two-step extraction, evaporation, lyophilization, and storage at −20 °C. For the murine osteoblasts, doxorubicin reduced survival by −65%, dexamethasone by −32% and −60% after 24 and 48 h of incubation time, respectively. The extract was effective in preventing the osteoblast count-reduction induced by dexamethasone; it was also effective at preventing the inhibition of mineralization induced by dexamethasone. Doxorubicin and cisplatin caused a significant reduction in cell growth by −77% for bone marrow cells, −43% for irinotecan, and −60% for dexamethasone, but there was no evidence for the cytoprotective effects of the extract in these cells. Staurosporine and doxorubicin caused a fiber death rate of >−40% after 18 and 24 h of incubation, yet the extract was not effective at preventing these effects. The extract was effective in preventing the staurosporine-induced reduction of HEK293 proliferation and colony formation in the crystal violet DNA staining and the clonogenic assays. It was also effective for the cisplatin-induced reduction in HEK293 cell proliferation. The extract, however, failed to protect the SHSY5Y neurons against cisplatin and irinotecan-induced cytotoxicity. A UV/VIS spectroscopy analysis showed three peaks at the wavelengths of 350, 260, and 190 nm, which correspond to flavonoids, proanthocyanins, salicylates, and AA, constituting the extract. These data suggest the possible development of this extract for use against dexamethasone-induced bone loss and renal chemotherapy-induced damage.

Anti-Inflammatory Effects of a Novel Acetonitrile–Water Extract of Lens Culinaris against LPS-Induced Damage in Caco-2 Cells

“…The viability of Caco-2 cells subjected to LE concentrations ranging from 1 to 3 mg/mL for either 24 or 48 h remained unaltered when compared to the control group (p > 0.05). These data align with safety ranges documented in other studies [15,16], where LE has demonstrated safety within the concentration range from 1 to 3 mg/mL across various cell types. Co-incubation of LPS at concentrations of 100, 10, and 1 µg/mL with LE (2.5 mg/mL) exhibited a cytoprotective effect against LPS-induced damage in Caco-2 cells (Figure 1).…”

“…As previous research indicates, LE exhibits noteworthy health benefits and bioactive properties [15]. In vitro studies have demonstrated its potential to protect against cytotoxic damage induced by cisplatin, irinotecan, and doxorubicin in various cell lines [15,16]. Its antiinflammatory properties make it a promising candidate for managing inflammatory conditions.…”

“…LE was obtained from Lens culinaris supplied by Terre di Altamura Srl Bari, Italy. Based on these results and previous studies on various flavonoids in monolayer cell models [15,16], 2.5 mg/mL of LE was selected for the subsequent experiments. The concentration of LE at 2.5 mg/mL was chosen due to its cytoprotective activity in different cells observed in previous in vitro experiments [15,16].…”

“…Among flavonoids, the Lens culinaris Medik extract (LE), prepared according to the method proposed by Di Turi et al [15], stands out for its notable health benefits and bioactive properties; its liquid chromatography-mass spectrometry (LC-MS) analysis revealed a high content of flavonoids. LE at a concentration range of 0.1-5 mg/mL in in vitro studies has provided evidence of its potential to protect against cytotoxic damage induced by cisplatin, irinotecan, and doxorubicin on different cell lines [15,16]. The anti-inflammatory potential of LE positions it as a promising candidate for managing inflammatory conditions.…”