Chitosan Elicitation Impacts Flavonolignan Biosynthesis in Silybum marianum (L.) Gaertn Cell Suspension and Enhances Antioxidant and Anti-inflammatory Activities of Cell Extracts

Abstract

Silybum marianum (L.) Gaertn is a rich source of antioxidants and anti-inflammatory flavonolignans with great potential for use in pharmaceutical and cosmetic products. Its biotechnological production using in vitro culture system has been proposed. Chitosan is a well-known elicitor that strongly affects both secondary metabolites and biomass production by plants. The effect of chitosan on S. marianum cell suspension is not known yet. In the present study, suspension cultures of S marianum were exploited for their in vitro potency to produce bioactive flavonolignans in the presence of chitosan. Established cell suspension culture was maintained on the same hormonal media supplemented with 0.5 mg/L BAP (6-benzylaminopurine) and 1.0 mg/L NAA (α-naphthalene acetic acid) under photoperiod 16/8 h (light/dark) and exposed to various treatments of chitosan (ranging from 0.5 to 50.0 mg/L). The highest biomass production was observed for cell suspension treated with 5.0 mg/L chitosan, resulting in 123.3 g/L fresh weight (FW) and 17.7 g/L dry weight (DW) productions. Chitosan treatment resulted in an overall increase in the accumulation of flavonoids, phenolic compounds and silymarin. High accumulation levels of silybin B, silydianin and silybin A were recorded by HPLC analysis. The corresponding extracts displayed interesting antioxidant and anti-inflammatory capacities. In particular, high ABTS antioxidant activity (741.5 μM Trolox C equivalent antioxidant capacity) was recorded in extracts obtained in presence of 0.5 mg/L of chitosan. On the opposite, highest inhibitions of cyclooxygenase 2 (COX-2, 30.5 %), secretory phospholipase A2 (sPLA2, 33.9 %) and 15-lipoxygenase (15-LOX-2, 31.6 %) enzymes involved in inflammation process were measured in extracts obtained in presence of 5.0 mg/L of chitosan. Taken together, these results highlight the high potential of the chitosan elicitation of the S. marianum cell suspension for enhanced production of antioxidant and anti-inflammatory silymarin-rich extracts.

Plant‐Based Synthesis of Zinc Oxide Nanoparticles (ZnO‐NPs) Using Aqueous Leaf Extract of Aquilegia pubiflora: Their Antiproliferative Activity against HepG2 Cells Inducing Reactive Oxygen Species and Other In Vitro Properties

“…In the experiment, acarbose was used as a positive control and carried out in triplicate and repeated twice. [34,35]. To assess the antioxidant capacity of test samples, DPPH reagents at concentrations ranging from 12.5 mL to 400 mL were employed.…”

Copper oxide (CuO) and manganese oxide (MnO) nanoparticles induced biomass accumulation, antioxidants biosynthesis and abiotic elicitation of bioactive compounds in callus cultures of Ocimum basilicum (Thai basil)

“…The leaf explant (0.5 cm 2 ) was used from in vitro germinated plantlets (28 days old) and innocluated on hormonal media. The BAP (5 mg/L and NAA (1 mg/L) was used in media to induce optimum callogenesis as optimised previously [33]. Prior autoclaving, pH of the growth media was adjusted to 5.2-5.6 with 0.1 N NaOH and 0.1 N HCL.…”

“…The callus cultures were weighed for fresh weight analysis while for dry weight analysis the cultures were placed in an incubator to dry. For phytochemical analysis, calli powder was used to prepare the methanolic extracts by following the protocol of [33]. Briefly, 10 mL methanol (99.9%) was poured in 100 mg of ground samples and kept for 24 h at room temperature on rotatory shaker.…”

Phytochemical analysis and versatile in vitro evaluation of antimicrobial, cytotoxic and enzyme inhibition potential of different extracts of traditionally used Aquilegia pubiflora Wall. Ex Royle

“…The protocol reported previously by [44,45], was adopted with minor changes. The possible free radical scavenging ability of test samples was investigated for their antioxidant potential using DPPH reagent at concentrations ranging from 12.5 μL to 400 μL.…”