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Abstract Background Prunella vulgaris is medicinally important plant containing high-valued chemical metabolites like Prunellin which belong to family Lamiaceae and it is also known as self-heal. In this research, calli culture were exposed to differential ratios of gold (Au) and silver (Ag) nanoparticles (1:1, 1:2, 1:3, 2:1 and 3:1) along with naphthalene acetic acid (2.0 mg NAA) to investigate its antimicrobial potential. A well diffusion method was used for antimicrobial properties. Results Here, two concentrations (1 and 2 mg/6 µl) of all treated calli cultures and wild plants were used against Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Bacillus atrophaeus, Bacillus subtilis, Agrobacterium tumefaciens, Erwinia caratovora and Candida albicans. Dimethyl sulfoxide (DMSO) and antibiotics were used as negative and positive controls. Here, the calli exposed to gold (Au) nanoparticles (NPs) and 2.0 mg naphthalene acetic acid (NAA) displayed the highest activity (25.7 mm) against Salmonella typhi than other extracts, which was considered the most susceptible species, while Agrobacterium tumefaciens and Candida albicans was the most resistance species. A possible mechanism of calli induced nanoparticles was also investigated for cytoplasmic leakage. Conclusion From the above data it is concluded that Prunella vulgaris is medicinally important plant for the development of anti-microbial drugs using nanotechnology and applicable in various pharmaceutical research.

In this study, poly(isobutylene-alt-maleic anhydride) (PMA)-coated spinel ferrite (MFe2O4, where M = Fe, Co, Ni, or Zn) nanoparticles (NPs) were developed as carriers of the anticancer drugs doxorubicin (DOX) and methotrexate (MTX). Physical characterizations confirmed the formation of pure cubic structures (14–22 nm) with magnetic properties. Drug-loaded NPs exhibited tumor specificity with significantly higher (p < 0.005) drug release in an acidic environment (pH 5.5). The nanoparticles were highly colloidal (zeta potential = −35 to −26 mV) in deionized water, phosphate buffer saline (PBS), and sodium borate buffer (SBB). They showed elevated and dose-dependent cytotoxicity in vitro compared to free drug controls. The IC50 values ranged from 0.81 to 3.97 μg/mL for HepG2 and HT144 cells, whereas IC50 values for normal lymphocytes were 10 to 35 times higher (18.35–43.04 µg/mL). Cobalt ferrite (CFO) and zinc ferrite (ZFO) NPs were highly genotoxic (p < 0.05) in cancer cell lines. The nanoparticles caused cytotoxicity via oxidative stress, causing DNA damage and activation of p53-mediated cell cycle arrest (significantly elevated expression, p < 0.005, majorly G1 and G2/M arrest) and apoptosis. Cytotoxicity testing in 3D spheroids showed significant (p < 0.05) reduction in spheroid diameter and up to 74 ± 8.9% of cell death after two weeks. In addition, they also inhibited multidrug resistance (MDR) pump activity in both cell lines suggesting effectivity in MDR cancers. Among the tested MFe2O4 NPs, CFO nanocarriers were the most favorable for targeted cancer therapy due to excellent magnetic, colloidal, cytotoxic, and biocompatible aspects. However, detailed mechanistic, in vivo cytotoxicity, and magnetic-field-assisted studies are required to fully exploit these nanocarriers in therapeutic applications.

Aquilegia pubiflora leaves are reported against hepatitis, for treatment of skin burns, wound healing, jaundice, gynea, circulatory and cardiovascular disease in several studies. Therefore, green synthesis of silver nanoparticles with A. pubiflora leaves extract, as an alternative to chemical synthesis, is beneficial from its biological applications point of view. The formation and size distribution of Ag-NPs were confirmed by UV-spectroscopy, Scanning electron microscopy (SEM), X-ray powder diffraction (XRD), Transmission electron microscopy (TEM), Energy Dispersive X-ray (EDX), Fourier transform infrared spectroscopy (FTIR) and HPLC. Silver nanoparticles with a particle size of 19 nm were used in various assays including antimicrobial, antioxidant, anti-Alzheimer, anti-diabetic, anti-aging, anti-cancer and biocompatibility studies. Our findings revealed that A. pubiflora mediated Ag-NPs were highly active against both bacterial and fungal strains with Escherichia coli and Aspergillus niger displaying the highest zone of inhibition 10.9 ± 0.29 mm and 13.11 ± 0.33 mm respectively. Dose dependent cytotoxic activity was shown by NPs against Leishmania Tropica with significant IC50 of 175 μg/mL for promastigote and 169 μg/mL for amastigote forms. The Ag-NPs proved to be highly toxic for the HepG2 cell line and inhibit cell viability up to 52.06 ± 2.9% after 24 h of Ag-NPs treatment. Ag-NPs also showed excellent inhibitory potential against the enzymes acetylcholinesterase (AChE: 71.38 ± 1.75) and butyrylcholinesterase (BChE: 73.61 ± 1.96) which are involved in Alzheimer's disease. Overall, the enzymes involved in aging, diabetes, and inflammation showed a moderate inhibitory response to Ag-NPs, led to the conclusion that they are a promising choice for various biological applications.

Abstract Ajuga integrifolia Buch. Ham. ex D.Don, a member of Lamiaceae family is pharmaceutically an active perennial herb widely spread in China, Afghanistan and Pakistan Himalayan region. The application of biotic elicitors is a promising approach to cover limitations of in vitro cell technology and challenges faced by pharmaceuticals industry for bulk up production. The current study involved the induction of agitated micro-shoot cultures with the aim to investigate the growth-promoting as well as phytochemicals enhancement role of yeast extract (YE) and pectin (PE). The results showed that both elicitors induced a considerable physiological response. Biomass accumulation was observed maximum (DW: 18.3 g/L) against PE (10 mg/L) compared to YE and control. Eleven secondary phytocompounds were quantified using high-performance liquid chromatography. PE (50 mg/L) was found to be effective in elicitation of rosmarinic acid (680.20 µg/g), chlorogenic acid (294.12 µg/g), apigenin (579.61 µg/g) and quercetin (596.89 µg/g). However, maximum caffeic acid (359.52 µg/g) and luteolin (546.12 µg/g accumulation was noted in PE (1 mg/L) treatment. Harpagide, aucubin, harpagoside and 8-O-acetyl-harpagoside production was suppressed by both elicitors except for YE (100 mg/L). Catalpol accumulation in micro-shoot cultures was also downregulated except in response to YE (50 and 100 mg/L). Antioxidant activity and anti-inflammatory activity remained higher under PE (50 mg/L) and YE (100 mg/L) respectively. Therefore, results suggested that Ajuga integrifolia micro-shoot cultures treated with yeast extract and pectin might be an efficient bio-factory to produce commercially potent specific secondary metabolites.

Abstract Background Himalayan Columbine (Aquilegia pubiflora Wall. Ex Royle) is a medicinal plant and have been used as traditional treatments for various human diseases including skin burns, jaundice, hepatitis, wound healing, cardiovascular and circulatory diseases. Till now there is no report available on phytochemical investigation of Himalayan Columbine and to the best of our knowledge, through present study we have reported for the first time, the phytochemical analysis and pharmacological potentials of different leaf extracts of Aquilegia pubiflora. Methods Four types of extracts were prepared using solvent of different polarities (Distilled water APDW, Methanol APM, Ethanol APE and Ethyl acetate APEA), and were evaluated to determine the best candidate for potent bioactivity. Phytochemical constituents in prepared extracts were quantified through HPLC analysis. Subsequently, all four types of leaf extracts were then evaluated for their potential bioactivities including antimicrobial, protein kinase inhibition, anti-inflammatory, anti-diabetic, antioxidant, anti-Alzheimer, anti-aging and cytotoxic effect. Results HPLC analysis demonstrated the presence of dvitexin, isovitexin, orientin, isoorientin, ferulic acid, sinapic acid and chlorogenic acid in varied proportions in all plant extracts. Antimicrobial studies showed that, K. pneumonia was found to be most susceptible to inhibition zones of 11.2 ± 0.47, 13.9 ± 0.33, 12.7 ± 0.41, and 13.5 ± 0.62 measured at 5 mg/mL for APDW, APM, APE and APEA respectively. A. niger was the most susceptible strain in case of APDW with the highest zone of inhibition 14.3 ± 0.32, 13.2 ± 0.41 in case of APM, 13.7 ± 0.39 for APE while 15.4 ± 0.43 zone of inhibition was recorded in case of APEA at 5 mg/mL. The highest antioxidant activity of 92.6 ± 1.8 μgAAE/mg, 89.2 ± 2.4 μgAAE/mg, 277.5 ± 2.9 μM, 289.9 ± 1.74 μM for TAC, TRP, ABTS and FRAP, respectively, was shown by APE. APM, APE and APEA extracts showed a significant % cell inhibition (above 40%) against HepG2 cells. The highest anti-inflammatory of the samples was shown by APE (52.5 ± 1.1) against sPLA2, (41.2 ± 0.8) against 15-LOX, followed by (38.5 ± 1.5) and (32.4 ± 0.8) against COX-1 and COX-2, respectively. Conclusions Strong antimicrobial, Protein Kinase potency and considerable α-glucosidase, α-amylase, and cytotoxic potential were exhibited by plant samples. Significant anti-Alzheimer, anti-inflammatory, anti-aging, and kinase inhibitory potential of each plant sample thus aware us for further detailed research to determine novel drugs.

Abstract Fagonia indica (L.) is an important medicinal plant with multitude of therapeutic potentials. Such application has been attributed to the presence of various pharmacological important phytochemicals. However, the inadequate biosynthesis of such metabolites in intact plants has hampered scalable production. Thus, herein, we have established an in vitro based elicitation strategy to enhance such metabolites in callus culture of F. indica. Cultures were exposed to various doses of UV radiation (UV-C) and grown in different photoperiod regimes and their impact was evaluated on biomass accumulation, biosynthesis of phytochemicals along antioxidant expression. Cultures grown under photoperiod (16L/8D h) after exposure to UV-C (5.4 kJ/m2) accumulated optimal biomass (438.3 g/L FW; 16.4 g/L DW), phenolics contents (TPC: 11.8 μgGAE/mg) and flavonoids contents (TFC: 4.05 μgQE/mg). Similarly, HPLC quantification revealed that total production (6.967 μg/mg DW) of phytochemicals wherein kaempferol (1.377 μg/mg DW), apigenin (1.057 μg/mg DW), myricetin (1.022 μg/mg DW) and isorhamnetin (1.022 μg/mg DW) were recorded highly accumulated compounds in cultures at UV-C (5.4 kJ/m2) dose than other UV-C radiations and light regimes.. The antioxidants activities examined as DPPH (92.8%), FRAP (182.3 µM TEAC) and ABTS (489.1 µM TEAC) were also recorded highly expressed by cultures under photoperiod after treatment with UV-C dose 5.4 kJ/m2. Moreover, same cultures also expressed maximum % inhibition towards phospholipase A2 (sPLA2: 35.8%), lipoxygenase (15-LOX: 43.3%) and cyclooxygenases (COX-1: 55.3% and COX-2: 39.9%) with 1.0-, 1.3-, 1.3- and 2.8-fold increased levels as compared with control, respectively. Hence, findings suggest that light and UV can synergistically improve the metabolism of F. indica and could be used to produce such valuable metabolites on commercial scale.

Zinc oxide nanoparticles (ZnO-NPs) have been produced by physical and chemical methods. Here, the comparative evaluation of both chemically-synthesised ZnO-NPs (C-ZNPs) and in-vitro cultured S. marianum mediated green-synthesised ZnO-NPs (G-ZNPs) were investigated on seed germination frequency, root and shoot growth, callus induction and biochemical profile of medicinally important plant Silybum marianum. Of all the treatments, callus-mediated ZnO-NPs gave optimum results for seed germination (65%), plantlet’s root length (4.3 cm), shoot length (5.3 cm) and fresh and dry weights (220.4 g L−1 and 21.23 g L−1, respectively). Similarly, the accumulation of phenolic (12.3 µg/mg DW) and flavonoid (2.8 µg/mg DW) contents were also enhanced in callus cultures treated with G-ZNPs. We also observed maximum antioxidant activity (99%) in callus cultures treated with G-ZNPs, however, in case of plantlets, these activities were found highest for in-vitro whole plant-mediated ZnO-NPs. Moreover, G-ZNPs also enhanced total protein content (265.32 BSAE/20g FW) in callus cultures. G-ZNPs were further assessed for their effects on several multidrug resistant bacterial strains and human liver carcinoma (HepG2) cells and our findings revealed that callus extracts treated with G-ZNPs show ameliorated antibacterial (highest zone of inhibition (19 mm) against Klebsiella pneumonia) and anticancer (highest cytotoxicity of 64%) activities.

Nano-elicitation is one among the prioritised strategies considered globally for sustainable and uniform production of industrially important medicinal compounds. Ocimum basilicum (Thai basil), a renowned medicinal species is a reservoir of commercially vital metabolites and proved for its health assuring effects in cancer, diabetes, microbial and cardiovascular diseases. However, its consumption and industrial demand raised intent to divert towards better alternates for ensuring sustainable production of medicinal compounds. Herein, we investigated the comparative potential of metal oxide [copper oxide (CuO) and manganese oxide (MnO)] nanoparticles to elicit the biosynthesis of bioactive metabolites and antioxidative capacity of O.basilicum callus cultures. Results showed that callus grown on MS media supplemented with 10 mg/L CuO-NPs resulted in the highest biomass accumulation (FW: 172.8 g/L, DW: 16.7 g/L), phenolic contents (TPC: 27.5 mg/g DW), and flavonoid contents (TFC: 9.1 mg/g DW) along with antioxidant activities (DPPH: 94%, ABTS: 881 μM TEAC, FRAP: 386 μM TEAC) compared with MnO-NPs and control. Likewise, the Superoxide dismutase (SOD: 1.28 nM/min/mg FW) and Peroxidase (POD: 0.48 nM/min/mg FW) activities were also recorded maximum in CuO-NPs elicited cultures than MnO-NPs and control. Moreover, the HPLC results showed that rosmarinic acid (11.4 mg/g DW), chicoric acid (16.6 mg/g DW), eugenol (0.21 mg/g DW) was found optimum in cultures at 10 mg/L CuO-NPs. Overall, it can be concluded that CuO nanoparticles can be effectively used as a elicitor for biosynthesis of metabolites in callus cultures of O. basilicum (Thai basil). The study is indeed a contribution to the field that will help decoding the mechanism of action of CuO NPs. However, further molecular investigations are needed to fully develop understanding about the metabolic potential of O. bascillicum and scalling up this protocol for bulkup production of bioactive compounds.

Prunella vulgaris L. is one of the therapeutic herbs containing various polyphenolics, which is used for multiple medicinal purposes. In this study, plant growth regulators (PGRs)-induced calli cultures from seed-derived leaf explants were exploited for the production of stress enzymes and polyphenolics. A growth curve was plotted for each PGR for 49 days period, which showed a distinct lag, log and decline phases. Here, the combination of naphthalene acetic acid (NAA) and 6-benzyleadenine (BA; 0.5 and 2.0 mg l−1) produced maximum fresh (6.32 FW-g/100 ml) and dry biomass (0.75 DW-g/100 ml) in contrast to control. The maximum synthesis of SOD (0.0154 FW-nM/min/mg) was detected on media comprising mixture of NAA and BA (1.5 mg l−1), while POD enzyme (0.366 FW-nM/min/mg) was higher at 0.5 mg l−1 NAA and 2, 4-dichlorophenoxy acetic acid. Further, NAA and BA (1.5 and 2.0 mg l−1) boosted up the synthesis of phenolics (18.83 GAE-mg/g-DW) and flavonoids content (18.05 RE-mg/g-DW) than control. Moreover, NAA of 1.0 and 2.0 mg l−1 were found supportive for maximum antioxidant activity (87.4%) and total protein (716 µg BSAE/mg-DW). This study will contribute in the development of cell culture in fermenter and synthesis of antioxidant secondary metabolites for commercial uses.

Virus-like particles (VLPs) are nanostructures that possess diverse applications in therapeutics, immunization, and diagnostics. With the recent advancements in biomedical engineering technologies, commercially available VLP-based vaccines are being extensively used to combat infectious diseases, whereas many more are in different stages of development in clinical studies. Because of their desired characteristics in terms of efficacy, safety, and diversity, VLP-based approaches might become more recurrent in the years to come. However, some production and fabrication challenges must be addressed before VLP-based approaches can be widely used in therapeutics. This review offers insight into the recent VLP-based vaccines development, with an emphasis on their characteristics, expression systems, and potential applicability as ideal candidates to combat emerging virulent pathogens. Finally, the potential of VLP-based vaccine as viable and efficient immunizing agents to induce immunity against virulent infectious agents, including, SARS-CoV-2 and protein nanoparticle-based vaccines has been elaborated. Thus, VLP vaccines may serve as an effective alternative to conventional vaccine strategies in combating emerging infectious diseases.

Moringa oleifera, also called miracle tree, is a pharmaceutically important plant with a multitude of nutritional, medicinal, and therapeutic attributes. In the current study, an in-vitro-based elicitation approach was used to enhance the commercially viable bioactive compounds in an in vitro callus culture of M. oleifera. The callus culture was established and exposed to different monochromatic lights to assess the potentially interactive effects on biomass productions, biosynthesis of pharmaceutically valuable secondary metabolites, and antioxidant activity. Optimum biomass production (16.7 g/L dry weight), total phenolic contents (TPC: 18.03 mg/g), and flavonoid contents (TFC: 15.02 mg/g) were recorded in callus cultures placed under continuous white light (24 h), and of other light treatments. The highest antioxidant activity, i.e., ABTS (550.69 TEAC µM) and FRAP (365.37 TEAC µM), were also noted under white light (24 h). The analysis of phytochemicals confirmed the significant impact of white light exposures on the enhanced biosynthesis of plant secondary metabolites. The enhanced levels of secondary metabolites, i.e., kaempferol (1016.04 µg/g DW), neochlorogenic acid (998.38 µg/g DW), quercetin (959.92 µg/g DW), and minor compounds including luteolin, apigenin, and p-coumaric acid were observed as being highest in continuous white light (24 h with respect to the control (photoperiod). Similarly, blue light enhanced the chlorogenic acid accumulation. This study shows that differential spectral lights demonstrate a good approach for the enhancement of nutraceuticals along with novel pharmacologically important metabolites and antioxidants in the in vitro callus culture of M. oleifera.

Flavonoids are secondary metabolites that represent a heterogeneous family of plant polyphenolic compounds. Recent research has determined that the health benefits of fruits and vegetables, as well as the therapeutic potential of medicinal plants, are based on the presence of various bioactive natural products, including a high proportion of flavonoids. With current trends in plant metabolite research, flavonoids have become the center of attention due to their significant bioactivity associated with anti-cancer, antioxidant, anti-inflammatory, and anti-microbial activities. However, the use of traditional approaches, widely associated with the production of flavonoids, including plant extraction and chemical synthesis, has not been able to establish a scalable route for large-scale production on an industrial level. The renovation of biosynthetic pathways in plants and industrially significant microbes using advanced genetic engineering tools offers substantial promise for the exploration and scalable production of flavonoids. Recently, the co-culture engineering approach has emerged to prevail over the constraints and limitations of the conventional monoculture approach by harnessing the power of two or more strains of engineered microbes to reconstruct the target biosynthetic pathway. In this review, current perspectives on the biosynthesis and metabolic engineering of flavonoids in plants have been summarized. Special emphasis is placed on the most recent developments in the microbial production of major classes of flavonoids. Finally, we describe the recent achievements in genetic engineering for the combinatorial biosynthesis of flavonoids by reconstructing synthesis pathways in microorganisms via a co-culture strategy to obtain high amounts of specific bioactive compounds

Elicited artificial in vitro cultures are gaining more interest due to their uniform biosynthesis of industrially valuable secondary metabolites. In this study, a unique methodology was applied, in which different ratios of gold (Au) and silver (Ag) nanoparticles (NPs) were supplemented to submerge cultures to investigate sustainable production of biomass and antioxidant secondary metabolites. Cell suspension cultures were exposed to Ag and AuNPs alone or different ratios of AgAuNPs (1:2; 1:3; 2:1; 3:1) in combination with NAA. The combination of AgAuNPs (3:1) with NAA enhanced fresh (9.25 g/100 ml) and dry biomass (0.64 g/100 ml) of suspended cells than control (6.67; 0.233 g/100 ml). AuNPs with NAA-augmented media enhanced biomass accumulation in lag, log and stationary phases in a period of 49 days. Furthermore, AgAu (3:1) and AgAuNPs (2:1; 1:2) with NAA enhanced protein contents, peroxidase and superoxide dismutase enzymes. However, maximum phenolics (TPC; 10.61 mg/g-DW) and flavonoids (7.62 mg/g-DW) were observed in cell cultures exposed to a combination of AgAuNPs (1:3) and NAA than control (6.27, 5.49 mg/g-DW). The combination of AgAuNPs (2:1) with NAA enhanced antioxidant activity (87.85%) in cell cultures. This study will help in illuminating the impact of NPs on cultures development and production of natural antioxidants.

Zinc oxide nanoparticles (NPs) have emerged as a novel elicitor for enhanced biosynthesis of secondary metabolites in in vitro plant cell cultures. The current study was aimed to explore elicitation abilities of ZnO-NPs for enhanced accumulation of lignans and neolignans in cell cultures of Linum usitatissimum. We optimized concentration of zinc oxide NPs before carrying out a full-fledged experiment. Subsequently, an optimum dose of 100 mg/l was introduced into the culture medium on day 0, days 0 and 15, and finally days 0 and 25. We observed that repeated elicitation stimulated various parameters and physiological responses in Linum usitatissimum cell cultures than one-time elicitation. Repeated elicitation of cell cultures on day 0 and 15 resulted in highest fresh weight (412.16 g/l) and lignans production (secoisolariciresinol diglucoside 284.12 mg/l: lariciresinol diglucoside 86.97 mg/l). Contrarily, repeated elicitation on day 0 and 25 resulted in highest DW (13.53 g/l), total phenolic production (537.44 mg/l), total flavonoid production (123.83 mg/l) and neolignans production (dehydrodiconiferyl alcohol glucoside 493.28 mg/l: guaiacylglycerol-β-coniferyl alcohol ether glucoside 307.69 mg/l). Enhancement in plant growth and secondary metabolites accumulation was several fold higher than controls. Furthermore, a linear relationship existed between total phenolic and flavonoid contents which in turn was correlated with higher antioxidant activities.

Liver and kidney diseases are the most frequently encountered problems around the globe. Damage to the liver and kidney may occur as a result of exposure to various drugs, chemicals, toxins, and pathogens, leading to severe disease conditions such as cirrhosis, fibrosis, hepatitis, acute kidney injury, and liver and renal failure. In this regard, the use of nanoparticles (NPs) such as silver nanoparticles (AgNPs), gold nanoparticles (AuNPs), and zinc oxide nanoparticles (ZnONPs) has emerged as a rapidly developing field of study in terms of safe delivery of various medications to target organs with minimal side effects. Due to their physical characteristics, NPs have inherent pharmacological effects, and an accidental buildup can have a significant impact on the structure and function of the liver and kidney. By suppressing the expression of the proinflammatory cytokines iNOS and COX-2, NPs are known to possess anti-inflammatory effects. Additionally, NPs have demonstrated their ability to operate as an antioxidant, squelching the generation of ROS caused by substances that cause oxidative stress. Finally, because of their pro-oxidant properties, they are also known to increase the level of ROS, which causes malignant liver and kidney cells to undergo apoptosis. As a result, NPs can be regarded as a double-edged sword whose inherent therapeutic benefits can be refined as we work to comprehend them in terms of their toxicity.

Platinum nanoparticles (PtNPs) are an attractive candidate for application in many areas of biotechnology, nanomedicine, and pharmacology, owing to their large surface area and biological properties. Different approaches, including physical, chemical, and biological (plants, bacteria, algae, and fungi) are presently being used for synthesis of PtNPs. However, these conventional methods (physical and chemical) present potential threat to health and environment because of the use of harsh chemicals and hazardous reaction conditions, except biological synthesis means which are substantially considered ecofriendly, economic and non-toxic. PtNPs are extensively studied due to their potent physicochemical and biological properties including antioxidant, antimicrobial and anticancer properties. This review offers a comprehensive assessment of the current knowledge about the synthesis of PtNPs using physical, chemical, and biological (plants, bacteria, algae, and fungi) approaches and their potential antimicrobial and biomedical applications especially in cancer and photothermal therapy. Interestingly, this review highlights role of PtNPs as nano-diagnostic and nanomedicine. Furthermore, nanotoxicity related to PtNPs, as well as the future prospects and opportunities of PtNPs based nano-therapeutics are also discussed. Overall, the emerging potential biological applications of PtNPs makes it viable to foresee their more promising outcomes in biomedical field in the upcoming future.

Brassica oleracea var. acephala is listed as the healthiest vegetable due to its high valued secondary metabolites content and antioxidant potential. This study was conducted to establish adventitious roots (ARs) culture as an alternative and feasible production of antioxidant secondary metabolites. ARs were induced from cotyledon explants in commercially available Murashige and Skoog (MS) plant nutrient media, gelled with 0.8% phyto-agar and supplemented with different concentration (0.1 ̶ 1.5 mg·L-1) of auxins (α-Naphthalene acetic acid; NAA, or Indole acetic acid; IAA, or Indole-3-butyric acid; IBA). AR formation responses in MS media at varying concentrations (0 ̶ 50 g·L-1) of sucrose and initial media pH (4, 5.0, 5.8, 7 & 8) were also studied. The bioprocessing of ARs were studied in liquid MS media containing NAA (1.5 mg·L-1) as growth regulator. The growth curve, important antioxidants (phenols & flavonoids), and free radical scavenging potential of ARs were studied for a period of 9-weeks. The ARs at stationary phase (7-week) attained highest accumulation of phenols and flavonoids, which ultimately showed the highest reactive species scavenging potential. This study provides the base for production of B. oleraceae var. acephala secondary metabolites on large scale to strengthen the bio-based economy of developing world.

Nanotechnology is an emerging area of research that deals with the production, manipulation, and application of nanoscale materials. Bio-assisted synthesis is of particular interest nowadays, to overcome the limitations associated with the physical and chemical means. The aim of this study was to synthesize ZnO nanoparticles (NPs) for the first time, utilizing the seed extract of Lepidium sativum. The synthesized NPs were confirmed through various spectroscopy and imagining techniques, such as XRD, FTIR, HPLC, and SEM. The characterized NPs were then examined for various in vitro biological assays. Crystalline, hexagonal-structured NPs with an average particle size of 25.6 nm were obtained. Biosynthesized ZnO NPs exhibited potent antioxidant activities, effective α-amylase inhibition, moderate urease inhibition (56%), high lipase-inhibition (71%) activities, moderate cytotoxic potential, and significant antibacterial activity. Gene expression of caspase in HepG2 cells was enhanced along with elevated production of ROS/RNS, while membrane integrity was disturbed upon the exposure of NPs. Overall results indicated that bio-assisted ZnO NPs exhibit excellent biological potential and could be exploited for future biomedical applications. particularly in antimicrobial and cancer therapeutics. Moreover, this is the first comprehensive study on Lepidium sativum-mediated synthesis of ZnO nanoparticles and evaluation of their biological activities.

Solanum virginianum L. (Solanum xanthocarpum) is an important therapeutic plant due to the presence of medicinally useful plant-derived compounds. S. virginianum has been shown to have anticancer, antioxidant, antibacterial, antiaging, and anti-inflammatory properties. This plant is becoming endangered due to overexploitation and the loss of its native habitat. The purpose of this research is to develop an ideal technique for the maximum biomass and phytochemical accumulation in S. virginianum leaf-induced in vitro cultures, as well as to evaluate their potential antiaging, anti-inflammatory, and antioxidant abilities. Leaf explants were grown on media (Murashige and Skoog (MS)) that were supplemented with various concentrations and combinations of plant hormones (TDZ, BAP, NAA, and TDZ + NAA) for this purpose. When compared with the other hormones, TDZ demonstrated the best response for callus induction, biomass accumulation, phytochemical synthesis, and biological activities. However, with 5 mg/L of TDZ, the optimal biomass production (FW: 251.48 g/L and DW: 13.59 g/L) was estimated. The highest total phenolic level (10.22 ± 0.44 mg/g DW) was found in 5 mg/L of TDZ, whereas the highest flavonoid contents (1.65 ± 0.11 mg/g DW) were found in 10 mg/L of TDZ. The results of the HPLC revealed that the highest production of coumarins (scopoletin: 4.34 ± 0.20 mg/g DW and esculetin: 0.87 ± 0.040 mg/g DW) was determined for 10 mg/L of TDZ, whereas the highest accumulations of caffeic acid (0.56 ± 0.021 mg/g DW) and methyl caffeate (18.62 ± 0.60 mg/g DW) were shown by 5 mg/L of TDZ. The determination of these phytochemicals (phenolics and coumarins) estimates that the results of our study on biological assays, such as antioxidant, anti-inflammatory, and antiaging assays, are useful for future cosmetic applications.

Over the last few decades, methods relating to plant tissue culture have become prevalent within the cosmetic industry. Forecasts predict the cosmetic industry to grow to an annual turnover of around a few hundred billion US dollars. Here we focused on Linum usitatissimum L., a plant that is well-known for its potent cosmetic properties. Following the a) establishment of cell cultures from three distinct initial explant origins (root, hypocotyl, and cotyledon) and b) selection of optimal hormonal concentrations, two in vitro systems (callus vs cell suspensions) were subjected to different light conditions. Phytochemical analysis by UPLC-HRMS not only confirmed high (neo)lignan accumulation capacity of this species with high concentrations of seven newly described (neo)lignans. Evaluation over 30 days revealed strong variations between the two different in vitro systems cultivated under light or dark, in terms of their growth kinetics and phytochemical composition. Additionally, antioxidant (i.e. four different in vitro assays based on hydrogen-atom transfer or electron transfer mechanism) and anti-aging (i.e. four in vitro inhibition potential of the skin remodeling enzymes: elastase, hyaluronidase, collagenase and tyrosinase) properties were evaluated for the two different in vitro systems cultivated under light or dark. A prominent hydrogen-atom transfer antioxidant mechanism was illustrated by the DPPH and ABTS assays. Potent tyrosinase and elastase inhibitory activities were also observed, which was strongly influenced by the in vitro system and light conditions. Statistical treatments of the data showed relationship of some (neo)lignans with these biological activities. These results confirmed the accumulation of flax (neo)lignans in different in vitro systems that were subjected to distinct light conditions. Furthermore, we showed the importance of optimizing these parameters for specific applications within the cosmetic industry.

Herein, we report a facile, economic, one-pot green synthesis of zinc oxide nanoparticles (ZnO-NPs) for diverse biomedical applications. In the study, ZnO-NPs were synthesized using an aqueous extract of Aquilegia pubiflora as an effective reducing and capping agent. The biomediated nanoparticles were characterized using various techniques including high-performance liquid chromatography (HPLC), X-ray diffraction, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy, Dynamic light scattering, photoluminescence, and RAMAN. The particles were highly pure, having an average size of 34.23 nm with spherical or elliptical morphology, and displayed good aqueous dispersion capability. FTIR and HPLC confirmed the successful capping of flavonoids and hydroxycinnamic acid derivatives. The characterized NPs were then explored for their antimicrobial and anti-leishmanial potential. Among the tested bacterial and fungal strains, ZnO-NPs were more potent toward Pseudomonas aeruginosa and Fusarium solani with inhibition zone of 10.3 ± 0.19 mm and 13 ± 1.4 mm, respectively. A dose-dependent cytotoxic effect was observed against Leishmania tropica (KWH23) with significant IC50 for both the promastigote (48 μg/mL) and amastigote form (51 μg/mL) of the parasite. In addition, bacterial kinase enzymes were inhibited by ZnO-NPs, thus allowing us to elaborate a possible action mechanism. Finally, the remarkable biocompatible nature of the particles was confirmed against freshly isolated human red blood cells (hRBCs). Altogether, these results affirmed the high antimicrobial and antiparasitic potential of ZnO-NPs obtained through a biogenic synthetic approach using aqueous extract of the Himalayan Columbine (Aquilegia pubiflora).

Eclipta alba L., also known as false daisy, is well known and commercially attractive plant with excellent hepatotoxic and antidiabetic activities. Light is considered a key modulator in plant morphogenesis and survival by regulating important physiological cascades. Current study was carried out to investigate growth and developmental aspects of E. alba under differential effect of multispectral lights. In vitro derived callus culture of E. alba was exposed to multispectral monochromatic lights under controlled aseptic conditions. Maximum dry weight was recorded in culture grown under red light (11.2 g/L) whereas negative effect was observed under exposure of yellow light on callus growth (4.87 g/L). Furthermore, red light significantly enhanced phenolics and flavonoids content (TPC: 57.8 mg/g, TFC: 11.1 mg/g) in callus cultures compared to rest of lights. HPLC analysis further confirmed highest accumulation of four major compounds i.e. coumarin (1.26 mg/g), eclalbatin (5.00 mg/g), wedelolactone (32.54 mg/g) and demethylwedelolactone (23.67 mg/g) and two minor compounds (β-amyrin: 0.38 mg/g, luteolin: 0.39 mg/g) in red light treated culture whereas stigmasterol was found optimum (0.22 mg/g) under blue light. In vitro based biological activities including antioxidant, antidiabetic and lipase inhibitory assays showed optimum values in cultures exposed to red light, suggesting crucial role of these phytochemicals in the enhancement of the therapeutic potential of E. alba. These results clearly revealed that the use of multispectral lights in in vitro cultures could be an effective strategy for enhanced production of phytochemicals.

Medicinal plants are an inevitable source of pharmaceutical drugs and most of the world population depends on these plants for health benefits. The increasing global demand for bioactive compounds from medicinal plants has posed a great threat to their existence due to overexploitation. Adventitious root and hairy root culture systems are an alternative approach to the conventional method for mass production of valuable compounds from medicinal plants owing to their rapid growth, biosynthetic and genetic stability. The main purpose of this review is to investigate the recent scientific research published worldwide on the application of adventitious and hairy root cultures to produce valuable compounds from medicinal plants. Furthermore, a comparison of adventitious root vs. hairy root cultures to produce valuable compounds has also been discussed. Various aspects such as medium composition, carbon source, pH, amount of macronutrients, optimization strategy, scale-up cultures, and use of biotic abiotic and nano-elicitors at various concentrations are the topic of discussion in this review. Several studies on adventitious and hairy root cultures of Polygonum multiflorum¸ Withania somnifera¸ Echinacea purpurea and Ajuga bracteosa have been discussed in detail which highlights the importance of elicitation strategies and bioreactor system, presenting commercial applications.

Nanotechnology is a new star in the science horizon with many valuable applications and promises to offer. It includes the synthesis and utilization of nanostructure materials ranging from 1 to 100 nm. Mostly these materials are generally (or “could be”) produced via the laborious and hazard-prone physical and chemical methods but the green synthesis approaches easier, safe and scalable have been recently developed. Among other metal oxides nanoparticles, Titanium oxide (TiO2) nanoparticles have been mostly exploited for their photocatalytic, antimicrobial and antiparasitic applications. A diverse set of biological entities are used to reduce the precursor metal salt into respective nanoparticles. The secondary metabolites present in organisms such as plants or microbes are involved in the bio-reduction and capping processes. This article will provide an overview of the green synthesis of TiO2 NPs from different biological extracts such as plants, microbes and biological products as well as their potential applications.

Withania somnifera L. is an endangered medicinal plant of higher market value. The in vitro callus cultures were established on Murashige and Skoog (MS) media augmented with different plant growth regulators. The MS medium containing 0.5 mg∙L−1 of each TDZ and NAA was found to be optimal for callus formation and growth. Further, callus cultures were raised in different light wavelengths to find the right wavelength carrying the photons for the ideal cell growth of W. somnifera. Among the different wavelengths, red light was best for maximum biomass accumulation in callus culture. However, violet light condition was proven to be favouring the phenols and flavonoids synthesis in the callus cultures. Compared to other wavelengths, red light grown callus extract showed significantly higher content of chlorogenic acid, and withaferin A. This study concludes that red light treatment was optimum for maximum biomass accumulation and anti-oxidant activity in calli of W. somnifera. Keywords: Withania, Light, Phenolic, Withanolides, Callus, Anti-oxidant

Stevia rebaudiana is an important medicinal plant that belongs to the Asteraceae family. The leaves of Stevia rebaudiana are a rich source of many health-promoting agents such as polyphenols, flavonoids, and steviol glycoside, which play a key role in controlling obesity and diabetes. New strategies such as the elicitation of culture media are needed to enhance the productivity of active components. Herein, the Cuscuta reflexa extracts were exploited as elicitors to enhance the productivity of active components. Cuscuta reflexa is one of the parasitic plants that has the ability to elongate very fast and cover the host plant. Consequently, it may be possible that the addition of Cuscuta reflexa extracts to adventitious root cultures (ADR) of Stevia rebaudiana may elongate the root more than control cultures to produce higher quantities of the desired secondary metabolites. Therefore, the main objective of the current study was to investigate the effect of Cuscuta reflexa extract as a biotic elicitor on the biomass accumulation and production of antioxidant secondary metabolite in submerged adventitious root cultures of Stevia rebaudiana. Ten different concentrations of Cuscuta reflexa were added to liquid media containing 0.5 mg/L naphthalene acetic acid (NAA). The growth kinetics of adventitious roots was investigated for a period of 49 days with an interval of 7 days. The maximum biomass accumulation (7.83 g/3 flasks) was observed on medium containing 10 mg/L extract of Cuscuta reflexa on day 49. As the concentration of extract increases in the culture media, the biomass gradually decreases after 49 days of inoculation. In this study, the higher total phenolics content (0.31 mg GAE/g-DW), total flavonoids content (0.22 mg QE/g-DW), and antioxidant activity (85.54%) were observed in 100 mg/L treated cultures. The higher concentration (100 mg/L) of Cuscuta reflexa extract considerably increased the total phenolics content (TPC), total phenolics production (TPP), total flavonoids content (TFC), total flavonoids production (TFP), total polyphenolics content (TPPC), and total polyphenolics production (TPPP). It was concluded that the extract of Cuscuta reflexa moderately improved biomass accumulation but enhanced the synthesis of phenolics, flavonoids, and antioxidant activities. Here, biomass’s independent production of secondary metabolites was observed with the addition of extract. The present study will be helpful to scale up adventitious roots culture into a bioreactor for the production of secondary metabolites rather than biomass accumulation in medicinally important Stevia rebaudiana.

Nanotechnology is a booming avenue in science and has a multitude of applications in health, agriculture, and industry. It exploits materials’ size at nanoscale (1–100 nm) known as nanoparticles (NPs). These nanoscale constituents are made via chemical, physical, and biological methods; however, the biological approach offers multiple benefits over the other counterparts. This method utilizes various biological resources for synthesis (microbes, plants, and others), which act as a reducing and capping agent. Among these sources, microbes provide an excellent platform for synthesis and have been recently exploited in the synthesis of various metallic NPs, in particular iron. Owing to their biocompatible nature, superparamagnetic properties, small size efficient, permeability, and absorption, they have become an integral part of biomedical research. This review focuses on microbial synthesis of iron oxide nanoparticles using various species of bacteria, fungi, and yeast. Possible applications and challenges that need to be addressed have also been discussed in the review; in particular, their antimicrobial and anticancer potentials are discussed in detail along with possible mechanisms. Moreover, some other possible biomedical applications are also highlighted. Although iron oxide nanoparticles have revolutionized biomedical research, issues such as cytotoxicity and biodegradability are still a major bottleneck in the commercialization of these nanoparticle-based products. Addressing these issues should be the topmost priority so that the biomedical industry can reap maximum benefit from iron oxide nanoparticle-based products.

Nanotechnology is a fast-expanding and multidisciplinary field with many applications in science and technology [...]

The green synthesis of nanoparticles has emerged as a simple, safe, sustainable, reliable and eco-friendly protocol. Among different types of NPs, green-synthesized zinc oxide NPs (ZnONPs) show various promising biological uses due to their interesting magnetic, electrical, optical and chemical characteristics. Keeping in view the dependence of the therapeutic efficacy of NPs on their physico-chemical characteristics, the green synthesis of ZnONPs using Casuarina equisetifolia leaf extract under UV-A and UV-C light was carried out in this study. UV-irradiation helped to control the size and morphology of ZnONPs by exciting the electrons in the photoactive compounds of plant extracts to enhance the bio-reduction of ZnO into ZnONPs. C. equisetifolia leaf extract was found enriched with phenolic (2.47 ± 0.12 mg GAE/g DW) and flavonoid content (0.88 ± 0.28 mg QE/g DW) contributing to its 74.33% free-radical scavenging activity. FTIR spectra showed the involvement of polyphenols in the bio-reduction, stabilization and capping of ZnONPs. Moreover, SEM-EDX and XRD analyses showed great potential of UV-C light in yielding smaller (34–39 nm) oval-shaped ZnONPs, whereas UV-A irradiation resulted in the formation of fairly spherical 67–71 nm ZnONPs and control ZnONPs were of mixed shape and even larger size (84–89 nm). Green-synthesized ZnONPs, notably CE-UV-C-ZnONPs, showed promising anti-bacterial activities against Bacillus subtilis, Pseudomonas fluorescens and Pseudomonas aeruginosa. Moreover, ZnONPs also enhanced ROS production which led to a significant loss of mitochondrial membrane potential and activated caspase-3 gene expression and caspase-3/7 activity in human hepatocellular carcinoma (HepG2) cells. CE-UV-C-ZnONP treatment reduced HepG2 cell viability to as low as 36.97% owing to their unique shape and smaller size. Lastly, ZnONPs were found to be highly biocompatible towards brine shrimp and human red blood cells suggesting their bio-safe nature. This research study sheds light on the plausible role of UV radiation in the green synthesis of ZnONPs with reasonable control over their size and morphology, thus improving their biological efficacy.

Fagonia indica is a rich source of pharmacologically active compounds. The variation in the metabolites of interest is one of the major issues in wild plants due to different environmental factors. The addition of chemical elicitors is one of the effective strategies to trigger the biosynthetic pathways for the release of a higher quantity of bioactive compounds. Therefore, this study was designed to investigate the effects of chemical elicitors, aluminum chloride (AlCl3) and cadmium chloride (CdCl2), on the biosynthesis of secondary metabolites, biomass, and the antioxidant system in callus cultures of F. indica. Among various treatments applied, AlCl3 (0.1 mM concentration) improved the highest in biomass accumulation (fresh weight (FW): 404.72 g/L) as compared to the control (FW: 269.85 g/L). The exposure of cultures to AlCl3 (0.01 mM) enhanced the accumulation of secondary metabolites, and the total phenolic contents (TPCs: 7.74 mg/g DW) and total flavonoid contents (TFCs: 1.07 mg/g DW) were higher than those of cultures exposed to CdCl2 (0.01 mM) with content levels (TPC: 5.60 and TFC: 0.97 mg/g) as compared to the control (TPC: 4.16 and TFC: 0.42 mg/g DW). Likewise, AlCl3 and CdCl2 also promoted the free radical scavenging activity (FRSA; 89.4% and 90%, respectively) at a concentration of 0.01 mM, as compared to the control (65.48%). For instance, the quantification of metabolites via high-performance liquid chromatography (HPLC) revealed an optimum production of myricetin (1.20 mg/g), apigenin (0.83 mg/g), isorhamnetin (0.70 mg/g), and kaempferol (0.64 mg/g). Cultures grown in the presence of AlCl3 triggered higher quantities of secondary metabolites than those grown in the presence of CdCl2 (0.79, 0.74, 0.57, and 0.67 mg/g). Moreover, AlCl3 at 0.1 mM enhanced the biosynthesis of superoxide dismutase (SOD: 0.08 nM/min/mg-FW) and peroxidase enzymes (POD: 2.37 nM/min/mg-FW), while CdCl2 resulted in an SOD activity up to 0.06 nM/min/mg-FW and POD: 2.72 nM/min/mg-FW. From these results, it is clear that AlCl3 is a better elicitor in terms of a higher and uniform productivity of biomass, secondary cell products, and antioxidant enzymes compared to CdCl2 and the control. It is possible to scale the current strategy to a bioreactor for a higher productivity of metabolites of interest for various pharmaceutical industries.

In vitro cultures of scarlet flax (Linum grandiflorum L.), an important ornamental flax, have been established as a new possible valuable resource of lignans and neolignans for antioxidant and anti-inflammatory applications. The callogenic potential at different concentrations of α-naphthalene acetic acid (NAA) and thidiazuron (TDZ), alone or in combinations, was evaluated using both L. grandiflorum hypocotyl and cotyledon explants. A higher callus induction frequency was observed on NAA than TDZ, especially for hypocotyl explants, with a maximum frequency (i.e., 95.2%) on 1.0 mg/L of NAA. The presence of NAA (1.0 mg/L) in conjunction with TDZ tended to increase the frequency of callogenesis relative to TDZ alone, but never reached the values observed with NAA alone, thereby indicating the lack of synergy between these two plant growth regulators (PGRs). Similarly, in terms of biomass, NAA was more effective than TDZ, with a maximum accumulation of biomass registered for medium supplemented with 1.0 mg/L of NAA using hypocotyls as initial explants (DW: 13.1 g). However, for biomass, a synergy between the two PGRs was observed, particularly for cotyledon-derived explants and for the lowest concentrations of TDZ. The influence of these two PGRs on callogenesis and biomass is discussed. The HPLC analysis confirmed the presence of lignans (secoisolariciresinol (SECO) and lariciresinol (LARI) and neolignan (dehydrodiconiferyl alcohol [DCA]) naturally accumulated in their glycoside forms. Furthermore, the antioxidant activities performed for both hypocotyl- and cotyledon-derived cultures were also found maximal (DPPH: 89.5%, FRAP 866: µM TEAC, ABTS: 456 µM TEAC) in hypocotyl-derived callus cultures as compared with callus obtained from cotyledon explants. Moreover, the anti-inflammatory activities revealed high inhibition (COX-1: 47.4% and COX-2: 51.1%) for extract of hypocotyl-derived callus cultures at 2.5 mg/L TDZ. The anti-inflammatory action against COX-1 and COX-2 was supported by the IC50 values. This report provides a viable approach for enhanced biomass accumulation and efficient production of (neo)lignans in L. grandiflorum callus cultures.

Healthcare, as a basic human right, has often become the focus of the development of innovative technologies. Technological progress has significantly contributed to the provision of high-quality, on-time, acceptable, and affordable healthcare. Advancements in nanoscience have led to the emergence of a new generation of nanostructures. Each of them has a unique set of properties that account for their astonishing applications. Since its inception, nanotechnology has continuously affected healthcare and has exerted a tremendous influence on its transformation, contributing to better outcomes. In the last two decades, the world has seen nanotechnology taking steps towards its omnipresence and the process has been accelerated by extensive research in various healthcare sectors. The inclusion of nanotechnology and its allied nanocarriers/nanosystems in medicine is known as nanomedicine, a field that has brought about numerous benefits in disease prevention, diagnosis, and treatment. Various nanosystems have been found to be better candidates for theranostic purposes, in contrast to conventional ones. This review paper will shed light on medically significant nanosystems, as well as their applications and limitations in areas such as gene therapy, targeted drug delivery, and in the treatment of cancer and various genetic diseases. Although nanotechnology holds immense potential, it is yet to be exploited. More efforts need to be directed to overcome these limitations and make full use of its potential in order to revolutionize the healthcare sector in near future.

Plant secondary metabolites are known to have a variety of biological activities beneficial to human health. They are becoming more popular as a result of their unique features and account for a major portion of the pharmacological industry. However, obtaining secondary metabolites directly from wild plants has substantial drawbacks, such as taking a long time, posing a risk of species extinction owing to over-exploitation, and producing a limited quantity. Thus, there is a paradigm shift towards the employment of plant tissue culture techniques for the production of key secondary metabolites in vitro. Elicitation appears to be a viable method for increasing phytochemical content and improving the quality of medicinal plants and fruits and vegetables. In vitro culture elicitation activates the plant’s defense response and increases the synthesis of secondary metabolites in larger proportions, which are helpful for therapeutic purposes. In this respect, light has emerged as a unique and efficient elicitor for enhancing the in vitro production of pharmacologically important secondary metabolites. Various types of light (UV, fluorescent, and LEDs) have been found as elicitors of secondary metabolites, which are described in this review.

Nanotechnology has emerged as a promising multidisciplinary field. It has shown several applications including diagnostics, imaging and structural design. Nanoparticles can be synthesized via chemical and physical approaches, carrying many threats to the ecosystem. To overcome these threats, sustainable routes for the synthesis of nanoparticles were implemented. Green synthesis is the most fascinating and attractive alternative to chemical synthesis as it offers more advantages. Nontoxic and eco-friendly secondary metabolites from plants are used as reducing and capping agents. This process is comparatively simple and cost-effective. A gold salt is simply reduced by biomolecules (phenols, alkaloids, proteins, etc.) present in the extracts of these plants. In this review, we have emphasized the synthesis and antimicrobial potential of gold nanoparticles using various plant extracts and their proposed mechanisms. Abbreviation: Au: aurum; gold; DLS: dynamic light scattering; EDAX: energydispersive X-ray analysis; EDS: energy-dispersive spectroscopy; FTIR: Fourier transform infrared spectroscopy; HRTEM: highresolution transmission electron microscopy; NPs: nanoparticles; UV–VIS: ultra violet-visible spectroscopy; SEM: scanning electron microscopy; XRD: X-ray Diffraction

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 potential to produce bioactive flavonolignans in the presence of chitosan. Established cell suspension cultures were 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 ± 1.7 g/L fresh weight (FW) and 17.7 ± 0.5 g/L dry weight (DW) productions. All chitosan treatments resulted in an overall increase in the accumulation of total flavonoids (5.0 ± 0.1 mg/g DW for 5.0 mg/L chitosan), total phenolic compounds (11.0 ± 0.2 mg/g DW for 0.5 mg/L chitosan) and silymarin (9.9 ± 0.5 mg/g DW for 0.5 mg/L chitosan). In particular, higher accumulation levels of silybin B (6.3 ± 0.2 mg/g DW), silybin A (1.2 ± 0.1 mg/g DW) and silydianin (1.0 ± 0.0 mg/g DW) were recorded for 0.5 mg/L chitosan. The corresponding extracts displayed enhanced antioxidant and anti-inflammatory capacities: in particular, high ABTS antioxidant activity (741.5 ± 4.4 μM Trolox C equivalent antioxidant capacity) was recorded in extracts obtained in presence of 0.5 mg/L of chitosan, whereas highest inhibitions of cyclooxygenase 2 (COX-2, 30.5 ± 1.3 %), secretory phospholipase A2 (sPLA2, 33.9 ± 1.3 %) and 15-lipoxygenase (15-LOX-2, 31.6 ± 1.2 %) enzymes involved in inflammation process were measured in extracts obtained in the presence of 5.0 mg/L of chitosan. Taken together, these results highlight the high potential of the chitosan elicitation in the S. marianum cell suspension for enhanced production of antioxidant and anti-inflammatory silymarin-rich extracts.

Flavonoids represent a popular class of industrially important bioactive compounds. They possess valuable health-benefiting and disease preventing properties, and therefore they are an important component of the pharmaceutical, nutraceutical, cosmetical and medicinal industries. Moreover, flavonoids possess significant antiallergic, antihepatotoxic, anti-inflammatory, antioxidant, antitumor, antiviral, and antibacterial as well as cardio-protective activities. Due to these properties, there is a rise in global demand for flavonoids, forming a significant part of the world market. However, obtaining flavonoids directly from plants has some limitations, such as low quantity, poor extraction, over-exploitation, time consuming process and loss of flora. Henceforth, there is a shift towards the in vitro production of flavonoids using the plant tissue culture technique to achieve better yields in less time. In order to achieve the productivity of flavonoids at an industrially competitive level, elicitation is a useful tool. The elicitation of in vitro cultures induces stressful conditions to plants, activates the plant defense system and enhances the accumulation of secondary metabolites in higher quantities. In this regard, nanoparticles (NPs) have emerged as novel and effective elicitors for enhancing the in vitro production of industrially important flavonoids. Different classes of NPs, including metallic NPs (silver and copper), metallic oxide NPs (copper oxide, iron oxide, zinc oxide, silicon dioxide) and carbon nanotubes, are widely reported as nano-elicitors of flavonoids discussed herein. Lastly, the mechanisms of NPs as well as knowledge gaps in the area of the nano-elicitation of flavonoids have been highlighted in this review.

With the increase in biotechnological, environmental, and nutraceutical importance of algae, about 100 whole genomic sequences of algae have been published, and this figure is expected to double in the coming years. The phenotypic and ecological diversity among algae hints at the range of functional capabilities encoded by algal genomes. In order to explore the biodiversity of algae and fully exploit their commercial potential, understanding their evolutionary, structural, functional, and developmental aspects at genomic level is a pre-requisite. So forth, the algal genomic analysis revealed us that algae evolved through endosymbiotic gene transfer, giving rise to around eight phyla. Amongst the diverse algal species, the unicellular green algae Chlamydomonas reinhardtii has attained the status of model organism as it is an ideal organism to elucidate the biological processes critical to plants and animals, as well as commercialized to produce range of bio-products. For this review, an overview of evolutionary process of algae through endosymbiosis in the light of genomics, as well as the phylogenomic, studies supporting the evolutionary process of algae was reviewed. Algal genomics not only helped us to understand the evolutionary history of algae but also may have an impact on our future by helping to create algae-based products and future biotechnological approaches.

Algae have long been exploited commercially and industrially as food, feed, additives, cosmetics, pharmaceuticals, and fertilizer, but now the trend is shifting towards the algae-mediated green synthesis of nanoparticles (NPs). This trend is increasing day by day, as algae are a rich source of secondary metabolites, easy to cultivate, have fast growth, and are scalable. In recent era, green synthesis of NPs has gained widespread attention as a safe, simple, sustainable, cost-effective, and eco-friendly protocol. The secondary metabolites from algae reduce, cap, and stabilize the metal precursors to form metal, metal oxide, or bimetallic NPs. The NPs synthesis could either be intracellular or extracellular depending on the location of NPs synthesis and reducing agents. Among the diverse range of algae, the most widely investigated algae for the biosynthesis of NPs documented are brown, red, blue-green, micro and macro green algae. Due to the biocompatibility, safety and unique physico-chemical properties of NPs, the algal biosynthesized NPs have also been studied for their biomedical applications, which include anti-bacterial, anti-fungal, anti-cancerous, anti-fouling, bioremediation, and biosensing activities. In this review, the rationale behind the algal-mediated biosynthesis of metallic, metallic oxide, and bimetallic NPs from various algae have been reviewed. Furthermore, an insight into the mechanism of biosynthesis of NPs from algae and their biomedical applications has been reviewed critically.

Thai basil is a renowned medicinal plant and a rich source of bioactive antioxidant compounds with several health benefits, with actions to prevent of cancer, diabetes and cardiovascular disease. Plant cell and tissue culture technologies can be routinely established as an important, sustainable and low-cost biomass source to produce high-value phytochemicals. The current study aimed at developing an effective protocol to produce Thai basil leaf-derived callus cultures with sustainable and high production of biomass and antioxidants as an alternative of leaves production. MS basal medium with various concentrations of plant growth regulators (PGRs) compatible with nutraceutical applications (i.e., gibberellic acid (GA3) and 6-benzylaminopurine (BAP) either alone or in combination with naphthalene acetic acid (NAA)) were evaluated. Among all tested PGRs, the combination BAP:NAA (5 mg/L:1 mg/L) yields the maximum biomass accumulation (fresh weight (FW): 190 g/L and dry weight (DW): 13.05 g/L) as well as enhanced phenolic (346.08 mg/L) production. HPLC quantification analysis indicated high productions of chicoric acid (35.77 mg/g DW) and rosmarinic acid (7.35 mg/g DW) under optimized callus culture conditions. Antioxidant potential was assessed using both in vitro cell free and in vivo cellular antioxidant assays. Maximum in vitro antioxidant activity DPPH (93.2% of radical scavenging activity) and ABTS (1322 µM Trolox equivalent antioxidant capacity) was also observed for the extracts from callus cultures grown in optimal conditions. In vivo cellular antioxidant activity assay confirmed the effective protection against oxidative stress of the corresponding extract by the maximum inhibition of ROS and RNS production. Compared to commercial leaves, callus extracts showed higher production of chicoric acid and rosmarinic acid associated with higher antioxidant capacity. In addition, this biological system also has a large capacity for continuous biomass production, thus demonstrating its high potential for possible nutraceutical applications.

Linum usitatissimum biosynthesizes lignans and neolignans that are diet and medicinally valuable metabolites. In recent years, zinc oxide nanoparticles (ZnONPs) have emerged as potential elicitors for the enhanced biosynthesis of commercial secondary metabolites. Herein, we investigated the influence of biogenic ZnONPs on both seedlings and stem-derived callus of L. usitatissimum. Seedlings of L. usitatissimum grown on Murashige and Skoog (MS) medium supplemented with ZnONPs (1–1000 mg/L) presented the highest antioxidant activity, total phenolic content, total flavonoid content, peroxidase and superoxide dismutase activities at 500 mg/L, while the maximum plantlet length was achieved with 10 mg/L. Likewise, the high-performance liquid chromatography (HPLC) analysis revealed the enhanced production of secoisolariciresinol diglucoside, lariciresinol diglucoside, dehydrodiconiferyl alcohol glucoside and guaiacylglycerol-β-coniferyl alcohol ether glucoside in the plantlets grown on the 500 mg/L ZnONPs. On the other hand, the stem explants were cultured on MS media comprising 1-naphthaleneacetic acid (1 mg/L) and ZnONPs (1–50 mg/L). The highest antioxidant and other activities with an enhanced rooting effect were noted in 25 mg/L ZnONP-treated callus. Similarly, the maximum metabolites were also accumulated in 25 mg/L ZnONP-treated callus. In both systems, the dose-dependent production of reactive oxygen species (ROS) was recorded, resulting in oxidative damage with a more pronounced toxic effect on in vitro cultures. Altogether, the results from this study constitute a first comprehensive view of the impact of ZnONPs on the oxidative stress and antioxidant responses in seedlings vs. in vitro cultures.

Solanum xanthocarpum is considered an important traditional medicinal herb because of its unique antioxidant, and anti-diabetic, anti-aging, and anti-inflammatory potential. Because of the over exploitation linked to its medicinal properties as well as destruction of its natural habitat, S. xanthocarpum is now becoming endangered and its supply is limited. Plant in vitro culture and elicitation are attractive alternative strategies to produce biomass and stimulate biosynthesis of medicinally important phytochemicals. Here, we investigated the potential influence of seven different monochromatic light treatments on biomass and secondary metabolites accumulation in callus culture of S. xanthocarpum as well as associated biological activities of the corresponding extracts. Among different light treatments, highest biomass accumulation was observed in white light-treated callus culture. Optimum accumulation of total flavonoid contents (TFC) and total phenolic contents (TPC) were observed in callus culture kept under continuous white and blue light respectively than control. Quantification of phytochemicals through HPLC revealed that optimum production of caffeic acid (0.57 ± 0.06 mg/g DW), methyl-caffeate (17.19 mg/g ± 1.79 DW), scopoletin (2.28 ± 0.13 mg/g DW), and esculetin (0.68 ± 0.07 mg/g DW) was observed under blue light callus cultures. Compared to the classic photoperiod condition, caffeic acid, methyl-caffeate, scopoletin, and esculetin were accumulated 1.7, 2.5, 1.1, and 1.09-folds higher, respectively. Moreover, high in vitro cell free antioxidant, anti-diabetic, anti-aging, and anti-inflammatory activities were closely associated with the production of these secondary metabolites. These results clearly showed the interest to apply multispectral light as elicitor of in vitro callus cultures S. xanthocarpum to promote the production of important phytochemicals, and allow us to propose this system as an alternative for the collection of this endangered species from the wild.