Shake-flask in vitro culture of Buddleja cordata cells produces large amounts of biomass and synthetizes verbascoside (VB), linarin and hydroxycinnamic acids, bioactive phenolic secondary metabolites (PSMs). In this work, we determined the effect of stirring speed on the growth of and production of PSMs [total phenolic, phenylethanoid glycoside and flavonoid contents (PeC, PeGC and FC, respectively)] by B. cordata cells cultured in two bioreactors. Two different stirring speeds (120 and 400 rpm) were tested in two stirred-tank bioreactors: a 2 L bioreactor equipped with a ring diffuser (B2RD) and a 3 L bioreactor with a sintered diffuser (B3SD). Growth kinetics of B. cordata cells were measured in the bioreactors and shake-flask systems. The stirring speed and type of bioreactor affected phases, parameters of growth and production of PSMs. The highest production of biomass (13.62 g L −1) and PSMs [PeC of 64.63 mg gallic acid equivalents g −1 (mg GAE g −1); PeGC of 119.24 mg VB equivalents g −1 (mg VBE g −1); and FC of 5.02 mg quercetin equivalents g −1 (mg QE g −1)] occurred in B2RD at 400 rpm. These values were similar to the found in shake-flasks system. This work establishes the basis for bioprocess advances of B. cordata focused on the development of a sustainable strategy for the management of natural resources and as a source of bioactive PSMs on a large scale. Key message Buddleja cordata cells cultured in a mechanically agitated bioreactor possess an outstanding biosynthetic potential that represents a suitable biotechnological alternative for the production of bioactive phenolic secondary metabolites. Keywords Bioreactor • Buddleja cordata • Phenolic secondary metabolites • Diffuser • Stirring speed • Verbascoside Abbreviations µ Specific growth rate ANOVA Analysis of variance B2RD 2 L Bioreactor equipped with a ring diffuser B3SD 3 L Bioreactor equipped with a sintered diffuser CSCBc Cell suspension culture of B. cordata CV Cellular viability Communicated by Sergio J. Ochatt.

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Arnica montana cell suspension culture could be a sustainable source of a vegetal material producer of secondary metabolites (SMs) possessing biological effects. Different plant growth regulator concentrations (0–5 mg/L) were tested in foliar explants to induce a callus that was used to establish a cell suspension culture. Growth kinetics was carried out for 30 days. A methanolic extract obtained from biomass harvested at 30 days of growth kinetics was fractionated, and three fractions were tested for bioactivities. We induced a callus with 1 mg/L of picloram and 0.5 mg/L of kinetin in foliar explants, which allowed for the establishment of a cell suspension culture, and the latter had the highest total SMs contents at day 30. Three fractions showed differences in total SMs contents, with the highest values per gram as follows: 270 mg gallic acid equivalent for total phenolic content, 200 mg quercetin equivalent for total flavonoid content, 83 mg verbascoside equivalent for total phenolic acid content, and 396 mg parthenolide equivalent for total sesquiterpene lactone content. The best bioactivities were 2–6 µg/mL for the 50% inhibition of 2,2-diphenyl-1-picrylhydrazyl radical, 30% cellular viability of lymphoma cells at 40 µg/mL, 17% inhibition against Escherichia coli and Staphylococcus aureus at 8 µg/disk, and α-amylase inhibition at 12% with 10 µg/mL. The total SMs contents were correlated with bioactivities.

para preparar extractos acuosos con posible aplicación biotecnológica o farmacológica,

The research on compounds exhibiting photoprotection against ultraviolet radiation (UVR) is a matter of increasing interest. The methanolic extract of a cell culture of Buddleja cordata has potential photoprotective effects as these cells produce phenolic secondary metabolites (SMs). These metabolites are attributed with biological activities capable of counteracting the harmful effects caused by UVR on skin. In the present work, the methanolic extract (310–2500 µg/mL) of B. cordata cell culture showed a photoprotective effect on UVB-irradiated 3T3-Swiss albino fibroblasts with a significant increase in cell viability. The greatest photoprotective effect (75%) of the extract was observed at 2500 µg/mL, which was statistically comparable with that of 250 µg/mL verbascoside, used as positive control. In addition, concentrations of the extract higher than 2500 µg/mL resulted in decreased cell viability (≤83%) after 24 h of exposure. Phytochemical analysis of the extract allowed us to determine that it was characterized by high concentrations of total phenol and total phenolic acid contents (138 ± 4.7 mg gallic acid equivalents and 44.01 ± 1.33 mg verbascoside equivalents per gram of extract, respectively) as well as absorption of UV light (first and second bands peaking at 294 and 330 nm, respectively). Some phenylethanoid glycosides were identified from the extract.

Buddleja cordata is a species used by Mexican folk medicine for treating illnesses related to inflammation such as skin wounds and arthritis. It bio-synthesizes metabolites such as verbascoside that contributes to its ethno-therapeutic properties as antiinflammatory remedy. HPLC analysis showed that the methanolic extract from cell suspension cultures (Bc-Cc) and wild plant leaves (Bc-Wp) contained verbascoside, but concentration was higher in Bc-Cc (87.48 mg/g) than in wild plant (47.34 mg/g).In the acute toxicity model, none of the extracts generated any lethality or adverse effects. In acute inflammation model induced with TPA, Bc-Cc extract showed a greater edema inhibition at 2 mg/ear (61.72%), as well for carrageenan model at 200 mg/kg (48.87%). Bc-Wp showed lesser anti-inflammatory effect in both acute inflammation models than Bc-Cc. For Adjuvant-induced arthritis both extracts at 250 mg/kg generated a moderate inhibition over edema (≈ 33%) at day 28, and they were statistically no different to phenilbutazone. The culture in suspension of B. cordata obtained by biotechnological process contains greater amount of verbascosido and showed better anti-inflammatory activity; thus, representing a source for obtaining this type of secondary metabolite of pharmacological interest. Keywords: cell suspension culture, Buddleja cordata, verbascoside, anti-inflammatory activity, median lethal dose. ResumenBuddleja cordata es una planta medicinal usada en México para el tratamiento de enfermedades relacionadas con heridas en la piel y artritis. Esta especie bio-sintetiza metabolitos secundarios como el verbascósido, el cual contribuye a sus propiedades etno-medicinales. El análisis por HPLC mostró que el extracto metanólico del cultivo en suspensión celular (Bc-Cc) y de las hojas de la planta silvestre (Bc-Wp) contienen verbascósido, siendo su concentración mayor en el cultivo Bc-Cc (87.48 mg/g) que en la planta silvestre (47.34 mg/g). Ninguno de los extractos generó letalidad o efectos adversos en el ensayo de toxicidad aguda. En el modelo de inflamación tópica inducido con TPA, Bc-Cc mostró mayor actividad antiinflamatoria; a 2 mg/oreja generó 61.72% de inhibición y en el modelo de carragenina a 200 mg/kg generó 48.87%, mientras que Bc-Wp mostró menor actividad anti-inflamatoria en ambos modelos. En el modelo de artritis inducida con Adyuvante de Freud completo, ambos extractos a 250 mg/kg mostraron moderada actividad anti-inflamatoria, con inhibición de aprox. 33% al día 28. El cultivo en suspensión celular de B. cordata obtenido por proceso biotecnológico contiene mayor cantidad de verbascosido y mostró mejor actividad anti-inflamatoria; por lo que representa una fuente para la obtención de este tipo de metabolito secundario de interés

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