L ascorbic acid

2,2-Diphenyl-1-picrylhydrazyl (DPPH), Folin-Ciocalteu phenol reagent, L-ascorbic acid, di-sodium hydrogen phosphate de-hydrate, and sodium dihydrogen phosphate dehydrate were purchased from HiMedia Laboratories Pvt. Ltd. (India). HPLC grade water, sodium carbonate, and hydrogen peroxide were procured from Loba Chemie Pvt. Ltd. (India). Gallic acid, quercetin, ellagic acid, vanillic acid, catechin, epigallocatechin gallate (EGCG), and DCM were procured from Sigma-Aldrich (USA). LC–MS grade acetonitrile, methanol, and formic acid (of 98% purity) were procured from Fisher Chemicals (Hampton, NH, USA). Colistin was procured from HiMedia Laboratories Pvt. Ltd. (India). Labeled, CHROMAFIL Xtra poly ether sulfone (PES), 25 mm, and 0.20 μm syringe filters were purchased from Macherey-Nagel (Düren, Germany). UV-vis spectrophotometer (Evolution 201) was procured from Thermo Fisher Scientific- Shanghai (China). ORBITEK shaker was purchased from Scigenics Biotech Pvt. Ltd. (India). Refrigerated centrifuge C-24 Plus was from REMI Sales and Engineering Ltd. (India). Analytical balance Aczet, CY2202 was purchased from Mettler-Toledo Pvt. Ltd. (India). Rotary vacuum evaporator-RE-52 was purchased from SONAR (India). Micropipette (20-200, 100-1000, and 0.5-10 μL) of variable volume purchased from HiMedia Laboratories Pvt. Ltd. (India).

The UC-MSCs were cultured in complete DMEM for three weeks along with osteogenic induction medium (consisting of 10 mM β-glycerophosphate (Sigma Aldrich, Saint Louis, MO, USA), 100 nM dexamethasone (Himedia, Mumbai, India), and 50 µg/mL l-Ascorbic acid (Himedia, Mumbai, India)), or adipogenic induction medium containing 1μM dexamethasone, 0.5 mM 3-isobutyl-l-methyl xanthine, 200 μM indomethacine, and 10 μg/mL of insulin, or chondrogenic induction medium [34 (link)]. After three weeks of culturing in osteogenic induction medium, alkaline phosphatase (ALP) staining was performed using ALP kit (Sigma-Aldrich, Saint Louis, MO, USA) according to the manufacturer’s protocol. After adipo-induction for four weeks, the UC-MSCs were stained with Oil Red O. Briefly; the cells were fixed with 10% formalin for 15 min and then incubated with 60% isopropanol for 5 min, following which the cells were stained with Oil Red O for 10 min. The petri plates were observed under microscope and images were taken. The chondrogenic assay was performed using Alcian blue staining as described by Eswaramoorthy et al. [34 (link)].

Flowers of W. fruticosa were harvested from the forest of district Mandi located in Himachal Pradesh, India. Analytical reagent grade emulsifying and a surface-active agent such as Gum arabic (spray dried) and tween-80 were procured from Sigma Aldrich Co. St. Louis, MO, USA. From Hi-Media Laboratories Pvt. Ltd., Mumbai, India, ethanol, L-ascorbic acid, calcium chloride, sodium chloride, phosphate buffer, Muller Hinton Agar (MHA), and Sabouraud Dextrose Agar (SDA) were purchased. For the oil phase and dispersion of flower extract, sunflower seed oil was used and it was obtained from CDH Pvt. Ltd., Mumbai, India. For antimicrobial studies, standard pathogenic bacterial strains such as Staphylococcus aureus (MTCC 3160), Klebsiella pneumoniae (MTCC 3384), Pseudomonas aeruginosa (MTCC 2295), and Salmonella typhmurium (MTCC 1254) and fungal strain of Candida albicans (MTCC 183) were obtained from IMTECH, Chandigarh, India.