D pbs

Aurobindo Pharma, Hyderabad provided PIO racemic form and PIO-R and S for bioanalytical studies. The internal standard, Glimepiride, was obtained from Hetero Labs, Hyderabad. Merck, India supplied Ammonium acetate, Acetonitrile, and methanol HPLC grade for the study, while Millipore, MA, USA provided 0.45 µm pore size filters for filtering the mobile phase and solutions. The in vitro study utilized 3T3-L1-mouse embryo fibroblast cell lines (NCCS, Pune, Maharastra, India) and DMEM glucose-free medium (Himedia, Mumbai, India) for cell culture. The researchers also used adjustable multichannel pipettes and Fetal Bovine Serum (#RM10432, of Himedia, Mumbai, India), D-PBS (#TL1006, of Himedia, Mumbai, India), 2-NBDG (Invitrogen: Cat no. 11046, Cayman Chemical, Rajasthan, India), a 6-well cell culture plate (Biolite—Thermo, Bengaluru, Karnataka, India), 50 mL centrifuge tubes (# 546043 TORSON, Mysuru, Karnataka, India). A glucose reagent kit from AGAPPE (Mumbai, India) was used to assess glucose in rat plasma utilizing a semi-automated biochemistry analyzer (Labmate, Ranchi, India). In the study, all chemicals used were over 95% pure.

For the synthesis of SPIONs, the following analytical grade and extra pure chemicals were used: Ferrous sulphate heptahydrate (FeSO₄.7H₂O), extra pure, from HiMedia laboratories, Ferric chloride hexahydrate (FeCl_3. 6H₂O) from Thomas Baker PVT limited; Tetramethyl ammonium hydroxide (C₄H₁₃NO), and Formaldehyde solution (CH₂O) (41%), Acetone (C₃H₆O) from SDFCL,SDS (Sodium dodecyl Sulphate (C₁₂H₂₅NaO₄S) from Karnataka Fine Chem, Bangalore, and N, N Dimethylformamide (HCON(CH₃) from Fisher Scientific–Qualigens, Mumbai, Curcumin (C₂₁H₂₀O₆) from SDFCL, Chitosan (C₆H₁₁NO₄)_n (cell culture tested) from HiMedia, Pyridoxine hydrochloride (pure) (C₈H₁₁NO₃HCL) and Curcumin from SRL (Sisco Research Laboratories) Mumbai, RPMI 1640 medium (#AL199A, HiMedia), Fetal Bovine Serum (RM10432), HiMedia), Caspase 3 (Cat No: 560901, BD Biosciences) and D-PBS (#TL1006, HiMedia). Throughout the synthesis of nanoparticles, nitrogenized double distilled water was used. For functionalization, drug loading and final polymer coating freshly prepared double distilled water was used. For giving proper agitation during incubation time a cyclo-rotator was designed to fit our purpose. For cell culture experiments, HeLa cell line was obtained from Stellixer Biotec, Bengaluru, India.

CEO was purchased from WinLab, India. Tween 80 and Butanaol were procured from Sigma Aldrich, USA. Other materials such as A549 human alveolar lung adenocarcinoma cell line (NCCS, Pune), DMEM high-glucose media (Cat No: AL007, Himedia, Mumbai, India), Fetal Bovine Serum (#RM10432, Himedia), MTT Reagent (5 mg/mL) (# 4060 Himedia), D-PBS (#TL1006, Himedia), and 96-well plate for culturing the cells (from Corning, NY, USA) were directly procured from the company. There was no additional purification of the solvents and compounds of analytical grade.

Letrozole powder (HPLC purity 99.12%) was kindly provided by Hetero drugs Pvt. Ltd., Hyderabad. DMSO (#PHR1309), camptothecin (#C9911), ethanol, berberine chloride, ascorbic acid, and gold chloride (HAuCl₄.3H₂O) were purchased from Sigma Aldrich (St. Louis, MO, USA) and used without additional purification. The MDA-MB-231 human breast adenocarcinoma cell line (NCCS, Pune) was used. The cell culture medium—high-glucose DMEM media (Cat. #: AL007)—fetal bovine serum (#RM10432), MTT reagent (5 mg/mL) (# 4060), and D-PBS (#TL1006) were purchased from Himedia, India. A 96-well plate for culturing the cells (from Corning, Bedford, MA, USA) and T25 flask (# 12556009, Biolite-Thermo, Waltham, MA, USA) were also used.

Chitosan from crab shell, highly viscous was from sigma-Aldrich, glacial acetic acid from Merck, ethanol absolute 99.9% AR from Jebsen & Jessen GmbH & Co, Germany, Tripoly Phosphate (TPP) having >98% purity from sigma-Aldrich, Tween 80 for synthesis from Merck, 5-FU > 99% (HPLC) from sigma-Aldrich, Doxorubicin Hydrochloride 98.0–102.0% HPLC from sigma Aldrich, DMEM media from sigma Aldrich, Fetal Bovine Serum from Gibco, 0.25% Trypsinfrom Invitrogen and MTT from Sigma -Aldrich were used for the experiments. DMEM high glucose medium (#AL111, Himedia), Fetal Bovine Serum (#RM10432, Himedia), D-PBS (#TL1006, Himedia), Camptothecin-15 µM (Sigma), SDS 20% w/v (Himedia), Chloroform: isoamyl alcohol (24:1) (Himedia), Isopropanol (Himedia), Ethyl alcohol 70% v/v (Fisher Sceintific), Dihydroethidium (Hydroethidine) (Thermoscientific, Number: D-11347), DHE stock (5 mM, in DMSO), H₂O₂ (Fisher Scientific), Acridine orange - 20 µg/mL solution, Thermo Fischer, USA were used for the analyses. All experiments were carried out using triple distilled water.