Doxorubicin dox

Human OS cell lines U2OS, MG-63, Saos-2, HOS, and normal osteoblasts hFOB1.19 were obtained from American Type Culture Collection (ATCC), were cultured in DMEM or McCoy’s 5A medium (Gibco) with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Cat#C0222; Beyotime) in an incubator at 37ºC with 5% CO₂. Cisplatin, Sorafenib, and Doxorubicin (Dox) were purchased from MedChem Express (Shanghai, China). The control miRNA, miR-605-3p mimics and antagomir were provided by GenePharma (Shanghai, China). Cell transfection was conducted using lipofectamine 3000 according to the manufacturer’s protocol.

PLGA NPs were synthesized by a double emulsion solvent evaporation (W/O/W) technique as previously published, with modifications [33 (link)]. For this purpose, 25 mg of PLGA (acid terminated, lactide: glycolide 50:50, Mw 24–38 kDa) (Resomer RG 503 H. Sigma-Aldrich) was dissolved in 1.5 ml dichloromethane (DCM) alone (empty PLGA) or with coumarin-6 (Sigma-Aldrich) (37.5 µg), IR780 (1 mg) (Sigma-Aldrich), epirubicin (EPI) (2 mg) or doxorubicin (DOX) (2 mg) (MedChem Express). This organic solution was emulsified dropwise with 0.5 ml of polyvinyl alcohol (PVA) (0.001%) with a probe sonicator (100 W, 100% amplitude) for 3 min (UP100H, Hielscher). The obtained emulsion (W/O) was added dropwise to 24.5 ml of PVA (0.001%) and sonicated again (100 W, 100% amplitude) for 3 min to form a double emulsion (W/O/W). The DCM was evaporated by magnetic stirring for 2 h. For docetaxel (DOC) encapsulation, the above protocol was modified according to Cho et al. [34 ]. For this purpose, 6 mg of docetaxel (MedChem Express) was dissolved in 0.5 ml of DCM and subsequently, 25 mg of PLGA was added. Both solutions were mixed and sonicated (100 W, 100% amplitude) for 2 min. The obtained mixture was added to 10 ml of PVA (0.2%) and sonicated again (100 W, 100% amplitude) for 10 min. The DCM was evaporated by magnetic stirring for 2–3 h.

Propegal S.L. has supplied the skin shark fish wastes (BS—blue shark, Prionace glauca) to produce the gelatin that was extracted in the laboratory of Recycling and Valorisation of Waste Materials (REVAL) of the Instituto de Investigacións Mariñas de Vigo (IIM) CSIC (Spanish National Research Council) in Vigo, Galicia, Spain [85 (link)].
The blue shark skin type was dispersed in distilled water and heated slowly up to 60 °C to then cooled to room temperature (22 °C) to produce a gel upon cooling. The selected weight of Shark gelatin was measured by a Mettler AE-240 electronic balance, with an accuracy of 5·10⁻⁵ g. Thereafter, the gelatin is blended with a known volume of distilled water to obtain the targeted weight fraction gel composition (25% wt). Furthermore, the gelatin solution was agitated using an Ultrasonic Bath (J. P. Selecta.S.A., 120 W) for 30 min to dissolve the fish collagen adequately and thus, forming a homogenous gel.
The weight of the obtained gelatin was measured in a Mettler AE-240 balance with an accuracy of 5 10^–5 g. Afterwards, The weighted gelatin was blended in the selected volume of deionized water. Ionogels, with percent weight concentrations of 25% wt for the gelatin. An ultrasonic bath (J. P. Selecta.S.A., 120 W) was used to obtain the correct dispersion. The FIL employed 1-ethyl-3-methylpyridinium perfluorobutanesulfonate ([C₂C₁py][C₄F₉SO₃]) (>99% mass fraction purity) was obtained by Iolitec (Heilbronn, Germany). [C₂C₁py] [C₄F₉SO₃] features at 30 °C a density of 1.52 g·cm⁻³ with a viscosity of 150.3 mPa s, and molar volume of 279.01 cm³·mol⁻¹.
For the therapeutic agents, (MTM) was purchased from EntreChem SL and Doxorubicin (DOX) was purchased from MedChemExpress.