Resomer rg 504 h

Resomer® RG 504 H (PLGA, LA:GA = 50:50, Mw = 38–54 k), Resomer® RG 502 H (PLGA, LA:GA = 50:50, Mw = 7–17 k,), salicylic acid (SA, ≥99.0%), poly(vinyl alcohol) (PVA, Mw = 13–23 k, 87–89% hydrolyzed), sodium dodecyl sulfate (SDS, ≥98.5%), and dimethyl sulfoxide (DMSO, ≥99.9%) were purchased from Sigma Aldrich (ON, CA). Acetonitrile (ACN, HPLC grade) and formic acid (FA, LC/MS grade) were purchased from Fisher Chemical (ON, CA). Dichloromethane (DCM, HPLC grade) and methanol (HPLC grade) were purchased from Caledon Laboratories Ltd. (ON, CA). Olaparib (OLA, ≥99%) was purchased from MedKoo Bioscience (NC, US). Phosphate buffered saline (PBS, Gibco, pH = 7.4) was purchased from Thermofisher (Massachusetts, USA). Tween 80 (reagent grade) was purchased from BioShop (ON, CA).

Sodium hyaluronate (Mw 1.2 × 10⁶ Da) was purchased from Lifecore Biomedical, USA. Polyvinyl alcohol (Mw 13k–23k Da), 4‐(4.6‐dimethoxy‐1,3,5‐triazin‐2‐yl)‐4‐methylmorpholinium chloride (DMTMM), PLGA (Resomer RG 504H, poly(d,l‐lactide‐co‐glycolide); 50:50, average Mw ≈ 38k–54k Da), and chitosan (average Mw ≈ 50k–190k Da) were purchased from Sigma‐Aldrich. LPS, FITC‐dextran (Mw 4k Da), Alcian blue, Nuclear Fast Red were purchased from Sigma‐Aldrich. ELISA kits for IL‐6 and TNF‐α were purchased from Bio‐legend, Multiplex ELISA pro‐inflammatory panel was purchased from Meso Scale Discovery. E‐Cadherin monoclonal antibody (Alexa Fluor 488 conjugate; eBioscience, Thermo Scientific), recombinant anti‐beta catenin antibody (Alexa Fluor 568 conjugate; ab201823), Human/Mouse CD44 antibody (Alexa Fluor 488 conjugate; R&D systems), occludin, mouse IgG1 Alexa Fluor 488, β‐actin from Santa Cruz Biotechnology, and DAPI purchased from Life Technologies. Colitis grade DSS (Mw 36k–50k Da) was from MP Bio, and other reagents used were of analytical grade.

Resomer^® RG 504 H; Poly(D,L-lactide-co-glycolide) (PLGA; #719900), poly(vinyl alcohol) (PVA; #363138), dichloromethane (DCM; #650463), Verteporfin (VP; #SML0534), perfluorooctylbromide (PFOB, or 1-Bromoheptadecafluorooctane; #343862), acetone (#179124), dimethyl sulfoxide (DMSO; # D2650), human PDAC cell line (PANC-1), Dulbecco’s Modified Eagle’s Medium-high glucose without L-Glutamine (#D5671), Gentian violet (#G2039) were purchased from Sigma-Aldrich, North Ryde, NSW, Australia. Singlet Oxygen Sensor Green probe (SOSG; #S-36002), NucBlue™ Live ReadyProbes™ Reagent (#R37605), LIVE/DEAD™ Viability/Cytotoxicity Kit for mammalian cells (#L3224), MitoTracker™ Green FM (#M7514), LysoTracker™ Green DND-26 (#L7526), DPBS, with calcium, magnesium (#14040182), DPBS, with no calcium, no magnesium (#14190250), TrypLE™ Express Enzyme, with no phenol red (#12604021), L-Glutamine, 200 mM (#25030081) and Fetal Bovine Serum, qualified, US origin (#26140079) were purchased from Life Technologies, Mulgrave, VIC, Australia. CellTiter 96^® AQueous One Solution Assay was purchased from Promega, Sydney, NSW, Australia.

The MPs were prepared by the oil-in-water (O/W) emulsion-solvent evaporation procedure. Concisely, 30 mg of PLGA (Resomer® RG504H, molecular weight 38–54 kDa, Sigma-Aldrich) was dissolved in 2.5 mL of dichloromethane and vortexed. Next, 3 mg of KGN (Tocris Bioscience) in dimethyl sulfoxide (DMSO) was added to the PLGA solution.
The solution was emulsified with 5 mL of 1% (w/v) polyvinyl alcohol (PVA; 87–90% hydrolyzed, average molecular weight 30–70 kDa, Sigma-Aldrich) and one drop of Tween 20. This emulsion was homogenized at 5000 rpm and subsequently added to the aqueous phase that included 100 mL of 0.1% (w/v) PVA, and stirred at 300 rpm at 37 °C for 3.5 h to evaporate the organic solvent. The MP suspension was washed in distilled water and harvested by centrifugation at 9000 rpm for 20 min at 4 °C. Supernatants were collected to calculate the encapsulation efficiency of KNG according to the following formula:
$$\text{Encapsulation efficiency}\left(\%\right)=\left[\text{actual drug loading}/\text{total drug added}\right]\times 100$$
The KGN-free MPs were prepared based on the same procedure. The particles were freeze-dried and stored at 4 °C for subsequent experiments. To increase the cell attachment, PLGA MPs were coated with gelatin through dispersion in a gelatin solution (0.1 mg/mL, Type B, Sigma-Aldrich) for 3 h, after which they were collected by centrifugation prior to cell culture.