Poly l arginine hydrochloride

Calcium chloride, sodium carbonate, ethylenediaminetetraacetic acid disodium salt, dextran sulfate sodium salt (MW > 70,000), poly-L-arginine hydrochloride (MW > 70,000), Rhodamine B isothiocyanate, phosphate-buffered saline (PBS, 0.01 M), bovine serum albumin, trypsin from porcine pancreas (∼1500 U/mg), and tris(hydroxymethyl)aminomethane (≥9.8%) were purchased from Sigma-Aldrich (Taufkirchen, Germany). Hydrochloric acid was obtained from Reakhim (Moscow, Russia); dimethyl sulfoxide was purchased from Merck (Darmstadt, Germany); cyanine 7 NHS ester (Cy7) was obtained from Lumiprobe (Moscow, Russia). All chemicals were used without further purification. Deionized water produced with a water treatment system Milli-Q (Merck Millipore, Darmstadt, Germany) was used in all experimental stages. ELISA Kit For Anti-Tumor Necrosis Factor Alpha Antibody (Anti-TNFa) AEA133Hu from Cloud-Clone Corp. (Katy, TX, USA), 9K MWCO concentrators Thermo Fisher Scientific (Waltham, MA, USA).

All chemicals were of analytical grade and used as received without further purification. Calcium chloride dihydrate (CaCl₂·2H₂O), anhydrous sodium carbonate (Na₂CO₃), glycerol, ethylenediaminetetraacetic acid (EDTA), dextran sulfate sodium (DS) salt (MW 50 kDa), poly-L-arginine hydrochloride (Parg) with MW 15–70 kDa, sodium chloride (NaCl), phosphate buffered saline (PBS), isopropyl-β-D-1-thiogalactopyranoside, Rhodamine 6G (MW 479), Fluorescein isothiocyanate (FITC-) dextran (MW 40 kDa), doxorubicin hydrochloride (MW 580) and Hoechst 33258 were purchased from Sigma-Aldrich; DMSO (dimethyl sulfoxide, 99.5%), Trypan Blue solution, phosphate-buffered saline (PBS, pH 7.4), 0.25% (v/v) trypsin-EDTA solution, Versene solution, Dulbecco’s modified Eagle’s medium with Phenol Red (DMEM) and MTT (Thiazolyl Blue Tetrazolium Bromide, 98%) were purchased from PanEko (Moscow, Russia). Fetal bovine serum (FBS) was obtained from PAA Laboratories (Pasching, Austria). Fluorescent dyes sulfo-Cyanine3 maleimide and BDP FL maleimide were obtained from Lumiprobe (Moscow, Russia). Deionized water from a three-stage Milli-Q Plus purification system was used in the experiments.

Calcium chloride (CaCl₂, #223506), sodium carbonate (Na₂CO₃, #S7795), ethylenediaminetetraacetic acid disodium salt dihydrate (Na₂EDTA, #E5134), citric acid (HOC(COOH)(CH2COOH)₂, #C0759), cerium (III) chloride heptahydrate (CeCl₃·7H₂O, #22300), gadolinium(III) nitrate hexahydrate (#451134), dextran sulfate sodium salt (DS, MW ≈ 10 kDa, #D4911), 4-iodophenol (#I10201), and poly-L-arginine hydrochloride (PArg, MW ≈ 70 kDa, #P3892), were purchased from Sigma-Aldrich. All chemicals were used as received. Ultrapure water with a resistance greater than 18.2 MΩ cm⁻¹ was used for all experiments. MTT reagent and Hoechst 33,342 dye were purchased from PanEKO (Moscow, Russia). A live/dead assay kit, LDH assay kit, rhodamine B isothiocyanate, and phalloidin-FITC were purchased from Thermo Fisher Scientific, Cambridge, UK.

Krebs-Henseleit (KH) solution and the nominally Ca²⁺-free solution were prepared as previously described [24 (link)]. Membrane permeant acetoxymethyl (AM) ester forms of Fura-2 and Pluronic F127 were obtained from Invitrogen (Carlsbad, USA). Uridine 5′-triphosphate (UTP), uridine 5′-diphosphate (UDP), adenosine 5′-(γ-thio)triphosphate (ATPγS), forskolin, poly-l-arginine hydrochloride, SQ 22536, U73122, E₂, and G1 were obtained from Sigma-Aldrich (St. Louis, USA). G15 was obtained from Tocris (Bristol, UK). H89 dihydrochloride and MDL 12330A were obtained from Calbiochem (La Jolla, USA). All other general laboratory reagents were obtained from Sigma-Aldrich, and all cell culture reagents were obtained from Invitrogen.

Poly(DL-lactide-co-glycolide) 50:50 (PLGA, Mw 45 kDa) was obtained from Boehringer Ingelheim (Belgium); oligo-(l)-lactide (OLL, Mw 5 kDa), oligo-(D)-lactide (OLD, Mw 5 kDa) were synthesized from the respective lactic acid isomers as described earlier [14 (link)]; Dextran sulphate sodium salt (DS) (Mw~9–15 kDa), Poly-l-arginine hydrochloride (PAr, Mw~15–70 kDa), Polyallylamine hydrochloride (PAH, Mw~58 kDa), Poly(sodium 4-styrenesulfonate) (PSS) (Mw~70 kDa), Poly vinyl alcohol (PVA, Mw 70 kDa), Fluorescein isothiocyanate (FITC), Rhodamine B isothiocyanate (RBITC), Resazurin sodium salt, Crystal Violet, EDTA, zimozan were all obtained from Sigma-Aldrich (Saint Louis, MO, USA); Phosphate buffered Saline (PBS), DMEM/F12, trypsin, penicillin and streptomycin were purchased from PanEco (Moscow, Russia); Fetal calf serum (FCS) was obtained from Invitrogen (Carlsbad, CA, USA); Chloroform was obtained from Chimmed (Moscow, Russia); DMSO was obtained from Panreac (Castellar del Vallès, Spain).

Iron (III) chloride hexahydrate (99.8%), iron (II) chloride tetrahydrate (99.8%), sodium hydroxide (99.8%), citric acid (99.8%), sodium carbonate, calcium chloride, sodium chloride, poly-l-arginine hydrochloride (Parg, Mw = 15–70 kDa), dextran sulfate sodium salt (DS, Mw = 100 kDa), bovine serum albumin (BSA, lyophilized powder), poly(allylamine hydrochloride) (PAH, Mw = 50 kDa), poly(sodium 4-styrenesulfonate) (PSS, Mw = 70 kDa), ethylenediamine tetraacetic acid disodium salt (EDTA), phosphate-buffered saline (PBS, 0.01 M), ammonium rhodanide (NH4SCN), and Dulbecco’s modified Eagle’s medium (DMEM) with a high glucose content were obtained from Sigma-Aldrich (St. Louis, MO, USA). Glycerin was purchased from Reachem (Moscow, Russia). Fetal bovine serum (FBS), penicillin–streptomycin solution, 0.25% trypsin solution with 0.02% EDTA, DAPI Solution (1 mg/mL), Alexa Fluor 488 Phalloidin, and the 0.4% Trypan blue solution was obtained from Thermo Fisher Scientific (Waltham, MA, USA). All chemicals were used without additional purification. Deionized water (specific resistivity higher than 18.2 M∙Ω∙cm) from the Milli-Q Direct 8 (Millipore, Merck KGaA, Darmstadt, Germany) water purification system was used to prepare all solutions.