Sodium dihydrogen phosphate

The lyophilized powder of DNA oligonucleotide (5'- GTC GTA CTG ATA CAT GAG CC –'3; 6117 Da) was the product of Genomed (Warsaw, Poland) or Syntol (Moscow, Russia). It was used as an aqueous solution prepared in accordance with the producer recommendations and stored at –20 °C no longer than 1 month. Indazolium trans-[tetrachloridobis(1H-indazole)ruthenate(III)] was synthesized at the University of Vienna (see the Electronic Supplementary Material (ESM) for chemical formula). The stock solution of the Ru drug (1 mM) was prepared daily in 100 mM NaCl. Human transferrin (>98%), glutathione (>98%), and L-ascorbic acid (>99.5%) were purchased from Sigma-Aldrich (St. Louis, USA). Sodium chloride, sodium dihydrogen phosphate, disodium hydrogen phosphate, also from Sigma-Aldrich (St. Louis, USA), were of analytical reagent grade. Sodium hydroxide and citric acid (>99.5%) were obtained from Fluka (Buchs, Switzerland). High-purity water (18 MΩ cm^–1) used throughout this work was obtained from a Milli-Q water purification system (Millipore Elix 3; Saint-Quentin, France).

L-amino acid standards, norvaline, sarcosine, sodium dihydrogen phosphate (NaH₂PO₄), hydrochloric acid, sodium hydroxide, guanidine hydrochloride, 2,4-dinitrophenylhydrazine, iron (II) chloride, iron (III) chloride, ammonium thiocyanate, ortho-phthalaldehyde, 9-fluorenylmethoxycarbonyl chloride, iodoacetic acid, 3-mercaptopropionic acid, boric acid (H₃BO₃), trichloroacetic acid, and MS-grade ammonium acetate were from Sigma-Aldrich (Darmstadt, Germany). Standards for fatty acid analysis included Supelco 37 Component FAME mix (Supelco, St. Louis, MO, USA), 68D (Nu-Check-Prep, Elysian, MN, USA), and GLC-490 (Nu-Check-Prep, Elysian, MN, USA). Standards for the lipid class analysis were oleic acid, oleoyl monoacylglycerol, dioleoyl diacylglycerol, triolein, dioleoyl phosphatidylcholine, cholesteryl oleate and ethyl docosahexaenoic acid (Larodan, Solna, Sweden). All solvents used were at least HPLC grade. Water used was ultra-pure water (ELGA LabWater, High Wycombe, UK).

The following materials were used: sucrose (Sigma-Aldrich), polysorbate 80 (Tween 80, Sigma-Aldrich), inulin (4kD; Sensus, Roosendaal), dextran (5kD; Pharmacosmos Denmark), sodium dihydrogen phosphate (Sigma-Aldrich), disodium hydrogen phosphate (Sigma-Aldrich) and 5 mg/mL of Infliximab antibody bulk (5 mg/ml in 5% sucrose and 0.005% (polysorbate 80, pH 7.2) (obtained from AIMM therapeutics).

Reagents used in the various methods, 1,1-diphenyl-2-picrylhydrazyl hydrate (DPPH), Folin-Ciocalteu reagent and (±)-catechin were purchased from Sigma (Sigma-Aldrich Chemie GmbH, Sternheim, Germany). The following reagents were also purchased from Sigma-Aldrich Chemie: iron (III)-chloride, potassium hexacyanoferrate (III), sodium hydrogen phosphate anhydrous, sodium dihydrogen phosphate andtrichloroacetic acid. Gallic acid was purchased from Sigma (St. Louis, MO, USA). Maltodextrin of dextrose equivalent (DE) 16.5–19.5 (Sigma-Aldrich Chemie GmbH) was used as a carrier material. All other chemicals and reagents were of analytical grade.

Three types of extracellular matrix models were crafted for this study. Collagens solely comprised of rat tail monomers (4 g/l rat collagen type I, SERVA, Heidelberg, Germany), collagens containing only bovine skin monomers (4 g/l bovine collagen type I, Biochrom, Berlin, Germany) and collagens composed of a mixture of both collagen monomer types (rat tail and bovine skin) in a mass fraction of 1:2 were used for all collagen related experiments (Fischer et al., 2017 (link), 2019 (link), 2020 (link); Kunschmann et al., 2019 (link)). For the polymerization of the monomer solution, a 1 M phosphate buffered solution containing disodium hydrogen phosphate (Sigma Aldrich, Cat. No. 71636), sodium dihydrogen phosphate (Sigma Aldrich, Cat. No. 71507) and ultrapure water were mixed keeping stable conditions (pH value 7.4, ionic strength 0.7, final phosphate concentration 200 mM). The components were kept at 0°C for mixing and finally added to 6-well plates for invasion assays, Petri dishes for elasticity measurements, 96-well plates for investigating the polymerization dynamics, or ibidi 24-well μ-plates for studying the pore size characteristics. Due to differences in the polymerization times (Supplementary Figure S5) R and RB collagen matrices were incubated for 2 h whereas B collagens were incubated for 5 h under normal conditions.

Analytical grade rosuvastatin calcium (>99%), sodium dihydrogen phosphate (≥99.9%), formic acid (>98%), leucine enkephalin (>98%), and isopropanol (>99%) were obtained from Sigma-Aldrich (St. Louis, MO, USA). High purity nitrous oxide was purchased from Messer Croatia Plin (Zaprešić, Croatia) and acetonitrile (≥99.9%) from Merck Millipore (Burlington, MA, USA). Ultrapure water (18 MΩ·cm) was generated in-house using a Milli-Q System from Merck Millipore (Burlington, MA, USA).