Direct q1 ultrapure water system

Silk from Bombyx mori was sourced from the cocoons of sericulture silkworms maintained in the facilities of IMIDA (Murcia, Spain) and fed on fresh Morus alba L. leaves. The cocoons were treated to remove the sericin by boiling in an aqueous solution of Na₂CO₃ (0.05 N) twice for 60 min. After washing with pure water and air-drying, the resulting substance had a bright white, cotton-like appearance. The silk was dissolved in the ionic liquid 1-ethyl-3-methylimidazolium acetate [emim+][acetate−] by high-power ultrasound, as described in a previous report [12 (link)]. The ionic liquid (97% purity) was obtained from Io-LiTec GmbH (Frankfurt, Germany) and was applied without any further purification. Ibrutinib (IB) [38 (link)] (Figure S1 of Supporting Information) was provided by AdooQ Bioscience (Irvine, CA, USA). Highly purified water (18.2 MΩ·cm at 25 °C; from a Millipore Direct-Q1 ultrapure water system, Billerica, MA, USA) was used throughout. N-Hydroxysuccinimide (NHS) and N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) were purchased from Merck. All other chemicals and solvents were of analytical grade and were used without any extra purification.

The silk fibroin (SF) used in this work was extracted from Bombyx mori silk cocoons kindly provided by Instituto Murciano de investigación y Desarrollo Agrario y Alimentario (Murcia, Spain). The SF was purified as reported elsewhere [47 (link)]. In brief, cocoons were shredded in a mill and filter through a 1 mm stainless still mesh and later boiled for 2 h in a 0.2 N Na₂CO₃ solution to remove sericin, waxes and other impurities. Following this, the SF was collected in a 0.1 mm pore size strainer and washed thoroughly with warm water to remove any unbounded sericin. Finally, the SF was air-dried in a fume hood until constant weight was achieved. NAR was purchased from Sigma-Aldrich (Madrid, Spain) and was used as received. Purified water from a Millipore Direct-Q1 ultrapure water system (18.2 MΩ·cm at 25 °C; Billerica, MA, USA) was used throughout the experimentation.

Bombyx mori silk cocoons were reared in the sericulture facilities of IMIDA (Murcia, Spain) and raised on a diet of natural fresh leaves of Morus alba L. To extract the SF, the raw silk cocoons were shredded in a mill to a particle size of 1 mm, and then boiled in a 0.1 M Na₂CO₃ aqueous solution for 120 min to remove sericin, waxes, and impurities. The remaining water-insoluble SF was rinsed thoroughly with ultrapure water and air-dried in a fume hood until constant weight (approximately 24 h). Hyaluronic acid sodium was purchased from Monteloeder Digital Nutracetics (MW 8 × 10⁵ Da, >90% purity). Curcumin (99% purity) was purchased from ChromaDex (Irvine, CA, USA). Purified water (18.2 MΩ·cm at 25 °C; from a Millipore Direct-Q1 ultrapure water system, Billerica, MA, USA) was used throughout. All other chemicals and solvents were of analytical grade and were used without further purification.