Spectra por

Proteins from the control (i.e., no sonication), ultrasonic cleaner bath, or probe-type ultrasound-treated brans were hydrolyzed with either Flavourzyme, Papain, or Alcalase. In each case, 1.5 g of proteins was dissolved in 20 mL of water. Appropriate amounts of protease were added to ensure a 2% enzyme to substrate ratio (w/w). The pH and temperature conditions were as follows: Flavourzyme: pH 7.0 and 50 °C; Papain: pH 7.0 and 60 °C; Alcalase: pH 8.0 and 60 °C. All the digestions were performed for 3 h and terminated by heating in a hot water bath at 85 °C for 5 min. Once cooled to room temperature, they were centrifuged at 8000× g (15 min, 4 °C) to collect the hydrolysates (i.e., supernatants).
The dialysis was performed using a cellulose ester Spectra/Por^® (Fisher Scientific, Nepean, ON, Canada) membrane with a 100–500 Da molecular weight cut-off. The membranes were prepared by soaking in water to remove the storage buffer (0.05% sodium azide). The prepared membranes containing hydrolyzed protein solutions were immersed in water and gently stirred overnight (approximately 14 h). Solutions within the membranes were freeze dried, and their protein content was determined as previously reported [12 (link)].

Silkworm (Bombyx mori) cocoons were purchased from Mulberry Farms as a source of silk fibroin protein (henceforth referred to as silk). Sodium carbonate (Na₂CO₃), and lithium bromide (LiBr) were purchased from Millipore Sigma for silk fibroin extraction from silkworm cocoons. Dialysis bags, 3.5 kDa molecular weight cut‐off (Spectra/Por), were purchased from Fisher Scientific to facilitate purification of silk fibroin. ICG dye was purchased from MP Biomedicals (#ICN15502050) and stored at 4°C. All solutions were freshly prepared in nanopure water (NPW; resistivity ~18.2 MΩ cm; Millipore Filtration System). BALB/c mice were purchased at ~10 weeks from Charles River Laboratories. Commercially available 4‐0 Monosof™ Monofilament Nylon Sutures (Medtronic) were purchased from esutures.com.

dPSA was produced by heating a solution (10 mL) of 100 mg of colominic acid (MilliporeSigma) and 10 mg sodium cyanoborohydride (MilliporeSigma) in 2 M NaOH to 100 °C in a sealed hydrolysis tube (ThermoFisher Scientific) for 40 min. The solution was cooled, neutralized with 2 M HCl, dialyzed (Spectra/Por™, Fisher Scientific) two times with 4 L of water and lyophilized. dPSA containing 35% de-N-acetyl neuraminic acid residues, as determined by a modified resorcinol assay [30 (link)], was conjugated to biotin as described previously [28 (link)]. The ELISA was performed with serial dilutions of nucleolin or SEAM 3 as in the absence or presence of 50 μg/mL of colominic acid or unmodified dPSA as described previously [28 (link)].

The powdered pulps and seeds (60 g, each) were defatted with 95% ethanol and petroleum ether with continuous stirring for 24 h. The residues were dried and then extracted with hot water at 90°C for 5 h (three-time, 3 × 5 h). Following centrifugation at 4500 rpm for 10 min, the supernatants were mixed with 95% cold ethanol (3: 1, v/v) at 4°C overnight. Precipitates were dissolved in distilled water and deproteinized using Sevag reagent (chloroform/butanol 4: 1, v/v). The deproteinized mixture was dialyzed for 3 days (with 3500 Da cut-off, Spectra/Por™, Fisher Scientific, Illkirch, France) and lyophilized to obtain the water-soluble polysaccharides from C. azarolus seeds and pulps named, respectively, CAS and CAP.
Finally, the CAP and CAS extraction yields were calculated.

The acid-solubilized collagen using vinegar extraction was done according to the modification method of Tamilmozhi et al. [21 (link)]. The pretreated tuna-tail tendons (from 2.1) were soaked in 5% vinegar (Kewpie (Thailand) Co., Ltd., Bangkok, Thailand) with the ratio of tendon: 5% vinegar 1:20 (w/v) at 10 ± 1 °C using shaking incubator (LSI-3016R, Daihan Labtech Co., Ltd., Gyeonggi-do, Republic of Korea) with 200 rpm for 72 h. The extracted solution was filtrated using a double layer of cheesecloth and kept at 4 °C. The pellet was re-extracted with the same manner. The extracted solutions were mixed and precipitated using NaCl adding with a final concentration of 2.6 M for collagen obtaining. The resulting precipitate was obtained by centrifugation at 10,000× g for 30 min at 4 °C. Thereafter, the pellet was resuspended in a minimum volume of 5% vinegar (1:10, w/v), followed by dialysis in 50 volumes of distilled water for 24 h using the 100 kDa dialysis membrane (Spectra/Por, Thermo Fisher Scientific Inc., Gothenburg, Sweden). The resulting dialysate was lyophilized using a freeze-dryer (GFD-3H, Grisrianthong, Samut Sakhon, Thailand) to obtain acid-solubilized collagen using vinegar extraction (VTC) and kept at −18 to −20 °C until further use.

Iron (III) chloride hexahydrate (≥99%), iron (II) chloride tetrahydrate (98%), oleic acid (≥90%) (OA), potassium dihydrogen phosphate (≥98%), N-hydroxysuccinimide (NHS) (98%), N-(3-dimethyl aminoproyl)-N’-ethylcarbodiimide hydrochloride (EDC), N,N′-dicyclohexylcarbodiimide (DCC) (≥99.0%), poly(ethylene glycol diamine) (NH₂-PEG-NH₂, Mn = 3000), (≥98.0%), polyvinyl alcohol (PVA, Mowiol 4–88), deuterated chloroform (≥99%) and oxaliplatin (OXA) (≥99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dialysis bags (Spectra/Por, MWCO: 3.5 and 12–14 kDa) and a monoclonal anti-CD133-TMP4 Ab conjugated to Alexa Fluor 488 were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Commercial PLGA Mn of 10 kDa (Purasorb PDLG 7502A, 70/30 LA/GA) was kindly donated by Corbion (The Netherlands). Sodium hydroxide, chloroform, ethyl ether, tetrahydrofuran (THF), dichloromethane (DCM), ethanol, and ammonia p.a. were purchased from Penta (The Czech Republic). Media used for cell culture Eagle’s Minimum Essential Medium (EMEM) and Dulbecco’s Modified Eagles’ medium (DMEM) were bought from Thermo Fisher Scientific (Waltham, MA, USA). Microscopy dishes with glass bottom (ø35 mm, 1.5#) were supplied by MatTek Life Sciences (Boston, MA, USA). Filters with a 0.45-µm pore size were purchased from JetBiofil (China). For all experiments, Millipore water was used.