2 hydroxy 4 2 hydroxyethoxy 2 methylpropiophenone irgacure 2959

All chemicals were used without treatment, as received. The LCE precursor included RM257 (monomer, >98.0%, TCI), PETMP (crosslinker, >95.0%, Sigma-Aldrich), EDDET (flexible chain extender, >95.0%, Sigma-Aldrich), dipropylamine (thermal catalyst, 99.0%, Sigma-Aldrich), 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure-2959, photocatalyst, 98.0%, Sigma-Aldrich), 5CB (nematic liquid crystal dopant, 98%, Sigma-Aldrich), 2,6-di-tert-butyl-4-methylphenol (thermal inhibitor, ≥99.0%, Sigma-Aldrich), 2-(N-ethyl-4-((4-nitrophenyl)diazenyl)anilino)ethyl prop-2-enoate (Disperse Red 1 acylate, photothermal dye, 95.0%, Sigma-Aldrich) and chloroform (solvent, ≥99.0%, Sigma-Aldrich). IP-S (Nanoscribe) was used as the inactive photoresist for fabricating the passive microstructures, and 2-propanol (≥99.0%, Sigma-Aldrich) was used as the solvent for the development.

PEG (Mn = 10 kg/mol), tin(II) 2-ethylhexanoate (Sn(Oct)₂), hydroquinone (HQ), triethylamine (TEA), methacrylic anhydride (MAAh), deuterated chloroform, 1,6-hexanediol, 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959), tris(hydroxymethyl)aminomethane (TRIS), Matrigel, verapamil hydrochloride (CAS RN: 52-53-9) and 3,4-dihydroxy-L-phenylalanine (L-DOPA) were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands). Propylene carbonate (PC), ethanol absolute and calcium hydride were obtained from Merck Millipore (Darmstadt, Germany). Methanol, dichloromethane (DCM) and diethyl ether were purchased from VWR (Amsterdam, The Netherlands). Trimethylene carbonate (TMC) was obtained from Huizhou ForYou Medical Devices Co (Huizou, China). Essential 8 medium was purchased from Thermo Fisher (Naarden, The Netherlands), Wnt activator CHIR99021 (1.5 µmol/L) from Axon Medchem (Groningen, The Netherlands), Activin-A (20 ng/mL) from Miltenyi Biotec (Bergisch Gladbach, Germany) and BMP4 (20 ng/mL) and XAV939 (5 µmol/L) from R&D Systems (Abingdon, UK).

Anti-HPV protein (bovine β-lactoglobulin modified with 3-hydroxyphthalic anhydride) was supplied by Jingbo Biological Pharmaceutical Co., Ltd. (Taiyuan, China). A stock solution was prepared in deionized (DI) water at a concentration of 26.9 mg/mL. 2-Hydroxy-4-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959) and [2-(acryloyloxy)ethyl]trimethylammonium chloride solution (AETAC) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium alginate (ALG, (C₆H₉O₇Na)_n, Mn = 200,000, G blocks: M blocks = 39:61, low viscosity) was obtained from JiSiEnBei International Trade Co., Ltd. (Hongkong, China). Polyethylene glycol diacrylate (PGEDA, Mn = 6 kDa) was purchased from Huateng Pharmaceutical Co., Ltd. (Changsha, China). NIH 3T3 cells (American Type Culture Collection, Manassas, VA, USA) and human umbilical vein endothelial cells (HUVECs, China Infrastructure of Cell Line Resources, Beijing, China) were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Grand Island, NY, USA), 1% nonessential amino acid solution (NEAA, Gibco, Grand Island, NY, USA), 1% GlutaMAXTM-I (Gibco, Grand Island, NY, USA), and 1% antibiotics (penicillin and streptomycin, Gibco, Grand Island, NY, USA) at 37 °C in a humidified atmosphere containing 5% CO₂.

Glycerol, 4-dimethylaminopyridine and triethylamine were obtained from Fisher Scientific (Loughborough, Leicestershire, UK). Sebacic acid was obtained from Alfa Aesar (Ward Hill, MA, USA). Dichloromethane, polyethylene glycol (M_w 8000), stannous octanoate, p-toluenesulfonic acid monohydrate, lactic acid (85%), polycaprolactone (M_w 80,000), 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), polyethylene glycol diacrylate (M_w 8000) (PEGDA₈₀₀₀), 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959) and anhydrous magnesium sulphate (MgSO₄) were obtained from Sigma Aldrich (Burlington, MA, USA).

Gellan gum (Gelzan), glycerol (Gly), methacrylic anhydride (MAA), triethylamine (TEA), anhydrous dimethyl sulfoxide (DMSO), deuterated water (D₂O), 4-dimethylaminopyridine (4-DMAP), phosphate buffer saline (10 mM phosphate and 150 mM NaCl, pH = 7.4, which was labelled PBS), polyethylenglycole (PEG 20,000 Da), 2-hydroxy-4-(2-hydroxyethoxy)-2-methyl-propiophenone (Irgacure 2959) and dialysis tubes (cut-off 12–14 kDa) were purchased from Sigma Aldrich (Milan, Italy). Silver nitrate (AgNO₃), monobasic potassium phosphate (KH₂PO₄, used to prepare 40 mM phosphate buffer, pH = 7.4, which was labelled PB), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were purchased from Carlo Erba (Milan, Italy). Finally, 1-methyl-2-pyrrolidinone from Fluka (St. Louis, MO, USA). Triptone Soy Agar (TSA) and Brain Heart Infusion broth (BHI) were purchased from Oxoid (Basingstoke, UK).