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7 protocols using l lactide

1

Polymer Scaffold for Cell Culture

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Ethylene brassylate (EB) monomer was supplied by Sigma Aldrich (Spain), while D,L-lactide and L-lactide monomers were provided by Corbion (The Netherlands). The triphenyl bismuth (Ph3Bi) catalyst was obtained from Gelest (USA). Dichloromethane and n-Hexane were supplied by Labbox (Spain). Multi-Walled Carbon Nanotubes (MWCNT), having an average diameter of 10–15 nm with 5–15 walls and 1–10 μm in length, were supplied by Arkema (France). Dulbecco’s Modified Eagle Medium (DMEM), Hanks’ Balanced Salt Solution (HBSS), Penicillin Streptomycin (P/S), Fetal Bovine Serum (FBS), AlamarBlue cell viability reagent and rhodamine-phalloidin were supplied by Fisher Scientific (Spain). Laminin, PBS (Phosphate-buffered saline), Diiodomethane, Fluoroshield with DAPI, Triton X-100 and Tween 20 were supplied by Sigma Aldrich (Spain), and 16% Formaldehyde solution was supplied by Thermo Fisher Scientific (USA). Chloroform was supplied by PanReac (Spain).
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2

Synthesis of Poly(Lactic Acid) Polymers

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L-lactide (L-LA) and D-Lactide (D-LA) were purchased from Corbion (Amsterdam, The Netherlands). Tin(II)bis(2-ethylhexanoate) (Sn(Oct)2) (Sigma Chemical Co., St. Louis, MO, USA, purity ≥ 99%) and 1-dodecanol (DoOH) (Sigma-Aldrich, St. Louis, MO, USA, purity ≥ 99.5%) were purified by distillation under reduced pressure and dissolved in dry toluene. The toluene was dried by refluxing over a benzophenone-Na complex and distilled in a nitrogen atmosphere just prior to use. High molecular weight PLLA (Mn = 181,493 g/mol, Mw = 425,561 g/mol, PDI = 2.345) was synthesized by ring-opening polymerization of L-lactide (Corbion, Amsterdam, The Netherlands) at 130 °C for 24 h. Other PLA homopolymers for nucleating agent materials: PDLA (Mn = ~87,000 g/mol, Mw = ~125,000 g/mol, PDI = 1.437) and PLLA (Mn = ~87,000 g/mol, Mw = 153,000 g/mol, PDI = 1.759) were also synthesized by a similar method. Dichloromethane (JT Baker, HPLC grade) and CO2 (Purity ≥ 99.9%) were used as received.
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3

Sunitinib-Loaded Polymeric Nanoparticles

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Sunitinib malate was obtained from LC laboratories, USA. Poly(vinyl alcohol) (PVA; M w 30,000-70,000; 88% hydrolyzed), Tween 20, stannous octoate and dimethyl sulfoxide (DMSO) were obtained from Sigma Aldrich, Germany. Dichloromethane (DCM), acetonitrile and tetrahydrofuran (THF) were purchased from Biosolve (Valkenswaard, The Netherlands). PEG1000, 1,4-butanediol and 1,4 dioxane were purchased from Acros (Geel, Belgium), 1,4butanediisocyanate was purchased from Bayer (Leverkusen, Germany) and DL-lactide and L-lactide were purchased from Corbion Purac (Gorinchem, The Netherlands). Polystyrene standards were purchased from Agilent technologies (Santa Clara, USA). Fertilized white Leghorn eggs were obtained from Het Anker company (Ochten, The Netherlands).
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4

Synthesis of High-Molecular-Weight PLLA

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L-lactide (18 g, 0.12 mol) (Corbion, Amsterdam, The Netherlands), 0.09 mL stannous octoate (Sigma-Aldrich, St. Louis, MO, USA), and 0.54 g (0.003 mol) of the initiator 1-dodecanol (Sigma-Aldrich) were placed in a 1 L double-jacketed reactor at room temperature. After oxygen and moisture removal, nitrogen was injected into the reactor for 5 min. The mixture was gradually heated to 120 °C for 1 h, and then stirred at 120 °C for 4 h. The synthesized PLLA was collected via vacuum filtration, and the unreacted substances were removed using ethanol. PLLA was dissolved at room temperature in 220 mL dichloromethane (99.5%; Samchun Chemical Co., Ltd., Seoul, Republic of Korea). The PLLA solution was diluted via the slow, dropwise addition of 320 mL of 1% polyvinyl alcohol (87–90%, mol wt 30,000–70,000; Sigma-Aldrich) using a glass pipette while stirring it at 2000 rpm with a shear mixer. The solution was filtered under a vacuum for 3 h at room temperature and centrifuged at 3000 rpm for 10 min at room temperature to separate PLLA. After removing the supernatant, polyvinyl alcohol that remained in PLLA was redispersed in 50 mL distilled water and centrifuged at 3000 rpm for 10 min at room temperature. Finally, PLLA polymer was obtained by drying it in an oven at 45 °C for 24 h [31 ,32 ,33 (link),34 (link)].
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5

Synthesis and Characterization of Biodegradable PEG Hydrogels

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d,l-Lactide, d-lactide, and l-lactide were purchased from Corbion
(Gorinchem, The Netherlands). Hydroxyl-terminated eight-armed poly(ethylene
glycol) (tripentaerythritol) (PEG8arm10k, Mw = 10 000 g mol–1 and PEG8arm20k, Mw = 20 000 g mol–1) was purchased from JenKem Technology Co., Ltd (Beijing,
China). Tin(II) 2-ethylhexanoate (Sn(Oct)2), toluene, dichloromethane
(DCM), N,N-dimethylformamide (DMF),
triethylamine (Et3N), methacryloyl chloride, fluorescein
isothiocyanate (FITC), rhodamine B isothiocyanate, Celite 545, diethyl
ether (Et2O), ethanol (EtOH), tetrahydrofuran (THF), and
acetone were purchased from Sigma-Aldrich (St Quentin Fallavier, France).
All chemicals were used without further purification with the exception
of toluene and DCM, which were dried over calcium hydride and freshly
distilled before use.
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6

3D Printable Poloxamer-based Biomaterials

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Poloxamer 407 triblock copolymer (length of the PEG segments equal to 91 repeating units and the PPG segment segment is 56 units long (NMR)). was acquired from BASF (Ludwigshafen, Germany). D,L-lactide, glycolide and L-lactide were purchased from Corbion Purac (Gorinchem, The Netherlands). Irgacure 2959 was obtained from Ciba Specialty Chemicals (Basel, Switzerland). Solvents, unless indicated otherwise, were acquired from Biosolve (Valkenswaard, The Netherlands). Calcein acetoxymethyl ester (calcein-AM), Alamar Blue Cell viability reagent, Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin, streptomycin and ethidium homodimer were acquired from Fisher Scientific (Landsmeer, The Netherlands). Deuterated chloroform (CDCl3), є-caprolactone, fluorescein isothiocyanate (FITC), gelatin (type A from porcine skin, 175 g bloom), methacrylic anhydride, sodium azide, stannous octoate (Sn(Oct)2) and triethylamine (TEA) were all provided by Sigma Aldrich (Zwijndrecht, The Netherlands). Dialysis membranes (Spectra/Por 2, upper molecular weight cutoff 12-14 kDa) were obtained from Carl Roth (Karlsruhe, Germany). Cartridges and extrusion nozzles for 3D printing were obtained from Nordson EFD (Maastricht, The Netherlands). Biopsy punches were acquired from Milltex (Zaventem, Belgium).
All percentages concerning solutions are presented as %w/w, unless stated otherwise.
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7

Synthesis of Biodegradable Polyester-Urethanes

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l-Lactide was purchased from Corbion (Gorinchem, The Netherlands). 2,6-Diisocyanato ethyl caproate (l-lysine diisocyanate ethyl ester, LDI) (Infine Chemicals, Shanghai, China) and ε-caprolactone (Sigma-Aldrich, Taufkirchen, Germany) were purified by vacuum distillation. Dichloromethane, chloroform, acetone, heptane and cyclohexane were purchased from Fisher Scientific (Schwerte, Germany), meso-erythritol (Acros, Geel, Belgium), 1,4-Diazabicyclo[2.2.2]octane (DABCO, Sigma-Aldrich, Germany), DMSO (Carl Roth, Karlsruhe, Germany) and stannous octoate (ABCR, Karlsruhe, Germany) were used as received. Phosphate buffered saline tablets (Sigma-Aldrich, Taufkirchen, Germany) were dissolved in water, filtered and sterilized before use.
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