Nanostructure of Fully Injectable Hydrazone-Thiosuccinimide Interpenetrating. Polymer Network Hydrogels Assessed by Small-Angle Neutron Scattering and
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1 SUPPORTING INFORMATION Nanostructure of Fully Injectable Hydrazone-Thiosuccinimide Interpenetrating Polymer Network Hydrogels Assessed by Small-Angle Neutron Scattering and dstorm Single-Molecule Fluorescence Microscopy Trevor Gilbert 1, Richard J. Alsop 2, Mouhanad Babi 3, José Moran-Mirabal 3, Maikel C. Rheinstädter 2 and Todd Hoare* 1 1 Department of Chemical Engineering, McMaster University, 1280 Main St. W, Hamilton, Ontario, Canada L8S 4L7 2 Department of Physics and Astronomy, McMaster University, 1280 Main St. W, Hamilton, Ontario, Canada L8S 4M1 3 Department of Chemistry and Chemical Biology, Mc Master University, 1280 Main St. W, Hamilton, Ontario, Canada L8S 4M1 * To whom correspondence should be addressed: hoaretr@mcmaster.ca
2 Table S1: GPC hardware, solvents and protocols for acidic, basic and DMF instruments. Solvent Acidic 1M acetate buffer, ph 4.7 Basic 0.5M sodium nitrate, 25mM CHES, ph 10.0 DMF 50mM LiBr in DMF Temp. Pump & Flow rate (ml/min) 30 C 515, 0.8mL/min 30 C 515, 0.8mL/min 35 C 590, 0.5 ml/min Detector Autosampler 717 Plus 717 Plus Manual Columns Ultrahydrogel -120, -250, - 500; 7.8x300mm; 6μm particles Ultrahydrogel- 120, -250, - 500; 7.8x300mm; 6μm particles HR-2, -3, -4; 7.8x300mm; 5μm particles Figure S1: Greater than order-of-magnitude suppression of low q scattering in contrast matched single network hydrogels: (a) PNIPAM as a function of temperature; (b) PVP
3 Table S2: d7-pnipam single network at match point to suppress its scattering (confirmation of scattering suppression) squared Lorentzian model fitting parameters Fitting Parameter 25 C 32 C 37 C 45 C scale (A) correlation length (γ; Å) Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (C; 1/cm) Χ^2 (Chi^2) Sqrt(X^2/N) Table S3: PVP single network at match point to suppress its scattering (confirmation of scattering suppression) Porod model fitting parameters Fitting Parameter 25 C Porod term scale (A) 2.78E-07 Porod term exponent (m) 2.70 Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) 112 Fluid term Lorentzian exponent (n) 1.92 Background (C; 1/cm) Χ^2 250 Sqrt(X^2/N) 1.02
4 Table S4: Porod model fitting parameters for thiosuccinimide-cross-linked PVP single network (9 wt%) in D2O (25 C) Fitting Parameter 25 C Porod term scale (A) 6.54E-05 Porod term exponent (m) 2.39 Fluid term scale (B) 2.74 Fluid term Lorentzian Screening Length (ε; Å) 37.2 Fluid term Lorentzian exponent (n) 1.71 Background (C; 1/cm) Χ^ Sqrt(X^2/N) 2.57 Figure S2: Raw scattering curve and Porod model fit of thiosuccinimide-cross-linked PVP single network (9 wt%) in D 2 O (25 C)
5 Table S5: Porod model fitting parameters for semi-ipn of free PNIPAM in thiosuccinimide-cross-linked PVP network (9 wt%) in D2O Fitting Parameter 25 C 60 C Porod term scale (A) 8.25E E-07 Porod term exponent (m) Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (C; 1/cm) Χ^ Sqrt(X^2/N) Figure S3: Raw scattering curve and Porod model fit of semi-ipn of free PNIPAM in thiosuccinimide-cross-linked PVP network (9 wt%) in D 2 O. Porod model fits reasonably at 25 C, but poorly at low q for high temperature.
6 Table S6 Squared Lorentzian model fitting parameters for hydrazone-cross-linked PNIPAM single network hydrogels in D 2 O Fitting Parameter 25 C 32 C 37 C 45 C scale (A) 2.43E E E E+04 correlation length (γ; Å) Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (C; 1/cm) Χ^ Sqrt(X^2/N) Figure S4: Raw scattering curves and squared Lorentzian model fits of hydrazone-crosslinked PNIPAM single network hydrogels in D 2 O as a function of temperature: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C.
7 Table S7: Squared Lorentzian model fitting parameters for semi-ipn of unfunctionalized PVP in a cross-linked PNIPAM network Fitting Parameter 25 C 32 C 37 C 45 C scale (A) 8.90E E E E+05 correlation length (γ; Å) Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (C; 1/cm) Χ^ Sqrt(X^2/N) Figure S5: Raw scattering curves and squared Lorentzian model fits of the semi-ipn control of unfunctionalized PVP trapped inside a cross-linked PNIPAM network hydrogels in D 2 O as a function of temperature: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C.
8 Table S8: Hybrid model fitting parameters for PNIPAM-PVP IPN in D 2 O Fitting Parameter 25 C 32 C 37 C 45 C Porod term scale (A) 2.39E E E E-05 Porod term exponent (m) scale (B) correlation length (γ; Å) Fluid term scale (C) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (D; 1/cm) Χ^ Sqrt(X^2/N) Figure S6: Raw scattering curves and hybrid model fits (with component breakdowns) of PNIPAM-PVP IPN hydrogels in D 2 O as a function of temperature: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C.
9 Table S9: Porod model fitting parameters for IPN using d7-pnipam, with deuterated PNIPAM scattering contrast-suppressed Fitting Parameter 25 C 32 C 37 C 45 C Porod term scale (A) 2.09E E E E-05 Porod term exponent (m) Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (C; 1/cm) Χ^ Sqrt(X^2/N) Figure S7: Raw scattering curves and Porod model fits of IPN hydrogels with d7- PNIPAM, with deuterated PNIPAM scattering contrast-suppressed, at temperatures: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C. The Porod model does not describe the small but distinct feature seen below q=0.01 Å -1.
10 Table S10: Squared Lorentzian model fitting parameters of IPN hydrogels using d7- PNIPAM, with PVP scattering contrast-suppressed Fitting Parameter 25 C 32 C 37 C 45 C scale (A) 5.30E E E E+03 correlation length (γ; Å) Fluid term scale (B) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (C; 1/cm) Χ^ Sqrt(X^2/N) Figure S8: Raw scattering curves and squared Lorentzian model fits of IPN hydrogels using d7-pnipam, with PVP scattering suppressed by contrast matching, at temperatures: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C. This model assumes a plateau feature below the q range of the NG30 SANS instrument to fit the data collected.
11 Table S11: Hybrid model fitting parameters for IPN hydrogels using d7-pnipam, with PNIPAM scattering contrast-suppressed Fitting Parameter 25 C 32 C 37 C 45 C Porod term scale (A) 1.62E E E E-05 Porod term exponent (m) scale (B) correlation length (γ; Å) Fluid term scale (C) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (D; 1/cm) Χ^ Sqrt(X^2/N) Figure S9: Raw scattering curves and hybrid model fits (with component breakdowns) of IPN hydrogels, with d7-pnipam scattering suppressed by contrast matching, at temperatures: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C.
12 Table S12: Hybrid model fitting parameters for IPN using d7-pnipam, with PVP scattering contrast-suppressed Fitting Parameter 25 C 32 C 37 C 45 C Porod term scale (A) 3.14E E E E-05 Porod term exponent (m) scale (B) correlation length (γ; Å) Fluid term scale (C) Fluid term Lorentzian Screening Length (ε; Å) Fluid term Lorentzian exponent (n) Background (D; 1/cm) Χ^ Sqrt(X^2/N) Figure S10: Raw scattering curves and hybrid model fits (with component breakdowns) of IPN hydrogels with PVP scattering suppressed by contrast matching as a function of temperature: (a) 25 C; (b) 32 C; (c) 37 C; (d) 45 C.
13 Figure S11: Attempted fits of PNIPAM single network scattering data to a hybrid model considering Guinier and Porod components (along with the Lorentzian-type fluid scattering term conserved from other models, Eq. S1) more commonly used in the literature to fit conventional PNIPAM hydrogels. (Eq. S1) I(Q) = A + R 2 Q m Bexp( Q2 ) + C 3 1+(ξQ) n + D Note that poor fits were observed at low q in particular, with lesser (but still significant) fitting errors also observed at high q. This lack of fit supports our choice of model based on the higher heterogeneity of our in situ-gelled IPN hydrogel versus conventional monomer-derived single network hydrogels.
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