Poly(L-lysine) hydrobromide (PLL), poly(ethylene imine) (PEI), poly(L-glutamic) acid (PGA) were from Sigma-Alrich (St Quentin Fallavier, France), hyaluronic acid (HA) from Lifecore medical (USA). For biological functionalization, PGA-RGD was synthesized as previously described [36 ]. PLL, HA and PEI at respectively 0.5, 1 and 2 mg/mL were dissolved in a HEPES-NaCl buffer (20 mM Hepes at pH 7.4, 0.15 M NaCl). A first layer of PEI was always deposited. All rinsing steps were performed with 0.15 M NaCl at pH ~6.5. The films were chemically crosslinked using 1-Ethyl-3-(3-Dimethylamino-propyl)Carbodiimide (EDC, final concentration of 5, 10, 30 or 70 mg/mL depending of the films) and N-Hydrosulfosuccinimide sodium salt (Sulfo-NHS, 11 mg/mL) as catalyzer, as previously described [32 (link), 42 (link)]. BMP loading in the films was done following an established protocol [31 (link), 33 (link)].
Protocol detail
Multilayer Biomaterial Scaffold Fabrication
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Biological
Buffer
Carbodiimide
Hepes
Hyaluronic acid
L glutamic acid
L lysine
Poly
Poly ethylene imine
Salt
Sodium
Sulfo nhs
Corresponding Organization : Institut polytechnique de Grenoble
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Variable analysis
independent variables
- EDC concentration (5, 10, 30 or 70 mg/mL)
- Crosslinking using EDC and Sulfo-NHS
- BMP loading in the films
- PLL, HA and PEI concentrations (0.5, 1 and 2 mg/mL respectively)
- HEPES-NaCl buffer (20 mM Hepes at pH 7.4, 0.15 M NaCl)
- Rinsing steps with 0.15 M NaCl at pH ~6.5
- PGA-RGD synthesis as previously described [36]
- Not explicitly mentioned
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