**2. Materials and methods**

#### **2.1 Materials**

Disks of 316L SS (0.74 cm2, from Outokumpu Stainless AB, ARC) were used. The bulk chemical composition of 316L SS was given previously (Landoulsi et al., 2008a). All chemicals (NaCl, Na2SO4, NaNO3 NaHCO3, NaHPO4, Na2PO4) were provided by Prolabo (VWR, France) and ensured 99% minimum purity. 3-aminopropyl(triethoxysilane) (APTES, 99%), glucose oxidase (Gox, EC 1.1.3.4, 47200 U.g-1) from *Aspergillus Niger* and *D*-glucose were purchased from Sigma-Aldrich (France). Bis(sulfosuccinimidyl) suberate (BS) was purchased from Pierce (Rockford, IL, USA).

#### **2.2 Stainless steel surface preparation**

104 Biomaterials – Physics and Chemistry

Poly(dimethylsiloxane) SSMCC DNA Vaidya & Norton, 2004

Polystyrene BS3, GMBS IgG North et al., 2010

Cellulose SMP RGDC peptide Bartouilh de Taillac et

al., 2004

d. AFM: increase of surface roughness, presence of nanostructures presumably due to

d. XPS: elemental composition and peak shapes, demonstration of silane deposition.

d. Fluorescence labeling: increase of amount of immobilized proteins by silanization +

b. In dry toluene under argon, 1.5 h; rinsing with dry toluene; outgassing. c. Increase of osteoprogenitor cells attachment and proliferation. Blank = native. d. XPS: qualitative consistency with expectations but probable Si contamination on final product.

proteins or peptide.

a. Plasma treatment.

a. Drying, outgassing.

BSA = bovine serum albumin BS3 = bis(sulfosuccinimidyl) suberate CDI = 1,1'-carbonyldimidazole DSC = N,N'-disuccinimidyl-carbonate DSS = N,N'-disuccinimidyl-suberate DS3 = N,N'-disulphosuccinimidyl-suberate

ECM = extracellular matrix EGF = epidermal growth factor

QCM = quartz crystal microbalance

**2. Materials and methods** 

**2.1 Materials** 

TNBS = 2,4,6-trinitrobenzenesulfonic acid

GA = glutaraldehyde

LbL = layer-by-layer

c. -

linker step.

a. Surface plasma oxidation.

b. Vapor deposition, 30 min, 100 °C.

c. Hybridization of attached DNA. No blank.

b. In ethanol+acetic acid; rinsing in methanol.

EDC = 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide.

GMBS = 4-maleimidobutyric acid N-hydrosuccinimide ester

RGDC = arginine - glycine - aspartic acid – cysteine SMP = N-succinimidyl-3-maleimidopropionate

meaning of a, b, c, and d is given in the upper box.

LC-SPDP = Succinimydil 6-[30-(2-pyridildithio)-propionamido]hexanoate

SSMCC = sulfosuccinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxylate

Table 1. Illustration of the use of APTES for retaining biomolecules on surfaces. The

Disks of 316L SS (0.74 cm2, from Outokumpu Stainless AB, ARC) were used. The bulk chemical composition of 316L SS was given previously (Landoulsi et al., 2008a). All chemicals (NaCl, Na2SO4, NaNO3 NaHCO3, NaHPO4, Na2PO4) were provided by Prolabo (VWR, France) and ensured 99% minimum purity. 3-aminopropyl(triethoxysilane) (APTES, 99%), glucose oxidase (Gox, EC 1.1.3.4, 47200 U.g-1) from *Aspergillus Niger* and *D*-glucose The samples (both faces and perimeter) were polished with 1 µm diamond suspension (Struers, Denmark), rinsed in binary mixture of milliQ water/ethanol (1/1, v/v) in a sonication bath (70W, 40 kHz, Branson, USA) and dried under nitrogen gas flow.The samples were then immediately immersed for 48 h in synthetic aqueous medium (NaCl 0.46 mmol.L-1, Na2SO4 0.26 mmol.L-1, NaNO3 0.2 mmol.L-1, NaHCO3 3.15 mmol.L-1, pH about 8), abundantly rinsed with milliQ water (Millipore, Molsheim, France) and dried under nitrogen gas flow. These samples are considered as native SS and designated as "nat".
