Dental implants with titanium grade 4, size 3.75 by 6 mm, external hexagon platform (P-I Brånemark, São Paulo, Brazil) were hydroxylated in a solution of H2SO4/30%H2O2 (dilution ratio 1:1) for 1 h prior to being silanized by dipping in a 10% APTES (Sigma-Aldrich Corp., St. Louis, MO, USA) in boiling anhydrous toluene (Merck SA, Rio de Janeiro, Brazil) for 4 h under reflux [8 (link),19 (link)]. It is critical to use an anhydrous solvent to control the increase the aminosilane polymerization that occurs on the surface [29 (link)]. Besides, the hydrogen bonds can be disturbed with high temperatures which then can reduce the number of infirm bonded silane molecules [30 (link)].
The titanium implants were subsequently cleaned in toluene and acetone (Merck SA/ Brazil, Rio de Janeiro, Brazil) during 10 min each in an ultrasonic bath to remove unbonded molecules. The implants were dried overnight at 110 °C in a vacuum oven to condensate the hydrogen-bonded silanols into siloxane bonds [29 (link)]. The control group only received titanium screws treated with acid (Ti) to present the same surface topography as the test group. All implants were sterilized with 25 kGy gamma radiation before use (CBE Embrarad, SP, Brazil).
The titanium implants were subsequently cleaned in toluene and acetone (Merck SA/ Brazil, Rio de Janeiro, Brazil) during 10 min each in an ultrasonic bath to remove unbonded molecules. The implants were dried overnight at 110 °C in a vacuum oven to condensate the hydrogen-bonded silanols into siloxane bonds [29 (link)]. The control group only received titanium screws treated with acid (Ti) to present the same surface topography as the test group. All implants were sterilized with 25 kGy gamma radiation before use (CBE Embrarad, SP, Brazil).
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