Four mineralization buffers were prepared, solutions A, B, C, and D [25 , 26 (link)].
Mineralization buffer A was prepared with 2.12 mM calcium chloride dihydrate (CaCl2·2H2O: Fujifilm Wako Pure Chemical Co., Osaka, Japan), 1.27 mM K-PO4, which is a mixture of potassium dihydrogenphosphate and dipotassium hydrogen phosphate (KH2PO4, K2HPO4: Kanto Chemical Co., Inc., Tokyo, Japan). Mineralization buffer B was prepared with 2.24 mM CaCl2·2H2O and 1.34 mM K-PO4. Mineralization buffer C was prepared with 2.35 mM CaCl2·2H2O and 1.41 mM K-PO4. The final pH of mineralization buffers A, B, and C was adjusted to 7.4 by 10 mM HEPES–KOH [prepared from 10 mM hydroxyethyl piperazine ethane sulfonic acid (HEPES: Nacalai Tesque, Inc., Kyoto, Japan) mixed with potassium hydroxide (KOH: Nacalai Tesque, Inc.), and 150 mM potassium chloride (KCl: Nacalai Tesque, Inc.)].
Mineralization buffer D was prepared with 2.58 mM CaCl2·2H2O, 1.55 mM KH2PO4, and 180 mM sodium chloride (NaCl); the pH was adjusted to 7.6 with 50 mM Tris–HCl.
With all mineralization buffers, pH adjustment was carried out using a pH meter (F-22: Horiba Ltd., Kyoto, Japan). All of the mineralization buffers used in the present study had a Ca/P molar ratio of 1.67; the degrees of solution saturations with respect to HAp were (A) 2.58 × 107, (B) 3.85 × 107, (C) 5.50 × 107, and (D) 8.32 × 107 [25 ].
Mineralization buffer A was prepared with 2.12 mM calcium chloride dihydrate (CaCl2·2H2O: Fujifilm Wako Pure Chemical Co., Osaka, Japan), 1.27 mM K-PO4, which is a mixture of potassium dihydrogenphosphate and dipotassium hydrogen phosphate (KH2PO4, K2HPO4: Kanto Chemical Co., Inc., Tokyo, Japan). Mineralization buffer B was prepared with 2.24 mM CaCl2·2H2O and 1.34 mM K-PO4. Mineralization buffer C was prepared with 2.35 mM CaCl2·2H2O and 1.41 mM K-PO4. The final pH of mineralization buffers A, B, and C was adjusted to 7.4 by 10 mM HEPES–KOH [prepared from 10 mM hydroxyethyl piperazine ethane sulfonic acid (HEPES: Nacalai Tesque, Inc., Kyoto, Japan) mixed with potassium hydroxide (KOH: Nacalai Tesque, Inc.), and 150 mM potassium chloride (KCl: Nacalai Tesque, Inc.)].
Mineralization buffer D was prepared with 2.58 mM CaCl2·2H2O, 1.55 mM KH2PO4, and 180 mM sodium chloride (NaCl); the pH was adjusted to 7.6 with 50 mM Tris–HCl.
With all mineralization buffers, pH adjustment was carried out using a pH meter (F-22: Horiba Ltd., Kyoto, Japan). All of the mineralization buffers used in the present study had a Ca/P molar ratio of 1.67; the degrees of solution saturations with respect to HAp were (A) 2.58 × 107, (B) 3.85 × 107, (C) 5.50 × 107, and (D) 8.32 × 107 [25 ].
Free full text: Click here
