In vitro tests that verify the ability of a material to form bone-like apatite precipitated (HAp) use a solution that imitates the inorganic part of blood plasma, simulated body fluid (SBF). During such tests, it is necessary to maintain a neutral pH, for which purpose the International Standards Organization recommends the TRIS (tris(hydroxymethyl)aminomethane) buffer (ISO 23,317:2014). To do this, TRIS buffer must remain inert, but, as we have previously reported, TRIS interacts with highly bioreactive materials (such as 45S5 Bioglass-derived scaffolds), thereby accelerating scaffold dissolution. In the search for an alternative to TRIS, we have also recently published results for the 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 3-(N-morpholino) propanesulfonic acid (MOPS) buffers, which were also unable to maintain a neutral pH in SBF. Thus, we here continue our search for a more suitable Good’s buffer by comparing BES (N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid) and TES (2-[(2-hydroxy-1,1-bis(hydroxymethyl) ethyl) amino] ethanesulfonic acid) with TRIS. 45S5-derived glass–ceramic scaffolds were tested under static–dynamic conditions for bone-like apatite formation using SBF + BES and SBF + TES solutions. The pH measurements, leachate analysis [atomic absorption spectrophotometry (AAS) (Ca²⁺, Si^IV), UltraViolet-Visible (UV-VIS) spectrophotometry (PO₄)³⁻], and scaffold analysis [X-Ray powder diffraction analysis (XRD), X-ray fluorescent analysis (XRF), scanning electron microscopy (SEM)/energy dispersive spectroscopy (EDS), Brunauer.Emmett.Teller specific surface area analysis (BET)] all showed that both BES and TES quickly interacted with the tested material. Moreover, the kinetics of the glass–ceramic dissolution affected the crystallinity and morphology of the precipitated HAp. Based on our previous results and the present data, it appears that Good’s buffers are not suitable for the in vitro testing of bioactivity of highly bioreactive materials.

体外验证材料形成骨状磷灰石沉淀 (HAp) 的能力的测试使用模拟血浆无机部分的溶液，模拟体液 (SBF)。在此类测试期间，必须保持中性 pH 值，为此国际标准组织推荐使用 TRIS（三（羟甲基）氨基甲烷）缓冲液 (ISO 23,317:2014)。为此，TRIS 缓冲液必须保持惰性，但正如我们之前报道的那样，TRIS 会与高生物反应性材料（例如 45S5 生物玻璃衍生支架）相互作用，从而加速支架溶解。在寻找替代 TRIS 的过程中，我们最近还发布了 4-(2-羟乙基)-1-哌嗪乙磺酸 (HEPES) 和 3-(N-吗啉代) 丙磺酸 (MOPS) 缓冲液的结果，它们也是无法在 SBF 中保持中性 pH。N，N-双（2-羟乙基）-2-氨基乙磺酸）和TES（2-[（2-羟基-1,1-双（羟甲基）乙基）氨基]乙磺酸）与TRIS。使用 SBF + BES 和 SBF + TES 溶液在静态-动态条件下测试了 45S5 衍生的玻璃陶瓷支架的骨样磷灰石形成。pH 测量、渗滤液分析 [原子吸收分光光度法 (AAS) (Ca ²⁺ , Si ^IV )、紫外-可见 (UV-VIS) 分光光度法 (PO ₄ ) ³⁻]、支架分析[X射线粉末衍射分析(XRD)、X射线荧光分析(XRF)、扫描电子显微镜(SEM)/能量色散光谱(EDS)、Brunauer.Emmett.Teller比表面积分析(BET) )] 都表明 BES 和 TES 都与测试材料快速相互作用。此外，玻璃陶瓷溶解的动力学影响沉淀的 HAp 的结晶度和形态。根据我们之前的结果和目前的数据，Good 的缓冲液似乎不适合对高生物反应性材料的生物活性进行体外测试。