Hohenbild Frederike, Arango-Ospina Marcela, Moghaddam Arash, Boccaccini Aldo R, Westhauser Fabian
Center of Orthopedics, Traumatology and Spinal Cord Injury, Heidelberg University Hospital, Schlierbacher Landstraße 200a, 69118 Heidelberg, Germany.
Institute of Biomaterials, University of Erlangen-Nuremberg, Cauerstraße 6, 91058 Erlangen, Germany.
Methods Protoc. 2020 May 9;3(2):38. doi: 10.3390/mps3020038.
Due to their high bioreactivity, the in-vitro analysis of bioactive glasses (BGs) can be challenging when it comes to maintaining a physiological pH. To improve BG biocompatibility, a heterogenic spectrum of preconditioning approaches, such as "passivation" of the BGs by incubation in cell culture medium, are used but have never been directly compared. In this study, the effect of passivation periods of up to 72 h on pH alkalization and viability of human bone marrow-derived mesenchymal stromal cells was evaluated to determine a time-efficient passivation protocol using granules based on the 45S5-BG composition (in wt%: 45.0 SiO, 24.5 NaO, 24.5 CaO, 6.0 PO) in different concentrations. pH alkalization was most reduced after passivation of 24 h. Cell viability continuously improved with increasing passivation time being significantly higher after passivation of at least 24 h compared to non-passivated 45S5-BG and the necessary passivation time increased with increasing BG concentrations. In this setting, a passivation period of 24 h presented as an effective approach to provide a biocompatible cell culture setting. In conclusion, before introduction of BGs in cell culture, different passivation periods should be evaluated in order to meet the respective experimental settings, e.g., by following the experimental protocols used in this study.
由于生物活性玻璃（BGs）具有高生物反应活性，在体外分析时，要维持生理pH值颇具挑战性。为提高BG的生物相容性，人们采用了多种预处理方法，如将BGs置于细胞培养基中进行“钝化”，但这些方法从未被直接比较过。在本研究中，评估了长达72小时的钝化时间对人骨髓间充质基质细胞pH值碱化和活力的影响，以确定一种高效的钝化方案，该方案使用基于45S5 - BG成分（重量百分比：45.0 SiO、24.5 NaO、24.5 CaO、6.0 PO）的不同浓度颗粒。钝化24小时后，pH值碱化程度降低最为明显。细胞活力随着钝化时间的增加而持续提高，至少钝化24小时后的细胞活力显著高于未钝化的45S5 - BG，且所需的钝化时间随BG浓度的增加而增加。在这种情况下，24小时的钝化期是提供生物相容性细胞培养环境的有效方法。总之，在将BGs引入细胞培养之前，应评估不同的钝化时间，以满足各自的实验设置，例如遵循本研究中使用的实验方案。
