Wolint Petra, Näf Lukas, Schibler Désirée, Hild Nora, Stark Wendelin J, Giovanoli Pietro, Calcagni Maurizio, Buschmann Johanna
Division of Plastic and Hand Surgery, University Hospital Zurich, 8091 Zurich, Switzerland.
Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.
Biology (Basel). 2021 Jul 16;10(7):675. doi: 10.3390/biology10070675.
Amorphous calcium phosphate (aCaP) nanoparticles may trigger the osteogenic commitment of adipose-derived stem cells (ASCs) in vitro. The ASCs of three human donors are investigated using basal culture medium DMEM to either 5 or 50 µg/mL aCaP nanoparticles suspension (control: no nanoparticles). After 7 or 14 days, stem cell marker genes, as well as endothelial, osteogenic, chondrogenic, and adipogenic genes, are analyzed by qPCR. Free calcium and phosphate ion concentrations are assessed in the cell culture supernatant. After one week and 5 µg/mL aCaP, downregulation of osteogenic markers ALP and Runx2 is found, and averaged across the three donors. Our results show that after two weeks, ALP is further downregulated, but Runx2 is upregulated. Endothelial cell marker genes, such as CD31 and CD34, are upregulated with 50 µg/mL aCaP and a 2-week exposure. Inter-donor variability is high: Two out of three donors show a significant upregulation of ALP and Runx2 at day 14 with 50 µg/mL aCaP compared to 5 µg/mL aCaP. Notably, all changes in stem cell commitment are obtained in the absence of an osteogenic medium. While the chemical composition of the culture medium and the saturation status towards calcium phosphate phases remain approximately the same for all conditions, gene expression of ASCs changes considerably. Hence, aCaP nanoparticles show the potential to trigger osteogenic and endothelial commitment in ASCs.
无定形磷酸钙(aCaP)纳米颗粒可能在体外触发脂肪来源干细胞(ASC)的成骨定向分化。使用基础培养基DMEM对三名人类供体的ASC进行研究,分别加入5或50μg/mL的aCaP纳米颗粒悬浮液(对照:无纳米颗粒)。7天或14天后,通过qPCR分析干细胞标记基因以及内皮、成骨、软骨生成和脂肪生成基因。评估细胞培养上清液中的游离钙和磷酸根离子浓度。在加入5μg/mL aCaP一周后,发现成骨标记物碱性磷酸酶(ALP)和Runx2下调,并对三名供体进行了平均。我们的结果表明,两周后,ALP进一步下调,但Runx2上调。内皮细胞标记基因,如CD31和CD34,在加入50μg/mL aCaP并暴露2周后上调。供体间变异性较高:与5μg/mL aCaP相比,三名供体中有两名在第14天使用50μg/mL aCaP时显示ALP和Runx2显著上调。值得注意的是,干细胞定向分化的所有变化都是在没有成骨培养基的情况下获得的。虽然所有条件下培养基的化学成分和磷酸钙相的饱和状态大致相同,但ASC的基因表达却有很大变化。因此,aCaP纳米颗粒显示出触发ASC成骨和内皮定向分化的潜力。