Bolander Johanna, Chai Yoke Chin, Geris Liesbet, Schrooten Jan, Lambrechts Dennis, Roberts Scott J, Luyten Frank P
Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Bus 813, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Bus 813, 3000 Leuven, Belgium.
Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Bus 813, 3000 Leuven, Belgium; Biomechanics Research Unit, University of Liege, Chemin des Chevreuils 1, BAT 52/3, 4000 Liege 1, Belgium; Biomechanics Section, KU Leuven, Celestijnenlaan 300C, Bus 2419, 3001 Leuven, Belgium.
Biomaterials. 2016 Apr;86:106-18. doi: 10.1016/j.biomaterials.2016.01.059. Epub 2016 Jan 29.
The development of osteoinductive calcium phosphate- (CaP) based biomaterials has, and continues to be, a major focus in the field of bone tissue engineering. However, limited insight into the spatiotemporal activation of signalling pathways has hampered the optimisation of in vivo bone formation and subsequent clinical translation. To gain further knowledge regarding the early molecular events governing bone tissue formation, we combined human periosteum derived progenitor cells with three types of clinically used CaP-scaffolds, to obtain constructs with a distinct range of bone forming capacity in vivo. Protein phosphorylation together with gene expression for key ligands and target genes were investigated 24 hours after cell seeding in vitro, and 3 and 12 days post ectopic implantation in nude mice. A computational modelling approach was used to deduce critical factors for bone formation 8 weeks post implantation. The combined Ca(2+)-mediated activation of BMP-, Wnt- and PKC signalling pathways 3 days post implantation were able to discriminate the bone forming from the non-bone forming constructs. Subsequently, a mathematical model able to predict in vivo bone formation with 96% accuracy was developed. This study illustrates the importance of defining and understanding CaP-activated signalling pathways that are required and sufficient for in vivo bone formation. Furthermore, we demonstrate the reliability of mathematical modelling as a tool to analyse and deduce key factors within an empirical data set and highlight its relevance to the translation of regenerative medicine strategies.
基于骨诱导磷酸钙（CaP）的生物材料的开发一直是骨组织工程领域的主要研究重点，并且仍将持续如此。然而，对信号通路时空激活的了解有限，阻碍了体内骨形成的优化以及后续的临床转化。为了进一步了解骨组织形成早期的分子事件，我们将人骨膜来源的祖细胞与三种临床使用的CaP支架相结合，以获得在体内具有不同骨形成能力范围的构建体。在体外细胞接种后24小时以及在裸鼠异位植入后3天和12天，研究了关键配体和靶基因的蛋白质磷酸化以及基因表达。采用计算建模方法推断植入后8周骨形成的关键因素。植入后3天，Ca(2+)介导的BMP、Wnt和PKC信号通路的联合激活能够区分有骨形成能力和无骨形成能力的构建体。随后，开发了一个能够以96%的准确率预测体内骨形成的数学模型。本研究说明了定义和理解体内骨形成所需且充分的CaP激活信号通路的重要性。此外，我们证明了数学建模作为一种工具来分析和推导经验数据集中关键因素的可靠性，并强调了其与再生医学策略转化的相关性。
