Othman Z, Mohren R J C, Cillero-Pastor B, Shen Z, Lacroix Y S N W, Guttenplan A P M, Tahmasebi Birgani Z, Eijssen L, Luider T M, van Rijt S, Habibovic P
MERLN Institute for Technology-Inspired Regenerative Medicine, Department of Instructive Biomaterials Engineering, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands.
The Maastricht Multimodal Molecular Imaging Institute (M4I), Division of Imaging Mass Spectrometry, Maastricht University, Universiteitssingel 50, 6229 ER, Maastricht, the Netherlands.
Mater Today Bio. 2020 Jun 24;7:100066. doi: 10.1016/j.mtbio.2020.100066. eCollection 2020 Jun.
In recent years, synthetic calcium phosphate (CaP) ceramics have emerged as an alternative to bone grafts in the treatment of large critical-sized bone defects. To successfully substitute for bone grafts, materials must be osteoinductive, that is, they must induce osteogenic differentiation and subsequent bone formation . Although a set of osteoinductive CaP ceramics has been developed, the precise biological mechanism by which a material directs cells toward osteogenesis and the role of individual chemical and physical properties in this mechanism remain incompletely understood. Here, we used proteomics to compare serum protein adsorption to two CaP ceramics with different osteoinductive potential, namely an osteoinductive β-tricalcium phosphate (TCP) and a non-osteoinductive hydroxyapatite (HA). Moreover, we analyzed the protein profiles of human mesenchymal stromal cells (hMSCs) cultured on these two ceramics. The serum protein adsorption experiments in the absence of cells highlighted the proteins that are highly abundant in the serum and/or have a high affinity to CaP. The extent of adsorption was suggested to be affected by the available surface area for binding and by the ion exchange dynamics on the surface. Several proteins were uniquely expressed by hMSCs on TCP and HA surfaces. Proteins identified as enriched on TCP were involved in processes related to wound healing, cell proliferation, and the production of extracellular matrix. On the other hand, proteins that were enriched on HA were involved in processes related to protein production, translation, localization, and secretion. In addition, we performed a separate proteomics analysis on TCP, HA, and two biphasic calcium phosphates with known osteoinductive potential and performed a clustering analysis on a combination of a set of proteins found to be enriched on osteoinductive materials with a set of proteins already known to be involved in osteogenesis. This yielded two protein networks potentially involved in the process of osteoinduction - one consisting of collagen fragments and collagen-related enzymes and a second consisting of endopeptidase inhibitors and regulatory proteins. The results of this study show that protein profiling can be a useful tool to help understand the effect of biomaterial properties on the interactions between a biomaterial and a biological system. Such understanding will contribute to the design and development of improved biomaterials for (bone) regenerative therapies.
近年来，合成磷酸钙（CaP）陶瓷已成为治疗大型临界尺寸骨缺损时骨移植的替代物。为了成功替代骨移植材料，该材料必须具有骨诱导性，也就是说，它们必须诱导成骨细胞分化并随后形成骨组织。尽管已经开发出了一组具有骨诱导性的CaP陶瓷，但材料引导细胞向成骨方向分化的确切生物学机制以及各个化学和物理性质在该机制中的作用仍未完全了解。在这里，我们使用蛋白质组学来比较血清蛋白在两种具有不同骨诱导潜力的CaP陶瓷上的吸附情况，这两种陶瓷分别是具有骨诱导性的β-磷酸三钙（TCP）和无骨诱导性的羟基磷灰石（HA）。此外，我们分析了在这两种陶瓷上培养的人间充质基质细胞（hMSCs）的蛋白质谱。在无细胞情况下进行的血清蛋白吸附实验突出了血清中高度丰富和/或对CaP具有高亲和力的蛋白质。吸附程度被认为受结合可用表面积和表面离子交换动力学的影响。hMSCs在TCP和HA表面上独特地表达了几种蛋白质。被确定在TCP上富集的蛋白质参与了与伤口愈合、细胞增殖和细胞外基质产生相关的过程。另一方面，在HA上富集的蛋白质参与了与蛋白质产生、翻译、定位和分泌相关的过程。此外，我们对TCP、HA以及两种已知具有骨诱导潜力的双相磷酸钙进行了单独的蛋白质组学分析，并对一组在具有骨诱导性的材料上富集的蛋白质与一组已知参与成骨的蛋白质进行了聚类分析。这产生了两个可能参与骨诱导过程的蛋白质网络——一个由胶原蛋白片段和胶原蛋白相关酶组成，另一个由内肽酶抑制剂和调节蛋白组成。这项研究的结果表明，蛋白质谱分析可以成为帮助理解生物材料特性对生物材料与生物系统之间相互作用影响的有用工具。这种理解将有助于设计和开发用于（骨）再生治疗的改良生物材料。
