UMR 7365 CNRS, Université de Lorraine, IMoPA, Bâtiment Biopôle - Faculté de Médecine, Nancy, France.
J Tissue Eng Regen Med. 2018 Mar;12(3):e1757-e1766. doi: 10.1002/term.2454. Epub 2017 Jul 14.
Cartilage tissue engineering is making progress, but the competing available strategies still leave room for improvement and consensual overviews regarding the best combinations of scaffolds and cell sources are limited by the capacity to compare them directly. In addition, because most strategies involve autologous cell transfer, once these are optimized, the resulting implants require individual quality control prior to grafting in order to emphasize patient-to-patient differential responsiveness to engineering processes. Here, cartilage substitutes prepared from human mesenchymal stem cells undergoing chondrogenic differentiation within distinct scaffolds were used as pilot samples to investigate the pertinence of a novel method with the aim of characterizing the implants. The limits and advantages of analysing, by label-free liquid chromatography-coupled matrix-assisted laser desorption and ionization (LC-MALDI) mass spectrometry, the secreted proteome released into culture medium by engineered cartilage tissues were investigated and compared with more classically used methods for biomaterial characterization. This method did not require sacrificing the biomaterials and robustly evidenced their chondrogenic statuses. In more detail, the method highlighted differences between batches prepared from distinct donors. It was adapted to distinct scaffolds and allowed a comparison of the influence of individual engineering steps, such as growth factor combinations and oxygen tension. Finally, it evidenced subtle changes between replicate substitutes within a series, thereby distinguishing the least and most accomplished ones. We conclude that relative quantification of secreted proteins through label-free LC-MALDI will be useful, not only to orientate engineering methodologies, but also to ultimately provide non-invasive quality control of engineered tissue substitutes for the repair of cartilage and possibly other connective tissues.
软骨组织工程学正在取得进展，但现有的竞争策略仍有改进的空间，而且由于大多数策略都涉及自体细胞转移，因此一旦这些策略得到优化，所得到的植入物在移植前需要进行个体质量控制，以强调患者对工程过程的差异性反应。在这里，使用从人类间充质干细胞在不同支架中进行软骨分化制备的软骨替代物作为试点样本，以研究一种新方法的相关性，目的是对植入物进行特征分析。本研究通过无标记液相色谱-基质辅助激光解吸电离（LC-MALDI）质谱法分析工程化软骨组织分泌到培养基中的分泌蛋白质组，并与更经典的生物材料特性分析方法进行了比较。该方法不需要牺牲生物材料，并且可以强有力地证明其软骨生成状态。更详细地说，该方法突出了不同供体来源的批次之间的差异。它适用于不同的支架，并允许比较单个工程步骤的影响，例如生长因子组合和氧张力。最后，它证明了一系列替代物中重复替代物之间的细微变化，从而区分了最差和最成功的替代物。我们得出结论，通过无标记 LC-MALDI 进行分泌蛋白的相对定量不仅有助于指导工程方法，而且最终可以对用于软骨修复和可能其他结缔组织修复的工程化组织替代物进行非侵入性质量控制。
