Biomineralizing three-dimensional bioprinted hydrogels for bone tissue replacement using dedicated peptides in high local concentrations presented on biotechnologically modified plant virus nanoparticles (PlantVirusBone)

使用生物技术修饰的植物病毒纳米粒子（PlantVirusBone）上呈现的高局部浓度的专用肽，生物矿化三维生物打印水凝胶，用于骨组织替代

• 批准号: 403762164
• 负责人: Professor Dr.-Ing. Horst Fischer
• 金额: --
• 依托单位: Lehr- und Forschungsgebiet Zahnärztliche Werkstoffkunde und Biomaterialforschung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/403762164?language=en
• 关键词: Biomineralizing; three; dimensional; bioprinted; hydrogels

A suitable biomaterial for tissue engineering must meet certain challenges such as biodegradability, cytocompatibility and bioactivity, but most hydrogels lack crucial biochemical cues for cell interaction. Current research therefore focuses on novel hydrogel blends which resemble the structure of the extracellular matrix and allow biochemical attachment of the encapsulated cells. In the first project phase, Potato virus X (PVX) was engineered to present mineralization- and osteogenesis-inducing peptides (MIPs), mimicking non-collagenous proteins (NCPs), which improved the human mesenchymal stem cells (hMSCs) osteogenesis and biomineralization in both 2D and 3D environments. Plant virus nanoparticles (VNPs) displaying different coverages of MIPs, demonstrated that the mineralization effect and cell interaction is the result of the high local density of presented MIPs, highlighting VNPs as interesting platforms for biochemical cues. Moreover, they showed a good cytocompatibility and a high retention inside the hydrogels.Another requirement for a biofunctional tissue replacement is a sufficient vascularization. Thus, the proposed second project phase aims to develop a regenerative bone tissue with multiple physicochemical properties to simultaneously provide oxygen and nutrient supply and promote osteogenesis. A synergistic osteogenic and vasculogenic effect in both 2D cell culture and 3D cell-embedded hydrogels can be achieved by employing different VNP modifications with various peptides derived from NCPs and vascular endothelial growth factor. VNPs will be modified with novel strategies, including different ribosomal skipping sequences, and covalent plug-and-display systems. The latter enables VNPs to interconnect to large bundles, which can produce a hydrogel itself consisting of a network of synergistically acting functional peptides. A capillary-like network will be induced by co-culturing hMSCs and human umbilical vein endothelial cells on 2D VNP-coated surfaces as well as in VNP-laden 3D hydrogels. Osteogenic and angiogenic capacities of the cells will be evaluated especially by real-time PCR, migration assay and fluorescence imaging. The mechanical properties, vital factors for influencing the cell behavior, will be tuned by changing the physical or chemical composition of hydrogels. Viscosity and stiffness of the VNP-laden hydrogels will be determined for different VNPs and hydrogels. Finally, bioprinting technology will be applied to generate a tissue substitute with spatially defined VNPs, cells and materials organization. The influence on the cell response will be studied in detail.

一种适合组织工程的生物材料必须满足一定的挑战，如生物可降解性、细胞相容性和生物活性，但大多数水凝胶缺乏细胞相互作用的关键生化线索。因此，目前的研究重点是新的水凝胶混合物，它类似于细胞外基质的结构，并允许被包裹细胞的生化附着。在第一个项目阶段，马铃薯病毒X （PVX）被改造成矿化和成骨诱导肽（MIPs），模仿非胶原蛋白（ncp），在2D和3D环境下改善人间充质干细胞（hMSCs）的成骨和生物矿化。植物病毒纳米颗粒（VNPs）显示不同的MIPs覆盖率，表明矿化效应和细胞相互作用是高局部密度的MIPs的结果，突出了VNPs作为生化线索的有趣平台。此外，它们还表现出良好的细胞相容性和水凝胶内的高保留率。生物功能组织替代的另一个要求是充分的血管化。因此，拟建的第二阶段项目旨在开发具有多种物理化学特性的再生骨组织，同时提供氧气和营养供应，促进成骨。在2D细胞培养和3D细胞包埋的水凝胶中，通过使用不同的VNP修饰，从ncp和血管内皮生长因子中衍生出不同的肽，可以实现协同成骨和血管生成效应。VNPs将采用新的策略进行修饰，包括不同的核糖体跳跃序列和共价插入-展示系统。后者使VNPs能够与大束相互连接，这可以产生由协同作用功能肽网络组成的水凝胶。将hMSCs和人脐静脉内皮细胞在二维vnp包被表面以及负载vnp的三维水凝胶中共培养，可诱导出毛细血管样网络。细胞的成骨和血管生成能力将通过实时PCR、迁移试验和荧光成像进行评估。机械性能是影响细胞行为的重要因素，可以通过改变水凝胶的物理或化学成分来调节。将对不同的vnp和水凝胶测定负载vnp的水凝胶的粘度和刚度。最后，生物打印技术将应用于生成具有空间定义的VNPs、细胞和材料组织的组织替代品。对细胞反应的影响将被详细研究。