Dewey Marley J, Kolliopoulos Vasiliki, Ngo Mai T, Harley Brendan A C
Dept. of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Dept. of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Materialia (Oxf). 2021 Aug;18. doi: 10.1016/j.mtla.2021.101149. Epub 2021 Jun 18.
Effective design of biomaterials to aid regenerative repair of craniomaxillofacial (CMF) bone defects requires approaches that modulate the complex interplay between exogenously added progenitor cells and cells in the wound microenvironment, such as osteoblasts, osteoclasts, endothelial cells, and immune cells. We are exploring the role of the glycosaminoglycan (GAG) content in a class of mineralized collagen scaffolds recently shown to promote osteogenesis and healing of craniofacial bone defects. We previously showed that incorporating chondroitin-6-sulfate or heparin improved mineral deposition by seeded human mesenchymal stem cells (hMSCs). Here, we examine the effect of varying scaffold GAG content on hMSC behavior, and their ability to modulate osteoclastogenesis, vasculogenesis, and the immune response. We report the role of hMSC-conditioned media produced in scaffolds containing chondroitin-6-sulfate (CS6), chondroitin-4-sulfate (CS4), or heparin (Heparin) GAGs on endothelial tube formation and monocyte differentiation. Notably, endogenous production by hMSCs within Heparin scaffolds most significantly inhibits osteoclastogenesis via secreted osteoprotegerin (OPG), while the secretome generated by CS6 scaffolds reduced pro-inflammatory immune response and increased endothelial tube formation. All conditioned media down-regulated many pro- and anti-inflammatory cytokines, such as IL6, IL-1β, and CCL18 and CCL17 respectively. Together, these findings demonstrate that modifying mineralized collagen scaffold GAG content can both directly (hMSC activity) and indirectly (production of secreted factors) influence overall osteogenic potential and mineral biosynthesis as well as angiogenic potential and monocyte differentiation towards osteoclastic and macrophage lineages. Scaffold GAG content is therefore a powerful stimulus to modulate reciprocal signaling between multiple cell populations within the bone healing microenvironment.
有效设计生物材料以辅助颅颌面（CMF）骨缺损的再生修复，需要采用一些方法来调节外源性添加的祖细胞与伤口微环境中的细胞（如成骨细胞、破骨细胞、内皮细胞和免疫细胞）之间复杂的相互作用。我们正在探索糖胺聚糖（GAG）含量在一类最近显示可促进颅面骨缺损骨生成和愈合的矿化胶原支架中的作用。我们之前表明，掺入硫酸软骨素-6或肝素可改善接种的人间充质干细胞（hMSC）的矿物质沉积。在此，我们研究了不同支架GAG含量对hMSC行为的影响，以及它们调节破骨细胞生成、血管生成和免疫反应的能力。我们报告了在含有硫酸软骨素-6（CS6）、硫酸软骨素-4（CS4）或肝素的GAG支架中产生的hMSC条件培养基对内皮管形成和单核细胞分化的作用。值得注意的是，肝素支架内hMSC的内源性产生通过分泌骨保护素（OPG）最显著地抑制破骨细胞生成，而CS6支架产生的分泌蛋白组减少了促炎免疫反应并增加了内皮管形成。所有条件培养基均分别下调了许多促炎和抗炎细胞因子，如IL6、IL-1β以及CCL18和CCL17。总之，这些发现表明，改变矿化胶原支架的GAG含量可直接（hMSC活性）和间接（分泌因子的产生）影响整体成骨潜力和矿物质生物合成，以及血管生成潜力和单核细胞向破骨细胞和巨噬细胞谱系的分化。因此，支架GAG含量是调节骨愈合微环境中多个细胞群体之间相互信号传导的有力刺激因素。
