Osteoclasts degrade fibrinogen scaffolds and induce mesenchymal stem/stromal osteogenic differentiation
Osteoclasts degrade fibrinogen scaffolds and induce mesenchymal stem/stromal osteogenic differentiation
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DOI：
10.1002/jbm.a.36863
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发表时间：
2019-12-24
影响因子：
4.9
通讯作者：
Santos, Susana G.
中科院分区：
文献类型：
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作者：
Almeida, Ana R.;Bessa-Goncalves, Mafalda;Santos, Susana G.
纤维蛋白原（Fg）是具有促愈合特性的促炎蛋白。之前的工作表明纤维蛋白原3D支架（Fg-3D）促进骨再生，但尚未确定细胞参与者。破骨细胞是一种骨吸收细胞，与成骨细胞密切相关，可促进骨重建。在此，在体外评估在Fg-3D上分化的破骨细胞降解支架和促进成骨细胞分化的能力。采用冷冻干燥法制备Fg-3D支架，并从原代人外周血单核细胞中诱导分化破骨细胞。获得的结果显示破骨细胞表达的酶组织蛋白酶K和抗酒石酸酸性磷酸酶定植Fg-3D支架。破骨细胞能够显著降解Fg-3D，减少支架的面积，并增加培养基中Fg降解产物D-二聚体的浓度。破骨细胞条件培养基从分化的第一周促进显着更强的人原代间充质干细胞/基质细胞（MSC）成骨分化，碱性磷酸酶活性评价。此外，第1周破骨细胞条件培养基促进早期MSC成骨分化，比化学成骨诱导剂。发现TGF-β 1在破骨细胞条件培养基中从分化第1周起与分化第3周相比增加。总之，我们的结果表明，破骨细胞能够分化和降解Fg-3D，产生促进MSC成骨分化的TGF-β 1等因子。
Fibrinogen (Fg) is a pro-inflammatory protein with pro-healing properties. Previous work showed that fibrinogen 3D scaffolds (Fg-3D) promote bone regeneration, but the cellular players were not identified. Osteoclasts are bone resorbing cells that promote bone remodeling in close crosstalk with osteoblasts. Herein, the capacity of osteoclasts differentiated on Fg-3D to degrade the scaffolds and promote osteoblast differentiation was evaluated in vitro. Fg-3D scaffolds were prepared by freeze-drying and osteoclasts were differentiated from primary human peripheral blood monocytes. Results obtained showed osteoclasts expressing the enzymes cathepsin K and tartrate resistant acid phosphatase colonizing Fg-3D scaffolds. Osteoclasts were able to significantly degrade Fg-3D, reducing the scaffold's area, and increasing D-dimer concentration, a Fg degradation product, in their culture media. Osteoclast conditioned media from the first week of differentiation promoted significantly stronger human primary mesenchymal stem/stromal cell (MSC) osteogenic differentiation, evaluated by alkaline phosphatase activity. Moreover, week 1 osteoclast conditioned media promoted earlier MSC osteogenic differentiation, than chemical osteogenesis inductors. TGF-beta 1 was found increased in osteoclast conditioned media from week 1, when compared to week 3 of differentiation. Taken together, our results suggest that osteoclasts are able to differentiate and degrade Fg-3D, producing factors like TGF-beta 1 that promote MSC osteogenic differentiation.