Alinejad Yasaman, Lauzon Marc-Antoine, Grenier Guillaume, Balg Frédéric, Faucheux Nathalie
Laboratory of Endovascular Biomaterials (LBeV), Centre de recherche du CHUM (CRCHUM), 900 Saint-Denis Street, Montreal, QC H2X 0A9, Canada.
Department of Mechanical Engineering, École de Technologie Supérieure (ETS), 1100 Notre-Dame West, Montreal, QC H3C 1K3, Canada.
J Clin Med. 2020 Apr 19;9(4):1175. doi: 10.3390/jcm9041175.
Hematoma and skeletal muscles play a crucial role in bone fracture healing. The muscle resident mesenchymal stromal cells (mrSCs) can promote bone formation by differentiating into osteoblasts upon treatment by bone morphogenetic proteins (BMP), such as BMP9. However, the influence of hematoma fracture extracts (Hema) on human mrSC (hmrSC) response to BMP9 is still unknown. We therefore determined the influence of Hema, human healthy serum (HH), and fetal bovine serum (FBS, control) on BMP9-induced osteoblast commitment of hmrSC by measuring alkaline phosphatase activity. Multiplex assays of 90 cytokines were performed to characterize HH and Hema composition and allow their classification by a multivariate statistical approach depending on their expression levels. We confirmed that BMP9 had a greater effect on osteoblastic differentiation of hmrSCs than BMP2 in presence of FBS. The hmrSCs response to BMP9 was enhanced by both Hema and HH, even though several cytokines were upregulated (IL-6, IL-8, MCP-1, VEGF-A and osteopontin), downregulated (BMP9, PDGF) or similar (TNF-alpha) in Hema compared with HH. Thus, hematoma may potentiate BMP9-induced osteogenic differentiation of hmrSCs during bone fracture healing. The multivariate statistical analyses will help to identify the cytokines involved in such phenomenon leading to normal or pathological bone healing.
血肿和骨骼肌在骨折愈合过程中发挥着关键作用。肌肉驻留间充质基质细胞（mrSCs）在受到骨形态发生蛋白（BMP）（如BMP9）处理后可分化为成骨细胞，从而促进骨形成。然而，血肿骨折提取物（Hema）对人mrSC（hmrSC）对BMP9反应的影响尚不清楚。因此，我们通过测量碱性磷酸酶活性，确定了Hema、人健康血清（HH）和胎牛血清（FBS，作为对照）对BMP9诱导的hmrSC成骨细胞定向分化的影响。对90种细胞因子进行了多重分析，以表征HH和Hema的成分，并根据它们的表达水平通过多变量统计方法对其进行分类。我们证实，在存在FBS的情况下，BMP9对hmrSCs的成骨细胞分化作用比BMP2更大。尽管与HH相比，Hema中有几种细胞因子上调（IL-6、IL-8、MCP-1、VEGF-A和骨桥蛋白）、下调（BMP9、PDGF）或相似（TNF-α），但Hema和HH均增强了hmrSCs对BMP9的反应。因此，在骨折愈合过程中，血肿可能增强BMP9诱导的hmrSCs成骨分化。多变量统计分析将有助于识别参与这种导致正常或病理性骨愈合现象的细胞因子。
