雷利度胺是否影响骨形成？：人骨髓间充质干细胞和成骨细胞的体外研究。
Does Riluzole Influence Bone Formation?: An In Vitro Study of Human Mesenchymal Stromal Cells and Osteoblast.
机构信息
Department of Orthopaedic Surgery, The Rothman Institute at Thomas Jefferson University, Philadelphia, PA.
AO Research Institute Davos, Davos, Switzerland.
出版信息
Spine (Phila Pa 1976). 2019 Aug 15;44(16):1107-1117. doi: 10.1097/BRS.0000000000003022.
STUDY DESIGN
A post-test design biological experiment.
OBJECTIVE
The aim of this study was to evaluate the osteogenic effects of riluzole on human mesenchymal stromal cells and osteoblasts.
SUMMARY OF BACKGROUND DATA
Riluzole may benefit patients with spinal cord injury (SCI) from a neurologic perspective, but little is known about riluzole's effect on bone formation, fracture healing, or osteogenesis.
METHODS
Human mesenchymal stromal cells (hMSCs) and human osteoblasts (hOB) were obtained and isolated from healthy donors and cultured. The cells were treated with riluzole of different concentrations (50, 150, 450 ng/mL) for 1, 2, 3, and 4 weeks. Cytotoxicity was evaluated as was the induction of osteogenic differentiation of hMSCs. Differentiation was evaluated by measuring alkaline phosphatase (ALP) activity and with Alizarin red staining. Osteogenic gene expression of type I collagen (Col1), ALP, osteocalcin (Ocn), Runx2, Sox9, Runx2/Sox9 ratio were measured by qRT-PCR.
RESULTS
No cytotoxicity or increased proliferation was observed in bone marrow derived hMSCs and primary hOBs cultured with riluzole over 7 days. ALP activity was slightly increased in hMSCs after treatment for 2 weeks with riluzole 150 ng/mL and slightly upregulated by 150% (150 ng/mL) and 90% (450 ng/mL) in hMSCs at 3 weeks. In hOBs, ALP activity almost doubled after 2 weeks of culture with riluzole 150 ng/mL (P < 0.05). More pronounced 2.6-fold upregulation was noticed after 3 weeks of culture with riluzole at both 150 ng/mL (P = 0.05) and 450 ng/mL (P = 0.05). No significant influence of riluzole on the mRNA expression of osteocalcin (OCN) was observed.
CONCLUSION
The effect of riluzole on bone formation is mixed; low-dose riluzole has no effect on the viability or function of either hMSCs or hOBs. The activity of ALP in both cell types is upregulated by high-dose riluzole, which may indicate that high-dose riluzole can increase osteogenic metabolism and subsequently accelerate bone healing process. However, at high concentrations, riluzole leads to a decrease in osteogenic gene expression, including Runx2 and type 1 collagen.
LEVEL OF EVIDENCE
N/A.
研究设计
一个测试后设计的生物实验。
目的
本研究旨在评估利鲁唑对人基质干细胞和成骨细胞的成骨作用。
背景资料概要
利鲁唑可能对脊髓损伤（SCI）患者从神经学角度有益，但对其对骨形成、骨折愈合或成骨的影响知之甚少。
方法
从健康供体中获得并分离出人基质干细胞（hMSCs）和人成骨细胞（hOB），并进行培养。用不同浓度（50、150、450ng/ml）的利鲁唑处理细胞 1、2、3 和 4 周。评估细胞毒性和成骨细胞向 hMSCs 的诱导分化。通过碱性磷酸酶（ALP）活性和茜素红染色来评估分化。通过 qRT-PCR 测量Ⅰ型胶原（Col1）、ALP、骨钙素（Ocn）、Runx2、Sox9、Runx2/Sox9 比值的成骨基因表达。
结果
骨髓来源的 hMSCs 和原代 hOB 在利鲁唑培养 7 天内没有观察到细胞毒性或增殖增加。在利鲁唑处理 2 周后，hMSCs 的 ALP 活性略有增加，在 3 周时，利鲁唑 150ng/ml 组上调 150%，利鲁唑 450ng/ml 组上调 90%。在 hOBs 中，用利鲁唑培养 2 周后，ALP 活性几乎翻倍（P＜0.05）。在利鲁唑培养 3 周时，用利鲁唑 150ng/ml（P=0.05）和 450ng/ml（P=0.05），ALP 的表达量分别上调 2.6 倍。用利鲁唑处理对骨钙素（OCN）的 mRNA 表达没有显著影响。
结论
利鲁唑对骨形成的影响是混合的；低剂量利鲁唑对 hMSCs 或 hOBs 的活力或功能均无影响。两种细胞类型的 ALP 活性均被高剂量利鲁唑上调，这可能表明高剂量利鲁唑可以增加成骨代谢，从而加速骨愈合过程。然而，在高浓度时，利鲁唑会导致成骨基因表达减少，包括 Runx2 和Ⅰ型胶原。
证据水平
无。
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