生物信息学分析和实验鉴定慢性骨不连的血液生物标志物。
Bioinformatic analysis and experimental identification of blood biomarkers for chronic nonunion.
机构信息
Department of Orthopedics, Xuanwu Hospital, Capital Medical University, Beijing, 100053, People's Republic of China.
出版信息
J Orthop Surg Res. 2020 Jun 5;15(1):208. doi: 10.1186/s13018-020-01735-1.
BACKGROUND
Incomplete fracture healing may lead to chronic nonunion; thus, determining fracture healing is the primary issue in the clinical treatment. However, there are no validated early diagnostic biomarkers for assessing chronic nonunion. In this study, bioinformatics analysis combined with an experimental verification strategy was used to identify blood biomarkers for chronic nonunion.
METHODS
First, differentially expressed genes in chronic nonunion were identified by microarray data analysis. Second, Dipsaci Radix (DR), a traditional Chinese medicine for fracture treatment, was used to screen the drug target genes. Third, the drug-disease network was determined, and biomarker genes were obtained. Finally, the potential blood biomarkers were verified by ELISA and qPCR methods.
RESULTS
Fifty-five patients with open long bone fractures (39 healed and 16 nonunion) were enrolled in this study, and urgent surgical debridement and the severity of soft tissue injury had a significant effect on the prognosis of fracture. After the systems pharmacology analysis, six genes, including QPCT, CA1, LDHB, MMP9, UGCG, and HCAR2, were chosen for experimental validation. We found that all six genes in peripheral blood mononuclear cells (PBMCs) and serum were differentially expressed after injury, and five genes (QPCT, CA1, MMP9, UGCG, and HCAR2) were significantly lower in nonunion patients. Further, CA1, MMP9, and QPCT were markedly increased after DR treatment.
CONCLUSION
CA1, MMP9, and QPCT are biomarkers of nonunion patients and DR treatment targets. However, HCAR2 and UGCG are biomarkers of nonunion patients but not DR treatment targets. Therefore, our findings may provide valuable information for nonunion diagnosis and DR treatment.
TRIAL REGISTRATION
ISRCTN, ISRCTN13271153. Registered 05 April 2020-Retrospectively registered.
背景
骨折愈合不全会导致慢性骨不连，因此，判断骨折愈合情况是临床治疗的首要问题。然而，目前还没有经过验证的早期诊断生物标志物来评估慢性骨不连。在这项研究中，采用生物信息学分析结合实验验证策略来鉴定慢性骨不连的血液生物标志物。
方法
首先，通过微阵列数据分析鉴定慢性骨不连中的差异表达基因。其次，采用治疗骨折的中药续断（DR）筛选药物靶基因。然后，确定药物-疾病网络，获取生物标志物基因。最后，通过 ELISA 和 qPCR 方法验证潜在的血液生物标志物。
结果
本研究共纳入 55 例开放性长骨骨折患者（39 例愈合，16 例不愈合），急诊清创和软组织损伤严重程度对骨折预后有显著影响。经过系统药理学分析，选择了 6 个基因，包括 QPCT、CA1、LDHB、MMP9、UGCG 和 HCAR2，进行实验验证。我们发现，损伤后外周血单核细胞（PBMCs）和血清中的这 6 个基因均表达差异，且不愈合患者的 5 个基因（QPCT、CA1、MMP9、UGCG 和 HCAR2）表达明显降低。进一步研究发现，DR 治疗后 CA1、MMP9 和 QPCT 显著升高。
结论
CA1、MMP9 和 QPCT 是骨不连患者的生物标志物和 DR 治疗靶点。而 HCAR2 和 UGCG 是骨不连患者的生物标志物，但不是 DR 治疗靶点。因此，我们的研究结果可能为骨不连的诊断和 DR 治疗提供有价值的信息。
试验注册
ISRCTN，ISRCTN13271153。于 2020 年 4 月 5 日注册-回顾性注册。
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