骨不连患者炎症相关基因的表达水平及BMP-2感染间充质干细胞联合nHA/PA66对股骨骨不连大鼠炎症水平的影响

IF 1.9 4区医学 Q3 PHYSIOLOGY

Physiological research Pub Date : 2024-11-19

Y Huang, Q Zhang, Q Jing, X Li, F Dong

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引用次数: 0

摘要

骨不连会延迟骨折端修复，并与炎症有关。虽然在临床实践中骨不连可以有效修复，但失败的病例很多。研究证实，BMP-2 和 nHA/PA66 可成功修复骨缺损。关于 BMP-2 和 NHA/PA66 联合应用对骨不连成骨作用和炎症的影响的研究很少。我们旨在研究骨不连患者炎症相关基因的表达水平，以及 BMP-2 感染间充质干细胞与 NHA/PA66 联合应用对股骨不连大鼠炎症水平的影响。我们在GEO公共数据库中搜索了与骨不连炎症相关的基因表达谱（GSE93138）。培养并通过 SD 大鼠的骨髓间充质干细胞（MSCs）。我们用携带 BMP-2 的慢病毒感染第三代间充质干细胞，并诱导受感染的间充质干细胞进行骨定向。我们用 RT-PCR 检测了 BMP-2 的表达水平，用 CCK8 检测了细胞活力和碱性磷酸酶（ALP）活性，然后分析了细胞粘附能力。最后，我们检测了非椎体内髁大鼠相关炎症因子的水平，包括 C 反应蛋白（CRP）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和红细胞沉降率（ESR）。我们的研究结果非腱鞘炎患者有 26 个不同炎症基因的表达上调。这些基因主要集中在先天性免疫反应、细胞外区域、钙离子结合、泛酸和 CoA 生物合成途径。Lenti-BMP-2 组的 BMP-2 表达水平更高（与空慢病毒载体组相比：t=5.699；与未感染组相比 t=3.996）。间充质干细胞 + BMP-2 + nHA/PA66 组的细胞活性逐渐增加。转染了 BMP-2 的间充质干细胞与 nHA/PA66 结合后，布满了 nHA/PA66 的表面，并向材料孔中生长。间充质干细胞+BMP-2+nHA/PA66细胞的ALP染色呈阳性，且ALP的OD值最高。间充质干细胞 + BMP-2 + nHA/PA66 组的 CRP、IL-6、TNF-α 和 ESR 水平低于间充质干细胞组和间充质干细胞 + nHA/PA66 组，但高于间充质干细胞 + BMP-2 组。以上比较的 P

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The Expression Level of Inflammation-Related Genes in Patients With Bone Nonunion and the Effect of BMP-2 Infected Mesenchymal Stem Cells Combined With nHA/PA66 on the Inflammation Level of Femoral Bone Nonunion Rats.

Bone nonunion delays fracture end repair and is associated with inflammation. Although bone nonunion can be effectively repaired in clinical practice, many cases of failure. Studies have confirmed that BMP-2 and nHA/PA66 repaired bone defects successfully. There are few studies on the effects of the combined application of BMP-2 and NHA/PA66 on bone nonunion osteogenesis and inflammation. We aimed to investigate the expression level of inflammation-related genes in patients with bone nonunion and the effect of BMP-2-infected mesenchymal stem cells combined with nHA/PA66 on the level of inflammation in femur nonunion rats. We searched for a gene expression profile related to bone nonunion inflammation (GSE93138) in the GEO public database. Bone marrow mesenchymal stem cells (MSCs) of SD rats were cultured and passed through. We infected the third generation of MSCs with lentivirus carrying BMP-2 and induced the infected MSCs to bone orientation. We detected the expression level of BMP-2 by RT-PCR and the cell viability and alkaline phosphatase (ALP) activity by CCK8 and then analyzed the cell adhesion ability. Finally, the levels of related inflammatory factors, including C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and Erythrocyte Sedimentation Rate (ESR), were detected in nonunion rats. Our findings: The patients with nonunion had up-regulated expression of 26 differentially inflammatory genes. These genes are mainly enriched in innate immune response, extracellular region, calcium ion binding, Pantothenate and CoA biosynthesis pathways. The expression level of BMP-2 in the Lenti-BMP-2 group was higher (vs. empty lentivirus vector group: t=5.699; vs. uninfected group t=3.996). The cell activity of the MSCs + BMP-2 + nHA/PA66 group increased gradually. After being combined with nHA/PA66, MSCs transfected with BMP-2 spread all over the surface of nHA/PA66 and grew into the material pores. MSCs + BMP-2 + nHA/PA66 cells showed positive ALP staining, and the OD value of ALP was the highest. The levels of CRP, IL-6, TNF-alpha, and ESR in the MSCs + BMP-2 + nHA/PA66 group were lower than those in the MSCs and MSCs + nHA/PA66 group but higher than those in MSCs + BMP-2 group. The above comparisons were all P<0.05. The findings demonstrated that the expression level of inflammation-related genes increased in the patients with bone nonunion. The infection of MSCs by BMP-2 could promote the directed differentiation of MSCs into osteoblasts in the bone marrow of rats, enhance the cell adhesion ability and ALP activity, and reduce inflammation in rats with bone nonunion.

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来源期刊

Physiological research 医学-生理学

CiteScore

4.00

自引率

4.80%

发文量

108

审稿时长

3 months

期刊介绍： Physiological Research is a peer reviewed Open Access journal that publishes articles on normal and pathological physiology, biochemistry, biophysics, and pharmacology. Authors can submit original, previously unpublished research articles, review articles, rapid or short communications. Instructions for Authors - Respect the instructions carefully when submitting your manuscript. Submitted manuscripts or revised manuscripts that do not follow these Instructions will not be included into the peer-review process. The articles are available in full versions as pdf files beginning with volume 40, 1991. The journal publishes the online Ahead of Print /Pre-Press version of the articles that are searchable in Medline and can be cited.