Leeds Institute of Rheumatic and Musculoskeletal Medicine, School of Medicine, University of Leeds, Leeds, UK.
NIHR Leeds Biomedical Research Centre, Chapel Allerton Hospital, Leeds, UK.
Sci Rep. 2019 Nov 22;9(1):17340. doi: 10.1038/s41598-019-53927-3.
Bone marrow-Multipotential stromal cells (BM-MSCs) are increasingly used to treat complicated fracture healing e.g., non-union. Though, the quality of these autologous cells is not well characterized. We aimed to evaluate bone healing-related capacities of non-union BM-MSCs. Iliac crest-BM was aspirated from long-bone fracture patients with normal healing (U) or non-united (NU). Uncultured (native) CD271highCD45low cells or passage-zero cultured BM-MSCs were analyzed for gene expression levels, and functional assays were conducted using culture-expanded BM-MSCs. Blood samples were analyzed for serum cytokine levels. Uncultured NU-CD271highCD45low cells significantly expressed fewer transcripts of growth factor receptors, EGFR, FGFR1, and FGRF2 than U cells. Significant fewer transcripts of alkaline phosphatase (ALPL), osteocalcin (BGLAP), osteonectin (SPARC) and osteopontin (SPP1) were detected in NU-CD271CD45 cells. Additionally, immunoregulation-related markers were differentially expressed between NU- and U-CD271CD45 cells. Interestingly, passage-zero NU BM-MSCs showed low expression of immunosuppressive mediators. However, culture-expanded NU and U BM-MSCs exhibited comparable proliferation, osteogenesis, and immunosuppression. Serum cytokine levels were found similar for NU and U groups. Collectively, native NU-BM-MSCs seemed to have low proliferative and osteogenic capacities; therefore, enhancing their quality should be considered for regenerative therapies. Further research on distorted immunoregulatory molecules expression in BM-MSCs could potentially benefit the prediction of complicated fracture healing.
骨髓-多能基质细胞（BM-MSCs）越来越多地用于治疗复杂骨折愈合，例如骨不连。尽管这些自体细胞的质量尚未得到很好的描述。我们旨在评估骨不连 BM-MSCs 的骨愈合相关能力。从正常愈合（U）或未愈合（NU）的长骨骨折患者的髂嵴骨髓中抽吸未培养（天然）CD271highCD45low 细胞或传代零培养的 BM-MSCs，分析基因表达水平，并使用培养扩增的 BM-MSCs 进行功能测定。分析血样中的血清细胞因子水平。未培养的 NU-CD271highCD45low 细胞与 U 细胞相比，生长因子受体 EGFR、FGFR1 和 FGRF2 的转录本表达显著减少。在 NU-CD271CD45 细胞中，碱性磷酸酶（ALPL）、骨钙素（BGLAP）、骨粘连蛋白（SPARC）和骨桥蛋白（SPP1）的转录本显著减少。此外，NU-和 U-CD271CD45 细胞之间的免疫调节相关标志物表达存在差异。有趣的是，传代零的 NU BM-MSCs 表现出免疫抑制介质的低表达。然而，培养扩增的 NU 和 U BM-MSCs 表现出相似的增殖、成骨和免疫抑制能力。NU 和 U 组的血清细胞因子水平相似。总的来说，天然 NU-BM-MSCs 似乎增殖和成骨能力较低；因此，应考虑增强其质量，以用于再生治疗。进一步研究 BM-MSCs 中免疫调节分子表达的扭曲可能有助于预测复杂骨折愈合。
