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[不同解剖部位骨髓基质细胞的比较,以评估其在潜在临床应用中的适用性]

[Comparison of Bone Marrow Stromal Cells from Different Anatomical Locations for Evaluation of Their Suitability for Potential Clinical Applications].

作者信息

NeckaŘ P, Havlas V, LykovÁ D, BraniŠ J, KvÍzovÁ J, Bauer P O

机构信息

Ortopedické oddělení, oddělení sportovní medicíny, Krajská zdravotní a.s., Masarykova nemocnice, Ústí nad Labem.

出版信息

Acta Chir Orthop Traumatol Cech. 2020;87(3):183-190.

Abstract

PURPOSE OF THE STUDY In patients older than 40 years of age, treatment of chondral lesions of the knee employing microfractures does not provide satisfactory outcomes. One of the contributing factors may be the age-related lack of sufficient mesenchymal stromal cells able to efficiently migrate into the desired site of the lesion. Concomitant application of mononuclear cells (MNCs) or cultured mesenchymal stem cells (MSCs) isolated from bone marrow and seeded on a 3D scaffold could provide an alternative enhancing therapeutic efficacy in these patients. The aim of our study was to assess two different sources of bone marrow for isolation of progenitor cells. To be specific, material obtained from proximal tibia and iliac crest (the commonly used bone marrow source) was compared in terms of quantity and quality of cells and their suitability for cellbased treatment of chondral lesions. MATERIAL AND METHODS Bone marrow was collected during a total knee replacement surgery from the iliac crest and the proximal metaphysis of the tibia from ten volunteers older than 40 years of age, using aspiration biopsy needles. The MNCs from the obtained material were isolated, cultured and analyzed for their phenotypic features. Both sources were compared as to the yield and viability of MNCs and MSCs as well as the ability of MSCs of chondrogenic differentiation. RESULTS The MNCs concentration/yield was significantly higher in samples from the iliac crests. Similar results were obtained with cultured MSCs after the first passage when the MSCs/MNCs ratio was compared. Nevertheless, the qualitative analysis that included MSCs immuno-phenotyping, viability and population doubling time showed no difference between the two tested bone marrow sources. DISCUSSION Particularly in older patients, therapies of chondral defects employing bone marrow-stimulating techniques result in unsatisfactory outcomes. Therefore, orthopedic surgeons have turned their attention to cell-based treatments. Bone marrow located in pelvic bone and metaphysis of long bones (distal femur, proximal tibia) contains mononuclear cells that possess features of MSCs. In accordance with other authors, quantitative differences between the cells from two anatomical locations were found in our cohort of patients. Qualitatively, however, there was no significant variability observed. We also confirmed that the metaphysis of proximal tibia is a suitable source for cultured MSCs. Moreover, in contrast to several reports, the quality of these cells does not appear to decrease with the patients age. CONCLUSIONS The iliac crest represents a superior bone marrow source for the MNCs and MSCs yield when compared with tibia. However, there was no qualitative difference between the isolated and cultured cells. The population doubling time analysis showed that the tibia is a good alternative source of MSCs which can be obtained at therapeutically relevant numbers for example for the treatment of chondral lesions of the knee. Key words: bone marrow, mesenchymal stem cells, MSC, mononuclear cells, chondral defects, osteoarthritis.

摘要

研究目的 在40岁以上的患者中,采用微骨折技术治疗膝关节软骨损伤无法获得令人满意的效果。其中一个影响因素可能是与年龄相关的间充质基质细胞不足,无法有效迁移至损伤的预期部位。同时应用从骨髓中分离并接种于三维支架上的单核细胞(MNCs)或培养的间充质干细胞(MSCs),可能为提高这些患者的治疗效果提供一种替代方法。本研究的目的是评估两种不同的骨髓来源以分离祖细胞。具体而言,比较了从胫骨近端和髂嵴(常用的骨髓来源)获取的材料在细胞数量和质量以及它们对软骨损伤细胞治疗的适用性方面的差异。

材料与方法 在全膝关节置换手术期间,使用穿刺活检针从10名40岁以上志愿者的髂嵴和胫骨近端干骺端采集骨髓。对所获材料中的MNCs进行分离、培养并分析其表型特征。比较了两种来源在MNCs和MSCs的产量及活力以及MSCs软骨分化能力方面的差异。

结果 髂嵴样本中的MNCs浓度/产量显著更高。在比较MSCs/MNCs比例时,第一代培养的MSCs也得到了类似结果。然而,包括MSCs免疫表型分析、活力和群体倍增时间在内的定性分析显示,两种测试的骨髓来源之间没有差异。

讨论 特别是在老年患者中,采用骨髓刺激技术治疗软骨缺损效果不理想。因此,骨科医生将注意力转向了基于细胞的治疗方法。位于骨盆骨和长骨干骺端(股骨远端、胫骨近端)的骨髓含有具有MSCs特征的单核细胞。与其他作者一致,在我们的患者队列中发现了两个解剖部位细胞的数量差异。然而,在质量上未观察到显著差异。我们还证实胫骨近端干骺端是培养MSCs的合适来源。此外,与一些报道相反,这些细胞的质量似乎不会随着患者年龄的增长而下降。

结论 与胫骨相比,髂嵴是MNCs和MSCs产量更高的骨髓来源。然而,分离和培养的细胞在质量上没有差异。群体倍增时间分析表明,胫骨是MSCs的良好替代来源,例如可获得治疗膝关节软骨损伤所需数量的细胞。

关键词

骨髓;间充质干细胞;MSCs;单核细胞;软骨缺损;骨关节炎

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