Ghebes Corina Adriana, Braham Maaike Vera Jasmijn, Zeegers Adelgunde Veronica Clemens Maria, Renard Auke Jan Sijbe, Fernandes Hugo, Saris Daniel B F
MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Drienerlolaan 5, 7522NB, Enschede, The Netherlands.
Department of Orthopaedics, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands.
BMC Biotechnol. 2016 Dec 12;16(1):89. doi: 10.1186/s12896-016-0318-1.
The human body has an extensive capacity to regenerate bone tissue after trauma. However large defects such as long bone fractures of the lower limbs cannot be restored without intervention and often lead to nonunion. Therefore, the aim of the present study was to assess the pool and biological functions of human mesenchymal stromal cells (hMSCs) isolated from different bone marrow locations of the lower limbs and to identify novel strategies to prime the cells prior to their use in bone fracture healing. Following, bone marrow from the ilium, proximal femur, distal femur and proximal tibia was aspirated and the hMSCs isolated. Bone marrow type, volume, number of mononuclear cells/hMSCs and their self-renewal, multilineage potential, extracellular matrix (ECM) production and surface marker profiling were analyzed. Additionally, the cells were primed to accelerate bone fracture healing either by using acoustic stimulation or varying the initial hMSCs isolation conditions.
We found that the more proximal the bone marrow aspiration location, the larger the bone marrow volume was, the higher the content in mononuclear cells/hMSCs and the higher the self-renewal and osteogenic differentiation potential of the isolated hMSCs were. Acoustic stimulation of bone marrow, as well as the isolation of hMSCs in the absence of fetal bovine serum, increased the osteogenic and ECM production potential of the cells, respectively.
We showed that bone marrow properties change with the aspiration location, potentially explaining the differences in bone fracture healing between the tibia and the femur. Furthermore, we showed two new priming methods capable of enhancing bone fracture healing.
人体在创伤后具有广泛的骨组织再生能力。然而，诸如下肢长骨骨折等大的骨缺损若不进行干预则无法恢复，且常常导致骨不连。因此，本研究的目的是评估从下肢不同骨髓部位分离的人间充质基质细胞（hMSCs）的储备及生物学功能，并确定在将这些细胞用于骨折愈合之前对其进行预处理的新策略。随后，抽取来自髂骨、股骨近端、股骨远端和胫骨近端的骨髓，并分离出hMSCs。分析了骨髓类型、体积、单个核细胞/hMSCs数量及其自我更新能力、多向分化潜能、细胞外基质（ECM）产生情况以及表面标志物谱。此外，通过使用声刺激或改变初始hMSCs分离条件对细胞进行预处理以加速骨折愈合。
我们发现骨髓抽取部位越靠近近端，骨髓体积越大，单个核细胞/hMSCs含量越高，分离出的hMSCs的自我更新能力和成骨分化潜能也越高。对骨髓进行声刺激以及在无胎牛血清的情况下分离hMSCs，分别提高了细胞的成骨能力和ECM产生潜能。
我们表明骨髓特性随抽取部位而变化，这可能解释了胫骨和股骨骨折愈合的差异。此外，我们展示了两种能够增强骨折愈合的新预处理方法。
