Abstract
BackgroundThe 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.ResultsWe 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.ConclusionWe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0318-1) contains supplementary material, which is available to authorized users.
Highlights
The human body has an extensive capacity to regenerate bone tissue after trauma
Inter-donor variability in bone marrow aspirate The volume of BM aspirated from the different locations varied significant, with larger BM volumes obtained from the ilium (22 ± 7.6 ml) than the proximal femur (6 ± 3.3 ml), distal femur (5 ± 2.9 ml) or proximal tibia (4 ± 2.6 ml)
BM volumes from the ilium yielded a higher concentration of mononuclear cells (MNCs) for volumes close to 10 ml (2.6 x 107 MNC/ml), while volumes close and larger than 20 ml yielded a lower concentration of MNCs (1.4 x 107 MNC/ml), not statistical significant (p = 0.15)
Summary
The human body has an extensive capacity to regenerate bone tissue after trauma Large defects such as long bone fractures of the lower limbs cannot be restored without intervention and often lead to nonunion. Volume, number of mononuclear cells/hMSCs and their self-renewal, multilineage potential, extracellular matrix (ECM) production and surface marker profiling were analyzed. Ghebes et al BMC Biotechnology (2016) 16:89 production potential of hMSCs isolated from bone marrow (BM) aspirated from the lower limbs, such as the ilium, proximal femur, distal femur and proximal tibia. We hypothesize that BM located at the fracture site might play an important role in the fracture healing rate, due to differences in cell number, self-renewal-, proliferative-, ECM production- and multilineage differentiation potential
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