Laboratory of Biocompatibility, Technological Innovation and Advanced Therapy, Rizzoli RIT, IRCCS Rizzoli Orthopedic Institute, Bologna, Italy.
Department of Medical and Surgical Sciences, University of Bologna, Bologna, Italy.
Spine (Phila Pa 1976). 2018 Oct 15;43(20):1401-1410. doi: 10.1097/BRS.0000000000002626.
Laboratory study.
Mesenchymal stem cells (MSCs) derived from whole bone marrow aspirate (BMA) and MSCs derived from density-gradient centrifugation were isolated from vertebral bodies and cultured under either hypoxic or normoxic conditions to evaluate their biological characteristics and HOX and TALE signature able to improve spinal surgery procedures.
The use of spinal fusion procedures has increased over the last decades; however, failed fusion still remains an important problem. Clinician and researchers focused their attention on the therapeutic potential of bone marrow MSCs and several methods for their isolation and cultivation have been developed. However, the best source and techniques are still debated.
MSCs morphology, surface markers, colony-forming-units, and three lineage differentiation through quantitative real-time PCR (qPCR) were evaluated. Additionally, gene expression analysis of HOX and TALE signatures during osteogenic differentiation were analyzed.
Our study showed that MSCs derived from whole BMA were successfully isolated and when cultured under hypoxic condition presented greater proliferation, larger colonies, and differentiated onto osteogenic and chondrogenic lineage with greater ability, while adipogenic differentiation was less efficient. Results also revealed that MSCs, differently isolated and cultured, expressed different level of HOX and TALE signatures and that HOXB8 were up-regulated with greater efficiency in MSCs derived from whole BMA under hypoxia.
Our data indicated that hypoxic preconditioning of MSCs derived from whole BMA exhibited more suitable biological characteristics and different level of HOX and TALE gene activation. We, therefore, concluded that vertebral body MSCs derived from whole BMA may provide alternative sources of MSCs for tissue engineering applications for spine surgery.
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实验室研究。
从椎体中分离出源自全骨髓抽吸物（BMA）的间充质干细胞（MSCs）和源自密度梯度离心的 MSCs，并在低氧或常氧条件下培养，以评估其生物学特性和 HOX 和 TALE 特征，从而改善脊柱手术程序。
在过去几十年中，脊柱融合手术的应用有所增加；然而，融合失败仍然是一个重要问题。临床医生和研究人员将注意力集中在骨髓 MSCs 的治疗潜力上，并且已经开发出几种分离和培养它们的方法。然而，最佳来源和技术仍存在争议。
评估 MSCs 形态、表面标志物、集落形成单位以及通过实时定量 PCR（qPCR）的三系分化。此外，还分析了成骨分化过程中 HOX 和 TALE 特征的基因表达分析。
我们的研究表明，成功地从全 BMA 中分离出 MSCs，并且在低氧条件下培养时，表现出更高的增殖能力、更大的集落、向成骨和成软骨谱系分化的能力更强，而脂肪形成分化的效率较低。结果还表明，不同分离和培养的 MSCs 表达不同水平的 HOX 和 TALE 特征，并且在低氧条件下，源自全 BMA 的 MSCs 中 HOXB8 的表达被更有效地上调。
我们的数据表明，源自全 BMA 的 MSCs 的低氧预处理表现出更适合的生物学特性和不同水平的 HOX 和 TALE 基因激活。因此，我们得出结论，源自全 BMA 的椎体 MSC 可能为脊柱手术的组织工程应用提供替代的 MSC 来源。
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