Orthopaedic Biotechnology Laboratory, IRCCS Istituto Ortopedico Galeazzi, Via R. Galeazzi 4, 21061, Milano, Italy.
Department of Biomedical Sciences, Humanitas University, Via Rita Levi Montalcini 4, 20090, Pieve Emanuele, Milan, Italy.
Int Orthop. 2022 Feb;46(2):391-400. doi: 10.1007/s00264-021-05243-7. Epub 2021 Nov 2.
The aims of the present study were: (1) to characterize the bone-marrow aspirate (BMA) obtained with a centrifuge-free process, employing a dedicated aspiration device; (2) to test the in vitro efficacy of BMA in a model of cartilage inflammation; and (3) to report the preliminary clinical results in a small cohort of patients affected by knee OA.
Ten patients (4 M, 6 W; mean age: 51.9 ± 9.2 yy) affected by mild to moderate unicompartmental knee OA (KL grade 2-3) were treated by intra-articular and subchondral injections of BMA obtained by a centrifuge-free process. To evaluate the effectiveness of the device in harvesting mesenchymal stem cells (MSCs), samples of the obtained BMA were tested by flow cytometry before and after subculture; BMA ability to counteract inflammation was also tested in an in vitro model of cartilage cell inflammation, evaluating the expression of MMP1, MMP3, TGFβ and TIMP-1 by real-time PCR. Patients were also evaluated up to two years' follow-up by using: VAS for pain, IKDC-subjective and KOOS scores.
The laboratory analysis showed that BMSCs accounted for 0.011% of BMA cells, similar to what had been expected in native bone marrow. The paracrine activity of BMA was able to reduce in vitro the catabolic response of human chondrocyte, as shown by the decrease in metalloproteases concentration and increase in anti-inflammatory mediators. Moreover, the clinical evaluation showed significant improvements in all scores adopted, with stable results up to two years.
The present data showed the effectiveness of the study device to harvest pure bone marrow with minimal peripheral blood contamination. The relevant content of MSCs resulted in the ability to counteract the catabolic cascade through a paracrine action. The clinical outcomes in patients affected by unicompartmental knee OA were encouraging in terms of pain reduction and functional improvement up to mid-term evaluation.
本研究的目的是：（1）描述一种使用专用抽吸装置，无需离心即可获得骨髓抽吸物（BMA）的方法；（2）在软骨炎症模型中测试 BMA 的体外疗效；（3）报告一项小队列膝关节骨关节炎（OA）患者的初步临床结果。
10 名（4 男，6 女；平均年龄：51.9±9.2 岁）单侧膝关节 OA（KL 分级 2-3）患者接受了 BMA 关节内和软骨下注射治疗，BMA 由无需离心的方法获得。为评估该设备获取间充质干细胞（MSCs）的有效性，对获得的 BMA 样本进行了流式细胞术检测，包括在亚培养前后的检测；还通过体外软骨细胞炎症模型测试了 BMA 对抗炎症的能力，通过实时 PCR 评估了 MMP1、MMP3、TGFβ和 TIMP-1 的表达。患者还在 2 年的随访中接受了 VAS 疼痛评分、IKDC 主观评分和 KOOS 评分的评估。
实验室分析表明，BMSCs 占 BMA 细胞的 0.011%，与骨髓中的预期值相似。BMA 的旁分泌活性能够减少体外人软骨细胞的分解代谢反应，表现为金属蛋白酶浓度降低和抗炎介质增加。此外，临床评估显示，所有采用的评分均有显著改善，2 年内结果稳定。
本研究数据表明，研究设备能够有效采集纯骨髓，且外周血污染最小。MSCs 的含量与通过旁分泌作用拮抗分解代谢级联反应的能力有关。单侧膝关节 OA 患者的临床结果在减轻疼痛和改善功能方面令人鼓舞，中期评估结果稳定。
