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A single step, centrifuge-free method to harvest bone marrow highly concentrated in mesenchymal stem cells: results of a pilot trial

A Correction to this article was published on 19 January 2022

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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.

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Data are available from the corresponding Author upon reasonable request.

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The authors wish to thank Dr. Federico Valli, Dr. Paola De Luca, Dr. Elena De Vecchi and Dr. Agostino La Porta for their support to the study.


The present research was supported by: the Italian Ministry of Health “Ricerca Corrente”, Geistlich Biomaterials Italia Srl, and a grant from the ‘ON Foundation’, Switzerland (ON project number: 18–054).

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Correspondence to Berardo Di Matteo.

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The pilot clinical study was approved by the Hospital Ethics Committee and Internal Review Board of IRCCS Humanitas Research Center (protocol number: 530/18, approved on 18th September 2018).

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The authors declare no personal conflict of interests. Geistlich Biomaterials Srl provided part of the funding for the present research.

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Viganò, M., Ragni, E., Di Matteo, B. et al. A single step, centrifuge-free method to harvest bone marrow highly concentrated in mesenchymal stem cells: results of a pilot trial. International Orthopaedics (SICOT) 46, 391–400 (2022).

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  • Bone marrow aspirate
  • Knee osteoarthritis
  • Mesenchymal stem cells
  • Bone marrow harvesting