Assessment of the Monocyte Subpopulations and M1/M2 Macrophage Ratio in Concentrated Bone Marrow Aspirate.

IF 2.7 4区医学 Q1 ORTHOPEDICS CARTILAGE Pub Date : 2024-12-09 DOI:10.1177/19476035241304308

James J Butler, John F Dankert, Laura E Keller, Mohammad T Azam, Jari Dahmen, Gino M M J Kerkhoffs, John G Kennedy

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Abstract

Objective: The purpose of this study was to determine the M1/M2 macrophage ratio in concentrated bone marrow aspirate (cBMA) in patients undergoing surgical intervention augmented with cBMA for osteochondral lesions of the talus (OLTs).

Design: Samples of peripheral blood (PB), bone marrow aspirate (BMA), and cBMA were collected during the procedure. The samples were analyzed by automated cell counting and multicolor fluorescence-activated cell sorting with specific antibodies recognizing monocytes (CD14+ CD16+) and the M1 (CD86+) and M2 (CD163+CD206+) populations within that monocyte population. Cytokine concentrations within the samples were evaluated with enzyme-linked immunosorbent assay (ELISA). The composition of cBMA was compared between 2 commercially available BMA concentration systems.

Results: Thirty-eight patients with a mean age of 43.2 ± 10.1 years old undergoing a surgical procedure for the treatment of OLTs involving the use of cBMA were included. cBMA had a mean fold increase of 4.7 for all white blood cells, 6.1 for monocytes, 7.9 for lymphocytes, 2.4 for neutrophils, and 9.6 for platelets when compared to BMA. The mean M1/M2 ratio for PB, BMA, and cBMA was 15.2 ± 12.0, 20.8 ± 13.3, and 22.1 ± 16.0, respectively. There was a statistically significant higher concentration of interleukin-1 receptor antagonist (IL-1Ra) in the cBMA sample (8243.3 ± 14,837.4 pg/mL) compared to both BMA (3143.0 ± 2218.5 pg/mL) and PB (1847.5 ± 1520.4 pg/mL) samples. The IL-1Ra/IL-1β ratio for PB, BMA, and cBMA was 790.6 ± 581.9, 764.7 ± 675.2, and 235.7 ± 192.1, respectively. There was no difference in the cBMA M1/M2 ratio (19.0 ± 11.1 vs 24.0 ± 18.3) between the Magellan (Isto Biologics, Hopkinton, Massachusetts) and Angel systems (Arthrex Inc, Naples, Florida).

Conclusion: This prospective study found that the M1/M2 ratio in cBMA was 22.1 ± 16.0, with significant patient to patient variation observed. Overall, there was no statistically significant difference in the M1/M2 ratio across PB, BMA, and cBMA samples. This is the first study to characterize the macrophage subpopulation within cBMA, which may have significant clinical implications in future studies.

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骨髓浓缩液中单核细胞亚群和M1/M2巨噬细胞比值的评估。

目的：本研究的目的是测定在距骨软骨病变（OLTs）中接受cBMA强化手术干预的患者骨髓浓缩液（cBMA）中M1/M2巨噬细胞的比例。设计：在手术过程中采集外周血（PB）、骨髓抽液（BMA）和cBMA样本。使用特异性抗体识别单核细胞（CD14+ CD16+）和该单核细胞群中的M1 （CD86+）和M2 （CD163+CD206+）群体，通过自动细胞计数和多色荧光激活细胞分选对样品进行分析。采用酶联免疫吸附试验（ELISA）评估样品中的细胞因子浓度。比较了两种市售BMA浓缩体系中cBMA的组成。结果：38例患者，平均年龄43.2±10.1岁，接受手术治疗包括使用cBMA的olt。与BMA相比，所有白细胞的cBMA平均增加4.7倍，单核细胞增加6.1倍，淋巴细胞增加7.9倍，中性粒细胞增加2.4倍，血小板增加9.6倍。PB、BMA和cBMA的平均M1/M2比值分别为15.2±12.0、20.8±13.3和22.1±16.0。与BMA样品（3143.0±2218.5 pg/mL）和PB样品（1847.5±1520.4 pg/mL）相比，cBMA样品中白细胞介素-1受体拮抗剂（IL-1Ra）的浓度（8243.3±14837.4 pg/mL）具有统计学意义。PB、BMA和cBMA的IL-1Ra/IL-1β比值分别为790.6±581.9、764.7±675.2和235.7±192.1。Magellan （Isto Biologics, Hopkinton, Massachusetts）和Angel系统（Arthrex Inc, Naples, Florida）的cBMA M1/M2比值（19.0±11.1 vs 24.0±18.3）无差异。结论：本前瞻性研究发现cBMA M1/M2比值为22.1±16.0，患者间差异显著。总体而言，在PB、BMA和cBMA样本中，M1/M2比率没有统计学意义上的差异。这是第一个描述cBMA内巨噬细胞亚群的研究，可能在未来的研究中具有重要的临床意义。

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来源期刊

CARTILAGE ORTHOPEDICS-

CiteScore

6.90

自引率

7.10%

发文量

期刊介绍： CARTILAGE publishes articles related to the musculoskeletal system with particular attention to cartilage repair, development, function, degeneration, transplantation, and rehabilitation. The journal is a forum for the exchange of ideas for the many types of researchers and clinicians involved in cartilage biology and repair. A primary objective of CARTILAGE is to foster the cross-fertilization of the findings between clinical and basic sciences throughout the various disciplines involved in cartilage repair. The journal publishes full length original manuscripts on all types of cartilage including articular, nasal, auricular, tracheal/bronchial, and intervertebral disc fibrocartilage. Manuscripts on clinical and laboratory research are welcome. Review articles, editorials, and letters are also encouraged. The ICRS envisages CARTILAGE as a forum for the exchange of knowledge among clinicians, scientists, patients, and researchers. The International Cartilage Repair Society (ICRS) is dedicated to promotion, encouragement, and distribution of fundamental and applied research of cartilage in order to permit a better knowledge of function and dysfunction of articular cartilage and its repair.