High-speed centrifugation reduces immune rejection by removing bone marrow elements from fresh osteochondral allografts

IF 5.9 1区医学 Q1 ORTHOPEDICS

Journal of Orthopaedic Translation Pub Date : 2025-01-22 DOI:10.1016/j.jot.2024.12.008

Yongsheng Ma , Wenming Yang , Qitai Lin , Meiming Li , Zehao Li , Yugang Xing , Lei Wei , Wangping Duan , Xiaochun Wei

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Abstract

Background

Fresh osteochondral allografts (OCAs) contain numerous immunogenic components in the subchondral bone (SB). Whether high-speed centrifugation (HSC) reduces immune rejection by removing bone marrow elements (BMEs), compared to methods without HSC, remains unknown. This study aimed to validate the efficacy and safety of HSC in reducing immune rejection by removing allogeneic BMEs.

Methods

OCAs were obtained from the femoral condyles of the stifle joint in 18 pigs. Gross observations, histological staining, weight measurements, and DNA extraction were performed to assess the effects of centrifugation speed and duration on BMEs removal in OCAs. The effect of HSC on OCAs preservation was determined in vitro using microbiological testing, live/dead cell staining, and histological staining. Moreover, the co-culture effect of RAW264.7 cells and OCAs with or without HSC in vitro was evaluated using enzyme-linked immunosorbent assay (ELISA), histological staining, and immunohistochemical staining. The transplantation effect of OCAs with or without HSC was examined in vivo using a subcutaneous mouse model. Finally, the residues in the centrifuge tubes were analysed using ELISA, haematoxylin and eosin (HE) staining, and metabolomic analysis.

Results

Centrifugal speeds of 12000 rpm for 1 min were sufficient to reduce BMEs by over 90 %. HSC had a protective effect on chondrocytes and the extracellular matrix during the in vitro preservation of OCAs. In addition, OCAs using the HSC method exhibited reduced recognition by the host immune system compared with OCAs without HSC, thereby reducing immune rejection. Lipids were the most abundant and difficult-to-remove antigenic components and are the most likely to affect host macrophage polarisation, playing an important role in immune rejection.

Conclusion

Our study demonstrated that HSC method significantly reduces immune rejection by removing BMEs from OCAs.

The translational potential of this article

Our study demonstrated that HSC is a simple, efficient, and safe physical method for removing antigenic components from OCAs, effectively reducing immune rejection and highlighting its clinical potential.

Abstract Image

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高速离心通过从新鲜骨软骨移植物中去除骨髓元素来减少免疫排斥反应

新鲜骨软骨同种异体移植物（OCAs）在软骨下骨（SB）中含有许多免疫原性成分。与不含HSC的方法相比，高速离心（HSC）是否通过去除骨髓元素（BMEs）来减少免疫排斥反应尚不清楚。本研究旨在验证HSC通过去除异体BMEs减少免疫排斥反应的有效性和安全性。方法从18头猪的膝关节股骨髁处获得soca。通过大体观察、组织学染色、重量测量和DNA提取来评估离心速度和持续时间对oca中BMEs去除的影响。通过体外微生物检测、活/死细胞染色和组织学染色，确定HSC对OCAs保存的影响。此外，采用酶联免疫吸附试验（ELISA）、组织学染色和免疫组织化学染色评估RAW264.7细胞与oca体外共培养HSC或不含HSC的效果。采用小鼠皮下模型，研究了oca加或不加HSC的移植效果。最后，对离心管中的残留物进行ELISA、血红素和伊红（HE）染色和代谢组学分析。结果12000 rpm / min的离心转速可使bme降低90%以上。在oca体外保存过程中，HSC对软骨细胞和细胞外基质有保护作用。此外，与未使用HSC的oca相比，使用HSC方法的oca对宿主免疫系统的识别降低，从而减少了免疫排斥反应。脂质是最丰富和最难去除的抗原成分，最可能影响宿主巨噬细胞极化，在免疫排斥中起重要作用。结论本研究表明，HSC法通过去除oca中的BMEs可显著减少免疫排斥反应。我们的研究表明，HSC是一种简单、高效、安全的物理方法，可从oca中去除抗原成分，有效减少免疫排斥反应，突出其临床潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Orthopaedic Translation Medicine-Orthopedics and Sports Medicine

CiteScore

11.80

自引率

13.60%

发文量

审稿时长

29 days

期刊介绍： The Journal of Orthopaedic Translation (JOT) is the official peer-reviewed, open access journal of the Chinese Speaking Orthopaedic Society (CSOS) and the International Chinese Musculoskeletal Research Society (ICMRS). It is published quarterly, in January, April, July and October, by Elsevier.

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