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在大鼠股骨缺损模型中，通过预先在膜上接种 CD8 淋巴细胞耗竭的骨髓单核细胞，增强一步法膜技术治疗大骨缺损。

Enhancement of a one-step membrane technique for the treatment of large bone defects by pre-seeding the membrane with CD8 lymphocyte depleted bone marrow mononuclear cells in a rat femoral defect model.

机构信息

Department of Trauma Surgery and Orthopedics, University Hospital, Goethe University Frankfurt, Frankfurt am Main, Germany.

Vascular Research Center, Goethe University Frankfurt, Frankfurt am Main, Germany.

出版信息

Front Immunol. 2024 Oct 23;15:1488611. doi: 10.3389/fimmu.2024.1488611. eCollection 2024.

DOI:10.3389/fimmu.2024.1488611
PMID:39507531
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11537973/
Abstract

BACKGROUND

The one-step membrane technique, using a human acellular dermal matrix (hADM), is an experimental method for treating large bone defects. This eliminates the need for the Masquelet membrane induction step, shortening the procedure while maintaining effectiveness. However, previous studies showed that colonizing hADM with bone marrow mononuclear cells (BMC) worsens healing, likely due to the presence of CD8+ lymphocytes, which negatively affect bone regeneration. This study aims to investigate whether the negative impact of BMC on bone healing in this technique is due to the CD8+ cell population.

MATERIALS AND METHODS

A 5 mm femoral defect was created in 25 male Sprague-Dawley rats, divided into three groups (G1-G3). BMC were isolated from syngenic donor rats, with CD8+ lymphocytes removed magnetically from the BMC fraction in one group. The defects were filled with bone chips and wrapped with differently treated hADM: G1 received native hADM, G2 received hADM+BMC, and G3 received hADM+BMC-CD8. After 8 weeks, the femurs were evaluated through radiological, biomechanical, and histological examinations.

RESULTS

Bone defects and bone mineral density (BMD) were significantly improved in G3 (hADM+BMC-CD8) compared to G2 (hADM+BMC). Bone volume, bone formation, and median bending stiffness were higher in G3. Immunohistological analysis showed a significant decrease in CD8 cell count in G3, with a lower percentage of IFNγ-producing cells compared to G2.

CONCLUSION

Depleting CD8+ cells from BMC before colonizing hADM significantly improved bone healing, likely due to changes in the local mediator environment. This suggests that preoperative colonization with CD8+-depleted BMC could enhance the one-step membrane technique.

摘要

背景

一步法膜技术使用人脱细胞真皮基质（hADM）是治疗大骨缺损的实验方法。该方法消除了 Masquelet 膜诱导步骤的需要，缩短了手术时间，同时保持了疗效。然而，先前的研究表明，用骨髓单核细胞（BMC）殖民 hADM 会加重愈合，这可能是由于存在 CD8+淋巴细胞，它们对骨再生产生负面影响。本研究旨在探讨该技术中 BMC 对骨愈合的负面影响是否归因于 CD8+细胞群。

材料和方法

在 25 只雄性 Sprague-Dawley 大鼠中创建 5mm 股骨缺损，分为三组（G1-G3）。从同基因供体大鼠中分离 BMC，并用磁性从 BMC 部分去除 CD8+淋巴细胞。用不同处理的 hADM 填充缺陷：G1 接受天然 hADM，G2 接受 hADM+BMC，G3 接受 hADM+BMC-CD8。8 周后，通过放射学、生物力学和组织学检查评估股骨。

结果

与 G2（hADM+BMC）相比，G3（hADM+BMC-CD8）的骨缺损和骨密度（BMD）显著改善。G3 的骨体积、骨形成和中位数弯曲刚度较高。免疫组织化学分析显示 G3 中的 CD8 细胞计数显著减少，与 G2 相比，IFNγ产生细胞的百分比更低。

结论

在殖民 hADM 之前从 BMC 中耗尽 CD8+细胞可显著改善骨愈合，这可能是由于局部介质环境的变化。这表明术前用 CD8+细胞耗尽的 BMC 殖民可能会增强一步法膜技术。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/6b5be10d280d/fimmu-15-1488611-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/30306a303e74/fimmu-15-1488611-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/e99127197d0e/fimmu-15-1488611-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/3fc6a0cba63c/fimmu-15-1488611-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/6b5be10d280d/fimmu-15-1488611-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/30306a303e74/fimmu-15-1488611-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/e99127197d0e/fimmu-15-1488611-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/3fc6a0cba63c/fimmu-15-1488611-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79cd/11537973/6b5be10d280d/fimmu-15-1488611-g004.jpg

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