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损伤骨组织中的巨噬细胞特征

Macrophages Characterization in an Injured Bone Tissue.

作者信息

Nikovics Krisztina, Durand Marjorie, Castellarin Cédric, Burger Julien, Sicherre Emma, Collombet Jean-Marc, Oger Myriam, Holy Xavier, Favier Anne-Laure

机构信息

Imagery Unit, Department of Platforms and Technology Research, French Armed Forces Biomedical Research Institute, 91223 Brétigny-sur-Orge, France.

Osteo-Articulary Biotherapy Unit, Department of Medical and Surgical Assistance to the Armed Forces, French Armed Forces Biomedical Research Institute, 91223 Brétigny-sur-Orge, France.

出版信息

Biomedicines. 2022 Jun 11;10(6):1385. doi: 10.3390/biomedicines10061385.

DOI:10.3390/biomedicines10061385
PMID:35740407
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9219779/
Abstract

Biomaterial use is a promising approach to facilitate wound healing of the bone tissue. Biomaterials induce the formation of membrane capsules and the recruitment of different types of macrophages. Macrophages are immune cells that produce diverse combinations of cytokines playing an important role in bone healing and regeneration, but the exact mechanism remains to be studied. Our work aimed to identify in vivo macrophages in the Masquelet induced membrane in a rat model. Most of the macrophages in the damaged area were M2-like, with smaller numbers of M1-like macrophages. In addition, high expression of IL-1β and IL-6 cytokines were detected in the membrane region by RT-qPCR. Using an innovative combination of two hybridization techniques (in situ hybridization and in situ hybridization chain reaction (in situ HCR)), M2b-like macrophages were identified for the first time in cryosections of non-decalcified bone. Our work has also demonstrated that microspectroscopical analysis is essential for macrophage characterization, as it allows the discrimination of fluorescence and autofluorescence. Finally, this work has revealed the limitations of immunolabelling and the potential of in situ HCR to provide valuable information for in vivo characterization of macrophages.

摘要

生物材料的使用是促进骨组织伤口愈合的一种有前景的方法。生物材料会诱导膜性囊的形成以及不同类型巨噬细胞的募集。巨噬细胞是免疫细胞，可产生多种细胞因子组合，在骨愈合和再生中发挥重要作用，但其确切机制仍有待研究。我们的工作旨在在大鼠模型的Masquelet诱导膜中鉴定体内巨噬细胞。受损区域的大多数巨噬细胞呈M2样，M1样巨噬细胞数量较少。此外，通过RT-qPCR在膜区域检测到IL-1β和IL-6细胞因子的高表达。使用两种杂交技术（原位杂交和原位杂交链式反应（原位HCR））的创新组合，首次在未脱钙骨的冰冻切片中鉴定出M2b样巨噬细胞。我们的工作还表明，显微光谱分析对于巨噬细胞表征至关重要，因为它可以区分荧光和自发荧光。最后，这项工作揭示了免疫标记的局限性以及原位HCR为巨噬细胞体内表征提供有价值信息的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72eb/9219779/20dfc2194546/biomedicines-10-01385-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72eb/9219779/20dfc2194546/biomedicines-10-01385-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72eb/9219779/270a053014c3/biomedicines-10-01385-g005.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72eb/9219779/d0a4059b7dc7/biomedicines-10-01385-g007.jpg
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