Molecular mapping of articular cartilage CXCR4 expression after ACL injury via a novel small molecule-based probe

IF 3.5 2区医学 Q2 ENDOCRINOLOGY & METABOLISM

Bone Pub Date : 2025-03-17 DOI:10.1016/j.bone.2025.117463

Michael D. Newton , Mackenzie M. Fleischer , Howard W.T. Matthew , Tristan Maerz

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引用次数: 0

Abstract

Purpose

Molecular imaging is a powerful modality to spatially resolve molecular changes across tissues, but application to articular cartilage remains limited. CXCR4 is an established marker of chondrocyte hypertrophy and potential therapeutic target for osteoarthritis. The purpose of this study was to develop and apply a CXCR4-targeted, near-infrared fluorescent (NIR) probe to a rat model of post-traumatic osteoarthritis (PTOA).

Methods

A CXCR4-targeted, small molecule-based NIR probe (“Cy7-AMD”) was synthesized. Sensitivity and specificity of Cy7-AMD to CXCR4 was validated in vitro (HUVECs), and ex vivo (rat osteochondral explants). To induce PTOA, female Lewis rats underwent noninvasive anterior cruciate ligament (ACL) rupture. At 7- and 28-days post-injury, injured/contralateral femora and tibiae were dissected, incubated in Cy7-AMD vs a non-targeting control, and imaged via NIR imaging, as well as conventional and contrast-enhanced micro-computed tomography. Imaging datasets were co-registered, cartilage tissue volumes were segmented, and paired cartilage thickness and NIR signal maps were generated and analyzed for PTOA-relevant changes.

Results

Compared to a non-targeting control probe, in vitro and ex vivo assays confirm sensitivity and specificity of Cy7-AMD to CXCR4. Flow cytometry confirmed high correspondence between Cy7-AMD- and antibody-based measurement of CXCR4 expression. Cy7-AMD rapidly equilibrated within cartilage, and fluorescent histology confirmed full-thickness penetration. Injured femoral cartilage exhibited heterogeneous CXCR4 expression, with increased signal deviation compared to contralateral femora. Spatial CXCR4 expression patterns correlated to cartilage thickness patterns; high CXCR4 expression at boundaries of low-thickness lesions suggests an association between CXCR4 expression and cartilage loss.

Conclusions

Small molecule-based probes are advantageous for mapping spatial patterns of molecular expression in rodent articular cartilage, deepening our understanding of PTOA progression.

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来源期刊

Bone 医学-内分泌学与代谢

CiteScore

8.90

自引率

4.90%

发文量

264

审稿时长

30 days

期刊介绍： BONE is an interdisciplinary forum for the rapid publication of original articles and reviews on basic, translational, and clinical aspects of bone and mineral metabolism. The Journal also encourages submissions related to interactions of bone with other organ systems, including cartilage, endocrine, muscle, fat, neural, vascular, gastrointestinal, hematopoietic, and immune systems. Particular attention is placed on the application of experimental studies to clinical practice.