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Bio-Design and Manufacturing  2023 Vol.6 No.6 P.704-717

http://doi.org/10.1007/s42242-023-00252-4


Highly specific characterization and discrimination of monosodium urate crystals in gouty arthritis based on aggregation-induced emission luminogens


Author(s):  Wenjuan Wang, Guiquan Zhang, Ziyi Chen, Hanlin Xu, Bohan Zhang, Rong Hu, Anjun Qin & Yinghui Hua

Affiliation(s):  Department of Sports Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China; more

Corresponding email(s):   hurong@usc.edu.cn, msqinaj@scut.edu.cn, hua_cosm@aliyun.com

Key Words:  Gout, Monosodium urate, Hydroxyapatite, TPE-Ketoalkyne, Aggregation-induced emission, Confocal laser scanning microscope imaging


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Wenjuan Wang, Guiquan Zhang, Ziyi Chen, Hanlin Xu, Bohan Zhang, Rong Hu, Anjun Qin & Yinghui Hua. Highly specific characterization and discrimination of monosodium urate crystals in gouty arthritis based on aggregation-induced emission luminogens[J]. Journal of Zhejiang University Science D, 2023, 6(6): 704-717.

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Abstract: 
Existing technologies used to detect monosodium urate (MSU) crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation. The purpose of this study was to explore whether aggregation-induced emission luminogens (AIEgens) can be used for highly specific imaging of MSU crystals to assist in the diagnosis of gout. First, we developed a series of luminogens (i.e., tetraphenyl ethylene (TPE)-NH2, TPE-2NH2, TPE-4NH2, TPE-COOH, TPE-2COOH, TPE-4COOH, and TPE-Ketoalkyne), each of which was then evenly mixed with MSU crystals. Next, optimal fluorescence imaging of each of the luminogens was characterized by a confocal laser scanning microscope (CLSM). This approach was used for imaging standard samples of MSU, hydroxyapatite (HAP) crystals, and mixed samples with 1:1 mass ratio of MSU/HAP. We also imaged samples from mouse models of acute gouty arthritis, HAP deposition disease, and comorbidities of interest. Subsequently, CLSM imaging results were compared with those of compensated polarized light microscopy, and we assessed the biosafety of TPE-Ketoalkyne in the RAW264.7 cell line. Finally, CLSM time series and three-dimensional imaging were performed on MSU crystal samples from human gouty synovial fluid and tophi. As a promising candidate for MSU crystal labeling, TPE-Ketoalkyne was found to detect MSU crystals accurately and rapidly in standard samples, animal samples, and human samples, and could precisely distinguish gout from HAP deposition disease. This work demonstrates that TPE-Ketoalkyne is suitable for highly specific and timely imaging of MSU crystals in gouty arthritis and may facilitate future research on MSU crystal-related diseases.

复旦大学附属华山医院华英汇&华南理工大学秦安军课题组联合发表 | 基于聚集诱导发光技术高度特异性识别痛风性关节炎中的尿酸钠晶体

本研究论文聚焦基于聚集诱导发光技术高度特异性识别痛风性关节炎中的尿酸钠晶体。目前,国内外用于诊断痛风性关节炎的技术具有操作复杂、灵敏度较低的局限性,本研究目的是利用聚集诱导发光(AIE)技术探索高度特异性识别尿酸钠(MSU)晶体的探针分子,并协助痛风性关节炎的临床诊断。首先,基于MSU晶体结构的独特性,7种TPE探针被设计。它们分别与标准样品MSU晶体混合后,通过共聚焦激光扫描显微镜(CLSM)对每种混合样品进行荧光成像,TPE-Ketoalkyne被筛选为荧光成像效果最佳的AIE探针。此探针被用于MSU晶体、羟基磷灰石(HAP)晶体和MSU/HAP等比例混合样品的CLSM成像。结果显示TPE-Ketoalkyne可以快速、高效地标记MSU晶体,并且能够特异性区分MSU和HAP晶体,同时其良好的生物安全性被验证。此外,与金标准补偿偏振光显微镜(CPLM)成像方法相比,TPE-Ketoalkyne探针标记样本后的荧光成像操作更为简便,只需根据紫外光照射下的样本有无荧光聚集发光现象即可判断。最后,研究进一步通过CLSM时间序列和三维体外成像验证了TPE-Ketoalkyne动态标记痛风性关节炎患者关节中MSU晶体的过程。总的来说,我们发现了AIE探针分子TPE-Ketoalkyne可以高度特异性标记MSU晶体并且辅助痛风性关节炎的诊断,同时可能为未来MSU晶体相关疾病的研究提供帮助。

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