ACS Applied Materials & Interfaces
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2013
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IF 5.008
Shi Peng,Qiang Zheng,Xin Zhang,Linghui Dai,Jingxian Zhu,Yanbin Pi,Xiaoqing Hu,Wenqing Cheng,Chunyan Zhou,Yinlin Sha,Yingfang Ao
A disintegrin and metalloproteinase with thrombospondin motif-4 (ADAMTS-4) plays a pivotal role in degrading aggrecan, which is an early event in cartilage degrading joint diseases such as osteoarthritis (OA). Detection of ADAMTS-4 activity could provide useful clinical information for early diagnosis of such diseases and disease-modifying therapy. Therefore, we developed a ADAMTS-4 detective fluorescent turn-on AuNP probe (ADAMTS-4-D-Au probe) by conjugating gold nanoparticles with a FITC-modified ADAMTS-4-specific peptide (DVQEFRGVTAVIR). When the ADAMTS-4-D-Au probe was incubated with ADAMTS-4, the fluorescence recovered and fluorescence intensity markedly increased in proportion to concentrations of ADAMTS-4 and the probe. A nearly 3-fold increase in fluorescent intensity in response to only 3.9 pM of ADAMTS-4 was detected, whereas almost no fluorescence recovery was observed when the probe was incubated with matrix metalloproteinase (MMP)-1, -3, and -13. These results indicate a relative high sensitivity and specificity of the probe. Moreover, ADAMTS-4-D-Au probe was used to detect ADAMTS-4 activity in synovial fluid from 11 knee surgery patients. A substantial increase in fluorescent intensity was observed in the acute joint injury group as compared to the chronic joint injury and end-stage OA groups, indicating that this simple and low-cost sensing system might serve as a new detection method for ADAMTS-4 activity in biological samples and in screens for inhibitors for ADAMTS-4-related joint diseases. Additionally, this probe could be a potential biomarker for early diagnosis of cartilage-degrading joint diseases.
研究领域:
生物医学
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分子生物学
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纳米医学
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DOI
PubMed
高影响因子
American Journal of Translational Research
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2018
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IF 3.061
Zhenlong Liu,Xiaoqing Hu,Peng Yang,Jiying Zhang,Chunyan Zhou,Yingfang Ao
Using a rabbit model of early-stage osteoarthritis (OA) and a sampling patients with OA, we evaluated the diagnostic utility of a fluorogenic peptide-conjugated gold nanoparticle (AuNP) probe in detecting mild cartilage injury, based on the a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) enzyme. Synthesis of this fluorescent turn-on probe (or AU-probe) required conjugation of AuNPs with a fluorescein isothiocyanate (FITC)-modified ADAMTS-4-specific peptide (DVQEFRGVTAVIR). Synovial fluid samples were then collected from 48 adult rabbits and 100 patients for comparative testing (ADAMTS-4 ELISA and AU-probe). Rabbit and patient MRI images were also evaluated and scored. Receiver operating characteristic (ROC) curve analysis was applied to various diagnostic methods (MRI, ELISA, AU-probe, and arthroscopy), performing comparisons via logistic regression. In rabbits, the AU-probe proved nonsuperior to MRI T2 mapping and ELISA (fluorescence cutpoint > 864.965 au). In patient groups, logistic regression analysis indicated that combined AU-probe/MRI testing outperformed MRI alone, thus offsetting low MRI sensitivity and low AU-probe specificity for improved detection of mild cartilage injury (sensitivity, 82.5%; specificity, 80.0%). We have consequently confirmed the efficacy of this AU-probe, using ADAMTS-4 activity in synovial fluid to diagnose mild cartilage injury. Combining the AU-probe with conventional MRI assessment proved optimal in this setting.
研究领域:
医学
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生物技术
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诊断学
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DOI
PubMed
高影响因子
CYTOTECHNOLOGY
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2021
·Vol. 73
·期 5
·715-726
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IF 2.058
Xu Haiqiao,Lin Shibang,Huang Haizhou
CCL22, which could induce chondrocyte apoptosis, was identified to be overexpressed in damaged cartilage. This study was conducted with the aim of investigating the effects of CCL22 interference on chondrocyte injury. The osteoarthritis model was established by stimulating chondrocytes with LPS. The expressions of CCL22, CCR4, matrix metallopeptidase (MMP) 3, MMP9, MMP13, (a disintegrin and metalloproteinase with thrombospondin-like motifs) ADAMTS-4, collagen II and inflammatory cytokines were measured using quantitative reverse transcription PCR (RT-qPCR) and western blot. Besides, immunoprecipitation (IP) was employed to verify the binding of CCL22 and CCR4. After CCR4 was overexpressed, cell viability was observed using Cell Counting Kit-8 (CCK-8). Additionally, cell apoptosis as well as its related proteins was detected by TUNEL and western blot, respectively. ng What’s more, glycosaminoglycan (GAG) level was detected using GAG kits. CCL22 and CCR4 expression increased noticeably in LPS-stimulated ATDC5 chondrocytes. CCL22 inhibition could suppress the expression of CCR4 in LPS-induced ATDC5 cells. Likewise, CCL22 inhibition could revive the activation of LPS-induced ATDC5 cells by regulating CCR4. In addition, CCL22 knockdown alleviated inflammatory response and cell apoptosis through CCR4. Furthermore, the cartilage degradation of ADTC5 cells could be relieved by CCL22 silence via regulating CCR4. CCL22/CCR4 expression was increased in osteoarthritic cartilage injury and participated in the inflammation and cartilage degradation of chondrocytes.
研究领域:
风湿病学
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细胞生物学
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分子生物学
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免疫学
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DOI
PubMed
高影响因子
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