光学氧化还原成像可预测加载后软骨线粒体膜电位。

Optical Redox Imaging Predicts Post-Loading Cartilage Mitochondrial Membrane Potential.

作者信息

Wang Jingyi, Scheidt Greta E, Henak Corinne R

机构信息

Department of Mechanical Engineering, University of Wisconsin-Madison, 3031 Mechanical Engineering Building, 1513 University Ave, Madison, WI, 53706, USA.

Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.

出版信息

Ann Biomed Eng. 2025 Jun 27. doi: 10.1007/s10439-025-03784-1.

DOI:10.1007/s10439-025-03784-1

PMID:40576853

Abstract

PURPOSE

Disrupted cellular redox balance is associated with various diseases, including osteoarthritis. Although mitochondrial (MT) membrane potential is a proxy for redox balance, the translational potential of this method is limited by exogenous dye. Therefore, the objective of this study was to predict changes in MT membrane potential in response to mechanical loading using dye-free optical redox imaging (ORI). A secondary objective was to determine the effect of loading on ORI metrics.

METHODS

Full-thickness porcine cartilage strips were subjected to tensile loading at one of two strain rates (1.00 s or 0.10 s). ORI was done before, immediately after, and 30 minutes after loading. MT membrane potential was then measured using fluorescent dye. A generalized linear mixed-effects model (GLMM) tested main effects (ORI metrics, loading vs. control, loading rate, post-loading time, zone) and their interactions in prediction of MT membrane potential. Significant predictors were retained in a new GLMM that was trained using 70% of the dataset and evaluated using the remaining 30%. Two separate GLMMs evaluated the main effects on ORI metrics.

RESULTS

In the GLMM using MT red/green ratio as the dependent variable, ORI metrics, loading rate, and loading vs. control were significant main effect. GLMMs to predict MT red/green from ORI that retained significant main effects resulted in an average difference between predicted and actual values of 7.07%. When analyzing the effect of loading vs. control, loading rate, and zone on ORI metrics, only loading rate showed significance.

CONCLUSION

ORI can predict MT membrane potential measured by fluorescent dye and has the possibility to be developed as a clinical tool to evaluate cartilage redox balance.

摘要

目的

细胞氧化还原平衡紊乱与包括骨关节炎在内的多种疾病相关。尽管线粒体（MT）膜电位是氧化还原平衡的一个指标，但该方法的转化潜力受到外源性染料的限制。因此，本研究的目的是使用无染料光学氧化还原成像（ORI）预测MT膜电位对机械负荷的变化。第二个目的是确定负荷对ORI指标的影响。

方法

将全层猪软骨条以两种应变率之一（1.00 s或0.10 s）进行拉伸负荷。在负荷前、负荷后立即和负荷后30分钟进行ORI。然后使用荧光染料测量MT膜电位。采用广义线性混合效应模型（GLMM）测试主要效应（ORI指标、负荷组与对照组、负荷率、负荷后时间、区域）及其在预测MT膜电位中的相互作用。显著预测因子保留在一个新的GLMM中，该模型使用70%的数据集进行训练，并使用其余30%进行评估。两个独立的GLMM评估对ORI指标的主要效应。