Department of Chemical Engineering, McGill University, Montreal, Quebec, Canada.
Biophys J. 2013 Feb 19;104(4):943-50. doi: 10.1016/j.bpj.2012.12.036.
Electrolyte filtration arises due to the presence of fixed charges in cartilage extracellular matrix glycosaminoglycans (GAGs). Commonly assumed negligible, it can be important for design and interpretation of streaming potential measurements and modeling assumptions. To quantify the scale of this phenomenon, chloride ion concentration in exudate of compressed cartilage was measured by Mohr's titration and explant GAG content was colorimetrically assayed. Pilot studies indicated that an appropriate strain rate for experiments was 8 × 10(-3) s(-1) to eliminate concerns of exudate evaporation and explant damage (at low and high strain rates, respectively). Exudate chloride concentration of explants equilibrated in 1× PBS was significantly (p < 0.05) lower than the bath chloride concentration at strains of 37.5, 50, and 62.5%, with clear dependence on strain magnitude. Exudate chloride concentration was also significantly lower than that of the bath when 50% strain was applied after equilibration in 0.5, 1, and 2× PBS, with a trend for an increase in this relative difference with decreasing bath concentration (p = 0.065 between 0.5 and 2× PBS). Decreasing exudate chloride concentration correlated negatively with increasing postcompression GAG concentration. No difference between exudate chloride concentration and bath chloride concentration was ever observed for compression of uncharged agarose gel controls. Findings show that exudate from compressed cartilage is dilute relative to the bath due to the presence of matrix fixed charges, and this difference can generate diffusion potentials external to the explant, which may affect streaming potential measurements particularly under conditions of low strain rates and high strains.
由于软骨细胞外基质糖胺聚糖 (GAG) 中存在固定电荷,因此会出现电解质滤过。通常认为这种作用可以忽略不计,但对于流动电势测量和建模假设的设计和解释而言,它可能很重要。为了量化这种现象的规模,通过莫尔滴定法测量了压缩软骨渗出液中的氯离子浓度,并通过比色法测定了外植体 GAG 含量。初步研究表明,实验的适当应变速率为 8×10(-3) s(-1),以消除渗出液蒸发和外植体损伤的担忧(分别在低应变速率和高应变速率下)。在 1×PBS 中平衡的外植体渗出液氯离子浓度明显(p<0.05)低于浴氯离子浓度,在 37.5、50 和 62.5%应变下,且明显依赖于应变幅度。当在 0.5、1 和 2×PBS 中平衡后施加 50%应变时,外植体渗出液氯离子浓度也明显低于浴液,并且随着浴液浓度降低,这种相对差异呈增加趋势(在 0.5 和 2×PBS 之间,p=0.065)。外泌体氯离子浓度的降低与压缩后 GAG 浓度的增加呈负相关。对于不带电荷的琼脂糖凝胶对照物的压缩,从未观察到外泌体氯离子浓度与浴液氯离子浓度之间的差异。研究结果表明,由于基质固定电荷的存在,压缩软骨的渗出液相对于浴液是稀释的,这种差异会在外植体外部产生扩散电势,特别是在低应变速率和高应变条件下,这可能会影响流动电势测量。