Cumming G J, Handley C J, Preston B N
Biochem J. 1979 Aug 1;181(2):257-66. doi: 10.1042/bj1810257a.
A model connective-tissue system was developed that is amenable to the determination of permeability coefficients of diffusing solutes. The system involves the culture of 13-day chick-embryo chondrocytes on a Millipore filter (HA:0.45 micron pore size) to form what is, in effect, a confluent, extremely thin cartilage slice of uniform thickness. These cultured chondrocyte membranes were used to measure the steady-state flux of radioactively labelled low-molecular-weight solutes and micro-ions. Similarly, the permeability coefficients of either radioactively labelled or enzymically active proteins across the membranes were determined. The membrane was found to have no marked effects on the diffusional behaviour of low-molecular-weight non-electrolytes (water, proline, ribose, glucose, sorbitol, raffinose). For micro-ions (Na+, SO42-, Cl-, glutamate, glucuronate,), the diffusive behaviour was found to be markedly affected by the ionic strength of the solvent used in a manner which was consistent with a Donnan distribution resulting from the immobilized proteoglycans. Globular proteins permeated the membrane at rates which decreased as the molecular size of the diffusing solute increased. The apparent diffusion rates of fibrinogen and of collagen through the membranes were greater than would be expected on the basis of their diffusion coefficients in free solution. Reasons for this behaviour are discussed.
开发了一种模型结缔组织系统,该系统适用于测定扩散溶质的渗透系数。该系统包括在微孔滤膜(HA:孔径0.45微米)上培养13天的鸡胚软骨细胞,以形成实际上是厚度均匀的汇合极薄软骨切片。这些培养的软骨细胞膜用于测量放射性标记的低分子量溶质和微离子的稳态通量。同样,还测定了放射性标记或具有酶活性的蛋白质跨膜的渗透系数。发现该膜对低分子量非电解质(水、脯氨酸、核糖、葡萄糖、山梨醇、棉子糖)的扩散行为没有显著影响。对于微离子(Na +、SO42 -、Cl -、谷氨酸、葡萄糖醛酸),发现扩散行为受到所用溶剂离子强度的显著影响,其方式与固定化蛋白聚糖产生的唐南分布一致。球状蛋白透过膜的速率随着扩散溶质分子大小的增加而降低。纤维蛋白原和胶原蛋白通过膜的表观扩散速率大于根据它们在自由溶液中的扩散系数所预期的速率。讨论了这种行为的原因。