Lectin binding studies on C-28/I2 and T/C-28a2 chondrocytes provide a basis for new tissue engineering and drug delivery perspectives in cartilage research.
Lectin binding studies on C-28/I2 and T/C-28a2 chondrocytes provide a basis for new tissue engineering and drug delivery perspectives in cartilage research.
复制标题
DOI：
10.1016/j.jconrel.2006.10.004
复制
发表时间：
2007-01
期刊：
影响因子：
--
通讯作者：
S. Toegel;N. Harrer;V. E. Plattner;Frank M. Unger;H. Viernstein;M. Goldring;Franz Gabor;M. Wirth
中科院分区：
文献类型：
--
作者：
S. Toegel;N. Harrer;V. E. Plattner;Frank M. Unger;H. Viernstein;M. Goldring;Franz Gabor;M. Wirth
本研究旨在评估植物凝集素作为生物粘附介质在软骨研究中的适用性，使用人软骨细胞系C-28/I2和T/C-28 a2。用流式细胞术研究了不同糖特异性的荧光素标记凝集素的生物粘附特性。特异性的凝集素-细胞相互作用的竞争性抑制使用互补的碳水化合物被确定。与其他凝集素相比，麦胚凝集素（WGA）和软骨细胞之间的相互作用的特点是显着的细胞粘附，足够的结合强度和高度的特异性N-乙酰基-葡萄糖胺中所含的透明质酸链。因此，我们建议WGA是一个很有前途的候选人介导的生物粘附低粘附性支架软骨组织工程。此外，C-28/I2和T/C-28 a2细胞的WGA结合率依赖于温度，表明细胞对膜结合WGA的摄取。通过共聚焦显微镜证实细胞内富集。用莫能菌素平衡细胞内pH梯度导致淬灭作用逆转，表明软骨细胞的酸隔室内积累了WGA。因此，WGA可能通过CD 44受体介导的内吞作用途径与透明质酸一起内化到软骨细胞中，并在溶酶体内积累。这种生理过程可能代表了将WGA功能化药物递送装置靶向软骨细胞的可行途径。
The present study was performed to evaluate the applicability of plant lectins as mediators of bioadhesion in cartilage research using human chondrocyte cell lines C-28/I2 and T/C-28a2. The bioadhesive properties of fluorescein-labelled lectins with different carbohydrate specificities were investigated by flow cytometry. Specificity of the lectin–cell interactions was ascertained by competitive inhibition using complementary carbohydrates. As compared to that of other lectins, the interaction between wheat germ agglutinin (WGA) and chondrocytic cells was characterised by remarkable cytoadhesion, adequate binding strength and a high degree of specificity for N-acetyl-glucosamine as contained in hyaluronan chains. We therefore suggest WGA to be a promising candidate for mediating bioadhesion to low-adhesive scaffolds in cartilage tissue engineering. Moreover, the WGA-association rate of C-28/I2 and T/C-28a2 cells was dependent on temperature indicating cellular uptake of membrane-bound WGA. Intracellular enrichment was confirmed by confocal microscopy. Equilibration of intracellular pH gradients with monensin resulted in the reversal of quenching effects indicating accumulation of WGA within acid compartments of chondrocytic cells. Thus, WGA might be internalised into chondrocytes together with hyaluronan via the CD44 receptor-mediated endocytosis pathway and accumulated within lysosomes. This physiological process could represent a feasible pathway to target WGA-functionalised drug delivery devices into chondrocytes.