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.
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.
Toegel
S
S
Department of Pharmaceutical Technology and Biopharmaceutics, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
Harrer
N
N
Plattner
V E
VE
Unger
F M
FM
Viernstein
H
H
Goldring
M B
MB
Gabor
F
F
Wirth
M
M
D000117:Acetylglucosamine; D001667:Binding, Competitive / Q000187:drug effects; D002241:Carbohydrates / Q000737:chemistry; D002356:Cartilage / Q000166:cytology / Q000502:physiology*; D002448:Cell Adhesion; D002460:Cell Line; D002470:Cell Survival; D019902:Chondrocytes / Q000378:metabolism*; D016503:Drug Delivery Systems; D005434:Flow Cytometry; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry; D037102:Lectins; D018613:Microscopy, Confocal; D023822:Tissue Engineering; D014909:Wheat Germ Agglutinins
D002241:Carbohydrates; D037102:Lectins; D014909:Wheat Germ Agglutinins; D006820:Hyaluronic Acid; D000117:Acetylglucosamine