The role of Galectin-1 in the interaction between chondrocytes and a lactose-modified chitosan.
Evidences for the involvement of the Galectin-1 in the interaction of pig chondrocytes with a lactose-modified chitosan, namely Chitlac, are reported. The Chitlac glycopolymer has been shown to promote pig chondrocyte aggregation and to induce extracellular matrix production. Highly pure Galectin-1 was obtained from pig spleen by affinity chromatography and its identity was determined by ion spray mass spectrometry analysis of tryptic peptide fragments obtained after in-gel digestion. The complete sequence of pig Galectin-1 CDS was obtained by screening a pig EST database using human Galectin-1 sequence as template. The Galectin-1 cDNA was cloned into a pGEX-4T-1 expression vector and the recombinant protein was purified, characterized and used to produce a rabbit anti-serum. Recombinant Galectin-1 interacts in a dose-dependent manner with Chitlac as determined with ELISA assay. Expression level of galectin-1 gene, quantified by real-time PCR, was significantly higher in chondrocytes cultivated on Chitlac. In the same way, the presence of Chitlac stimulates secretion of Galectin-1 in culture medium that, by immunohistochemical analysis, revealed to be clustered on the surface of Chitlac-induced aggregates. These data indicate the role of Galectin-1 as a bridging agent between Chitlac and chondrocyte aggregates.
Marcon
Patrizia
P
Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Via L. Giorgieri 1, I-34127 Trieste, Italy.
Marsich
Eleonora
E
Vetere
Amedeo
A
Mozetic
Pamela
P
Campa
Cristiana
C
Donati
Ivan
I
Vittur
Franco
F
Gamini
Amelia
A
Paoletti
Sergio
S
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology*; D037483:Galectin 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007785:Lactose / Q000737:chemistry*; D008422:Materials Testing; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013552:Swine
D001672:Biocompatible Materials; D037483:Galectin 1; D011994:Recombinant Proteins; D048271:Chitosan; D007785:Lactose