Cell adhesion and cytotoxicity studies over polyanionic collagen surfaces with variable negative charge and wettability.
This work describes the cytotoxicity, and the cell adhesion behavior of K562 cell line from human erythroleukemia transfected with the DNA for the alpha(2)beta(1) integrin over type-I collagen matrices with variable degree of carboxyl group and wettability. The results showed that type-I collagen materials with variable degree of carboxyl group prepared by selective hydrolysis of carboxyamide side chains of Asn and Gln residues present in the protein, independently from the extent of side chain hydrolysis, was characterized by preserved triple helix structure for materials with a carboxyl group content up to 87 +/- 17. Imbibition and wettability increased linearly with increasing carboxyl group content from 46 +/- 12 to 87 +/- 17, and no signs of cytotoxicity were detected. Nevertheless, in comparison to native collagen, K562 cell adhesion to PACMs was significantly improved by factors ranging from 1.60 to 1.47x, with the reduction in cell adhesion observed with increasing carboxyl content attributed to a balance between the inhibition of increasing negative charge and the stimulation by increased wettability. On the other hand, the overall improvement of K562 cell adhesion to polyanionic collagen was attributed to the introduction of new distinct motifs described as the minimal active recognition sequence for alpha(2)beta(1) integrins binding with type-I collagen produced as a result of Asn-Gly Glu-Ala alpha2(I)294-297, and Gly Gln-Arg-Gly Val-Val carboxyamide side chains hydrolysis.
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Department of Chemistry and Molecular Physics, Institute of Chemistry, Universidade de São Paulo, SP Av Trabalhador Sãocarlense 400, São Carlos, São Paulo, CP 780-CEP 13560-970, Brazil.
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D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity*; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D024042:Collagen Type I / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity*; D004563:Electrochemistry; D006801:Humans; D066298:In Vitro Techniques; D038982:Integrin alpha2beta1 / Q000235:genetics / Q000378:metabolism; D020014:K562 Cells; D008422:Materials Testing; D008567:Membranes, Artificial; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013499:Surface Properties; D014162:Transfection
D001672:Biocompatible Materials; D024042:Collagen Type I; D038982:Integrin alpha2beta1; D008567:Membranes, Artificial; D011994:Recombinant Proteins