Alginate-poly-L-lysine microcapsule biocompatibility: a novel RT-PCR method for cytokine gene expression analysis in pericapsular infiltrates.
Transplantation of microencapsulated islets of Langerhans is impaired by a pericapsular host reaction that eventually induces graft failure. We are studying the role of cytokines in the pathogenesis of this reaction, using the model of alginate-polylysine microcapsule implantation in rat epididymal fat pads. The objectives were: (1) to develop a method to measure, by semiquantitative PCR, TGF-beta1 gene expression in fat pad pericapsular infiltrates, and (2) to use this method to evaluate TGF-beta1 gene expression 14 days after microcapsule implantation. TGF-beta1 mRNA level was significantly higher in pericapsular infiltrate cells than in nonimplanted tissue cells and saline-injected tissue cells (p < 0.0001 and p < 0.01, respectively). There was no significant difference between the TGF-beta1 mRNA levels of the two types of controls (p = 0.0945). These results suggest that TGF-beta1 plays a role in the pathogenesis of the pericapsular reaction. The method developed can be used to study the role of other fibrogenic cytokines potentially involved. This will shed light on the mechanisms underlying the pericapsular reaction and will serve as a basis for the development of strategies to control this reaction.
Robitaille
R
R
Centre de Recherche Guy-Bernier, Hôpital Maisonneuve-Rosemont, 5415, boulevard de l'Assomption, Montréal, Québec, H1T 2M4, Canada.
Leblond
F A
FA
Henley
N
N
Prud'homme
G J
GJ
Drobetsky
E
E
Hall
J P
JP
D000273:Adipose Tissue / Q000201:enzymology / Q000276:immunology; D000464:Alginates / Q000633:toxicity*; D000818:Animals; D001672:Biocompatible Materials / Q000633:toxicity*; D002214:Capsules; D002452:Cell Count; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006789:Host vs Graft Reaction; D007111:Immunity, Cellular / Q000187:drug effects; D008297:Male; D008422:Materials Testing; D011107:Polylysine / Q000031:analogs & derivatives* / Q000633:toxicity; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics
D000464:Alginates; D001672:Biocompatible Materials; D002214:Capsules; D016207:Cytokines; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; C051546:alginate-polylysine-alginate; D011107:Polylysine