Advanced glycation of type I collagen and fibronectin modifies periodontal cell behavior.
BACKGROUND
Advanced glycation end products (AGEs) have been linked to pathogenic mechanisms of diabetes mellitus. However, little is known about the contribution of protein glycation to periodontal disease in patients with diabetes. Therefore, this study investigated whether glycation of type I collagen (COLI) and fibronectin (FN) modified the behavior of human gingival fibroblasts (hGFs) and periodontal ligament fibroblasts (hPDLs).


METHODS
Procedures for rapid in vitro glycation of COLI and FN used methylglyoxal (MG). Formation of AGEs was analyzed by changes in protein migration using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antibodies specific for MG-glycated proteins. Experiments then characterized the effects of glycated FN and COLI on the behavior of hGFs and hPDLs.


RESULTS
MG glycated COLI and FN in <6 hours. Confirming the specificity of the reactions, antibodies specific for MG-induced AGEs reacted with glycated FN and COLI but not with control proteins. In cell culture experiments, glycated FN was significantly less efficient in supporting the attachment of hGFs and hPDLs (P <0.05). Moreover, the morphologic parameters, including length, area, perimeter, and shape factor, were altered (P <0.001) for cells on both glycated proteins. Finally, cell migration was reduced on glycated FN and COLI (P <0.001).


CONCLUSIONS
MG treatment efficiently glycated COLI and FN, providing a new tool to study the effects of diabetes on periodontal disease. The substantial effects of glycated COLI and FN on hGF and hPDL behavior indicated that protein glycation contributed to the pathogenesis and altered periodontal wound healing observed in patients with diabetes.
Murillo
Jesse
J
Department of Periodontics, University of Texas Health Science Center, San Antonio, TX, USA.
Wang
Yao
Y
Xu
Xiaoping
X
Klebe
Robert J
RJ
Chen
Zhihua
Z
Zardeneta
Gustavo
G
Pal
Sanjay
S
Mikhailova
Margarita
M
Steffensen
Bjorn
B
D024042:Collagen Type I / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D005881:Gingiva / Q000166:cytology / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006801:Humans; D010513:Periodontal Ligament / Q000166:cytology / Q000378:metabolism*; D011765:Pyruvaldehyde / Q000378:metabolism; D018709:Statistics, Nonparametric
D024042:Collagen Type I; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D017127:Glycation End Products, Advanced; D011765:Pyruvaldehyde