Hyperglycaemic conditions hamper keratinocyte locomotion via sequential inhibition of distinct pathways: new insights on poor wound closure in patients with diabetes.
BACKGROUND
Diabetes mellitus (DM) is characterized by impaired insulin signalling, elevated plasma glucose, and predisposition towards complications involving several organs. A major complication of DM is impairment of wound healing. In the re-epithelialization process during wound healing, migration of keratinocytes is a crucial step. Our previous report demonstrated that keratinocytes cultured in hyperglycaemic media showed decreased cell mobility.


OBJECTIVE
The current study aimed to explore the effects of high glucose on keratinocyte migration after different treatment durations.


METHODS
Keratinocytes were cultivated for indicated time periods under various concentrations of glucose. Relevant assays including Transwell migration and in vitro wound scratch assays, flow cytometric analysis, matrix metalloproteinase-1 (MMP-1) activity assay, determination of mRNA expression and Western blotting were performed.


RESULTS
We demonstrated that (i) keratinocyte motility progressively and significantly decreased; (ii) the keratinocyte activation marker K16 was significantly suppressed; (iii) expression of alpha2beta1 integrin and MMP-1, both crucial for keratinocyte locomotion on collagen type I, was significantly downregulated; and (iv) expression of the phosphorylated signal transducer and activator of transcription-1 significantly decreased after hyperglycaemic treatment. More specifically, different pathways become involved after prolonged duration of high glucose cultivation to reduce keratinocyte locomotion further.


CONCLUSIONS
We have demonstrated that high glucose treatment results in progressive suppression of keratinocyte locomotion and elucidated the molecular mechanisms involved. These results provide a reasonable explanation for the poor wound healing seen in patients with DM.
Lan
C-C E
CC
Department of Dermatology, Chung-Ho Memorial Hospital and College of Medicine, Kaohsiung Medical University, Taiwan.
Wu
C-S
CS
Kuo
H-Y
HY
Huang
S-M
SM
Chen
G-S
GS
D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D003920:Diabetes Mellitus / Q000378:metabolism / Q000503:physiopathology*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000502:physiology; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D013223:Statistics as Topic; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D014945:Wound Healing
D019715:Tissue Inhibitor of Metalloproteinase-1; D020781:Matrix Metalloproteinase 1; D005947:Glucose