Hyperglycemia-Induced Changes in Hyaluronan Contribute to Impaired Skin Wound Healing in Diabetes: Review and Perspective

Figure 2

Skin fibroblasts with a larger HA glycocalyx are more responsive to TGF-β and produce more collagen when cultured in a high glucose medium. Skin fibroblasts from either wild type (WT) or Has1/3 null mice were cultured in normal glucose (1 g/L) or in high glucose (4.5 g/L) in the absence or presence of TGF-β1 (2 ng/mL) in DMEM media with 1% FBS for 48 hours. (a) Erythrocyte exclusion assay to visualize the pericellular coat on WT and Has1/3 null fibroblasts [51]. Cells were labeled with calcein to delineate the cell bodies. The HA glycocalyx is indicated between the white and yellow lines. (b) Western blot of protein from WT or Has1/3 null fibroblasts, probed with an antibody to type 1 collagen. GAPDH, loading control. (c) Quantification of protein band intensities from two independent western analysis experiments, performed under the conditions shown in panel (b) and normalized to GAPDH. White bars, WT fibroblasts. Gray bars, Has1/3 null fibroblasts. Error bars, mean ± half-range.

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