sulfosuccinimidyl-6-((4-azido-2-nitrophenyl)amino)hexanoate and tetramethylrhodamine-isothiocyanate

Cell migration is critically important for the repair of chronic wounds, which cost billions of dollars each year to treat and can lead to serious complications, including amputation and death. Growth factors, including epidermal growth factor (EGF) and insulin-like growth factor-1 (IGF-1), are known to be deficient in chronic wounds; unfortunately, traditional delivery of soluble growth factors to wounds is expensive and complicated by their degradation. We have previously shown that directed and accelerated keratinocyte migration could be achieved by creating immobilized gradients of EGF. In this work, we have optimized EGF gradients for cell migration, synthesized and characterized gradient patterns of IGF-1, and tested for migration synergy upon combination of EGF and IGF-1 patterns. An optimal EGF concentration and pattern was identified, resulting in migration that was almost 10-fold that achieved on unpatterned controls. Immobilization of IGF-1 gradients also accelerated and directed keratinocyte migration (p < 0.05), however, no difference in migration was found across various IGF-1 concentrations or gradient patterns. Although combining EGF with IGF-1 patterns did not accelerate migration beyond levels achieved using EGF alone, these methods can be applied to create other types of multi-component gradients that may ultimately be utilized to create bioactive wound dressings.

Topics: Azides; Cell Line, Transformed; Cell Movement; Cross-Linking Reagents; Culture Media, Serum-Free; Dose-Response Relationship, Drug; Drug Synergism; Epidermal Growth Factor; Fluorescent Dyes; Humans; Indoles; Insulin-Like Growth Factor I; Keratinocytes; Recombinant Proteins; Reproducibility of Results; Rhodamines; Succinimides; Wound Healing