Abstract
Abstract: :
Purpose:To investigate the effect of human serum on human corneal fibroblast migration, proliferation, & matrix contraction in order to improve our understanding of corneal wound healing & the therapeutic actions of serum for persistent epithelial defect. Methods:Explanted human corneal fibroblasts were cultured to confluence in DMEM 10% FCS. Test media were; DMEM 10% FCS, 100% human serum & hypromellose 0.3%. Fibroblasts were seeded in 96 well plates, cultured in test media & proliferation assessed using WST–1 reagent. Fibroblast migration through transwell membranes was assessed after 16 hours. Relaxed & stressed serum–free fibroblast populated type I collagen gel contraction was measured, after 7 days and 2 & 3 days respectively, using digital image analysis software. In dose response studies gels were fed with serum–free media supplemented with 0%, 10%, 50% & 100% human serum and with DMEM 10% FCS. Results:Fibroblast proliferation & migration were significantly lower in 100% human serum than in the positive control DMEM 10% FCS (p = 0.0001, p = 0.0001 respectively); there was no significant difference between 100% human serum & hypromellose (p = 0.141, p = 0.944 respectively). Fibroblast–mediated relaxed gel contraction occurred to the same degree for 10, 50, 100% human serum & DMEM 10% FCS (maximal contraction 96.70; 96.42; 94.61; 92.78% respectively; p > 0.05) but was different to that in hypromellose (max. contract. 11.78%; p = 0.0001) & in DMEM without serum supplementation (max. contract. 53.36%; p = 0.0001). Stressed contraction was the same for 100% human serum & DMEM 10% FCS (p = 0.299). Conclusions:Human serum stimulates fibroblast mediated matrix contraction but does not stimulate fibroblast migration or proliferation. Corneal fibroblasts have an important role in the repair of injured tissue; they maybe responsible for tectonic repair but may also produce vision–threatening corneal opacity. Serum maybe present during corneal wound healing if it is given as a therapeutic agent; following haemorrhage after trauma or surgery, or due to leaky corneal new vessels. Our data suggest that human serum may modulate fibroblast contraction & possibly promote scarring if activated corneal fibroblasts are already present during wound healing. This should be considered in attempts to modify the wound healing process; particularly when serum is used therapeutically. Design of clinical studies of serum therapy should include the assessment of the potential effect of serum on corneal scarring.
Keywords: cornea: basic science • wound healing • drug toxicity/drug effects