Nitrosoglutathione triggers collagen deposition in cutaneous wound repair.

The presence of nitric oxide (NO) is associated with enhanced wound fibroblast collagen synthesis; previous observations have focused on the effect of NO on wound collagen content. This article emphasizes the effect of nitrosothiols on wound collagen deposition and matrix-metalloproteinase activity, which is the primary breakdown pathway of collagen. We examined the effects of S-nitrosoglutathione (GSNO) and glutathione (GSH) on rat scar tissue. Hydroxyproline content, matrix metalloproteinase activity, total glutathione, and total nitrite of scar tissue were measured 3, 5, 7, and 10 days after wounding. It was observed that, at Day 5 and Day 10, wound collagen content was 52.0 percent and 47.5 percent higher, respectively, after GSNO administration than in controls (p<0.05). GSH administration decreased wound collagen deposition 76.5 percent by Day 5 (p<0.05). GSH lowered the matrix metalloproteinase activity 67 percent at Day 5 and 50 percent (p<0.05) at Day 10. Nitrite and nitrate levels were 55 percent higher in the GSNO treated rats than in the control group (p<0.05) at Day 3, whereas the GSH-treated groups showed no changes. GSNO increased systemic nitrite 53 percent 3 hours after intraperitoneal injection. Our findings suggest that collagen deposition increases in cutaneous wound healing after the administration of GSNO and that this nitrosothiol does not interfere with the collagenolytic pathway, thus maintaining the physiological conditions necessary for wound healing.

Fibroblasts cocultured with keloid keratinocytes: normal fibroblasts secrete collagen in a keloidlike manner.

Keloid scars represent a pathological response to cutaneous injury, reflecting a new set point between synthesis and degradation biased toward extracellular matrix (ECM) collagen accumulation. Using a serum-free two-chamber coculture model, we recently demonstrated a significant increase in normal fibroblast proliferation when cocultured with keloid-derived keratinocytes. We hypothesized that similar keratinocyte-fibroblast interactions might influence fibroblast collagen production and examined conditioned media and cell lysate from coculture for collagen I and III production by Western blot, allied with Northern analysis for procollagen I and III mRNA. Normal fibroblasts cocultured with keloid keratinocytes produced increased soluble collagen I and III with a corresponding increase in procollagen I and III mRNA transcript levels. This was associated with decreased insoluble collagen from cell lysate. When keloid fibroblasts were cocultured with keloid keratinocytes, both soluble and insoluble collagen were increased with associated procollagen III mRNA upregulation. Transmission electron microscopy of normal fibroblasts cocultured with keloid keratinocytes showed an ECM appearance similar to in vivo keloid tissue, an appearance not seen when normal fibroblasts were cocultured with normal keratinocytes.