Sequential changes in trace metal, metallothionein and calmodulin concentrations in healing skin wounds

Metalloenzymes have an important role in repair and regenerative processes in skin wounds. Demands for different enzymes vary according to the phase i n the healing cascade and constituent events. Sequential changes in the con centrations of calcium, copper, magnesium and zinc were studied in the inci sional wound model in the rat over a 10 d period. Copper levels remained lo w(< 10 mu g/g dry weight) throughout, but calcium, magnesium and zinc incre ased from wounding and peaked at about 5 d at a time of high inflammation, granulation tissue formation and epidermal cell proliferation. Metal concen trations declined to normal by 7 d when inflammation had regressed, re-epit helialisation of the wound site was complete and the 'normalisation' phase had commenced. Although the wound was overtly healed by 10 d, the epidermis was still moderately hyperplastic. In view of competitive binding of trace metals at membrane receptors and carrier proteins, the ratios or balance b etween these trace metals was examined and the significance is discussed. U sing immunocytochemistry, we demonstrated increases in metallothionein immu noreactivity as an indication of zinc and copper activity in the papillary dermis and in basal epidermal cells near the wound margin 1-5 d after wound ing. This is consistent with metalloenzyme requirements in inflammation and fibrogenesis. Calmodulin, a major cytosolic calcium binding protein was hi ghest in maturing keratinocytes and in sebaceous gland cells of normal skin ; it was notably more abundant in the epidermis near the wound margin and i n re-epithelialising areas at a time when local calcium levels were highest .