Skin Wound Healing: Some Biochemical Parameters in Guinea-pig

Abstract

Abstract Hairs were removed from the dorsal skin of guinea-pigs and 5–6 wounds (7 × 7 mm) were surgically induced by totally removing the epidermal and part of the dermal surface. They were then allowed to heal. The newly formed wound tissues were dissected at different times during the process and analysed by biochemical and histological methods. Hydroxyproline, proteins, DNA and semicarbazide-sensitive amine oxidase (SSAO) were measured, as were [14C]leucine and [3H]thymidine incorporation in some samples. The peroxidase-like activity of plasma albumin and the histology of wounds stained with haematoxylin-eosin were also studied. It was shown that SSAO enzymes, which are present in normal guinea-pig skin and have a high affinity for benzylamine are localized in fibroblasts. During skin healing in the newly formed tissue there was an increase in protein content which reached a maximum after 4–6 days; DNA content also increased. The rate of incorporation of [3H]thymidine and [14C]leucine paralleled DNA and protein content, respectively. The content of hydroxyproline had greatly decreased with respect to that in normal skin after 2–10 days. SSAO activity increased much less than DNA after 4 days whereas after 10–11 days it increased more than DNA, thus indicating that at this time it was probably produced by fibroblasts. No significant increase in the peroxidase-like activity of albumin was observed 4, 8 or 11 days after surgery. Treatment of the animals with methylprednisolone acetate (20 mg kg−1, i.m.) two days before surgery decreased the rate of skin healing but did not alter the level of albumin peroxidase activity of the plasma. Histology showed that in the animals treated with this drug the re-epithelialization was slower and after 11 days the wound appeared similar in appearance to the 8-day control wounds. In the methylprednisolone-treated animals a positive correlation was observed between the DNA content of regenerating tissue and the hydroxyproline content, whereas a negative correlation was observed in the control wounds. This correlation was in full agreement with the histological observation which showed an increased amount of cytogen collagen in the wounds of the treated animals. The simultaneous study of the biochemical parameters (DNA, proteins, SSAO, hydroxyproline) appears to be a good method for differentiating the pharmacological effects on the different cells which are responsible for wound healing.