A new wound healing agent--sphingosylphosphorylcholine.

Spingosylphosphorylcholine (lysosphingomyelin or SPC) is an effective and broad spectrum cell growth promoting agent and a candidate for evaluation on wound healing. The effect of SPC on full-thickness excision and incision wounds in genetically healing-impaired diabetic (db/db) mice was evaluated by measurement of wound area, skin strength, and tissue histology. The effect on cell proliferation was measured in vivo by incorporation of bromo-deoxyuridine and in vitro by [3H] thymidine incorporation. SPC increased the rate of wound closure, with a statistically significant improvement in measured wound areas (p < 0.02, compared with vehicle controls). The optimum concentration was 2-3 microM. SPC, alone and in combination with insulin, stimulated DNA synthesis in cells known to participate in wound healing, including microvascular endothelial cells. In vivo, SPC stimulated proliferation of keratinocytes, fibroblasts, endothelial cells, and cells around sebaceous glands and hair follicles at day 2-4 postwound, resulting in a complete re- epithelialization and profound granulation tissue formation in excisional and incisional wound sites of db.db and db/+ mice. Quantitative assessment of wound tissue section morphology indicated that SPC induced up to a 3-fold increase in the numbers of mitotic cells, resulted in smaller cross-sectional scar area, and led to more normalized tissue in the wound sites. SPC had no deleterious effect on wound skin strength. In conclusion, the acceleration of dermal wound healing animal models suggests that SPC could be an interesting candidate for clinical application.

Transfection with aFGF cDNA improves wound healing.

Somatic gene therapy is a potentially useful strategy for the delivery of growth factors or cytokines to enhance wound healing. Experimental excisional and incisional wounds in impaired-healing diabetic mice (db/db) were treated with aFGF and with a plasmid coding for aFGF. A eukaryotic expression plasmid composed of the Hst signal peptide sequence in-frame with the human aFGF sequence was used. Transfection of tissues was accomplished either by direct plasmid uptake or by uptake facilitated with cationic liposomes. The results show that the closure of excisional wounds was significantly accelerated (p < 0.05) by topical application of human recombinant aFGF or by transfection with the aFGF plasmid but not by vehicle or control plasmid not containing the aFGF sequence. In incisional wounds, aFGF or transfection with the plasmid significantly increased the wound-breaking strength compared to their corresponding controls (p < 0.05). Quantitative histology of the plasmid-treated incisional wound sections revealed improved wound quality. The transcription of mRNA from human aFGF cDNA in the incisional wound tissue extracts was confirmed by RT-PCR, and the expressed aFGF was detected by immune dot blot and immunohistochemistry assays. The transfection was a transient process with a peak at 9 d in db/+ (littermates of the diabetic mice) incisional wounds, at 36 d in db/db incisional wounds, and at 27 d in db/db excisional wounds. Cells transfected with human aFGF occupied up to 6.4% of the transectional area in the wound sites. Thus, aFGF gene delivery resulted in both gene expression and a functional improvement in healing.