Vitronectin: Growth Factor Complexes Hold Potential as a Wound Therapy Approach Zee Upton, Leila Cuttle, Anthony Noble, Margit Kempf, Gemma Topping, Jos.

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Vitronectin: Growth Factor Complexes Hold Potential as a Wound Therapy Approach Zee Upton, Leila Cuttle, Anthony Noble, Margit Kempf, Gemma Topping, Jos Malda, Yan Xie, Julie Mill, Damien G. Harkin, Olena Kravchuk, David I. Leavesley, Roy M. Kimble Journal of Investigative Dermatology Volume 128, Issue 6, Pages 1535-1544 (July 2008) DOI: 10.1038/sj.jid.5701148 Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Stimulation of migration of keratinocytes derived from human skin by VN, IGF-I, and/or IGFBP-3 or -5. The data, expressed as % stimulation of the control (serum-free media; SFM), were pooled from three replicate experiments in which each treatment was tested in duplicate. To dissect the importance of molecular interactions with the IGF-I and VN-binding cell surface receptors, an analog of IGF-I that binds poorly to the type-1 IGF-IR (Leu24-IGF-I) and an anti-αv integrin monoclonal antibody (αv-mAB) were used, respectively. Error bars=mean±SEM, n=9. Journal of Investigative Dermatology 2008 128, 1535-1544DOI: (10.1038/sj.jid.5701148) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Stimulation of keratinocyte proliferation in response to wound fluid in the presence and absence of the complexes. The data, expressed as % stimulation of the control (serum-free media; SFM), were pooled from three replicate experiments in which each treatment was tested in triplicate. Error bars=mean±SEM, n=9. Journal of Investigative Dermatology 2008 128, 1535-1544DOI: (10.1038/sj.jid.5701148) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Re-epithelialization of 3D in vitro skin equivalent model is enhanced by VN:growth factor complexes. (a) Histological analyses of epithelialization of a human skin equivalent (HSE) model in the presence of the VN:growth factor complexes or the control solution (serum-free media; SFM). The thickness of the viable and cornified epidermis was measured. (b) Histological analyses of a HSE model before and after application of a thermal lesion. The dashed vertical line denotes the margin of the burn. To the right of the line is the burnt area. (c) Schematic representation of the measurements of burn wound healing in the HSEs analyzed. HSEs were treated with either topical application (every 2 days) of the VN:growth factor complexes or the control solution (SFM). The dotted line illustrates the parameters measured: area, lateral migration, and net migration of keratinocytes in the (c) wound bed, (d) area, (e) lateral migration, and (f) net migration. Journal of Investigative Dermatology 2008 128, 1535-1544DOI: (10.1038/sj.jid.5701148) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Effect of topical application of the VN:growth factor complexes on porcine wounds. Burn wounds in juvenile Large White pigs were treated with Acticoat7™ (a standard burn dressing)±VN:growth factor complexes. (a) Re-epithelialization was significantly faster (P=0.031) over the duration of the experiment in the VN:growth factor-treated wounds (refer to scoring scale in Materials and Methods). (e) There was significantly less organizing granulation tissue (OGT) in the VN:growth factor complex-treated group (P=0.010). There were no significant differences observed in (b) total skin thickness, (c) dermal thickness, (d) epidermal thickness, or (f) hair follicle density. Error bars=mean±SEM, control group n=4 and VN:growth factor complex-treated group n=3. Journal of Investigative Dermatology 2008 128, 1535-1544DOI: (10.1038/sj.jid.5701148) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Photographic images of the control vs VN:growth factor complex-treated wounds. Photographs of wounds on both the left and right flanks of all animals after 6 weeks of treatment. Photographs on the left show wounds in the four control-treated animals (Acticoat7™), whereas those on the right show wounds from the three VN:growth factor-treated animals (Acticoat7™+VN:growth factor complex). Scale bar=2cm. Journal of Investigative Dermatology 2008 128, 1535-1544DOI: (10.1038/sj.jid.5701148) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions