Cell Suspensions of Autologous Keratinocytes or Autologous Fibroblasts Accelerate the Healing of Full Thickness Skin Wounds in a Diabetic Porcine Wound Healing Model  Patrik Velander, M.D., Christoph Theopold, M.D., Oliver Bleiziffer, M.D., Juri Bergmann, M.D., Henry Svensson, M.D., Ph.D., Yao Feng, Ph.D., Elof Eriksson, M.D., Ph.D.  Journal of Surgical Research  Volume 157, Issue 1, Pages 14-20 (November 2009) DOI: 10.1016/j.jss.2008.10.001 Copyright © 2009 Elsevier Inc. Terms and Conditions

Fig. 1 Transplantation of keratinocytes or fibroblasts accelerates epidermal healing in the diabetic pig. (A) Level of re-epithelialization of full thickness wounds treated with a suspension of 500,000 keratinocytes or normal saline. Wounds measuring 1.5 × 1.5 cm were created on the backs of pigs. On the day of wounding, an injection of either a suspension of keratinocytes or normal saline was done into each wound. On days 8, 12, and 14, biopsies were harvested, embedded in paraformaldehyde and stained with hematoxylin and eosin for analysis. The error bars refer to SD. P = 0.001 (B) Re-epithelialization on day 12 of full thickness wounds treated with a suspension of 650,000 fibroblasts or normal saline. Wounds measuring 1.5 × 1.5 × 0.8 cm were created on the backs of pigs. On the day of wounding, an injection of either a suspension of fibroblasts or normal saline was done into each wound. On day 12, biopsies were harvested, embedded in paraformaldehyde and stained with hematoxylin and eosin for analysis. The error bars refer to SD. P = 0.035. Journal of Surgical Research 2009 157, 14-20DOI: (10.1016/j.jss.2008.10.001) Copyright © 2009 Elsevier Inc. Terms and Conditions

Fig. 2 Transplantation of keratinocyte into full thickness wounds in diabetic pigs leads to the formation of keratinocyte colonies, which contribute to epidermal healing. Vertical wound biopsies 2 mm wide were taken from the middle of the wounds, which included unwounded skin at the sides and subcutaneous tissue at the bottom. Biopsies were collected on day 12 post-wounding, embedded in paraformaldehyde and stained with H and E for analysis. The arrows show the edge of re-epithelialization. (A) Wounds treated with a suspension of 500,000 keratinocytes. (B) Normal saline treated wound. Scale 6.5 mm for A and B. (Color version of figure is available online.) Journal of Surgical Research 2009 157, 14-20DOI: (10.1016/j.jss.2008.10.001) Copyright © 2009 Elsevier Inc. Terms and Conditions

Fig. 3 Keratinocyte colonies migrate upwards from the base of the wound to enhance epidermal healing. Histology of wounds on day 12 (H and E staining). Pictures shows the center of the wound. The arrows show the end of the epithelial tongues. (A) Wound treated with a suspension of 500,000 keratinocytes. (B) Normal saline treated wound Scale 500 μm. (Color version of figure is available online.) Journal of Surgical Research 2009 157, 14-20DOI: (10.1016/j.jss.2008.10.001) Copyright © 2009 Elsevier Inc. Terms and Conditions

Fig. 4 The effect of keratinocytes or fibroblasts transplantation on the contraction of full thickness wounds in the diabetic pig. Full thickness wounds measuring 1.5 × 1.5 × 0.8 cm were created on the backs of pigs, and were treated with either a suspension of 500,000 keratinocytes (panel A), a suspension of 650,000 fibroblasts (panel B), or normal saline (panels A and B). On day 3, 6, 9, and 12, wound contractions were measured using the permanent boundaries left by the tattooed margins and planimetry. The percentage of contraction was calculated using the following formula: (area at wounding day – area at biopsy day)/(area on wounding day) × 100. The error bars refer to SD. Journal of Surgical Research 2009 157, 14-20DOI: (10.1016/j.jss.2008.10.001) Copyright © 2009 Elsevier Inc. Terms and Conditions