AN EXPERIMENTAL STUDY ON REPAIR OF FULL THICKNESS SKIN WOUND WITH GRAFT OF AUTOLOGOUS KERATINOCYTE SUSPENSION IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

YINKai ,  SHENChuanan , MALi , SHANGYuru , LIDawei , LILongzhu , ZHAODongxu , CHENGWenfeng

Burns Institute, the First Hospital Affiliated to Chinese PLA General Hospital, Beijing, 100048, P. R. China;

Corresponding author：

SHENChuanan, Email: shenchuanan@126.com

Keywords：

Cell transplantation; Epidermal cells; Skin grafting; Wound repair; Rat

DOI：

10.7507/1002-1892.20160045

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the application of autologous keratinocyte suspension transplantation in skin reconstruction. Methods Forty adult Sprague Dawley rats (weighing, 210-230 g) were randomly divided into 4 groups (n=10): high density keratinocyte suspension transplantation (1×10⁶ cells/cm²) group (group A), middle density (1×10⁵ cells/cm²) group (group B), low density (1×10⁴ cells/cm²) group (group C), and control group (group D). Skin samples were harvested from the rats in groups A, B, C for the isolation of keratinocytes. The model of anti-contracture of full thickness skin wound was made in rats and autologous keratinocyte suspension was transplanted into the wound. The wound was covered by the allogeneic skin (allogeneic skin derived from Wistar rats). The survival of rats was observed after operation. The survival of allogeneic skin was observed at 7, 14, and 21 days after operation, and wound healing rate was calculated after allogeneic skin dropped off. Histological staining and immunohistochemical staining were carried out at 21 days after operation. Results All the rats survived to the end of the experiment. The allograft skins survived in all groups, dried and dropped off. The epithelium sheet could be seen in groups A and B at 21 days, a few very thin epithelium in group C, and no epithelization in group D. The wound healing rate of groups A (62.9%±9.6%) and B (64.2%±9.1%) were significantly higher than that of groups C (38.5%±5.7%) and D (22.7%±5.5%) (P<0.05), and significant difference was found between groups C and D (P<0.05), but there was no significant difference between groups A and B (P>0.05). The results of histological observation showed that squamous epithelial cells were observed in groups A, B, and C, but not in group D; obvious layers of epidermis were observed in groups A and B, thin epidermis and inflammatory cell infiltration in group C, and granulation tissue in group D. The immunohistochemistry staining showed that the expressions of collagen type IV and collagen type VII in groups A, B, and C; the percentage of collagen type IV and collagen type VII positive cells in groups A and B were significantly higher than that of group C (P<0.05), but there was no significant difference between groups A and B (P>0.05). The expressions of collagen type IV and collagen type VII in group D were negative. Conclusion The repair of full thickness skin wound with graft of autologous keratinocyte suspension can achieve reconstruction of the skin. The appropriate density of keratinocyte suspension for wound healing is 1×10⁵ cells/cm².

Citation： YINKai, SHENChuanan, MALi, SHANGYuru, LIDawei, LILongzhu, ZHAODongxu, CHENGWenfeng. AN EXPERIMENTAL STUDY ON REPAIR OF FULL THICKNESS SKIN WOUND WITH GRAFT OF AUTOLOGOUS KERATINOCYTE SUSPENSION IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(2): 219-223. doi: 10.7507/1002-1892.20160045 Copy

1.	Ali N, Hosseini M, Vainio S, et al. Skin equivalents:skin from reconstructions as models to study skin development and diseases. Br J Dermatol, 2015, 173(2):391-403.
2.	Park HH, Park NY, Kim SG, et al. Potential wound healing activities of galla rhois in human fibroblasts and keratinocytes. Am J Chin Med, 2015, 43(8):1625-1636.
3.	Frew Q, Philp B, Shelley O, et al. The use of Biobrane^® as a delivery method for cultured epithelial autograft in burn patients. Burns, 2013, 39(5):876-880.
4.	You HJ, Han SK, Lee JW, et al. Treatment of diabetic foot ulcers using cultured allogeneic keratinocytes-a pilot study. Wound Repair Regen, 2012, 20(4):491-499.
5.	Lantis JC, Marston WA, Farber A, et al. The influence of patient and wound variables on healing of venous leg ulcers in a randomized controlled trial of growth-arrested allogeneic keratinocytes and fibroblasts. J Vasc Surg, 2013, 58(2):433-439.
6.	尚玉茹, 申传安, 海恒林, 等. 大鼠全层皮肤缺损创面抗挛缩模型的建立. 中华损伤与修复杂志(电子版), 2012, 7(6):596-599.
7.	Brusselaers N, Pirayesh A, Hoeksema H, et al. Skin replacement in burn wounds. J Trauma, 2010, 68(2):490-501.
8.	Hackl F, Kiwanuka E, Philip J, et al. Moist dressing coverage supports proliferation and migration of transplanted skin micrografts in full-thickness porcine wounds. Burns, 2014, 40(2):274-280.
9.	Chen XL, Liang X, Sun L, et al. Microskin autografting in the treatment of burns over 70% of total body surface area:14 years of clinical experience. Burns, 2011, 37(6):973-980.
10.	Mahjour SB, Fu X, Yang X, et al. Rapid creation of skin substitutes from human skin cells and biomimetic nanofibers for acute full-thickness wound repair. Burns, 2015, 41(8):1764-1774.
11.	Siritientong T, Angspatt A, Ratanavaraporn J, et al. Clinical potential of a silk sericin-releasing bioactive wound dressing for the treatment of split-thickness skin graft donor sites. Pharm Res, 2014, 31(1):104-116.
12.	Kamel RA, Ong JF, Eriksson E, et al. Tissue engineering of skin. J Am Coll Surg, 2013, 217(3):533-555.
13.	Kircik LH, Dickerson JE Jr, Kitten C, et al. Allogeneic growth arrested keratinocytes and fibroblasts delivered in a fibrin spray accelerate healing in Mohs micrographic surgery wounds. J Drugs Dermatol, 2013, 12(5):558-561.
14.	Lönnqvist S, Emanuelsson P, Kratz G. Influence of acidic pH on keratinocyte function and re-epithelialisation of human in vitro wounds. J Plast Surg Hand Surg, 2015, 49(6):346-352.
15.	McHeik JN, Barrault C, Pedretti N, et al. Foreskin-isolated keratinocytes provide successful extemporaneous autologous paediatric skin grafts. J Tissue Eng Regen Med, 2013.[Epub ahead of print].
16.	Rinnerthaler M, Streubel MK, Bischof J, et al. Skin aging, gene expression and calcium. Exp Gerontol, 2015, 68:59-65.
17.	Lee H. Outcomes of sprayed cultured epithelial autografts for full-thickness wounds:a single-centre experience. Burns, 2012, 38(6):931-936.
18.	Sun Q, Li F, Li H, et al. Amniotic fluid stem cells provide considerable advantages in epidermal regeneration:B7H4 creates a moderate inflammation microenvironment to promote wound repair. Sci Rep, 2015, 5:11560.
19.	Hanifi N, Halim AS, Aleas CF, et al. Epidermal regeneration of cultured autograft, allograft, and xenograft keratinocytes transplanted on full-thickness wounds in rabbits. Exp Clin Transplant, 2015, 13(3):273-278.
20.	Chan TR, Stahl PJ, Li Y, et al. Collagen-gelatin mixtures as wound model, and substrates for VEGF-mimetic peptide binding and endothelial cell activation. Acta Biomater, 2015, 15:164-172.

1. Ali N, Hosseini M, Vainio S, et al. Skin equivalents:skin from reconstructions as models to study skin development and diseases. Br J Dermatol, 2015, 173(2):391-403.
2. Park HH, Park NY, Kim SG, et al. Potential wound healing activities of galla rhois in human fibroblasts and keratinocytes. Am J Chin Med, 2015, 43(8):1625-1636.
3. Frew Q, Philp B, Shelley O, et al. The use of Biobrane^® as a delivery method for cultured epithelial autograft in burn patients. Burns, 2013, 39(5):876-880.
4. You HJ, Han SK, Lee JW, et al. Treatment of diabetic foot ulcers using cultured allogeneic keratinocytes-a pilot study. Wound Repair Regen, 2012, 20(4):491-499.
5. Lantis JC, Marston WA, Farber A, et al. The influence of patient and wound variables on healing of venous leg ulcers in a randomized controlled trial of growth-arrested allogeneic keratinocytes and fibroblasts. J Vasc Surg, 2013, 58(2):433-439.
6. 尚玉茹, 申传安, 海恒林, 等. 大鼠全层皮肤缺损创面抗挛缩模型的建立. 中华损伤与修复杂志(电子版), 2012, 7(6):596-599.
7. Brusselaers N, Pirayesh A, Hoeksema H, et al. Skin replacement in burn wounds. J Trauma, 2010, 68(2):490-501.
8. Hackl F, Kiwanuka E, Philip J, et al. Moist dressing coverage supports proliferation and migration of transplanted skin micrografts in full-thickness porcine wounds. Burns, 2014, 40(2):274-280.
9. Chen XL, Liang X, Sun L, et al. Microskin autografting in the treatment of burns over 70% of total body surface area:14 years of clinical experience. Burns, 2011, 37(6):973-980.
10. Mahjour SB, Fu X, Yang X, et al. Rapid creation of skin substitutes from human skin cells and biomimetic nanofibers for acute full-thickness wound repair. Burns, 2015, 41(8):1764-1774.
11. Siritientong T, Angspatt A, Ratanavaraporn J, et al. Clinical potential of a silk sericin-releasing bioactive wound dressing for the treatment of split-thickness skin graft donor sites. Pharm Res, 2014, 31(1):104-116.
12. Kamel RA, Ong JF, Eriksson E, et al. Tissue engineering of skin. J Am Coll Surg, 2013, 217(3):533-555.
13. Kircik LH, Dickerson JE Jr, Kitten C, et al. Allogeneic growth arrested keratinocytes and fibroblasts delivered in a fibrin spray accelerate healing in Mohs micrographic surgery wounds. J Drugs Dermatol, 2013, 12(5):558-561.
14. Lönnqvist S, Emanuelsson P, Kratz G. Influence of acidic pH on keratinocyte function and re-epithelialisation of human in vitro wounds. J Plast Surg Hand Surg, 2015, 49(6):346-352.
15. McHeik JN, Barrault C, Pedretti N, et al. Foreskin-isolated keratinocytes provide successful extemporaneous autologous paediatric skin grafts. J Tissue Eng Regen Med, 2013.[Epub ahead of print].
16. Rinnerthaler M, Streubel MK, Bischof J, et al. Skin aging, gene expression and calcium. Exp Gerontol, 2015, 68:59-65.
17. Lee H. Outcomes of sprayed cultured epithelial autografts for full-thickness wounds:a single-centre experience. Burns, 2012, 38(6):931-936.
18. Sun Q, Li F, Li H, et al. Amniotic fluid stem cells provide considerable advantages in epidermal regeneration:B7H4 creates a moderate inflammation microenvironment to promote wound repair. Sci Rep, 2015, 5:11560.
19. Hanifi N, Halim AS, Aleas CF, et al. Epidermal regeneration of cultured autograft, allograft, and xenograft keratinocytes transplanted on full-thickness wounds in rabbits. Exp Clin Transplant, 2015, 13(3):273-278.
20. Chan TR, Stahl PJ, Li Y, et al. Collagen-gelatin mixtures as wound model, and substrates for VEGF-mimetic peptide binding and endothelial cell activation. Acta Biomater, 2015, 15:164-172.

Chinese Journal of Reparative and Reconstructive Surgery

AN EXPERIMENTAL STUDY ON REPAIR OF FULL THICKNESS SKIN WOUND WITH GRAFT OF AUTOLOGOUS KERATINOCYTE SUSPENSION IN RATS

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content