Application of amniotic membrane-living skin equivalent in repairing skin defect after removal of congenital giant nevus_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

YANG Lüjun ^1,2 , WU Hongjuan ³ , ZHANG Mingjun ¹ , XIE Sitian ¹ , YE Danyan ² ,  TANG Shijie ¹

1. Department of Burns and Plastic Surgery, the Second Affiliated Hospital, Shantou University Medical College, Shantou Guangdong, 510041, P.R.China;
2. Research Center for Translational Medicine, Shantou University Medical College, Shantou Guangdong, 510041, P.R.China;
3. Department of Burns and Plastic Surgery, Shaanxi Provincial People’s Hospital, Xi’an Shaanxi, 710068, P.R.China;

Corresponding author：

TANG Shijie, Email: tang2302@163.com

Keywords：

Amniotic membrane; living skin equivalent; congenital giant nevus; skin defect; wound repair

DOI：

10.7507/1002-1892.201707052

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the feasibility of human amniotic membrane-living skin equivalent (AM-LSE) in repairing the skin defect. Methods A 5-year-old boy with giant nevus at neck, shoulder, and back was admitted in July 2016. Normal skin tissue of the patient was harvested and keratinocytes and dermal fibroblasts were separated and expanded in vitro. Human AM was donated from a normal delivery and de-epithelialized for constructing an LSE as a matrix. Keratinocytes were seeded on the epithelial side of the AM which was previously seeded with fibroblasts on the stromal side and then the complex was lifted for air-liquid surface cultivation for 10 days and observed under naked eyes and sampled for histological study. The nevus was excised to deep fascia and the skin defect in size of 20 cm×15 cm was covered with artificial skin of collagen sponge for 2 weeks to enhance granulation tissue formation, and then the AM-LSE grafts of stamp size were grafted on. The dressing was changed until the wound healed. Results After 10 days of air-liquid surface cultivation, the AM-LSE developed a multilayered and differentiated epidermis with the fibroblasts-populated amnion as the dermal matrix. The LSE stamps survived and expanded to cover the whole wound. The grafted area showed normal skin color and soft contexture at 6 months after operation, and histological study showed well developed epidermis with compactly aligned basal cells, stratified and well differentiated squamous, granular layers and stratum corneum and well vascularized dermal compartment without inflammatory cells infiltration. Conclusion The cultivated AM-LSE with autologous cells can repair skin defect and survive for a long term without rejection.

Citation： YANG Lüjun, WU Hongjuan, ZHANG Mingjun, XIE Sitian, YE Danyan, TANG Shijie. Application of amniotic membrane-living skin equivalent in repairing skin defect after removal of congenital giant nevus. Chinese Journal of Reparative and Reconstructive Surgery, 2017, 31(12): 1495-1499. doi: 10.7507/1002-1892.201707052 Copy

1.	Zöller N, Valesky E, Butting M, et al. Clinical application of a tissue-cultured skin autograft: an alternative for the treatment of non-healing or slowly healing wounds? Dermatology, 2014, 229(3): 190-198.
2.	Killat J, Reimers K, Choi CY, et al. Cultivation of keratinocytes and fibroblasts in a three-dimensional bovine collagen-elastin matrix (Matriderm^®) and application for full thickness wound coverage in vivo. Int J Mol Sci, 2013,14(7): 14460-14474.
3.	刘钰, 沈冲, 孟琴. 体外构建 3D 人工皮肤. 中国组织工程研究, 2016, 20(46): 6915-6921.
4.	Rossi A, Appelt-Menzel A, Kurdyn S, et al. Generation of a three-dimensional full thickness skin equivalent and automated wounding. J Vis Exp, 2015, (96):1-7.
5.	Reuter C, Walles H, Groeber F. Preparation of a three-dimensional full thickness skin equivalent. Methods Mol Biol, 2017, 1612: 191-198.
6.	Yang L, Shirakata Y, Shudou M, et al. New skin-equivalent model from de-epithelialized amnion membrane. Cell Tissue Res, 2006, 326(1): 69-77.
7.	Kubo M, Sonoda Y, Muramatsu R, et al. Immunogenicity of human amniotic membrane in experimental xenotransplantation. Invest Ophthalmol Vis Sci, 2001, 42(7): 1539-1546.
8.	Park WC, Tseng SC. Modulation of acute inflammation and keratocyte death by suturing, blood, and amniotic membrane in PRK. Invest Ophthalmol Vis Sci, 2000, 41(10): 2906-2914.
9.	Herndon DN, Branski LK. Contemporary methods allowing for safe and convenient use of amniotic membrane as a biologic wound dressing for burns. Ann Plast Surg, 2017, 78(2 Suppl 1): S9-S10.
10.	Yang L, Shirakata Y, Tokumaru S, et al. Living skin equivalents constructed using human amnions as a matrix. J Dermatol Sci, 2009, 56(3): 188-195.
11.	Yang L, Shirakata Y, Tamai K, et al. Microbubble-enhanced ultrasound for gene transfer into living skin equivalents. J Dermatol Sci, 2005, 40(2): 105-114.
12.	Bell E, Sher S, Hull B, et al. The reconstitution of living skin. J Invest Dermatol, 1983, 81(1 Suppl): 2s-10s.
13.	Qi F, Yoshida T, Koike T, et al. Construction and characterization of human oral mucosa equivalent using hyper-dry amniotic membrane as a matrix. Arch Oral Bio, 2016, 65: 26-34.
14.	Vourc'h-Jourdain M, Martin L, Barbarot S. Large congenital melanocytic nevi: Therapeutic management and melanoma risk: a systematic review. J Am Acad Dermatol, 2013, 68(3): 493-498.e1-14.
15.	Kumbla PA, Yuen JC, Tait MA. Applying a dermal regenerative template in management of congenital melanocytic nevi of the hand. Plast Reconstr Surg Glob Open, 2015, 3(9): e515.
16.	余继超, 彭文要, 陈柏秋, 等. 结合应用皮耐克修复手部热压伤病例分析. 中华损伤与修复杂志(电子版), 2008, 3(4): 454-457.
17.	陈欣, 王浩, 戴允东, 等. 负压引流技术辅助人工真皮与自体皮移植修复关节开放和/或骨折处骨外露的临床研究. 中华烧伤杂志, 2015, 31(2): 93-97.
18.	Danielsen PL, Jorgensen LN, Jørgensen B, et al. Erythema persists longer than one year in split-thickness skin graft donor sites. Acta Derm Venereol, 2013, 93(3): 281-285.

1. Zöller N, Valesky E, Butting M, et al. Clinical application of a tissue-cultured skin autograft: an alternative for the treatment of non-healing or slowly healing wounds? Dermatology, 2014, 229(3): 190-198.
2. Killat J, Reimers K, Choi CY, et al. Cultivation of keratinocytes and fibroblasts in a three-dimensional bovine collagen-elastin matrix (Matriderm^®) and application for full thickness wound coverage in vivo. Int J Mol Sci, 2013,14(7): 14460-14474.
3. 刘钰, 沈冲, 孟琴. 体外构建 3D 人工皮肤. 中国组织工程研究, 2016, 20(46): 6915-6921.
4. Rossi A, Appelt-Menzel A, Kurdyn S, et al. Generation of a three-dimensional full thickness skin equivalent and automated wounding. J Vis Exp, 2015, (96):1-7.
5. Reuter C, Walles H, Groeber F. Preparation of a three-dimensional full thickness skin equivalent. Methods Mol Biol, 2017, 1612: 191-198.
6. Yang L, Shirakata Y, Shudou M, et al. New skin-equivalent model from de-epithelialized amnion membrane. Cell Tissue Res, 2006, 326(1): 69-77.
7. Kubo M, Sonoda Y, Muramatsu R, et al. Immunogenicity of human amniotic membrane in experimental xenotransplantation. Invest Ophthalmol Vis Sci, 2001, 42(7): 1539-1546.
8. Park WC, Tseng SC. Modulation of acute inflammation and keratocyte death by suturing, blood, and amniotic membrane in PRK. Invest Ophthalmol Vis Sci, 2000, 41(10): 2906-2914.
9. Herndon DN, Branski LK. Contemporary methods allowing for safe and convenient use of amniotic membrane as a biologic wound dressing for burns. Ann Plast Surg, 2017, 78(2 Suppl 1): S9-S10.
10. Yang L, Shirakata Y, Tokumaru S, et al. Living skin equivalents constructed using human amnions as a matrix. J Dermatol Sci, 2009, 56(3): 188-195.
11. Yang L, Shirakata Y, Tamai K, et al. Microbubble-enhanced ultrasound for gene transfer into living skin equivalents. J Dermatol Sci, 2005, 40(2): 105-114.
12. Bell E, Sher S, Hull B, et al. The reconstitution of living skin. J Invest Dermatol, 1983, 81(1 Suppl): 2s-10s.
13. Qi F, Yoshida T, Koike T, et al. Construction and characterization of human oral mucosa equivalent using hyper-dry amniotic membrane as a matrix. Arch Oral Bio, 2016, 65: 26-34.
14. Vourc'h-Jourdain M, Martin L, Barbarot S. Large congenital melanocytic nevi: Therapeutic management and melanoma risk: a systematic review. J Am Acad Dermatol, 2013, 68(3): 493-498.e1-14.
15. Kumbla PA, Yuen JC, Tait MA. Applying a dermal regenerative template in management of congenital melanocytic nevi of the hand. Plast Reconstr Surg Glob Open, 2015, 3(9): e515.
16. 余继超, 彭文要, 陈柏秋, 等. 结合应用皮耐克修复手部热压伤病例分析. 中华损伤与修复杂志(电子版), 2008, 3(4): 454-457.
17. 陈欣, 王浩, 戴允东, 等. 负压引流技术辅助人工真皮与自体皮移植修复关节开放和/或骨折处骨外露的临床研究. 中华烧伤杂志, 2015, 31(2): 93-97.
18. Danielsen PL, Jorgensen LN, Jørgensen B, et al. Erythema persists longer than one year in split-thickness skin graft donor sites. Acta Derm Venereol, 2013, 93(3): 281-285.

Chinese Journal of Reparative and Reconstructive Surgery

Application of amniotic membrane-living skin equivalent in repairing skin defect after removal of congenital giant nevus

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