Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold
Tissue-engineered composite skin is a promising therapy for the treatment of chronic and acute wounds, including burns. Providing the wound bed with a dermal scaffold populated by autologous dermal and epidermal cellular components can further entice host cell infiltration and vascularization to ach...
| Main Authors: | , , , , , |
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| Format: | Journal Article |
| Published: |
2015
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| Online Access: | http://hdl.handle.net/20.500.11937/56662 |
| _version_ | 1848759907524804608 |
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| author | Paul, M. Kaur, Pritinder Herson, M. Cheshire, P. Cleland, H. Akbarzadeh, S. |
| author_facet | Paul, M. Kaur, Pritinder Herson, M. Cheshire, P. Cleland, H. Akbarzadeh, S. |
| author_sort | Paul, M. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | Tissue-engineered composite skin is a promising therapy for the treatment of chronic and acute wounds, including burns. Providing the wound bed with a dermal scaffold populated by autologous dermal and epidermal cellular components can further entice host cell infiltration and vascularization to achieve permanent wound closure in a single stage. However, the high porosity and the lack of a supportive basement membrane in most commercially available dermal scaffolds hinders organized keratinocyte proliferation and stratification in vitro and may delay re-epithelization in vivo. The objective of this study was to develop a method to enable the in vitro production of a human skin equivalent (HSE) that included a porous scaffold and dermal and epidermal cells expanded ex vivo, with the potential to be used for definitive treatment of skin defects in a single procedure. A collagen-glycosaminoglycan dermal scaffold (Integra(®)) was populated with adult fibroblasts. A near-normal skin architecture was achieved by the addition of coagulated human plasma to the fibroblast-populated scaffold before seeding cultured keratinocytes. This resulted in reducing scaffold pore size and improving contact surfaces. Skin architecture and basement membrane formation was further improved by the addition of aprotinin (a serine protease inhibitor) to the culture media to inhibit premature clot digestion. Histological assessment of the novel HSE revealed expression of keratin 14 and keratin 10 similar to native skin, with a multilayered neoepidermis morphologically comparable to human skin. Furthermore, deposition of collagen IV and laminin-511 were detected by immunofluorescence, indicating the formation of a continuous basement membrane at the dermal-epidermal junction. The proposed method was efficient in producing an in vitro near native HSE using the chosen off-the-shelf porous scaffold (Integra). The same principles and promising outcomes should be applicable to other biodegradable porous scaffolds, combined with autologous cells, for use in wound treatment. |
| first_indexed | 2025-11-14T10:07:20Z |
| format | Journal Article |
| id | curtin-20.500.11937-56662 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T10:07:20Z |
| publishDate | 2015 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-566622018-01-09T02:59:14Z Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold Paul, M. Kaur, Pritinder Herson, M. Cheshire, P. Cleland, H. Akbarzadeh, S. Tissue-engineered composite skin is a promising therapy for the treatment of chronic and acute wounds, including burns. Providing the wound bed with a dermal scaffold populated by autologous dermal and epidermal cellular components can further entice host cell infiltration and vascularization to achieve permanent wound closure in a single stage. However, the high porosity and the lack of a supportive basement membrane in most commercially available dermal scaffolds hinders organized keratinocyte proliferation and stratification in vitro and may delay re-epithelization in vivo. The objective of this study was to develop a method to enable the in vitro production of a human skin equivalent (HSE) that included a porous scaffold and dermal and epidermal cells expanded ex vivo, with the potential to be used for definitive treatment of skin defects in a single procedure. A collagen-glycosaminoglycan dermal scaffold (Integra(®)) was populated with adult fibroblasts. A near-normal skin architecture was achieved by the addition of coagulated human plasma to the fibroblast-populated scaffold before seeding cultured keratinocytes. This resulted in reducing scaffold pore size and improving contact surfaces. Skin architecture and basement membrane formation was further improved by the addition of aprotinin (a serine protease inhibitor) to the culture media to inhibit premature clot digestion. Histological assessment of the novel HSE revealed expression of keratin 14 and keratin 10 similar to native skin, with a multilayered neoepidermis morphologically comparable to human skin. Furthermore, deposition of collagen IV and laminin-511 were detected by immunofluorescence, indicating the formation of a continuous basement membrane at the dermal-epidermal junction. The proposed method was efficient in producing an in vitro near native HSE using the chosen off-the-shelf porous scaffold (Integra). The same principles and promising outcomes should be applicable to other biodegradable porous scaffolds, combined with autologous cells, for use in wound treatment. 2015 Journal Article http://hdl.handle.net/20.500.11937/56662 10.1089/ten.TEC.2014.0667 restricted |
| spellingShingle | Paul, M. Kaur, Pritinder Herson, M. Cheshire, P. Cleland, H. Akbarzadeh, S. Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold |
| title | Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold |
| title_full | Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold |
| title_fullStr | Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold |
| title_full_unstemmed | Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold |
| title_short | Use of clotted human plasma and aprotinin in skin tissue engineering - A novel approach to engineering composite skin on a porous scaffold |
| title_sort | use of clotted human plasma and aprotinin in skin tissue engineering - a novel approach to engineering composite skin on a porous scaffold |
| url | http://hdl.handle.net/20.500.11937/56662 |