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Comparison of therapeutic antibiotic treatments on tissue-engineered human skin substitutes
Angela L Gibson1, Michael J Schurr, Sandy J Schlosser
1Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
Abstract:
For regenerative medicine to gain clinical acceptance, the effects of commonly used treatment regimens on bioengineered organs must be considered. The antibiotics mafenide acetate (mafenide) and neomycin plus polymyxin (neo/poly) are routinely used to irrigate postoperative skin grafts on contaminated wounds. The effects of these clinically used antibiotics were investigated using tissue-engineered human skin substitutes generated with primary human keratinocytes or the near-diploid human keratinocyte cell line, Near-diploid Immortal Keratinocytes. Following topical or dermal treatment, the skin substitutes were assayed for viability, tissue morphology, glycogen content, and the expression of active caspase 3. Mafenide, but not neo/poly, induced morphological and biochemical changes in tissue-engineered skin substitutes. Keratinocytes in all histological layers of mafenide-treated skin substitutes exhibited ballooning degeneration and glycogen depletion. Mafenide-treatment also triggered separation of basal keratinocytes from the underlying dermis. None of the antibiotic treatments induced apoptosis, as measured by active caspase 3 immunostaining. The results demonstrate that mafenide, but not neo/poly, is detrimental to the viability and structural integrity of tissue-engineered human skin substitutes. These findings highlight the need to identify treatment regimens that are compatible with and hence enable the therapeutic efficacy of first-generation bioengineered organs such as skin.
Insights
Mafenide acetate (mafenide) harms tissue-engineered skin, causing structural damage. Neomycin plus polymyxin (neo/poly) did not affect skin substitutes, indicating mafenide is unsuitable for regenerative medicine applications.
Area of Science:
- Regenerative Medicine
- Tissue Engineering
- Dermatology
Background:
- Clinical acceptance of regenerative medicine requires understanding treatment effects on bioengineered organs.
- Antibiotics like mafenide acetate (mafenide) and neomycin plus polymyxin (neo/poly) are used post-surgery for skin grafts.
Purpose of the Study:
- To investigate the effects of mafenide and neo/poly on tissue-engineered human skin substitutes.
- To assess the impact of these antibiotics on skin substitute viability, morphology, and cellular integrity.
Main Methods:
- Tissue-engineered human skin substitutes were created using primary keratinocytes or a keratinocyte cell line.
- Substitutes were treated topically or dermally with mafenide or neo/poly.
- Assays included viability, morphology, glycogen content, and active caspase 3 expression.
Main Results:
- Mafenide induced significant morphological changes, including ballooning degeneration and glycogen depletion in all keratinocyte layers.
- Mafenide treatment caused basal keratinocytes to detach from the dermis.
- Neo/poly did not induce significant adverse effects on the skin substitutes.
- No antibiotic treatment triggered apoptosis as indicated by active caspase 3 staining.
Conclusions:
- Mafenide acetate is detrimental to the viability and structural integrity of tissue-engineered human skin substitutes.
- Neomycin plus polymyxin does not appear to harm these engineered skin models.
- Findings emphasize the need for antibiotic regimens compatible with bioengineered organs for therapeutic efficacy.
