Cytotoxicity and Epidermal Barrier Function Evaluation of Common Antiseptics for Clinical Use in an Artificial Autologous Skin Model

Abstract

Bioengineered artificial skin substitutes (BASS) are the main treatment used in addition to autografts when skin injuries involve a large body surface area. Antiseptic/antibiotic treatment is necessary to prevent infections in the BASS implant area. This study aims to evaluate the effect of antiseptics and antibiotics on cell viability, structural integrity, and epidermal barrier function in BASS based on hyaluronic acid during a 28 day follow-up period. Keratinocytes (KTs) and dermal fibroblasts (DFs) were isolated from skin samples and used to establish BASS. The following antibiotic/antiseptic treatment was applied every 48 h: colistin (1%), chlorhexidine digluconate (1%), sodium chloride (0.02%), and polyhexanide (0.1%). Cell viability (LIVE/DEAD® assay), structural integrity (histological evaluation), and epidermal barrier function (trans-epidermal water loss, (TEWL), Tewameter®) were also evaluated. Cell viability percentage of BASS treated with chlorhexidine digluconate was significantly lower (p ≤ 0.001) than the other antiseptics at day 28. Compared to other treatments, chlorhexidine digluconate and polyhexanide significantly affected the epithelium. No significant differences were found regarding epidermal barrier. These results may be useful for treatment protocols after implantation of BASS in patients and evaluating them in clinical practice. BASS represent a suitable model to test in vitro the impact of different treatments of other skin wounds.

Cytotoxicity, Epidermal Barrier Function and Cytokine Evaluation after Antiseptic Treatment in Bioengineered Autologous Skin Substitute

“…These strategies, while they solve the problem of availability in conventional treatments, also have several disadvantages. In the case of cell suspensions and cocultures, they fail to achieve complete biological function, and they have risk of rejection if the cells used in the treatment are allogenic [ 8 ]. Rejection risk is also a problem associated with commercial allogenic skin substitutes.…”

“…Rejection risk is also a problem associated with commercial allogenic skin substitutes. Acellular matrixes lack of a cellular component limiting the skin regeneration and restricting their use to only temporary wound coverage [ 3 , 8 ].…”

“…Another skin substitute approach, which we will be focusing on, is the use of Bioengineered Autologous Skin Substitutes (BASS). BASS is an autologous graft consisting of two layers; a basal layer composed of fibroblasts immersed in a scaffold mimicking the extracellular matrix (ECM) to substitute the dermal stroma, and an epithelium of keratinocytes seeded on top [ 3 , 5 , 8 ]. The scaffold has the function of anchoring the cells to the graft.…”

“…The scaffold has the function of anchoring the cells to the graft. It has a role in cell survival and proliferation, participates in the control of homeostasis and provides an optimal environment for wound healing [ 3 , 8 ]. Hyaluronic acid (HA) is one of the most widely used ECM components in wound dressings, stimulating cellular proliferation, cellular migration, and angiogenesis [ 9 ].…”

“…Once a skin substitute is applied onto the patient’s wound, cutaneous treatment with antiseptics is crucial to prevent infection of the wound. However, there is little information about the impact of these treatments on the viability of the cells in BASS [ 8 , 10 ]. There are few studies analyzing the effect of antiseptic treatment in human keratinocytes (hKT) and dermal fibroblasts (hDF).…”