MamC, an integral protein of the magnetosome membrane, has recently been proposed as a strong candidate to produce biomimetic (magnetosome-like) magnetite nanoparticles that could be used as an alternative to magnetosomes in different applications such as nanocarriers. The secondary structure of the protein contains two helical transmembrane domains connected by an α-helical loop oriented toward the magnetosome lumen. In this loop, the residues Glu66 and Asp70 seem to be responsible for a template effect that controls the nucleation and/or growth of biomimetic nanoparticles in vitro. In the present study, we have introduced a double mutation, E66A and D70A, in the sequence of MamC while working, for the first time, with the full-length protein. Our results show that this double mutation does not affect either the conformation or the stability of MamC, but it indeed makes the protein lose its functionality in terms of controlling the process of magnetite biomineralization in vitro. The present study shows that the ionotropic effect is not enough to account for the effect of the wild type MamC on the formation of BMNPs, but the template effect seems to rule such a process. Also, it shows that no other region of MamC is involved in controlling the process of magnetite biomineralization. Moreover, the stability of MamC in solution is only marginal, probably due to the absence of contacts established with the membrane lipid bilayer.
In recent years, new therapies, such as skin cell lines injections, have emerged to promote re-epithelialization of damaged areas such as chronic ulcers or to treat patients with severe burns. Antiseptics are commonly used during wound clinical management to avoid serious infections, but they may delay the healing process due to their apparent cytotoxicity to skin cells. The cytotoxicity of ethanol, chlorhexidine digluconate, sodium hypochlorite, povidone iodine and polyhexanide was evaluated in this in vitro study on human fibroblasts and keratinocytes. Treatments were applied to each cell type culture every 48 h for 14 days. To determine the cytotoxic of antiseptics, cell viability (Live/Dead®) and cell proliferation (AlamarBlue™) assays were performed on cell monolayers. Cell migration capacity was evaluated with a wound closure assay. Results showed how chlorhexidine digluconate and ethanol significantly reduced the viability of keratinocytes and inhibited cell migration. Povidone iodine followed by chlorhexidine digluconate significantly reduced fibroblast cell viability. Povidone iodine also inhibited cell migration. Sodium hypochlorite was the least detrimental to both cell types. If epithelial integrity is affected, the wound healing process may be altered, so the information gathered in this study may be useful in selecting the least aggressive antiseptic after treatment with new emerging therapies.
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