Alginate is considered an exceptional biomaterial due to its hydrophilicity, biocompatibility, biodegradability, nontoxicity and low-cost in comparison with other biopolymers. We have recently demonstrated that the incorporation of 1% graphene oxide (GO) into alginate films crosslinked with Ca²⁺ cations provides antibacterial activity against Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis, and no cytotoxicity for human keratinocyte HaCaT cells. However, many other reports in literature have shown controversial results about the toxicity of GO demanding further investigation. Furthermore, the synergic effect of GO with other divalent cations with intrinsic antibacterial and cytotoxic activity such as Zn²⁺ has not been explored yet. Thus, here, two commercially available sodium alginates were characterised and utilized in the synthesis of zinc alginate films with GO following the same chemical route reported for the calcium alginate/GO composites. The results of this study showed that zinc release, water sorption/diffusion and wettability depended significantly on the type of alginate utilized. Furthermore, Zn²⁺ and GO produced alginate films with increased water diffusion, wettability and opacity. However, neither the combination of GO with Zn²⁺ nor the use of different types of sodium alginates modified the antibacterial activity and cytotoxicity of the zinc alginates against these Gram-positive pathogens and human cells respectively.

Due to the current global health problem of antibiotic resistant recently announced by the World Health Organization, there is an urgent necessity of looking for new alternative antibacterial materials able to treat and impede multidrug-resistant infections which are cost-effective and non-toxic for human beings. In this regard, carbon nanofibers (CNFs) possess currently much lower cost than other carbon nanomaterials, such as graphene oxide, and exhibit excellent chemical, mechanical and electric properties. Furthermore, here, the first report on the antibacterial activity of CNFs was demonstrated. Thus, these nanomaterials, in pure form or incorporated in a minuscule amount into calcium alginate composite films to reduce production costs as much as possible, showed to be new weapons against a globally spreading multidrug-resistant pathogen, the methicillin-resistant Staphylococcus epidermidis (MRSE). This Gram-positive bacterium is becoming one of the most dangerous pathogens, due to its abundance on skin. In this study, these hollow filamentous materials, in direct contact with cells and loaded in the low-cost calcium alginate composite films, showed no cytotoxicity for human keratinocyte HaCaT cells, which render them very promising for biomedical applications. The CNFs used in this work were characterized by Raman spectroscopy and observed by high-resolution transmission electron with energy-disperse X-ray spectroscopy.

Alginate-based materials hold great promise in bioengineering applications such as skin wound healing and scaffolds for tissue engineering. Nevertheless, cell adhesion of mammalian cells on these hydrophilic materials is very poor. In cases such as polycaprolactone, poly(hydroxy-3-butyrate-co-3-valerate) and gelatin, the incorporation of hydrophobic carbon nanofibers (CNFs) and hydrophilic graphene oxide (GO) has shown significant improvement of cell adhesion and proliferation. The incorporation of these carbon nanomaterials (CNMs) into alginate films can enhance their mechanical performance, wettability, water diffusion and antibacterial properties. Herein, we report the effect of adding these CNMs into alginate films on cell adhesion for the first time. Thus, the results of this study showed that these nanocomposites are non-cytotoxic in human keratinocyte HaCaT cells. Nevertheless, contrary to what has been reported for other polymers, cell adhesion on these advanced alginate-based composites was not improved. Therefore, both types of composite films possess similar biological behavior, in terms of cell adhesion and non-cytotoxicity, and enhanced physical and antibacterial properties in comparison to neat alginate for potential biomedical and bioengineering applications.

The use of ionic metals such as zinc (Zn²⁺) is providing promising results in regenerative medicine. In this study, human keratinocytes (HaCaT cells) were treated with different concentrations of zinc chloride (ZnCl₂), ranging from 1 to 800 µg/mL, for 3, 12 and 24 h. The results showed a time–concentration dependence with three non-cytotoxic concentrations (10, 5 and 1 µg/mL) and a median effective concentration value of 13.5 µg/mL at a cell exposure to ZnCl₂ of 24 h. However, the zinc treatment with 5 or 1 µg/mL had no effect on cell proliferation in HaCaT cells in relation to the control sample at 72 h. The effects of the Zn²⁺ treatment on the expression of several genes related to glycoprotein synthesis, oxidative stress, proliferation and differentiation were assessed at the two lowest non-cytotoxic concentrations after 24 h of treatment. Out of 13 analyzed genes (superoxide dismutase 1 (SOD1), catalase (CAT), matrix metallopeptidase 1 (MMP1), transforming growth factor beta 1 (TGFB1), glutathione peroxidase 1 (GPX1), fibronectin 1 (FN1), hyaluronan synthase 2 (HAS2), laminin subunit beta 1 (LAMB1), lumican (LUM), cadherin 1 (CDH1), collagen type IV alpha (COL4A1), fibrillin (FBN) and versican (VCAN)), Zn²⁺ was able to upregulate SOD1, CAT, TGFB1, GPX1, LUM, CDH1, FBN and VCAN, with relative expression levels of at least 1.9-fold with respect to controls. We found that ZnCl₂ promoted glycoprotein synthesis and antioxidant gene expression, thus confirming its great potential in biomedicine.