Molecular study of wound healing after using biosynthesized BNC/Fe3O4 nanocomposites assisted with a bioinformatics approach

Mona Moniri,1,2 Amin Boroumand Moghaddam,1,2 Susan Azizi,1 Raha Abdul Rahim,3 Wan Zuhainis Saad4,5 Mohammad Navaderi6,7 Palanisamy Arulselvan8 Rosfarizan Mohamad1,4 1Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; 2Young Researcher and Elite Club, Sabzevar Branch, Islamic Azad University, Sabzevar, Iran; 3Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; 4Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; 5Institute of Tropical Forestry and Forest Products, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; 6Young Research and Elite Club, Parand Branch, Islamic Azad University, Parand, Iran; 7Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran; 8Muthayammal Centre for Advanced Research, Muthayammal College of Arts and Science, Rasipuram, Tamilnadu, India Background: Molecular investigation of wound healing has allowed better understanding about interaction of genes and pathways involved in healing progression. Objectives: The aim of this study was to prepare magnetic/bacterial nanocellulose (Fe3O4/BNC) nanocomposite films as ecofriendly wound dressing in order to evaluate their physical, cytotoxicity and antimicrobial properties. The molecular study was carried out to evaluate expression of genes involved in healing of wounds after treatment with BNC/Fe3O4 films. Study design, materials, and methods: Magnetic nanoparticles were biosynthesized by using Aloe vera extract in new isolated bacterial nanocellulose (BNC) RM1. The nanocomposites were characterized using X-ray diffraction, Fourier transform infrared, and field emission scanning electron microscopy. Moreover, swelling property and metal ions release profile of the nanocomposites were investigated. The ability of nanocomposites to promote wound healing of human dermal fibroblast cells in vitro was examined. Bioinformatics databases were used to identify genes with important healing effect. Key genes which interfered with healing were studied by quantitative real time PCR. Results: Spherical magnetic nanoparticles (15–30 nm) were formed and immobilized within the structure of BNC. The BNC/Fe3O4 was nontoxic (IC50>500 μg/mL) with excellent wound healing efficiency after 48 hours. The nanocomposites showed good antibacterial activity ranging from 6±0.2 to 13.40±0.10 mm against Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. The effective genes for the wound healing process were TGF-B1, MMP2, MMP9, Wnt4, CTNNB1, hsa-miR-29b, and hsa-miR-29c with time dependent manner. BNC/Fe3O4 has an effect on microRNA by reducing its expression and therefore causing an increase in the gene expression of other genes, which consequently resulted in wound healing. Conclusion: This eco-friendly nanocomposite with excellent healing properties can be used as an effective wound dressing for treatment of cutaneous wounds. Keywords: wound healing, antibacterial activity, bioinformatics study, scratch assay, bacterial nanocellulose, green metho