Nanobubble Ozone Stored in Hyaluronic Acid Decorated Liposomes: Antibacterial, Anti-SARS-CoV-2 Effect and Biocompatibility Tests

Abstract

Purpose SARS-CoV-2-infected individuals may be asymptomatic, and therefore, the virus is highly contagious. We aimed to develop an agent to control viral replication in the upper respiratory tract and to prevent progression of the disease into the lower airways as well as inter-individual transmission. For this purpose, we investigated the antibacterial and antiviral activities of our novel nanobubble ozonated hyaluronic acid-decorated liposomal (NOHAL) solution, developed by using nanotechnology. Methods The MIC levels of NOHAL solution were determined on blood agar cultures of Staphylococcus aureus (ATCC 6538), Streptococcus pneumoniae (ATCC 49619) and Escherichia coli (ATCC 25922). The in vitro anti-viral activity of NOHAL solution was studied using recombinant SARS-CoV-2 copies of the original virus, grown in Vero cells generated by reverse genetic technology. Human primary lung epithelial cells obtained by bronchoscopy or lung resection were used for cell viability tests using flow cytometry analysis. The cytotoxicity testing was performed using the BALB/c 3T3 (CCL-163) cell line. Skin, oral, nasal and ocular irritation tests were performed using New Zealand albino rabbits, Syrian hamsters, BALB c mice and New Zealand albino rabbits of both sexes. Results Bacterial growth was prevented by NOHAL solution in a time-/dose-dependent manner. In vivo or in vitro experiments did not show any toxicity of NOHAL solution. No cytotoxicity was recorded on cell viability. No skin, oral, nasal or ocular toxicities were recorded. In addition, in a SARS-CoV-2 mouse infection model, NOHAL solution diminished the viral RNA levels effectively in nasopharyngeal and lung samples after its prophylactic intranasal application. Conclusion NOHAL solution has the potential to reduce or prevent the spread of SARS-CoV-2 through the nose and/or oral cavity. The clinical efficacy of this solution needs to be tested in order to determine its efficacy in the early phase of COVID-19.

Antibiofilm Activity of Different Washing Solutions against Biofilms in Cystic Fibrosis Patients' Nasal Cavities

“…Due to the water-retaining properties of hyaluronic acid, its healing effects, and ability to moisturize the mucosa, nanobubble ozone is stored in hyaluronic acid-decorated liposomes. 11 Our ozone solution (1,600 ppm) is produced as a nasal spray, and all biocompatibility and toxicity tests (skin, oral, nasal, eye) of the product have been performed. 11 Zerillo et al 26 reported that ozonated liposomal products inhibited bacterial viability and eradicated P. aeruginosa and S. aureus biofilms.…”

“…11 Our ozone solution (1,600 ppm) is produced as a nasal spray, and all biocompatibility and toxicity tests (skin, oral, nasal, eye) of the product have been performed. 11 Zerillo et al 26 reported that ozonated liposomal products inhibited bacterial viability and eradicated P. aeruginosa and S. aureus biofilms. Silva et al 10 described how ozonated oils had a moderate to high ability to eliminate adhered cells and a high capacity to eradicate 24 hours old biofilms produced by MRSA.…”

“…We used a commercial ozone solution (GEOXYN NANO-TECH GEOXYNORAL1600, nasal, oral spray), prepared at 1,600 ppm for the most potent antimicrobial effect. 11 Nasal-oral spray approved by the Turkish Ministry of Agriculture, containing ozone-coated nano molecules (Patent no: PCT/TR2022/050177), has been previously tested on the nasal mucosa.…”

“…Ozone-coated nanomolecules previously tested in the nasal mucosa were used in this study. 11 We aimed to evaluate the antibiofilm potential of streptomycin and ozone solutions against Staphylococcus aureus and Pseudomonas aeruginosa isolated from patients with CF.…”

Fabrication of hyaluronic acid‐based nanoparticles and studies on their radical scavenging activity

“…23 As a major component of the cellular matrix, HA is derived from glycosaminoglycan in animal connective tissues. 24 As a bio-material, HA exhibits many inherent functions, for example, water-retaining, 25 antibacterial, 26 scavenging free radical, 27 and bio-compatibility, 28 respectively. Due to these merits, HA is widely used as a food additive in fruit juice, 29 soya-bean milk, and jelly, respectively.…”