Chitosan, a natural polysaccharide derived from chitin, possesses biocompatibility, biodegradability, and mucoadhesive characteristics, making it an attractive material for the delivery of mRNA payloads to the nasal mucosa and promoting their uptake by target cells such as epithelial and immune cells (e.g., dendritic cells and macrophages). In this project, we aimed at developing novel lipid-based nanoformulations for mRNA delivery to counteract the pandemic caused by SARS-CoV-2 virus. The formulations achieved a mRNA encapsulation efficiency of ~80.2% with chitosan-lipid nanoparticles, as measured by the RiboGreen assay. Furthermore, the evaluation of SARS-CoV-2 Spike (S) receptor-binding domain (RBD) expression via ELISA for our vaccine formulations showed transfection levels in human embryonic kidney cells (HEK 293), lung carcinoma cells (A549), and dendritic cells (DC 2.4) equal to 9.9 ± 0.1 ng/mL (174.7 ± 1.1 fold change from untreated cells (UT)), 7.0 ± 0.2 ng/mL (128.1 ± 4.9 fold change from UT), and 0.9 ± 0.0 ng/mL (18.0 ± 0.1 fold change from UT), respectively. Our most promising vaccine formulation was also demonstrated to be amenable to lyophilization with minimal degradation of loaded mRNA, paving the way towards a more accessible and stable vaccine. Preliminary in vivo studies in mice were performed to assess the systemic and local immune responses. Nasal bronchoalveolar lavage fluid (BALF) wash showed that utilizing the optimized formulation resulted in local antibody concentrations and did not trigger any systemic antibody response. However, if further improved and developed, it could potentially contribute to the management of COVID-19 through nasopharyngeal immunization strategies.

Given the increasing aging population and the rising prevalence of musculoskeletal diseases due to obesity and injury, urgent research is needed to formulate new treatment alternatives, as current options remain inadequate. Viruses can exacerbate arthritis and worsen symptoms in patients with pre-existing osteoarthritis. Over the past decade, the chikungunya virus (CHIKV) has emerged as a significant public health concern, especially in Asia and South America. Exploring natural products, such as berberine, has shown promise due to its anticatabolic, antioxidative, and anti-inflammatory effects. However, berberine’s low stability and bioavailability limit its efficacy. We hypothesized that encapsulating berberine into a proniosome gel, known for its ease of preparation and stability, could enhance its bioavailability and efficacy when applied topically, potentially treating CHIKV infection. Our investigation focused on how varying berberine loads and selected excipients in the proniosome gel influenced its physical properties, stability, and skin permeability. We also examined the biological half-life of berberine in plasma upon topical administration in mice to assess the potential for controlled and sustained drug release. Additionally, we analyzed the antioxidant stress activity and cell viability of HaCaT keratinocytes and developed a lipopolysaccharide-stimulated cell culture model to evaluate anti-inflammatory effects using pro-inflammatory cytokines. Overall, the research aims to transform the treatment landscape for arthritis by leveraging berberine’s therapeutic potential.

Osteoarthritis is a common and chronic degenerative joint disease that leads to persistent symptoms such as pain, stiffness, swelling, and fatigue, all of which significantly impair mobility and reduce quality of life. Viral infections can further worsen joint inflammation in individuals with pre-existing osteoarthritis. Among these, the chikungunya virus (CHIKV) has become an increasing public health concern, particularly in Asia and South America, where post-viral arthritis frequently mimics or intensifies chronic osteoarthritic symptoms. Current treatments rely mainly on analgesics, corticosteroids, and biologics, which often provide only temporary relief and can cause systemic side effects. Poor bioavailability and limited drug penetration into joint tissues also remain major obstacles to effective long-term therapy. To address these challenges, this study focused on developing a proniosome-based topical gel capable of delivering therapeutic agents locally and in a sustained manner to inflamed joints. The formulation incorporated berberine hydrochloride, Coptis chinensis extract, and glucosamine, selected for their anti-inflammatory and cartilage-protective properties. Non-ionic surfactants and co-lipids were optimized to enhance skin permeation, improve vesicle stability, and support controlled release. Preclinical evaluations were conducted using both osteoarthritis and CHIKV-induced arthritis in vivo models. The optimized proniosome gel demonstrated strong therapeutic efficacy, including reductions in inflammatory markers, improved cartilage preservation and better subchondral bone structure. Importantly, no hepatic or renal toxicity was observed. These findings suggest that the proniosome-based gel offers a safe and effective topical approach for arthritis management, providing localized and sustained drug delivery while minimizing systemic exposure.