Development and characterization of nanostructured lipid carriers containing Brazilian red propolis in topical formulations

Author(s)

Bianca Bueno Fontanezi Dias Mello

• Document type: Master's Dissertation
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC)
• Defense date: 2022-02-22

Examining board members

Priscyla Daniely Marcato Gaspari; Andreia Patrícia Henriques Ascenso; Eliana Martins Lima

• Advisor: Priscyla Daniely Marcato Gaspari

Abstract

Brazilian red propolis (BRP) is known for its antioxidant, anti-inflammatory, antitumor and photoprotective effects and can be a great ally against solar radiation damage and natural aging of the skin. However, it has unfavorable physicochemical characteristics for topic applications. Nanotechnology can overcome these limitations. Therefore, our main objective was the biological evaluation of BRP and its fractions and the development and biological evaluation of a nanostructured lipid carrier (NLC) containing BRP aiming its topical application. The ethanolic extract of BRP and 3 fractions of BRP were evaluated: BRP and the hexane fraction proved to be phototoxic in fibroblast cultures and the dichloromethane fraction was considered the most cytotoxic. All fractions showed antioxidant activity, highlighting the hexane and BRP extract fractions. Ethyl acetate and dichloromethane fractions and BRP extract reduced the inflammatory marker nitric oxide to non-toxic concentrations. The evaluation of acute toxicity showed that the dichloromethane fraction was considered the most toxic in the Galleria mellonella model. The BRP extract was chosen to be encapsulated because, despite its phototoxicity, presented low in vivo toxicity and biological activity even at low concentrations. The NLC was optimized by the Box-Behnken factorial design. It had a diameter around 141.2 ± 2.7 nm, polydispersion index (PdI) of 0.131 ± 0.020, negative zeta potential (- 8.90 ± 0.25mV) and encapsulation of 5 bioactives greater than 96%, being stable for 420 days. The anti-inflammatory activity of NLC-BRP was superior to the BRP extract with the advantage of decreasing toxicity. Both BRP and NLCs with and without BRP did not show eye irritation and are compatible with the topical route, in addition to having high antioxidant activity. As the last stage of the project, the scaling of the NLC in a high-pressure homogenizer was proposed and the nanoparticles obtained presented low PdI and diameter close to the NLC developed in a sonicator. The NLC-BRP dispersion was incorporated into a quartenized guar gum gel that remained stable for 8 months. Thus, the developed NLC-BRP has the potential to be used in topical formulation, which has the benefit of reducing the cytotoxicity of BRP and enhancing its antioxidant and anti-inflammatory effect, in addition to being possible to produce it on a semi-industrial-scale. (AU)

• FAPESP's process: 18/16172-9 - Development and characterization of nanostructured lipid carriers using Brazilian red propolis in photoprotective formulations
• Grantee: Bianca Bueno Fontanezi Dias Mello
• Support Opportunities: Scholarships in Brazil - Master