RESUMO
The term postbiotics has acquired great interest in recent years. Numerous studies have shown a potential beneficial effect of its use in many inflammatory pathologies. However, it has not been much explored in ocular inflammatory diseases. The aims of this study were to develop and characterise an ophthalmic formulation with a postbiotic of Lactiplantibacillus plantarum CRL 759, and to evaluate its anti-inflammatory actions on murine macrophage stimulated with lipopolysaccharides (LPS) in vitro. First, we evaluated the ability of L. plantarum CRL 759 to generate a supernatant with anti-inflammatory property using different buffers. Then, we studied the stability at different temperatures and storage times of the generated postbiotic. In vitro assays showed that incubation of L. plantarum CRL 759 in modified phosphate buffer according to Sorensen (called POF-759), generated a supernatant that significantly reduced the production of interleukin-6, tumour necrosis factor-α, and nitric oxide by RAW 264.7 cells stimulated with LPS. Furthermore, POF-759 maintained its anti-inflammatory activity at room temperature, 4 and -20 °C, up to 30 days of storage. From the studies reported here, a postbiotic product with anti-inflammatory properties and optimal characteristics for the formulation of eye drops was obtained.
Assuntos
Lactobacillus plantarum , Probióticos , Camundongos , Animais , Lipopolissacarídeos , Macrófagos , Anti-Inflamatórios/farmacologiaRESUMO
Anti-inflammatory effect of soluble secreted compounds of probiotic bacteria was widely demonstrated as therapy for different inflammatory diseases, but was not investigated in inflammatory eye disorders. The aim of this study was to determine whether Lactiplantibacillus plantarum CRL759 cell-free supernatant reduced inflammatory parameters and clinical signs in ocular inflammations. First, we evaluated the effect of L. plantarum CRL759 supernatant in vitro on human retinal cell line, ARPE-19 cells, stimulated with lipopolysaccharide (LPS). Then, we investigated in vivo its capacity to decrease inflammation by local administration on the eyes of mice with endotoxin induced inflammation. In vitro assays demonstrated that L. plantarum CRL759 supernatant reduced the production of interleukin (IL)-6, IL-8, nitric oxide and thiobarbituric acid reactive substances in LPS-stimulated ARPE-19 cells. Our in vivo data proved that L. plantarum supernatant significantly reduced the clinical score of endotoxin treated mice and diminished levels of tumour necrosis factor alpha, interferon gamma and protein concentration in aqueous humour. Histological examination showed reduction of infiltrating inflammatory cells in the posterior segment of the eyes. As far as we know, this is the first report showing that Lactobacillus spp. supernatant administered as drops reduces some parameters of ocular inflammation. This promising strategy is safe and could alleviate symptoms and signs of ocular inflammation in people that are refractories to the conventional therapies.