Olive pomace phenolic compounds and extracts can inhibit inflammatory- and oxidative-related diseases of human ocular surface epithelium

Producción CientíficaOxidative- and inflammatory-related ocular surface diseases have high prevalence and are an emerging issue in ophthalmology. Olive pomace (OP) is the olive oil’s industry main by-product, and is potentially environmentally hazardous. Nevertheless, it contains phenolic compounds with important bioactivities, like oleuropein (OL) and hydroxytyrosol (HT). The antioxidant and anti-inflammatory effects of four OP extracts (CONV, OPT(1–3)), pure OL and HT, and mixtures thereof were screened on human corneal (HCE) and conjunctival epithelial (IM-ConjEpi) cells. CONV was conventionally extracted, while OPT(1–3) were produced by pressurized liquid extraction. Thanks to their improved activity, CONV and OPT3 (HT-enriched) were selected for dose-dependent studies. Cells were stimulated with tumor necrosis factor-α or ultraviolet-B radiation, measuring interleukin (IL)-1β, IL-6, IL-8, and IL-17A as well as interferon γ-induced protein [IP]-10 secretion or intracellular ROS production, respectively. On HCE, both extracts and HT inhibited the secretion of most measured ILs, demonstrating a strong anti-inflammatory effect; while in IM-ConjEpi, all samples decreased IP-10 secretion. Moreover, HT, OL, and both extracts showed strong dose-dependent antioxidant activity in both cell lines. Compared with CONV, OPT3 was active at lower concentrations, demonstrating that intensified extraction techniques are selective towards targeted biomarkers. Hence, a high-value application as potential ocular surface therapy was proposed for the OP valorization.European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Initial Training Network (ITN) “IT-DED3” (H2020-MSCA-ITN-2017) grant agreement No. 76560

Pressurized liquid extraction optimization from supercritical defatted olive pomace: a green and selective phenolic extraction process

Producción CientíficaOlive pomace (OP) is the main by-product of the olive oil industry produced in large quantities. Its valorization as a source of phenolic bioactive compounds is paramount for the sustainable growth of related industries. This work proposes an intensified process to maximize the recovery of phenolic compounds in dry extracts using hydroalcoholic mixtures. Supercritical carbon dioxide defatting pre-treatment was performed. Following this, pressurized liquid extraction was optimized through a circumscribed central composite design. The factors consisted of temperature (65.0–185.0 °C), ethanol percentage (8.0–92.0%), and solid/liquid ratio (0.2–0.8 gOP/mLSOLVENT). Besides the total phenolic content (TPC) and the total flavonoid content (TFC), the major phenolic compounds of OP [hydroxytyrosol (HT), tyrosol (TY), and oleuropein (OL)] were evaluated. Further, decarboxymethyl OL aglycone dialdehyde (3,4-DHPEA-DEDA) was identified by HPLC-DAD-MS/MS as the most abundant polyphenol and was studied for the first time for OP. Different conditions were found to optimize each key compound. In 67% shorter extraction time and 38% less solvent consumption compared to conventional extraction, an increase of 475% for OL, 428% for HT, 194% for TY, 373% for 3,4-DHPEA-DEDA, 89% for TPC, and 158% for TFC was observed. The antioxidant activity by oxygen radical absorbance capacity (ORAC) assay increased 89% (optimal conditions) and correlated with TPC, 3,4-DHPEA-DEDA, and TFC. Thus, an efficient, selective, scalable, and green extraction process was established.European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Initial Training Network (ITN) “IT-DED3” (H2020-MSCA-ITN-2017) grant agreement No. 765608Fundação para a Ciência e Tecnologia (FCT) and Portugal 2020 to the Portuguese Mass Spectrometry Network (Rede Nacional de Espectrometria de Massa RNEM; LISBOA 01 0145 FEDER 402 022125

Olive pomace phenolic compounds: from an agro-industrial by-product to a promising ocular surface protection for dry eye disease

Producción CientíficaDry eye (DED) is a prevalent disease with immune-mediated inflammation as the principal pathophysiological etiology. Olive pomace, the major by-product of the olive oil industry, is rich in high-value polyphenols. Their anti-inflammatory and immunomodulatory activities were determined on human CD4+ T cells (hTCD4+) and in a DED animal model. The viability of hTCD4+ cells isolated from peripheral blood and activated with phytohemagglutinin-M was evaluated after treatment for 48 h with an olive pomace extract (OPT3, 0.10–0.40 mg/mL) and its major compound, hydroxytyrosol (25–100 μM). Regarding the DED animal model, 100 μM hydroxytyrosol, 0.20 mg/mL OPT3, or vehicle (borate buffer) were topically administered to 14 days-desiccating stress-exposed (constant airflow/scopolamine administration) C57BL/6 mice. Tear volume, corneal fluorescein staining (CFS), CD4+, and CD8+ T cell count in lymph nodes (flow cytometry), and IP-10 and TNF-α gene expression (qRT-PCR) in the cornea, conjunctiva, and lacrimal glands were evaluated. OPT3 (0.2–0.4 mg/mL) and hydroxytyrosol (100 μM) significantly reduced hTCD4+ proliferation. In mice, both treatments reduced lacrimal gland IP-10 gene expression. OPT3 also decreased CFS, and conjunctival IP-10 and corneal TNF-α gene expression. In lymph nodes, hydroxytyrosol reduced CD3+, OPT3, and CD8+ count. Thus, a high-value application as a promising DED protection was proposed for olive pomace.European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Initial Training Network (ITN) “IT-DED3” (H2020-MSCA-ITN-2017) grant agreement No. 765608Ministerio de Ciencia e Innovación y Fondo Europeo de Desarrollo Regional (FEDER) (RTI2018-094071-B-C21