Abstract
The present study aimed to synthesize solid lipid nanoparticles to enhance liposome-assisted intracellular uptake of basil seed active components in adipocytes and vascular smooth muscle cells to attain increased bioavailability. To obtain solid lipid nanoparticle (SLNp), the water phase containing basil seed extract (BSE) was encapsulated with lipid matrix containing chia seed phospholipids using homogenization and cold ultra-sonication method. The physicochemical characterization of BSE loaded solid lipid nanoparticles (BSE-SLNp) has been analyzed using Zetasizer, FT-IR, and TEM. The BSE-SLNp showed an average diameter of 20–110 nm on the day of preparation and it remains the same after 60 days of storage. The cytotoxicity assay confirmed that the BSE-SLNp did not produce toxicity in hMSCs, preadipocytes, or human umbilical vein endothelial cells (HUVECs) until the tested higher dose up to 64 μg/ml. During effective dose determination, 4 μg/ml of BSE-SLNp confirmed non-toxic and enhanced metabolic function in hMSCs, preadipocytes, and HUVECs. Biosafety assay confirmed normal nuclear morphology in PI staining and high mitochondrial membrane potential in JC-1 assay within 48 h in hMSCs. The maturing adipocyte treated with 4 μg/ml of BSE-SLNp significantly increased the mitochondrial efficiency and fatty acid beta-oxidation (PPARγC1α, UCP-1, and PRDM-16) related gene expression levels. Oxidative stress induced HUVECs treated with 4 μg/ml of BSE-SLNp potentially enhanced antioxidant capacity, cell growth, and microtubule development within 48 h H2O2 induced oxidative stressed HUVECs have shown 39.8% viable cells, but treatment with BSE-SLNp has shown 99% of viable cells within 48 h confirmed by Annexin-V assay. In addition, mitochondrial membrane potential (Δψm) increased to 89.4% confirmed by JC-1 assay. The observed DNA integrity, cell viability was confirmed by increased antioxidant and tumor suppressor-related gene expression levels. VEGF expression has been significantly increased and pro-inflammation-related mRNA levels were decreased in BSE-SLNp treated cells. In conclusion, enhanced adipocyte fatty acid oxidation is directly associated with decreased adipocytokine secretion which arrests obesity-associated comorbidities. In addition, suppressing vascular cell oxidative stress and metabolic inflammation supports vascular cell proliferation and arrests ageing-related vascular diseases.
Highlights
Pathogenesis of age-related chronic complications is majorly dependent on the functional and structural modifications of the vascular system on metabolic stress
We studied its toxicity in human mesenchymal stem cells, lipolytic effect in maturing adipocytes, angiogenesis, and protective effect in oxidative stressed human umbilical vein endothelial cells (HUVECs)
FBS and penicillin-streptomycin were obtained from Hyclone Laboratories, U.S MTT [3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], propidium iodide, JC-1 stain, Oil Red O, Nile red, and all other chemicals related to molecular biology experiments were purchased from Sigma-Aldrich
Summary
Pathogenesis of age-related chronic complications is majorly dependent on the functional and structural modifications of the vascular system on metabolic stress. Altered endothelial cells develop proinflammatory cytokines such as eNOS, ICAM, and VCAM were associated with the development of age-associated chronic inflammatory diseases such as atherosclerosis, osteoarthritis, metabolic diseases, and gastrointestinal tract complications (Al-Khazraji et al, 2018). Active stem cells of subcutaneous adipose tissue derived from obese patients were found with a reduced proliferation ability, viability, change in telomerase activity, and reduced differentiation potential or loss of proangiogenic capacity (Perez et al, 2015). Chronic obesity impairs gene expression patterns related to cellular proliferation, differentiation, angiogenic potential, and multipotent capacity (Onate et al, 2013). Pharmacological agents or therapeutically functionalized nanoparticles target adipocyte mitochondrial efficiency and vascular cell proliferation attracts the aged obese population majorly
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