# Anti-inflammatory potential of plant-derived extracellular vesicles from Solanum nigrum L. integrated in gelatine-dopamine hydrogel on RAW 264.7 and MC3T3 cells

## TL;DR

This study explores using plant-derived vesicles from Solanum nigrum in a hydrogel to reduce inflammation in bone therapy, showing promising results in cell tests.

## Contribution

First study to characterize and incorporate lyophilized PDEVs from Solanum nigrum into Gel-Dop hydrogel for anti-inflammatory bone therapy.

## Key findings

- Lyophilized PDEVs from Solanum nigrum berries were efficiently internalized by MC3T3 and RAW 264.7 cells without toxicity.
- PDEVs incorporated into Gel-Dop hydrogel effectively released and reduced IL-6 expression in LPS-stimulated cells.
- The hydrogel system showed potential for targeted delivery and storage stability of PDEVs.

## Abstract

Plant-derived extracellular vesicles (PDEV) from Solanum nigrum L. fruit show promise as a cell-free regenerative and inflammatory therapy for bone defects due to their anti-inflammatory properties. However, challenges such as storage stability and targeted delivery efficiency remain in PDEV's applications. Strategies such as lyophilization and injectable hydrogel delivery systems offer potential solutions.

In this study, lyophilized PDEVs derived from Solanum nigrum L. berries were incorporated into a thermosensitive injectable gelatine-dopamine (Gel-Dop) hydrogel and evaluated by in vitro for their anti-inflammatory potential using MC3T3 pre-osteoblast cells and RAW 264.7 macrophage cells.

The isolated PDEVs show a spherical morphology, an average size of approximately 132.6 nm, a polydispersity index of 0.197, and a protein concentration of 509 μg mL-1. These PDEVs were efficiently internalized by MC3T3 and RAW 264.7 cells after 12 hours of incubation and showed no cytotoxic effects at concentrations up to 10 μg mL-1. The release profile confirmed that the hydrogel effectively released the PDEVs, which remained non-toxic and were internalized by cells after 12 hours of incubation. Subsequently, treatment of lipopolysaccharide (LPS) stimulated MC3T3 and RAW 264.7 cells with PDEVs led to a reduction in IL-6 protein expression.

These findings suggest that lyophilized PDEVs from Solanum nigrum L. berries, when incorporated into Gel-Dop hydrogel, hold promise for future development as an anti-inflammatory agent in bone therapy. This study is the first to characterize and incorporate lyophilized PDEVs from Solanum nigrum L. into thermosensitive injectable Gel-Dop hydrogel and demonstrate their anti-inflammatory potential through the suppression IL-6 expression in LPS-stimulated MC3T3 and RAW 264.7 cells.