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A

B C D

E F

Figure 6. Evaluating the impact of LGG-EVs on OA-like chondrocytes in cartilage-on-chip. (A) Timeline indicating the treatment of chondrocyte
cultures with LGG-EVs on day 7, followed by observation until day 10. The figure was created using BioRender (BioRender.com) (B) DLS displays
the size distribution histogram of LGG-EVs, with the peak indicating the most common vesicle size. (C) TEM image displaying the ultrastructural
morphology of a single LGG-EV. (D) Histological and immunofluorescence staining of chondrocyte cultures: comparison between IL-1β treatment and
treatment with LGG-EVs. Alcian blue and SO/FG staining reveal proteoglycan and overall cartilage matrix, respectively, whereas immunofluorescence
assays demonstrate the presence of Col II and MMP13. (E) Western blot analysis displaying the protein expression of Col II and MMP13 in chondrocyte
cultures, indicating the cellular response to IL-1β and LGG-EVs treatments. (F) Quantification of protein expression levels of Col II, SOX9, MMP13,
and Adamts5, highlighting the effect of LGG-EVs on these molecular markers associated with cartilage anabolic and catabolic processes in the context of
IL-1β-induced inflammation (n=3, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). The data suggests that LGG-EVs may modulate the expression of key
proteins involved in cartilage homeostasis. Scale bars: 50 nm (C); 200 μm (E).
Abbreviations: GelMA: Gelatin methacryloyl; LGG-EVs: Lactobacillus rhamnosus GG-derived extracellular vesicles; OA: Osteoarthritis.

IL-1β to simulate the inflammatory state characteristic model. Unlike previous models that focus on mechanical
of OA. Notably, while IL-1β is the primary inflammatory loading or bone-cartilage interactions, our model directly
factor used in this study, other factors can be substituted simulates inflammation-induced cartilage degradation,
or added in future experiments depending on the specific providing a foundation for the development of targeted

Volume 1 Issue 1 (2025) 15 doi: 10.36922/or.8461