Facile and scalable fabrication of exosome-mimicking nanovesicles through PEGylated lipid detergent-aided cell extrusion

Abstract

We report a scalable fabrication method to generate exosome-mimicking nanovesicles (ENVs) using a biocompatible, cell-binding lipid detergent during cell extrusion. A PEGylated mannosylerythritol lipid (MELPEG) detergent was rationally engineered to...

Anti-Inflammatory Artificial Extracellular Vesicles with Notable Inhibition of Particulate Matter-Induced Skin Inflammation and Barrier Function Impairment

“…Through these modifications, we demonstrated an improved production of AEV P‑MEL , increasing particle production approximately 2.7 times while maintaining its vesicular morphology at the nanometer scale (Figures c,d and S4a–c). A previous study confirmed through quartz crystal microbalance with dissipation (QCM-D) analysis that the P-MEL strongly associated with phospholipid membranes, resulting in increased cell membrane deformability and the AEV production yield . The biocompatibility investigation of MEL, P-MEL, and AEV P‑MEL at various molar concentrations revealed that both P-MEL obtained by the PEGylation of MEL and AEV P‑MEL obtained using P-MEL exhibited improved cell viability (Figure e).…”

“…31,32 Taking advantage of a glycolipid produced by the yeast Pseudozyma 33 as an effective inhibitor for inflammatory molecules, mannosylerythritol lipids (MELs) were conjugated with PEG (P-MEL) via the thiol-maleimide reaction. 34,35 Thereafter, anti-inflammatory artificial extracellular vesicles, referred to as AEV P-MEL , were fabricated using the cell extrusion method, in which the cell membrane is decomposed and self-assembled as primary human keratinocytes pass through sequential membrane filters. 35,36 Through in vitro and in vivo studies, we aimed to prove the therapeutic potential of AEV P-MEL in PM-induced skin inflammation, as illustrated in Figure 1.…”

“…34,35 Thereafter, anti-inflammatory artificial extracellular vesicles, referred to as AEV P-MEL , were fabricated using the cell extrusion method, in which the cell membrane is decomposed and self-assembled as primary human keratinocytes pass through sequential membrane filters. 35,36 Through in vitro and in vivo studies, we aimed to prove the therapeutic potential of AEV P-MEL in PM-induced skin inflammation, as illustrated in Figure 1. Our data demonstrated that AEV P-MEL decreased PM-induced ROS production and expression levels of inflammatory proteins while increasing the levels of skin moisturizing proteins essential for maintaining skin moisture.…”

“…Prior to the fabrication of AEV P-MEL , P-MEL was prepared as a biosurfactant that would help achieve a high yield of AEV and improve therapeutic efficacy. 35 P-MEL was synthesized using the thiol-maleimide reaction between MEL-maleimide and PEG, which was confirmed by 1 H nuclear magnetic resonance (NMR) analyses (Figure S1a−c). Subsequently, cell extrusion was performed using P-MEL to fabricate AEV P-MEL (Figure 2a).…”

“…A previous study confirmed through quartz crystal microbalance with dissipation (QCM-D) analysis that the P-MEL strongly associated with phospholipid membranes, resulting in increased cell membrane deformability and the AEV production yield. 35 The biocompatibility investigation of MEL, P-MEL, and AEV P-MEL at various molar concentrations revealed that both P-MEL obtained by the PEGylation of MEL and AEV P-MEL obtained using P-MEL exhibited improved cell viability (Figure 2e). Moreover, they also exhibited high levels of cellular uptake capacity with intrinsic cell-targeting capabilities (Figures 2f−g and S6).…”