CD9-enriched extracellular vesicles from chemically reprogrammed basal progenitors of salivary glands mitigate salivary gland fibrosis.

Park, Sunyoung; Yoon, Yeo-Jun; Hong, Yongpyo; Yu, Jianning; Cho, Jae-Min; Jeong, Ye Jin; Yu, Haeun; Jeong, Hyorim et al. · Bioact Mater · 2025

basic_science · Level V

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Abstract

Extracellular vesicles (EVs) derived from stem cells offer promising potential for cell-free therapy. However, refining their cargo for precise disease targeting and delivery remains challenging. This study employed chemical reprogramming via dual inhibition of transforming growth factor beta (TGFβ) and bone morphogenetic protein (BMP) to expand salivary gland basal progenitor cells (sgBPCs). CD9-enriched (CD9<sup>+</sup>) EVs were then isolated from the sgBPC secretome concentrate using a dual microfluidic chip. Notably, CD9<sup>+</sup> EVs demonstrated superior uptake by salivary epithelial cells compared to CD9-depleted (CD9<sup>-</sup>) EVs and total EVs. <i>In vivo</i> studies using a salivary gland (SG) obstruction mouse model and <i>ex vivo</i> studies in SG fibrosis organoids revealed that CD9<sup>+</sup> EVs significantly enhanced anti-fibrotic effects over CD9<sup>-</sup> EVs and control treatments. The presence of miR-3162 and miR-1290 in CD9<sup>+</sup> EVs supported their anti-fibrotic properties by downregulating ACVR1 expression. The chemical reprogramming culture method effectively expanded sgBPCs, enabling consistent and scalable EV production. Utilizing microfluidic chip-isolated CD9<sup>+</sup> EVs and ductal delivery presents a targeted and efficient approach for anti-fibrotic SG regeneration.