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Volume 14
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10.3390/biomedicines14010099
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Article

Characterization and Anti-Inflammatory Effects on Periodontal Ligament Cells of Citrus limon-Derived Exosome-like Nanovesicles Under Different Storage Temperatures

by
Yiming Ma
,
Chenhao Yu
,
Guojing Liu
,
Jia Liu
* and
Qingxian Luan
*
Department of Periodontology, Peking University School and Hospital of Stomatology, National Center of Stomatology and National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
*
Authors to whom correspondence should be addressed.
Biomedicines 2026, 14(1), 99; https://doi.org/10.3390/biomedicines14010099
Submission received: 28 November 2025 / Revised: 25 December 2025 / Accepted: 31 December 2025 / Published: 3 January 2026
(This article belongs to the Special Issue Periodontal Disease and Periodontal Tissue Regeneration—Second Edition)
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Abstract

Objectives: The purpose of this study is to compare the differences between lemon-derived exosome-like nanovesicles (LELNs) stored at −80 °C, −20 °C, and 4 °C for one month and freshly isolated LELNs, in terms of characterization and anti-inflammatory effects on periodontal ligament cells, aiming to identify suitable storage conditions for LELNs. Methods: Nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and micro bicinchoninic acid assay (BCA) were conducted to access the characterization differences. LPS-induced human periodontal ligament cells were used as an in vitro inflammatory model, and the changes in biological functions were examined by qRT-PCR and ELISA. Results: LELNs stored at −80 °C retained the highest particle and protein concentration and showed the least aggregation and heterogeneity in size on TEM images, while the average particle sizes shown by NTA were similar. And LELNs exhibited similar anti-inflammatory effects on periodontal ligament cells after one month of storage at −80 °C, −20 °C, and 4 °C. Conclusions: We found that LELNs can maintain in vitro anti-inflammatory ability when stored at either −80 °C, −20 °C, or 4 °C for one month, while storing at −80 °C maintains the concentration and uniform particle size best.
Keywords: Citrus limon L.; plant-derived exosome-like nanoparticles; storage temperature; stability; periodontal ligament cells; inflammatory cytokines Citrus limon L.; plant-derived exosome-like nanoparticles; storage temperature; stability; periodontal ligament cells; inflammatory cytokines

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MDPI and ACS Style

Ma, Y.; Yu, C.; Liu, G.; Liu, J.; Luan, Q. Characterization and Anti-Inflammatory Effects on Periodontal Ligament Cells of Citrus limon-Derived Exosome-like Nanovesicles Under Different Storage Temperatures. Biomedicines 2026, 14, 99. https://doi.org/10.3390/biomedicines14010099

AMA Style

Ma Y, Yu C, Liu G, Liu J, Luan Q. Characterization and Anti-Inflammatory Effects on Periodontal Ligament Cells of Citrus limon-Derived Exosome-like Nanovesicles Under Different Storage Temperatures. Biomedicines. 2026; 14(1):99. https://doi.org/10.3390/biomedicines14010099

Chicago/Turabian Style

Ma, Yiming, Chenhao Yu, Guojing Liu, Jia Liu, and Qingxian Luan. 2026. "Characterization and Anti-Inflammatory Effects on Periodontal Ligament Cells of Citrus limon-Derived Exosome-like Nanovesicles Under Different Storage Temperatures" Biomedicines 14, no. 1: 99. https://doi.org/10.3390/biomedicines14010099

APA Style

Ma, Y., Yu, C., Liu, G., Liu, J., & Luan, Q. (2026). Characterization and Anti-Inflammatory Effects on Periodontal Ligament Cells of Citrus limon-Derived Exosome-like Nanovesicles Under Different Storage Temperatures. Biomedicines, 14(1), 99. https://doi.org/10.3390/biomedicines14010099

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