Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity
Abstract Background Small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) are recognized for their therapeutic potential in immune modulation and tissue repair, especially in veterinary medicine. This study introduces an innovative sequential stimulation (IVES) technique, in...
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2025-01-01
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| Online Access: | https://doi.org/10.1186/s12917-024-04465-2 |
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| author | Saranyou Oontawee Parkpoom Siriarchavatana Watchareewan Rodprasert Irma Padeta Yudith Violetta Pamulang Poorichaya Somparn Trairak Pisitkun Boondarika Nambooppha Nattawooti Sthitmatee Daneeya Na Nan Thanaphum Osathanon Hiroshi Egusa Chenphop Sawangmake |
| author_facet | Saranyou Oontawee Parkpoom Siriarchavatana Watchareewan Rodprasert Irma Padeta Yudith Violetta Pamulang Poorichaya Somparn Trairak Pisitkun Boondarika Nambooppha Nattawooti Sthitmatee Daneeya Na Nan Thanaphum Osathanon Hiroshi Egusa Chenphop Sawangmake |
| author_sort | Saranyou Oontawee |
| collection | DOAJ |
| description | Abstract Background Small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) are recognized for their therapeutic potential in immune modulation and tissue repair, especially in veterinary medicine. This study introduces an innovative sequential stimulation (IVES) technique, involving low-oxygen gas mixture preconditioning using in vitro fertilization gas (IVFG) and direct current electrical stimulation (ES20), to enhance the anti-inflammatory properties of sEVs from canine adipose-derived MSCs (cAD-MSCs). Initial steps involved isolation and comprehensive characterization of cAD-MSCs, including morphology, gene expression, and differentiation potentials, alongside validation of the electrical stimulation protocol. IVFG, ES20, and IVES were applied simultaneously with a control condition. Stimulated cAD-MSCs were evaluated for morphological changes, cell viability, and gene expressions. Conditioned media were collected and purified for sEV isolation on Day1, Day2, and Day3. To validate the efficacy of IVES for sEV production, various analyses were conducted, including microscopic examination, surface marker assessment, zeta-potential measurement, protein quantification, nanoparticle tracking analysis, and determination of anti-inflammatory activity. Results We found that IVES demonstrated non-cytotoxicity and induced crucial genotypic changes associated with sEV production in cAD-MSCs. Interestingly, IVFG influenced cellular adaptation, while ES20 induced hypoxia activation. By merging these stimulations, IVES enhanced sEV stability and quality profiles. The cAD-MSC-derived sEVs exhibited anti-inflammatory activity in lipopolysaccharide-induced RAW264.7 macrophages, emphasizing their improved effectiveness without cytotoxicity or immunogenicity. These effects were consistent across day 3 collection, indicating the establishment of an effective protocol for sEV production. Conclusions This research established an innovative sequential stimulation method with positive impact on sEV characteristics including stability, quality, and anti-inflammatory activity. This study not only contributes to the enhancement of sEV production but also sheds light on their functional aspects for therapeutic interventions. |
| format | Article |
| id | doaj-art-428dbc463adc4f21a02239637bde33fb |
| institution | Kabale University |
| issn | 1746-6148 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | BMC Veterinary Research |
| spelling | doaj-art-428dbc463adc4f21a02239637bde33fb2025-01-26T12:38:29ZengBMCBMC Veterinary Research1746-61482025-01-0121112110.1186/s12917-024-04465-2Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activitySaranyou Oontawee0Parkpoom Siriarchavatana1Watchareewan Rodprasert2Irma Padeta3Yudith Violetta Pamulang4Poorichaya Somparn5Trairak Pisitkun6Boondarika Nambooppha7Nattawooti Sthitmatee8Daneeya Na Nan9Thanaphum Osathanon10Hiroshi Egusa11Chenphop Sawangmake12Second Century Fund (C2F), Chulalongkorn University for Post-doctoral Fellowship, Chulalongkorn UniversitySecond Century Fund (C2F), Chulalongkorn University for Post-doctoral Fellowship, Chulalongkorn UniversityCenter of Excellence for Veterinary Clinical Stem Cells and Bioengineering, Chulalongkorn UniversityCenter of Excellence for Veterinary Clinical Stem Cells and Bioengineering, Chulalongkorn UniversityCenter of Excellence for Veterinary Clinical Stem Cells and Bioengineering, Chulalongkorn UniversityCenter of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn UniversityCenter of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Veterinary Biosciences and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai UniversityDepartment of Veterinary Biosciences and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai UniversityCenter of Excellence for Dental Stem Cell Biology, Faculty of Dentistry, Chulalongkorn UniversityDepartment of Anatomy, Faculty of Dentistry, Chulalongkorn UniversityCenter for Advanced Stem Cell and Regenerative Research, Division of Molecular and Regenerative Prosthodontics, Tohoku University Graduate School of DentistryCenter of Excellence for Veterinary Clinical Stem Cells and Bioengineering, Chulalongkorn UniversityAbstract Background Small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) are recognized for their therapeutic potential in immune modulation and tissue repair, especially in veterinary medicine. This study introduces an innovative sequential stimulation (IVES) technique, involving low-oxygen gas mixture preconditioning using in vitro fertilization gas (IVFG) and direct current electrical stimulation (ES20), to enhance the anti-inflammatory properties of sEVs from canine adipose-derived MSCs (cAD-MSCs). Initial steps involved isolation and comprehensive characterization of cAD-MSCs, including morphology, gene expression, and differentiation potentials, alongside validation of the electrical stimulation protocol. IVFG, ES20, and IVES were applied simultaneously with a control condition. Stimulated cAD-MSCs were evaluated for morphological changes, cell viability, and gene expressions. Conditioned media were collected and purified for sEV isolation on Day1, Day2, and Day3. To validate the efficacy of IVES for sEV production, various analyses were conducted, including microscopic examination, surface marker assessment, zeta-potential measurement, protein quantification, nanoparticle tracking analysis, and determination of anti-inflammatory activity. Results We found that IVES demonstrated non-cytotoxicity and induced crucial genotypic changes associated with sEV production in cAD-MSCs. Interestingly, IVFG influenced cellular adaptation, while ES20 induced hypoxia activation. By merging these stimulations, IVES enhanced sEV stability and quality profiles. The cAD-MSC-derived sEVs exhibited anti-inflammatory activity in lipopolysaccharide-induced RAW264.7 macrophages, emphasizing their improved effectiveness without cytotoxicity or immunogenicity. These effects were consistent across day 3 collection, indicating the establishment of an effective protocol for sEV production. Conclusions This research established an innovative sequential stimulation method with positive impact on sEV characteristics including stability, quality, and anti-inflammatory activity. This study not only contributes to the enhancement of sEV production but also sheds light on their functional aspects for therapeutic interventions.https://doi.org/10.1186/s12917-024-04465-2Small extracellular vesiclesCanineMesenchymal stem cellsSequential stimulationAnti-inflammatory activity |
| spellingShingle | Saranyou Oontawee Parkpoom Siriarchavatana Watchareewan Rodprasert Irma Padeta Yudith Violetta Pamulang Poorichaya Somparn Trairak Pisitkun Boondarika Nambooppha Nattawooti Sthitmatee Daneeya Na Nan Thanaphum Osathanon Hiroshi Egusa Chenphop Sawangmake Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity BMC Veterinary Research Small extracellular vesicles Canine Mesenchymal stem cells Sequential stimulation Anti-inflammatory activity |
| title | Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity |
| title_full | Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity |
| title_fullStr | Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity |
| title_full_unstemmed | Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity |
| title_short | Small extracellular vesicles derived from sequential stimulation of canine adipose-derived mesenchymal stem cells enhance anti-inflammatory activity |
| title_sort | small extracellular vesicles derived from sequential stimulation of canine adipose derived mesenchymal stem cells enhance anti inflammatory activity |
| topic | Small extracellular vesicles Canine Mesenchymal stem cells Sequential stimulation Anti-inflammatory activity |
| url | https://doi.org/10.1186/s12917-024-04465-2 |
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