Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis?
Abstract Background Large extracellular vesicles (L-EV) with a diameter between 1 and 10 µm are released by various cell types. L-EV contain and transport active molecules which are crucially involved in cell to cell communication. We have shown that secretory products of human regulatory macrophage...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-01-01
|
| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12967-023-03900-6 |
| _version_ | 1811171571397558272 |
|---|---|
| author | Martin Albrecht Lars Hummitzsch Rene Rusch Katharina Heß Markus Steinfath Jochen Cremer Frank Lichte Fred Fändrich Rouven Berndt Karina Zitta |
| author_facet | Martin Albrecht Lars Hummitzsch Rene Rusch Katharina Heß Markus Steinfath Jochen Cremer Frank Lichte Fred Fändrich Rouven Berndt Karina Zitta |
| author_sort | Martin Albrecht |
| collection | DOAJ |
| description | Abstract Background Large extracellular vesicles (L-EV) with a diameter between 1 and 10 µm are released by various cell types. L-EV contain and transport active molecules which are crucially involved in cell to cell communication. We have shown that secretory products of human regulatory macrophages (Mreg) bear pro-angiogenic potential in-vitro and our recent findings show that Mreg cultures also contain numerous large vesicular structures similar to L-EV with so far unknown characteristics and function. Aim of this study To characterize the nature of Mreg-derived L-EV (L-EVMreg) and to gain insights into their role in wound healing and angiogenesis. Methods Mreg were differentiated using blood monocytes from healthy donors (N = 9) and L-EVMreg were isolated from culture supernatants by differential centrifugation. Characterization of L-EVMreg was performed by cell/vesicle analysis, brightfield/transmission electron microscopy (TEM), flow cytometry and proteome profiling arrays. The impact of L-EVMreg on wound healing and angiogenesis was evaluated by means of scratch and in-vitro tube formation assays. Results Mreg and L-EVMreg show an average diameter of 13.73 ± 1.33 µm (volume: 1.45 ± 0.44 pl) and 7.47 ± 0.75 µm (volume: 0.22 ± 0.06 pl) respectively. Flow cytometry analyses revealed similarities between Mreg and L-EVMreg regarding their surface marker composition. However, compared to Mreg fewer L-EVMreg were positive for CD31 (P < 0.01), CD206 (P < 0.05), CD103 (P < 0.01) and CD45 (P < 0.05). Proteome profiling suggested that L-EVMreg contain abundant amounts of pro-angiogenic proteins (i.e. interleukin-8, platelet factor 4 and serpin E1). From a functional point of view L-EVMreg positively influenced in-vitro wound healing (P < 0.05) and several pro-angiogenic parameters in tube formation assays (all segment associated parameters, P < 0.05; number of meshes, P < 0.05). Conclusion L-EVMreg with regenerative and pro-angiogenic potential can be reproducibly isolated from in-vitro cultured human regulatory macrophages. We propose that L-EVMreg could represent a putative therapeutic option for the treatment of chronic wounds and ischemia-associated diseases. |
| first_indexed | 2024-04-10T17:16:15Z |
| format | Article |
| id | doaj.art-7907c0c1082f45798af3193f2fbfb373 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-04-10T17:16:15Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-7907c0c1082f45798af3193f2fbfb3732023-02-05T12:22:33ZengBMCJournal of Translational Medicine1479-58762023-01-0121111010.1186/s12967-023-03900-6Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis?Martin Albrecht0Lars Hummitzsch1Rene Rusch2Katharina Heß3Markus Steinfath4Jochen Cremer5Frank Lichte6Fred Fändrich7Rouven Berndt8Karina Zitta9Department of Anesthesiology and Intensive Care Medicine, University Hospital of Schleswig-HolsteinDepartment of Anesthesiology and Intensive Care Medicine, University Hospital of Schleswig-HolsteinClinic of Cardiovascular Surgery, University Hospital of Schleswig-HolsteinDepartment of Pathology, University Hospital of Schleswig-HolsteinDepartment of Anesthesiology and Intensive Care Medicine, University Hospital of Schleswig-HolsteinClinic of Cardiovascular Surgery, University Hospital of Schleswig-HolsteinDepartment of Anatomy, University of KielClinic for Applied Cell Therapy, University Hospital of Schleswig-HolsteinClinic of Cardiovascular Surgery, University Hospital of Schleswig-HolsteinDepartment of Anesthesiology and Intensive Care Medicine, University Hospital of Schleswig-HolsteinAbstract Background Large extracellular vesicles (L-EV) with a diameter between 1 and 10 µm are released by various cell types. L-EV contain and transport active molecules which are crucially involved in cell to cell communication. We have shown that secretory products of human regulatory macrophages (Mreg) bear pro-angiogenic potential in-vitro and our recent findings show that Mreg cultures also contain numerous large vesicular structures similar to L-EV with so far unknown characteristics and function. Aim of this study To characterize the nature of Mreg-derived L-EV (L-EVMreg) and to gain insights into their role in wound healing and angiogenesis. Methods Mreg were differentiated using blood monocytes from healthy donors (N = 9) and L-EVMreg were isolated from culture supernatants by differential centrifugation. Characterization of L-EVMreg was performed by cell/vesicle analysis, brightfield/transmission electron microscopy (TEM), flow cytometry and proteome profiling arrays. The impact of L-EVMreg on wound healing and angiogenesis was evaluated by means of scratch and in-vitro tube formation assays. Results Mreg and L-EVMreg show an average diameter of 13.73 ± 1.33 µm (volume: 1.45 ± 0.44 pl) and 7.47 ± 0.75 µm (volume: 0.22 ± 0.06 pl) respectively. Flow cytometry analyses revealed similarities between Mreg and L-EVMreg regarding their surface marker composition. However, compared to Mreg fewer L-EVMreg were positive for CD31 (P < 0.01), CD206 (P < 0.05), CD103 (P < 0.01) and CD45 (P < 0.05). Proteome profiling suggested that L-EVMreg contain abundant amounts of pro-angiogenic proteins (i.e. interleukin-8, platelet factor 4 and serpin E1). From a functional point of view L-EVMreg positively influenced in-vitro wound healing (P < 0.05) and several pro-angiogenic parameters in tube formation assays (all segment associated parameters, P < 0.05; number of meshes, P < 0.05). Conclusion L-EVMreg with regenerative and pro-angiogenic potential can be reproducibly isolated from in-vitro cultured human regulatory macrophages. We propose that L-EVMreg could represent a putative therapeutic option for the treatment of chronic wounds and ischemia-associated diseases.https://doi.org/10.1186/s12967-023-03900-6Large extracellular vesiclesMacrophagesAngiogenesisWound healing |
| spellingShingle | Martin Albrecht Lars Hummitzsch Rene Rusch Katharina Heß Markus Steinfath Jochen Cremer Frank Lichte Fred Fändrich Rouven Berndt Karina Zitta Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis? Journal of Translational Medicine Large extracellular vesicles Macrophages Angiogenesis Wound healing |
| title | Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis? |
| title_full | Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis? |
| title_fullStr | Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis? |
| title_full_unstemmed | Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis? |
| title_short | Characterization of large extracellular vesicles (L-EV) derived from human regulatory macrophages (Mreg): novel mediators in wound healing and angiogenesis? |
| title_sort | characterization of large extracellular vesicles l ev derived from human regulatory macrophages mreg novel mediators in wound healing and angiogenesis |
| topic | Large extracellular vesicles Macrophages Angiogenesis Wound healing |
| url | https://doi.org/10.1186/s12967-023-03900-6 |
| work_keys_str_mv | AT martinalbrecht characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT larshummitzsch characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT renerusch characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT katharinaheß characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT markussteinfath characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT jochencremer characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT franklichte characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT fredfandrich characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT rouvenberndt characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis AT karinazitta characterizationoflargeextracellularvesicleslevderivedfromhumanregulatorymacrophagesmregnovelmediatorsinwoundhealingandangiogenesis |