Abstract
Background & Aim
There is a growing interest in the use of extracellular vesicles (EVs) for regenerative dermatology applications. This work sought to evaluate the influence of tissue origin on their prospective applications in regenerative dermatology.
Methods, Results & Conclusion
Mesenchymal stem cells (MSCs) from primary adult bone marrow (BM) and perinatal umbilical cord (UB) were cultured for 5-days, switching to StemMACS™ xeno-free media (human) for 48 hours prior to conditioned media collection. EVs were then isolated utilising ultracentrifugation to comparatively profile for regenerative dermatology applications. These in-house isolated EVs were then further compared with a commercially available EV product marketed for topical regenerative dermatological applications. Outcomes indicated the presence of EVs in preparations isolated both in-house and commercially obtained. Cryo-TEM showed characteristic EVs in preparations isolated in-house, contrasting to minimally observed EV-matter in the commercial EV preparation. Particle concentrations measured by nano-flow cytometry (NanoFCM) were significantly different across preparations; UB-MSC EVs (3.51x1010 ±9.09x108 per ml), BM-MSC EVs (2.27x1010 ±9.98x108 per ml) and commercial preparation (9.17x109 ±6.61x108 per ml). This was also true for purity, based on particle to protein ratio, with UB-MSC EVs (5.24x107±2.42x106 particles per µg protein) displaying the highest purity and the commercial preparation the lowest (7.46x106±1.08x106 particles per µg protein). NanoFCM also showed markers of biogenesis (CD9, CD63 and CD81) to be differentially expressed significantly across EV preparations. Raman spectroscopy revealed similar spectral profiles within the main biological fingerprint region (400-1800 cm-1), with variations in peak intensities and observed peaks reported in studies evaluating skin (e.g.1655cm-1 amide 1, α helix structures often associated with keratin). Mass spectrometry analysis predicted proteins associated with biological pathways related to wound healing and skin development using PANTHER gene ontology analysis and STRING, suggested to be comparatively upregulated in BM-MSC EVs. Overall, preliminary outcomes suggested the potential of MSC-EVs in regenerative dermatology applications, with further evaluation needed to define the impact of observed variations on the functionality of each EV preparation.
There is a growing interest in the use of extracellular vesicles (EVs) for regenerative dermatology applications. This work sought to evaluate the influence of tissue origin on their prospective applications in regenerative dermatology.
Methods, Results & Conclusion
Mesenchymal stem cells (MSCs) from primary adult bone marrow (BM) and perinatal umbilical cord (UB) were cultured for 5-days, switching to StemMACS™ xeno-free media (human) for 48 hours prior to conditioned media collection. EVs were then isolated utilising ultracentrifugation to comparatively profile for regenerative dermatology applications. These in-house isolated EVs were then further compared with a commercially available EV product marketed for topical regenerative dermatological applications. Outcomes indicated the presence of EVs in preparations isolated both in-house and commercially obtained. Cryo-TEM showed characteristic EVs in preparations isolated in-house, contrasting to minimally observed EV-matter in the commercial EV preparation. Particle concentrations measured by nano-flow cytometry (NanoFCM) were significantly different across preparations; UB-MSC EVs (3.51x1010 ±9.09x108 per ml), BM-MSC EVs (2.27x1010 ±9.98x108 per ml) and commercial preparation (9.17x109 ±6.61x108 per ml). This was also true for purity, based on particle to protein ratio, with UB-MSC EVs (5.24x107±2.42x106 particles per µg protein) displaying the highest purity and the commercial preparation the lowest (7.46x106±1.08x106 particles per µg protein). NanoFCM also showed markers of biogenesis (CD9, CD63 and CD81) to be differentially expressed significantly across EV preparations. Raman spectroscopy revealed similar spectral profiles within the main biological fingerprint region (400-1800 cm-1), with variations in peak intensities and observed peaks reported in studies evaluating skin (e.g.1655cm-1 amide 1, α helix structures often associated with keratin). Mass spectrometry analysis predicted proteins associated with biological pathways related to wound healing and skin development using PANTHER gene ontology analysis and STRING, suggested to be comparatively upregulated in BM-MSC EVs. Overall, preliminary outcomes suggested the potential of MSC-EVs in regenerative dermatology applications, with further evaluation needed to define the impact of observed variations on the functionality of each EV preparation.
| Original language | English |
|---|---|
| Pages (from-to) | S89 |
| Journal | Cytotherapy |
| Volume | 26 |
| Issue number | 6, Supplement |
| DOIs | |
| Publication status | Published - 22 May 2024 |
| Event | ISCT Annual Meeting 2024 - Vancouver, Canada Duration: 29 May 2024 → 1 Jun 2024 https://www.isctglobal.org/isct2024 |