Associated Proteins’ Proteomic Profiling of Equine Periodontal Ligament Stem Cell–Derived Exosomes for Diabetic Wound Healing Regeneration
Ruchianasari Deliaputri, Teguh Budipitojo, Aris Haryanto, Medania PurwaningrumBackground and Aim
Exosome‐based cell‐free therapies have shown significant potential to enhance diabetic ulcer healing. Exosomes derived from equine periodontal ligament stem cells (ePDLSCs) offer a viable alternative for regenerative wound‐healing treatments. This study aimed to characterize and analyze the protein composition of these exosomes and assess their efficacy in treating diabetic wounds in vivo.
Materials and Methods
This study employed RT‐qPCR and Nanoparticle Tracking Analysis (NTA) to isolate and characterize ePDLSCs–exosomes, western blot analysis of ePDLSCs–exosomes, proteomic protein profile analysis of ePDLSCs–exosomes using high‐resolution mass spectrometry (HRMS), western blot analysis for ACTB and VIM, potential protein profiling with proteomic analysis data associated with wound healing (Reactome, gene ontology [GO], KEGG pathways analysis), and assessment of ePDLSCs–exosome gel in vivo wound healing of diabetic ulcer wounds.
Results
The findings of this study reveal that exosomes derived from ePDLSCs, as shown by NTA, had a size distribution of approximately 114 ± 39 nm and expressed the exosome surface marker CD9. ePDLSCs–exo contained 35 proteins with potential involvement in wound healing, with the largest PSMs being annexin 2 (ANXA2), actin beta (ACTB), and vimentin (VIM), as shown in Western blot analysis. Functional enrichment analysis revealed the potential biological significance and signaling pathways of proteins supporting wound healing identified from ePDLSCs–exo. Furthermore, in vivo experiments showed ePDLSCs–exosomes impacted diabetic wound healing, with higher healing rates in the exosome‐treated group.
Conclusion
Exosomes derived from ePDLSCs have demonstrated regenerative and healing properties, particularly in the context of tissue damage associated with diabetes.