Effects of periodontal cells‐derived extracellular vesicles on mesenchymal stromal cell function

• Published in: Journal of periodontal research Vol. 58; no. 6; pp. 1188 - 1200
• Main Authors: Han, Pingping, Johnson, Nigel, Abdal‐hay, Abdalla, Moran, Corey S., Salomon, Carlos, Ivanovski, Sašo
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc 01-12-2023
• Subjects: Alveolar bone; Apoptosis; Bone growth; CD9 antigen; Cell culture; Cell migration; Cell proliferation; Extracellular vesicles; Fibroblasts; Gene expression; Mesenchymal stem cells; Osteogenesis; Periodontal ligament; Regeneration; Regenerative medicine; Stromal cells; Tissue engineering
• ISSN: 0022-3484; 1600-0765
• DOI: 10.1111/jre.13171

Abstract
Objective
To enrich and compare three extracellular vesicles—EV subtypes (apoptotic bodies, microvesicles and small EV) from three periodontal cells (periodontal ligament cells—PDLCs, alveolar bone‐derived osteoblasts—OBs and gingival fibroblasts—GFs), and assess uptake and cell function changes in buccal fat pad‐derived mesenchymal stromal cells (BFP‐MSCs).
Background
Periodontal cells such as PDLCs, OBs and GFs have the potential to enhance bone and periodontal regeneration, but face significant challenges, such as the regulatory and cost implications of in vitro cell culture and storage. To address these challenges, it is important to explore alternative ‘cell‐free’ strategies, such as extracellular vesicles which have emerged as promising tools in regenerative medicine, to facilitate osteogenic differentiation and bone regeneration.
Methods and Materials
Serial centrifuges at 2600 and 16 000 
g
were used to isolate apoptotic bodies and microvesicles respectively. Small EV—sEV was enriched by our in‐house size exclusion chromatography (SEC). The cellular uptake, proliferation, migration and osteogenic/adipogenic differentiation genes were analysed after EVs uptake in BFP‐MSCs.
Results
Three EV subtypes were enriched and characterised by morphology, particle size and EV‐associated protein expression—CD9. Cellular uptake of the three EVs subtypes was observed in BFP‐MSCs for up to 7 days. sEV from the three periodontal cells promoted proliferation, migration and osteogenic gene expression. hOBs‐sEV showed superior levels of osteogenesis markers compared to that hPDLCs‐sEV and hGFs‐sEV, while hOBs‐16k EV promoted adipogenic gene expression compared to that from hPDLCs and hGFs.
Conclusions
Our proof‐of‐concept data demonstrate that hOBs‐sEV might be an alternative cell‐free therapeutic for bone tissue engineering.