Potential neuroprotective effect of stem cells from apical papilla derived extracellular vesicles enriched by lab-on-chip approach during retinal degeneration

Retinal degeneration (RD) is recognized as a frequent cause of visual impairments, including inherited (Retinitis pigmentosa) and degenerative (age-related macular) eye diseases. Dental stem cells (DSCs) have recently demonstrated a promising neuroprotection potential for ocular diseases through a paracrine manner carried out by extracellular vesicles (EVs). However, effective isolation of EVs is still challenging, and isolation methods determine the composition of enriched EVs and the subsequent biological and functional effects. In the present study, we assessed two enrichment methods (micro-electromechanical systems and ultrafiltration) to isolate the EVs from stem cells from apical papilla (SCAP). The size distribution of the corresponding isolates exhibited the capability of each method to enrich different subsets of EVs, which significantly impacts their biological and functional effects. We confirmed the neuroprotection and anti-inflammatory capacity of the SCAP-EVs in vitro. Further experiments revealed the possible therapeutic effects of subretinal injection of SCAP-EVs in the Royal College of Surgeons (RCS) rat model. We found that EVs enriched by the micro-electromechanical-based device (MEMS-EVs) preserved visual function, reduced retinal cell apoptosis, and prevented thinning of the outer nuclear layer (ONL). Interestingly, the effect of MEMS-EVs was extended to the retinal ganglion cell/retinal nerve fiber layer (GCL/RNFL). This study supports the use of the microfluidics approach to enrich valuable subsets of EVs, together with the choice of SCAP as a source to derive EVs for cell-free therapy of RD.