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Background: Small extracellular vesicles (sEVs) are nanometer-sized membranous particles shed by many types of cells and can transfer a multitude of cargos between cells. Recent studies of sEVs have been focusing on their potential to be novel drug carriers due to natural composition and other promising characteristics. However, there are challenges in sEVs-based drug delivery, one of which is the inefficient loading of drugs into sEVs, especially for large biomolecules. Results: In this study, we proposed a membrane-associated protein, milk fat globule–epidermal growth factor 8 protein (MFG-E8), to produce αvβ3-targeted sEVs with high delivery efficiency of interested protein. MFG-E8 is a secreted protein with NH2-terminal epidermal growth factor (EGF)–like domains, containing an Arg-Gly-Asp(RGD) sequence that binds αvβ3 and αvβ5 integrins, and COOH terminal domains C1 and C2, which can bind to lipid membrane with strong affinity. Firstly, we transiently expressed MFG-E8 in HEK293F cells and found that this protein could be secreted and adhere to the cell membrane. The recombinant MFG-E8 is also found to locate at the outer membrane of sEVs. Then we generated engineered sEVs by expressing high levels of the EGFP fused to MFG-E8 in HEK293F cells and showed that MFG-E8 could increase the delivery efficiency of EGFP into sEVs. Further delivery of Gaussia luciferase (GL) by fusion expression with MFG-E8 in donor cells demonstrated that target proteins fused with MFG-E8 still kept their activity. Finally, we identified the sEVs' target to integrin αvβ3 by comparing the transfection efficiency with MFG-E8 loaded sEVs (MFG-E8-sEVs) in αvβ3 positive cells and αvβ3 negative cells. Analysis showed higher target protein could transfect into αvβ3 positive cells with MFG-E8-sEVs than with EGFP loaded sEVs (EGFP-sEVs), meaning the engineered sEVs with MFG-E8 not only could increase the delivery of target protein into sEVs, but also could target the αvβ3 positive cells. Conclusion: This study suggests that recombinant MFG-E8 is an ideal protein to increasingly deliver the drug into sEVs and give sEVs the ability to target the αvβ3 positive cells. [ABSTRACT FROM AUTHOR]

Background: Cervical ossification of the posterior longitudinal ligament (OPLL) is a progressive disease. Posterior decompression surgery is reported to be an effective and comparatively safe procedure with few complications for treatment of patients with myelopathy caused by OPLL. However, some patients require revision surgery because of late neurological deterioration due to OPLL progression or kyphotic changes in cervical alignment. This study reports preliminary clinical results of anterior controllable antidisplacement and fusion (ACAF), a novel revision surgery after initial posterior surgery for OPLL. Methods: From January 2017 to June 2018, ten patients with cervical OPLL who underwent ACAF revision surgery after initial posterior surgery were included in this study. The mean age was 62.1 ± 8.0 years (52–78), and the mean interval between initial posterior surgery and revision was 78.0 ± 48.2 months (5–180). The Japanese Orthopaedic Association (JOA) scales, Neck Disability Index (NDI), visual analog scale (VAS), and surgical complications were recorded. Results: The mean surgery time was 179.3 ± 41.8 min (120–240), and the mean blood loss was 432.5 ± 198.3 ml (225–850). The patients were followed up for at least 12 months. The JOA scores improved from 8.7 ± 2.8 to 13.4 ± 2.4; the mean improvement rate was 59.9% ± 16.1%. Postoperative NDI and VAS scores were 13.3 ± 3.7 and 2.0 ± 1.6, respectively, and were significantly improved compared to those before the procedure (P < 0.05). Cervical lordosis improved from 3.8 ± 4.3° to 17 ± 4.6° after revision surgery. There was only one instance of cerebrospinal fluid (CSF) leakage; no instances of postoperative hematoma, C5 root palsy, or hoarseness occurred. Conclusions: The present study demonstrates that excellent postoperative outcomes can be achieved with the ACAF technique for revision treatment of OPLL. Though further study is required to confirm the conclusion, this novel technique has the potential to serve as an alternative surgical technique for revision treatment of OPLL. [ABSTRACT FROM AUTHOR]