Separation, characterization, and standardization of extracellular vesicles for drug delivery applications

Buschmann, Dominik; Mussack, Veronika; Byrd, James Brian

doi:10.1016/j.addr.2021.04.027

Cited by 82 publications

(47 citation statements)

References 209 publications

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“…Furthermore, the isolated and purified exosomes are sometimes with a relatively broad size range, usually mixed with other EVs. Considering that size-dependent passive targeting may result in unpredictable biodistribution of exosomes, appropriate purification procedures should be included to separate exosome population in homogeneous size (Buschmann et al, 2021). Exosomes of narrow size distribution can be isolated and enriched through semi-continuous size exclusion chromatography technique (Moleirinho et al, 2019).…”

Section: Challenges and Potential Solutionsmentioning

confidence: 99%

Exosomes, a New Star for Targeted Delivery

Chen

Wang

Zeng

et al. 2021

Front. Cell Dev. Biol.

174

125

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Exosomes are cell-secreted nanoparticles (generally with a size of 30–150 nm) bearing numerous biological molecules including nucleic acids, proteins and lipids, which are thought to play important roles in intercellular communication. As carriers, exosomes hold promise as advanced platforms for targeted drug/gene delivery, owing to their unique properties, such as innate stability, low immunogenicity and excellent tissue/cell penetration capacity. However, their practical applications can be limited due to insufficient targeting ability or low efficacy in some cases. In order to overcome these existing challenges, various approaches have been applied to engineer cell-derived exosomes for a higher selectivity and effectiveness. This review presents the state-of-the-art designs and applications of advanced exosome-based systems for targeted cargo delivery. By discussing experts’ opinions, we hope this review will inspire the researchers in this field to develop more practical exosomal delivery systems for clinical applications.

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Section: Challenges and Potential Solutionsmentioning

confidence: 99%

Exosomes, a New Star for Targeted Delivery

Chen

Wang

Zeng

et al. 2021

Front. Cell Dev. Biol.

174

125

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“…Native EVs offer unique advantages for cellular regulation and the efficient delivery of therapeutic payloads. However, several studies have highlighted the intrinsic limitations of native EVs including long-term maintenance of parental cell culture with minimal metabolic/phenotypic variation ( Lambshead et al, 2018 ; Cherian et al, 2020 ; Escude Martinez de Castilla et al, 2021 ), bio-distribution (i.e., organ targeting) ( Escude Martinez de Castilla et al, 2021 ; Lazaro-Ibanez et al, 2021 ; Ullah et al, 2021 ; Witwer and Wolfram, 2021 ), clearance rates [complement and immune systems (RES-MPS system)] ( Wiklander et al, 2015 ; Ha et al, 2016 ; Kwon, 2020 ; Lara et al, 2020 ; Buschmann et al, 2021 ; Escude Martinez de Castilla et al, 2021 ; Lazaro-Ibanez et al, 2021 ), and crucially, difficulties associated with large scale generation (i.e., processing times, variable potency between batches, good manufacturing practice; GMP) ( Whitford and Guterstam, 2019 ; Cherian et al, 2020 ; Buschmann et al, 2021 ; Escude Martinez de Castilla et al, 2021 ; Ullah et al, 2021 ; Witwer and Wolfram, 2021 ). Moreover, the regulatory machinery of EVs and their distinct subtypes associated with production and cellular uptake remains largely unknown.…”

Section: Engineered Evsmentioning

confidence: 99%

“…M-NVs simulate the biophysical properties of native EVs, including size (50–200 nm), which has been reported to influence half-life in circulation (i.e., nanoparticles smaller than 200 nm are able to evade RES uptake, and nanoparticles larger than 30 nm to avoid rapid renal elimination) ( Ha et al, 2016 ; Murphy et al, 2019 ; Lara et al, 2020 ; Buschmann et al, 2021 ; Dooley et al, 2021 ; Escude Martinez de Castilla et al, 2021 ; Lazaro-Ibanez et al, 2021 ; Witwer and Wolfram, 2021 ). M-NVs can be generated by either top-down (extruding parental cells into nano-sized fragments) to obtain biological M-NVs, or bottom-up methods (selecting cargo and capsule materials i.e., liposomes or polymer nanoparticles) to obtain chemically-defined synEVs ( Nasiri Kenari et al, 2020 ).…”

Section: Engineered Evsmentioning

confidence: 99%

Development of Extracellular Vesicle Therapeutics: Challenges, Considerations, and Opportunities

Claridge

Lozano

Poh

et al. 2021

Front. Cell Dev. Biol.

111

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Extracellular vesicles (EVs) hold great promise as therapeutic modalities due to their endogenous characteristics, however, further bioengineering refinement is required to address clinical and commercial limitations. Clinical applications of EV-based therapeutics are being trialed in immunomodulation, tissue regeneration and recovery, and as delivery vectors for combination therapies. Native/biological EVs possess diverse endogenous properties that offer stability and facilitate crossing of biological barriers for delivery of molecular cargo to cells, acting as a form of intercellular communication to regulate function and phenotype. Moreover, EVs are important components of paracrine signaling in stem/progenitor cell-based therapies, are employed as standalone therapies, and can be used as a drug delivery system. Despite remarkable utility of native/biological EVs, they can be improved using bio/engineering approaches to further therapeutic potential. EVs can be engineered to harbor specific pharmaceutical content, enhance their stability, and modify surface epitopes for improved tropism and targeting to cells and tissues in vivo. Limitations currently challenging the full realization of their therapeutic utility include scalability and standardization of generation, molecular characterization for design and regulation, therapeutic potency assessment, and targeted delivery. The fields’ utilization of advanced technologies (imaging, quantitative analyses, multi-omics, labeling/live-cell reporters), and utility of biocompatible natural sources for producing EVs (plants, bacteria, milk) will play an important role in overcoming these limitations. Advancements in EV engineering methodologies and design will facilitate the development of EV-based therapeutics, revolutionizing the current pharmaceutical landscape.

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“…Over the past years, many biological involvements of EVs in physiological and pathological states have been established or postulated (3)(4)(5). Given the functional capabilities to transfer diverse cargo, EVs have attracted increasing interest as therapeutic agents for clinical applications such as drug delivery, immunotherapy, and cellular reprogramming (2,6,7). To date, several drug delivery systems have been developed and approved for clinical use, while some of them are also limited due to cytotoxicity, inefficiency, and immunogenicity (8).…”

Section: Introductionmentioning

confidence: 99%

Skimmed Bovine Milk-Derived Extracellular Vesicles Isolated via “Salting-Out”: Characterizations and Potential Functions as Nanocarriers

Tan

Fang

et al. 2021

Front. Nutr.

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Bovine milk-derived extracellular vesicles (BM-EVs) are recognized as promising nanoscale delivery vectors owing to their large availability. However, few isolation methods can achieve high purity and yield simultaneously. Therefore, we developed a novel and cost-effective procedure to separate BM-EVs via “salting-out.” First, BM-EVs were isolated from skimmed milk using ammonium sulfate. The majority of BM-EVs were precipitated between 30 and 40% saturation and 34% had a relatively augmented purity. The separated BM-EVs showed a spherical shape with a diameter of 60–150 nm and expressed the marker proteins CD63, TSG101, and Hsp70. The purity and yield were comparable to the BM-EVs isolated via ultracentrifugation while ExoQuick failed to separate a relatively pure fraction of BM-EVs. The uptake of BM-EVs into endothelial cells was dose- and time-dependent without significant cytotoxicity. The levels of endothelial nitric oxide syntheses were regulated by BM-EVs loaded with icariside II and miRNA-155-5p, suggesting their functions as delivery vehicles. These findings have demonstrated that it is an efficient procedure to isolate BM-EVs via “salting-out,” holding great promise toward therapeutic applications.

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Separation, characterization, and standardization of extracellular vesicles for drug delivery applications

Cited by 82 publications

References 209 publications

Exosomes, a New Star for Targeted Delivery

Exosomes, a New Star for Targeted Delivery

Development of Extracellular Vesicle Therapeutics: Challenges, Considerations, and Opportunities

Skimmed Bovine Milk-Derived Extracellular Vesicles Isolated via “Salting-Out”: Characterizations and Potential Functions as Nanocarriers

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