ABSTRACT
As nano-drug carriers, small extracellular vesicles (sEVs) have shown unique advantages, but their drug loading and encapsulation efficiency are far from being satisfied, especially for the loading of hydrophilic small-molecule drugs. Inspired by the strategies of active loading of liposomal nanomedicines, pre-drug design and immobilization enzyme, here we developed a new platform, named "Esterase-responsive Active Loading" (EAL), for the efficient and stable drug encapsulation of sEVs. Widely used ferulic acid ester derivatives were chosen as prodrugs based on the EAL of engineered sEVs to establish a continuous transmembrane ion gradient for achieving efficient loading of active molecule ferulic acid into sEVs. The EAL showed that the drug loading and encapsulation efficiency were around 6-fold and 5-fold higher than passive loading, respectively. Moreover, characterization by nano-flow cytometry and Malvern particle size analyzer showed that differential ultracentrifugation combined with multiple types of membrane filtration methods can achieve large-scale and high-quality production of sEVs. Finally, extracellular and intracellular assessments further confirmed the superior performance of the EAL-prepared sEVs-loaded ferulic acid preparation in terms of slow release and low toxicity. Taken together, these findings will provide an instructive insight into the development of sEV-based delivery systems.
ABSTRACT
Reconstructing enamel-like hydroxyapatite structures on damaged teeth remains a great challenge in the materials science and dentistry due to its highly ordered hierarchical microstructure. Inspired by the mineralization of mollusk nacre in nature, abalone nacre water-soluble organic matrix (WSM) was isolated successfully though an ultrasonic-assisted water extraction (UWE) strategy with nondestructive activity and high-quality extraction for simulating the process of tooth hard tissue mineralization. Results showed that the UWE strategy significantly increased the protein yield from 7.60% to 9.60% and improved the polysaccharide yield from 2.59% to 3.34%, respectively, indicating its excellent extraction efficiency of WSM. Noteworthily, the smallest averaged particle size (~155 nm) of WSM were obtained at an ultrasound time of 6 h, whereas the highest absolute values (~ -32 mV) of zeta potential was produced. Moreover, it was proved that WSM could induce the growth of enamel-like hydroxyapatite crystals to further facilitate biomimetic remineralization of the demineralized enamel and restore its continuous and smooth surface structure in vitro. Besides, the hardness (4.37 ± 0.07 GPa) and modulus of elasticity (84.80 ± 1.49 GPa) of the WSM-repaired enamel was similar to that of native enamel, indicating superior mechanical properties after repair. Herein, it provides a promising green, efficient strategy for the remineralization of damaged enamel and high value utilization of waste abalone shells.
