Advanced micro-/nanotechnologies for exosome encapsulation and targeting in regenerative medicine.

Exosomes, a subset of vesicles generated from cell membranes, are crucial for cellular communication. Exosomes' innate qualities have been used in recent studies to create nanocarriers for various purposes, including medication delivery and immunotherapy. As a result, a wide range of approaches has been designed to utilize their non-immunogenic nature, drug-loading capacity, or targeting ability. In this study, we aimed to review the novel methods and approaches in exosome engineering for encapsulation and targeting in regenerative medicine. We have assessed and evaluated each method's efficacy, advantages, and disadvantages and discussed the results of related studies. Even though the therapeutic role of non-allogenic exosomes has been demonstrated in several studies, their application has certain limitations as these particles are neither fully specific to target tissue nor tissue retainable. Hence, there is a strong demand for developing more efficient encapsulation methods along with more accurate and precise targeting methods, such as 3D printing and magnetic nanoparticle loading in exosomes, respectively.

Sustained release of growth hormone and sodium nitrite from biomimetic collagen coating immobilized on silicone tubes improves endothelialization.

Biocompatibility of biomedical devices can be improved by endothelialization of blood-contacting parts mimicking the vascular endothelium's function. Improved endothelialization might be obtained by using biomimetic coatings that allow local sustained release of biologically active molecules, e.g. anti-thrombotic and growth-inducing agents, from nanoliposomes. We aimed to test whether incorporation of growth-inducing nanoliposomal growth hormone (nGH) and anti-thrombotic nanoliposomal sodium nitrite (nNitrite) into collagen coating of silicone tubes enhances endothelialization by stimulating endothelial cell proliferation and inhibiting platelet adhesion. Collagen coating stably immobilized on acrylic acid-grafted silicone tubes decreased the water contact angle from 102° to 56°. Incorporation of 50 or 500nmol/ml nNitrite and 100 or 1000ng/ml nGH into collagen coating decreased the water contact angle further to 48°. After 120h incubation, 58% nitrite and 22% GH of the initial amount of sodium nitrite and GH in nanoliposomes were gradually released from the nNitrite-nGH-collagen coating. Endothelial cell number was increased after surface coating of silicone tubes with collagen by 1.6-fold, and with nNitrite-nGH-collagen conjugate by 1.8-3.9-fold after 2days. After 6days, endothelial cell confluency in the absence of surface coating was 22%, with collagen coating 74%, and with nNitrite-nGH-collagen conjugate coating 83-119%. In the absence of endothelial cells, platelet adhesion was stimulated after collagen coating by 1.3-fold, but inhibited after nNitrite-nGH-collagen conjugate coating by 1.6-3.7-fold. The release of anti-thrombotic prostaglandin I2 from endothelial cells was stimulated after nNitrite-nGH-collagen conjugate coating by 1.7-2.2-fold compared with collagen coating. Our data shows improved endothelialization and blood compatibility using nNitrite-nGH-collagen conjugate coating on silicone tubes suggesting that these coatings are highly suitable for use in blood-contacting parts of biomedical devices.

Flow Preconditioning of Endothelial Cells on Collagen-Immobilized Silicone Fibers Enhances Cell Retention and Antithrombotic Function.

Stability and antithrombotic functionality of endothelial cells on silicone hollow fibers (SiHFs) are critical in the development of biohybrid artificial lungs. Here we aimed to enhance endothelial cell retention and anti-thrombotic function by low (12 dyn/cm2 , 24 h) fluid shear stress ("flow") preconditioning of endothelial cells seeded on collagen-immobilized SiHFs. The response of endothelial cells without preconditioning (48 h static culture) and with preconditioning (24 h static culture followed by 24 h flow preconditioning) on hollow fibers to high fluid shear stress (30 dyn/cm2 , 1 h) was assessed in a parallel-plate flow chamber. Finite element (FE) modeling was used to simulate shear stress within the flow chamber. We found that collagen immobilization on hollow fibers using carbodiimide bonds provided sufficient stability to high shear stress. Flow preconditioning for 24 h before treatment with high shear stress for 1 h on collagen-immobilized hollow fibers increased cell retention (1.3-fold). The FE model showed that cell flattening due to flow preconditioning reduced maximum shear stress on cells by 32%. Flow preconditioning prior to exposure to high fluid shear stress enhanced the production of nitric oxide (1.3-fold) and prostaglandin I2 (1.2-fold). In conclusion, flow preconditioning of endothelial cells on collagen-immobilized SiHFs enhanced cell retention and antithrombotic function, which could significantly improve current biohybrid artificial lungs.

Nanoliposomal Growth Hormone and Sodium Nitrite Release from Silicone Fibers Reduces Thrombus Formation Under Flow.

Biocompatibility of artificial lungs can be improved by endothelialization of hollow fibers. Bioavailability of growth-inducing and anti-thrombotic agents on the hollow fiber-blood interface inhibits thrombosis. We investigated if nanoliposomal growth-inducing growth hormone (nGH) and anti-thrombotic sodium nitrite (nNitrite) incorporation into collagen-coating on silicone hollow fibers improves blood biocompatibility by increasing endothelial cell growth and nitrite bioavailability under flow. Nitrite production rate was assessed under varying flow conditions. Finite element (FE) modeling was used to simulate nitrite transport within the parallel-plate flow chamber, and nitrite bioavailability on the fiber-blood interface at 1-30 dyn/cm(2) shear stress. Endothelial cell number on fibers coated with nNitrite-nGH-collagen conjugate was 1.5-fold higher than on collagen-coated fibers. For collagen-coated fibers, nitrite production reached a maximum at 18 dyn/cm(2) shear stress. When fibers were coated with nNitrite-nGH-collagen conjugate, nitrite production increased continuously by increasing shear stress. FE modeling revealed that nitrite concentrations at the fiber-blood interface were affected by shear stress-induced nitrite production, and diffusion/convection-induced nitrite removal. Highest nitrite concentrations and lowest thrombus deposition were observed on fibers coated with nNitrite-nGH-collagen conjugate exposed to 6-12 dyn/cm(2) shear stress. In conclusion, our results suggest that nNitrite-nGH-Col conjugate coatings promote endothelialization of silicone hollow fibers in biohybrid artificial lungs.