[Exploration of the Suitable Culture Conditions for Extracellular Microvesicles Derived from Human Mesenchymal Stem Cells].

OBJECTIVE: To explore the appropriate procedures for preparing extracellular microvesicles (MV) derived from human mesenchymal stem cells (MSC). METHODS: Human MSCs from umbilical cords were cultured in a serum-free medium and maintained in a basal medium for 72 hours after the cell confluence reached to 80%. The supernatants of cultured cells were collected and MVs were enriched. MVs were identified by flow cytometry and electron microscopy. The total protein amount in MVs was used as a parameter for the content of MVs. The supernatants were adjusted to different pH values, and the output of MVs was detected. The supernatants were also collected for enriching the MV and detecting the protein content of MV after the cells were maintained in the basic medium for different time. RESULTS: Flow cytometric analysis showed that the MVs expressed CD9, CD63 and CD81, morphologically presented round under an electron microscope and the diameter of MV was around 100 nm. After enrichment of MV, the protein content of MVs in the supernatants was 416.8±128.1, 255.4±77.9 and 142.8±46.4 µg per 107 MSC，respectively at pH of supernatant 3, 7 and 9 (P＜0.05). The protein content of the supernatants per 107 MSC was 173.6±44.5, 262.4±49.6 and 364.2±37.8 µg respectively after starvation culture for 48, 72 and 96 hrs (P＜0.05). CONCLUSION: MVs can be readily collected after MSCs were starved for 96 hours, and the pH of the supernatants is adjusted at 3.0.

Demulsification of Kerosene/Water Emulsion in the Transparent Asymmetric Plate-Type Micro-Channel.

Asymmetric plate-type micro-channels (APM) have one hydrophobic wall and one hydrophilic wall. By flowing through APM, a kerosene-in-water emulsion can be de-emulsified in one second. To date, however, the demulsification process in the APM is still a black box. In order to observe the demulsification process directly, transparent asymmetric plate-type micro-channels (TAPM) were fabricated with two surface-modified glass plates. Emulsions with oil contents of 10%, 30%, and 50% were pumped through TAPM with heights of 39.2 µm and 159.5 µm. The movement and coalescence of oil droplets (the dispersed phase of a kerosene-in-water emulsion) in the TAPM were observed directly with an optical microscope. By analyzing videos and photographs, it was found that the demulsification process included three steps: oil droplets flowed against and were adsorbed on the hydrophobic wall, then oil droplets coalesced to form larger droplets, whereupon the oil phase was separated. The experimental results showed that the demulsification efficiency was approximately proportional to the oil content (30⁻50%) of the emulsions and increased when the micro-channel height was reduced.

Simultaneously high stiffness and damping in nanoengineered microtruss composites.

Materials combining high stiffness and mechanical energy dissipation are needed in automotive, aviation, construction, and other technologies where structural elements are exposed to dynamic loads. In this paper we demonstrate that a judicious combination of carbon nanotube engineered trusses held in a dissipative polymer can lead to a composite material that simultaneously exhibits both high stiffness and damping. Indeed, the combination of stiffness and damping that is reported is quite high in any single monolithic material. Carbon nanotube (CNT) microstructures grown in a novel 3D truss topology form the backbone of these nanocomposites. The CNT trusses are coated by ceramics and by a nanostructured polymer film assembled using the layer-by-layer technique. The crevices of the trusses are then filled with soft polyurethane. Each constituent of the composite is accurately modeled, and these models are used to guide the manufacturing process, in particular the choice of the backbone topology and the optimization of the mechanical properties of the constituent materials. The resulting composite exhibits much higher stiffness (80 times) and similar damping (specific damping capacity of 0.8) compared to the polymer. Our work is a step forward in implementing the concept of materials by design across multiple length scales.