A bioinspired and chemically defined alternative to dimethyl sulfoxide for the cryopreservation of human hematopoietic stem cells.

The cryopreservation of hematopoietic cells using dimethyl sulfoxide (DMSO) and serum is a common procedure used in transplantation. However, DMSO has clinical and biological side effects due to its toxicity, and serum introduces variation and safety risks. Inspired by natural antifreeze proteins, a novel class of ice-interactive cryoprotectants was developed. The corresponding DMSO-, protein-, and serum-free cryopreservation media candidates were screened through a series of biological assays using human cell lines, peripheral blood cells, and bone marrow cells. XT-Thrive-A and XT-Thrive-B were identified as lead candidates to rival cryopreservation with 10% DMSO in serum based on post-thaw cell survival and short-term proliferation assays. The effectiveness of the novel cryopreservation media in freezing hematopoietic stem cells from human whole bone marrow was assessed by extreme limiting dilution analysis in immunodeficient mice. Stem cell frequencies were measured 12 weeks after transplant based on bone marrow engraftment of erythroid, myeloid, B-lymphoid, and CD34+ progenitors measured by flow cytometry. The recovered numbers of cryopreserved stem cells were similar among XT-Thrive A, XT-Thrive B, and DMSO with serum groups. These findings show that cryoprotectants developed through biomimicry of natural antifreeze proteins offers a substitute for DMSO-based media for the cryopreservation of hematopoietic stem cells.

Combinatorial Peptide Microarray Synthesis Based on Microfluidic Impact Printing.

In this Research Article, a novel inkjet printing technique, micro impact printing (MI printing), is applied for the first time to combinatorial peptide microarray synthesis on amine functionalized microdisc arrays through standard Fmoc chemistry. MI printing shows great advantages in combinatorial peptide microarray synthesis compared with other printing techniques, including (1) a disposable cartridge; (2) a small spot size (80 µm) increases array density; (3) minimal loading volume (0.6 µL) and dead volume (<0.1 µL), reduce chemical waste; and (4) multiplexibility of 5 channels/cartridge and capacity of multiple cartridges. Using this synthesis platform, a tetrapeptide library with 625 permutations was constructed and then applied for the screening of ligands targeting α4ß1 integrin on Jurkat cells.

A polymer-free, biomimicry drug self-delivery system fabricated via a synergistic combination of bottom-up and top-down approaches.

Compared to conventional carrier-assistant drug delivery systems (DDSs), drug self-delivery systems (DSDSs) have advantages of unprecedented drug loading capacity, minimized carrier-related toxicity and ease of preparation. However, the colloidal stability and blood circulation time of DSDSs still need to be improved. Here we report on the development of a novel biomimicry drug self-delivery system by the integration of a top-down cell membrane complexing technique into our self-delivery multifunctional nano-platform made from bottom-up approach that contains 100% active pharmaceutical ingredients (API) of Pheophorbide A and Irinotecan conjugates (named PI). Compared to conventional cell membrane coated nanoparticles with polymer framework as core and relatively low drug loading, this system consisting of red blood cell membrane vesicles complexed PI (RBC-PI) is polymer-free with up to 50% API loading. RBC-PI exhibited 10 times higher area under curve in pharmacokinetic study and much lower macrophage uptake compared with the parent PI nanoparticles. RBC-PI retained the excellent chemophototherapeutic effects of the PI nanoparticles, but possessed superior anti-cancer efficacy with prolonged blood circulation, improved tumor delivery, and enhanced photothermal effects in animal models. This system represents a novel example of using cell membrane complexing technique for effective surface modification of DSDSs. This is also an innovative study to form a polymer-free cell membrane nanoparticle complexing with positive surface charged materials. This biomimicry DSDS takes advantages of the best features from both systems to make up for each other's shortcomings and posed all the critical features for an ideal drug delivery system.

Photopatternable PEDOT:PSS/PEG hybrid thin film with moisture stability and sensitivity.

Degradation and delamination resulting from environmental humidity have been technically challenging for poly (3,4-ethylenedioxythiophene): poly(styrene sulfonate) (PEDOT:PSS) thin-film processing. To overcome this problem, we introduced a one-step photolithographic method to both pattern and link a PEDOT:PSS film onto a poly (ethylene glycol) (PEG) layer as a hybrid thin film structure on a flexible substrate. This film exhibited excellent long-term moisture stability (more than 10 days) and lithographic resolution (as low as 2 µm). Mechanical characterizations were performed, including both stretching and bending tests, which illustrated the strong adhesion present between the PEDOT:PSS and PEG layers as well as between the hybrid thin film and substrate. Moreover, the hybrid moisture-absorbable film showed a quick response of its permittivity to environmental humidity variations, in which the patterned PEDOT:PSS layer served as an electrode and the PEG layer as a moisture-sensing element. Perspiration tracking over various parts of the body surface as well as breath rate measurement under the nose were successfully carried out as demonstrations, which illustrated the potential utility of this stable hybrid thin film for emerging flexible and wearable electronic applications.

Microfluidic-Enabled Print-to-Screen Platform for High-Throughput Screening of Combinatorial Chemotherapy.

Since the 1960s, combination chemotherapy has been widely utilized as a standard method to treat cancer. However, because of the potentially enormous number of drug candidates and combinations, conventional identification methods of the effective drug combinations are usually associated with significantly high operational costs, low throughput screening, laborious and time-consuming procedures, and ethical concerns. In this paper, we present a low-cost, high-efficiency microfluidic print-to-screen (P2S) platform, which integrates combinatorial screening with biomolecular printing for high-throughput screening of anticancer drug combinations. This P2S platform provides several distinct advantages and features, including automatic combinatorial printing, high-throughput parallel drug screening, modular disposable cartridge, and biocompatibility, which can potentially speed up the entire discovery cycle of potent drug combinations. Microfluidic impact printing utilizing plug-and-play microfluidic cartridges is experimentally characterized with controllable droplet volume and accurate positioning. Furthermore, the combinatorial print-to-screen assay is demonstrated in a proof-of-concept biological experiment which can identify the positive hits among the entire drug combination library in a parallel and rapid manner. Overall, this microfluidic print-to-screen platform offers a simple, low-cost, high-efficiency solution for high-throughput large-scale combinatorial screening and can be applicable for various emerging applications in drug cocktail discovery.

Conjugated polyelectrolyte materials for promoting progenitor cell growth without serum.

The discovery of new active biomaterials for promoting progenitor cell growth and differentiation in serum-free medium is still proving more challenging for the clinical treatments of degenerative diseases. In this work, a conjugated polyelectrolyte, polythiophene derivative (PMNT), was discovered to significantly drive the cell cycle progression from G1 to S and G2 phases and thus efficiently promote the cell growth without the need of serum. Furthermore, the fluorescent characteristic of PMNT makes it simultaneously able to trace its cellular uptake and localization by cell imaging. cDNA microarray study shows that PMNT can greatly regulate genes related to cell growth or differentiation. To the best of our knowledge, this is the first example of cell growth or differentiation promotion by polyelectrolyte material without the need of serum, thereby providing an important demonstration of degenerative biomaterial discovery through polymer design.

Multifunctional non-viral delivery systems based on conjugated polymers.

Multifunctional nanomaterials with simultaneous therapeutic and imaging functions explore new strategies for the treatment of various diseases. Conjugated polymers (CPs) are considered as novel candidates to serve as multifunctional delivery systems due to their high fluorescence quantum yield, good photostability, and low cytotoxicity. Highly sensitive sensing and imaging properties of CPs are well reviewed, while the applications of CPs as delivery systems are rarely covered. This feature article mainly focuses on CP-based multifunctional non-viral delivery systems for drug, protein, gene, and cell delivery. Promising directions for the further development of CP-based delivery systems are also discussed.

Aptamer-based polymerase chain reaction for ultrasensitive cell detection.

A new system was developed for sensitive and selective detection of tumor cells taking advantage of cell-attached aptamers amplified by PCR and output signals amplified by cationic conjugated polymers.

New conjugated polymers for photoinduced unwinding of DNA supercoiling and gene regulation.

Three cationic polythiophene derivatives (P1, P2, P3) were synthesized and characterized. Under white light irradiation (400-800 nm), they sensitize oxygen molecule in the surrounding to generate reactive oxygen species (ROS) that can efficiently unwind the supercoiled DNA in vitro. Further study shows that this relaxation of the DNA supercoiling results in the decrease of gene (pCX-EGFP plasmid) expression level. The ability of these conjugated polymers for regulating gene expression will add a new dimension to the function of conjugated polymers.

Conjugated polymers for light-activated antifungal activity.

A cationic polythiophene-porphyrin (PTP) dyad is shown to exhibit efficient light-activated antifungal activity. Higher singlet oxygen (¹O2) generation efficiency can be attained from PTP upon photoexcitation due to the light-harvesting properties of the polymer backbone and efficient energy transfer from the polythiophene to the porphyrin units. PTP can be used for treating fungal infections in lower doses of irradiation light and polymer concentration.

A convenient preparation of multi-spectral microparticles by bacteria-mediated assemblies of conjugated polymer nanoparticles for cell imaging and barcoding.

A novel technique was developed for preparing encoded multicolour microparticles based on the self-assembly of bacteria and conjugated polymer nanoparticles (CPNs) by a very simple and time-saving manner. These bacteria-CPNs microparticles show multicolor emissions by tuning FRET efficiencies among CPNs under single excitation wavelength and can be successfully applied for cell imaging and optical barcoding.