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This study was conducted to develop biodegradable cellulose scaffolds by oxidising porous cellulose sponges for tissue engineering applications. Cellulose powder was dissolved in ionic liquid using a salt leaching method, and porous cellulose scaffolds of various pore sizes were prepared. The scaffolds were oxidised with periodate to introduce aldehyde at a rate controlled by the periodate concentration. Oxidised scaffolds exhibited weight loss in cell culture medium, but not in phosphate buffer. Therefore, we confirmed that Schiff base formation between the aldehyde and amino groups through a Maillard reaction triggered cellulose molecular degradation. The degradation rate was controlled by the oxidation degree, whereas the aldehyde content controlled protein adsorption and cell proliferation. Additionally, in vivo implantation tests revealed that optimising the oxidation ratio not only improved biodegradability but also reduced inflammation. In conclusion, our results suggest that simple oxidised cellulose is useful as a low-toxicity biodegradable scaffold.

Quantum dots (QDs) can be delivered efficiently inside macrophages using a freeze-concentration approach. In this study, we introduced a new, facile, high concentration-based freezing technology of low toxicity. We also developed QD-conjugated new hydrophobic polyampholytes using poly-L-lysine (PLL), a naturally derived polymer, which showed sustained biocompatibility, stability over one week, and enhanced intracellular delivery. When freeze-concentration was applied, the QD-encapsulated hydrophobic polyampholytes showed a higher tendency to adsorb onto the cell membrane than the non-frozen molecules. Interestingly, we observed that the efficacy of adsorption of QDs on RAW 264.7 macrophages was higher than that on fibroblasts. Furthermore, the intracellular delivery of QDs using hydrophobic polyampholytes was higher than those of PLL and QDs. In vitro studies revealed the efficient endosomal escape of QDs in the presence of hydrophobic polyampholytes and freeze-concentration. Collectively, these observations indicated that the promising combination of freeze-concentration and hydrophobic polyampholytes may act as an effective and versatile strategy for the intracellular delivery of QDs, which can be used for biological diagnosis and therapeutic applications., identifier:https://dspace.jaist.ac.jp/dspace/handle/10119/17044

Wound healing is a complex process involving an intricate cascade of body responses. A composite dressing that would effectively target different stages of wound healing and regeneration is urgently needed. In the current study, we tested the efficacy of a previously prepared micelle-hydrogel composite loaded with two drugs, in full-thickness excision wound model in rat. We found that the composite elicited almost no inflammation and effectively enhanced healing at all stages of the healing process. An initial burst of the first drug, amphotericin B, eliminated any preliminary infection. This burst was followed by a gradual release of curcumin as the healing and anti-inflammatory agent. Better healing was observed in rats treated with the drug-loaded composites than in blank and control groups. Wounds showed up to 80% closure in the treated group, with high collagen deposition. Re-epithelialization and granulation were also better in the treated group than in the non-treated control and blank groups. Histopathological examination revealed that drug-loaded composites improved cutaneous wound healing and regeneration. In conclusion, the micelle-hydrogel composite is an effective dressing and might have major applications in wound healing.

Physical methodologies such as electroporation and the gene-gun technology have been widely used for transfection; however, their applicability is limited because they lead to cell damage and low cell viability. Therefore, to address these limitations we developed a new freeze concentration-based gene transfection system that provides enhanced in vitro gene delivery compared to that provided by the commercially available systems. The system employs a facile freeze concentration step, whereby cells are simply frozen to very low temperatures in the presence of polymer-pDNA complexes. As part of system development, we also synthesized a low toxicity polyethyleneimine (PEI)-based polyampholyte prepared through succinylation with butylsuccinic anhydride. In aqueous solution, this modified polyampholyte self-assembles to form small (20 nm diameter), positively charged (net surface charge of 35 mV), nanoparticles through a combination of hydrophobic and electrostatic interactions. Agarose gel electrophoresis analysis indicated that the polyampholyte nanoparticle was able to form a complex with pDNA that provided stability against nuclease degradation. Using transfection of HEK-293T cells, we demonstrated that a combination of polyampholyte: pDNA, at an appropriate ratio, and the freeze concentration method resulted in significant enhancement of GFP and luciferase expression compared to commercially available carriers. Endosomal escape of pDNA was also found to be increased when using the modified polyampholyte compared to branched PEI. This study suggests that the efficient combination of freeze concentration and the modified polyampholyte described here has great potential for in vitro gene therapy., identifier:https://dspace.jaist.ac.jp/dspace/handle/10119/15875