Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury.

BACKGROUND: Severe peripheral nerve damage always requires surgical treatment. Autologous nerve transplantation is a standard treatment, but it is not sufficient due to length limitations and extended surgical time. Even with the available artificial nerves, there is still large room for improvement in their therapeutic effects. Novel treatments for peripheral nerve injury are greatly expected. METHODS: Using a specialized microfluidic device, we generated artificial neurite bundles from human iPSC-derived motor and sensory nerve organoids. We developed a new technology to isolate cell-free neurite bundles from spheroids. Transplantation therapy was carried out for large nerve defects in rat sciatic nerve with novel artificial nerve conduit filled with lineally assembled sets of human neurite bundles. Quantitative comparisons were performed over time to search for the artificial nerve with the therapeutic effect, evaluating the recovery of motor and sensory functions and histological regeneration. In addition, a multidimensional unbiased gene expression profiling was carried out by using next-generation sequencing. RESULT: After transplantation, the neurite bundle-derived artificial nerves exerted significant therapeutic effects, both functionally and histologically. Remarkably, therapeutic efficacy was achieved without immunosuppression, even in xenotransplantation. Transplanted neurite bundles fully dissolved after several weeks, with no tumor formation or cell proliferation, confirming their biosafety. Posttransplant gene expression analysis highlighted the immune system's role in recovery. CONCLUSION: The combination of newly developed microfluidic devices and iPSC technology enables the preparation of artificial nerves from organoid-derived neurite bundles in advance for future treatment of peripheral nerve injury patients. A promising, safe, and effective peripheral nerve treatment is now ready for clinical application.

Shape-memory balloon offering simultaneous thermo/chemotherapies to improve anti-osteosarcoma efficacy.

This paper proposes a shape-memory balloon (SMB) to improve bone cement injection efficiency and postoperative thermo/chemotherapy for bone tumors. The SMB consists of biodegradable poly(ε-caprolactone) (PCL), an anticancer drug (doxorubicin, DOX), and heat-generating magnetic nanoparticles (MNPs). The balloon shape is fabricated in a mold by crosslinking PCL macromonomers with DOX and MNPs. The mechanical properties and shape-transition temperature (approximately 40 °C) of the SMB are modulated by adjusting the molecular weight of PCL and the crosslinking density. This allows safe inflation at the affected site with a 400% expansion rate by simple blow molding. The expanded shape is temporarily memorized at 37 °C, and the computed tomography image shows that the bone cement is successfully injected without extra pressure or leakage. The SMB releases DOX for over 4 weeks, allowing a prolonged effect at the local site. The local dosing is constant as the medication is continuously released, demonstrating an ON-OFF switchable heating/cooling response to alternating magnetic field irradiation. In vitro cytotoxic studies have demonstrated that heat generation/drug release and only drug release from the balloon kill approximately 99% and 60% of human osteosarcoma cells, respectively. The proposed SMB is promising in postoperative local thermo/chemotherapy for bone tumors.

Hyperthermia Nanofiber Platform Synergized by Sustained Release of Paclitaxel to Improve Antitumor Efficiency.

Effective cancer therapy can be achieved by designing a smart nanofiber system with the combination of chemotherapy and hyperthermia. This study demonstrates the in vivo antitumor effect of a nanofiber mesh that can deliver heat and antitumor drug in a controlled manner. The mesh is composed of biodegradable poly(ε-caprolactone) (PCL) with paclitaxel (PTX) and magnetic nanoparticles (MNPs). The PCL mesh releases PTX slowly for at least 6 weeks when tested in vitro. The prolonged therapeutic effect is observed in vivo as a continuous release of medication from the mesh over an extended period of time compared with direct injection of PTX into the tumor site. In addition, the synergistic anticancer effect is achieved upon excitation of the mesh with an alternating magnetic field because the MNPs within the nanofiber generate localized heat which causes heat-induced cell killing as well as enhanced chemotherapeutic effect of PTX. Based on these results, the smart nanofiber system may be very promising for cancer therapeutics in the future and may provide knowledge for new development of localized drug delivery.

A Nanofiber Sheet Incorporating Vitamin B12 Promotes Nerve Regeneration in a Rat Neurorrhaphy Model.

Outcomes of peripheral nerve repair after injury are often suboptimal. Therefore, developing biological approaches to augment nerve regeneration is important. In this in vivo study, we tested the hypothesis that augmentation with an electrospun nanofiber sheet incorporating methylcobalamin (MeCbl) would be effective for regeneration after peripheral nerve transection and repair. METHODS: Rats were divided into 3 groups that either underwent sciatic nerve repair with or without the MeCbl sheet, or a sham operation. At 4 and/or 8 weeks after the operation, sensory and motor functional recovery, along with histological findings, were compared among the groups using the toe-spreading test, mechanical and thermal algesimetry tests, tibialis anterior muscle weight measurements, electrophysiological analyses, which included nerve conduction velocity (NCV), compound muscle action potential (CMAP), and terminal latency (TL), and histological analyses involving the myelinated axon ratio, axon diameter, and total axon number. RESULTS: Compared with the repair group without the MeCbl sheet, the repair group with the MeCbl sheet showed significant recovery in terms of tibialis anterior muscle weight, NCV and CMAP, and also tended to improve in the toe-spreading test, mechanical and thermal algesimetry tests, and TL. Histological analyses also demonstrated that the myelinated axon ratios and axon diameters were significantly higher. Among these findings, the repair group with the MeCbl sheet demonstrated the same recovery in NCV as the sham group. CONCLUSION: This study demonstrated that electrospun nanofiber MeCbl sheets promoted nerve regeneration and functional recovery, indicating that this treatment strategy may be viable for human peripheral nerve injuries.

Alternating Magnetic Field-Triggered Switchable Nanofiber Mesh for Cancer Thermo-Chemotherapy.

We have developed a smart anti-cancer fiber mesh that is able to control tumor-killing activity against lung adenocarcinoma precisely. The mesh is capable of carrying large loads of chemotherapeutic drug, paclitaxel (PTX), as well as magnetic nanoparticles (MNPs). The mesh generates heat when the loaded MNPs are activated in an alternating magnetic field (AMF). The mesh is thermo-responsive, so the heat generated can be also used to trigger PTX release from the mesh. An electrospinning method was employed to fabricate the mesh using a copolymer of N-isopropylacrylamide and N-hydroxymethylacrylamide, the phase transition temperature of which was adjusted to the mild-hyperthermia temperature range around 43 °C. In vitro anti-tumor studies demonstrated that both MNP- and PTX-loaded mesh killed about 66% of cells, whereas only PTX-loaded mesh killed about 43% of cells. In a mouse lung cancer model, the thermo-chemotherapy combo displayed enhanced anti-tumor activity and the systemic toxic effects on mice were eliminated due to local release of the chemotherapeutic agents. The proposed fiber system might provide a blueprint to guide the design of the next generation of local drug delivery systems for safe and effective cancer treatment.

Hybridizing Poly(ε-caprolactone) and Plasmonic Titanium Nitride Nanoparticles for Broadband Photoresponsive Shape Memory Films.

Plasmonic nanoparticles can confine light in nanoscale and locally heat the surrounding. Here we use titanium nitride (TiN) nanoparticles as broadband plasmonic light absorbers and synthesized a highly photoresponsive hybrid cross-linked polymer from shape memory polymer poly(ε-caprolactone) (PCL). The TiN-PCL hybrid is responsive to sunlight and the threshold irradiance was among the lowest when compared with other photoresponsive shape memory polymers studied previously. Sunlight heating with TiN NPs can be applied to other heat responsive smart polymers, thereby contributing to energy-saving smart polymers research for a sustainable society.

Inactivated Sendai Virus (HVJ-E) Immobilized Electrospun Nanofiber for Cancer Therapy.

Inactivated Hemagglutinating Virus of Japan Envelope (HVJ-E) was immobilized on electrospun nanofibers of poly(ε-caprolactone) by layer-by-layer (LbL) assembly technique. The precursor LbL film was first constructed with poly-L-lysine and alginic acid via electrostatic interaction. Then the HVJ-E particles were immobilized on the cationic PLL outermost surface. The HVJ-E adsorption was confirmed by surface wettability test, scanning laser microscopy, scanning electron microscopy, and confocal laser microscopy. The immobilized HVJ-E particles were released from the nanofibers under physiological condition. In vitro cytotoxic assay demonstrated that the released HVJ-E from nanofibers induced cancer cell deaths. This surface immobilization technique is possible to perform on anti-cancer drug incorporated nanofibers that enables the fibers to show chemotherapy and immunotherapy simultaneously for an effective eradication of tumor cells in vivo.