A novel flexible nerve guidance conduit promotes nerve regeneration while providing excellent mechanical properties.

JOURNAL/nrgr/04.03/01300535-202507000-00029/figure1/v/2024-09-09T124005Z/r/image-tiff Autografting is the gold standard for surgical repair of nerve defects > 5 mm in length; however, autografting is associated with potential complications at the nerve donor site. As an alternative, nerve guidance conduits may be used. The ideal conduit should be flexible, resistant to kinks and lumen collapse, and provide physical cues to guide nerve regeneration. We designed a novel flexible conduit using electrospinning technology to create fibers on the innermost surface of the nerve guidance conduit and employed melt spinning to align them. Subsequently, we prepared disordered electrospun fibers outside the aligned fibers and helical melt-spun fibers on the outer wall of the electrospun fiber lumen. The presence of aligned fibers on the inner surface can promote the extension of nerve cells along the fibers. The helical melt-spun fibers on the outer surface can enhance resistance to kinking and compression and provide stability. Our novel conduit promoted nerve regeneration and functional recovery in a rat sciatic nerve defect model, suggesting that it has potential for clinical use in human nerve injuries.

Plasma Membrane-Derived Biomimetic Apoptotic Nanovesicles Targeting Inflammation and Cartilage Degeneration for Osteoarthritis.

Osteoarthritis (OA) is a degenerative whole-joint disease in which the synovium and joint cartilage become inflamed and damaged. The essential role of inflammation in the development of OA has been recognized recently. Accordingly, simultaneous regulation of local inflammation and tissue degeneration is proposed as a promising therapeutic strategy. Herein, multifunctional biomimetic apoptotic nanovesicles (Apo-NVs) are constructed with plasma membrane derived from apoptotic T cells. The anti-inflammatory microRNA-124 is further encapsulated into Apo-NVs in the hope of achieving an enhanced immunomodulatory effect. It is found that apoptotic nanovesicles, including Apo-NVs and Apo-NVs-miR-124, both efficiently promote the M2 repolarization of M1 macrophages and inhibit the degenerative phenotype of chondrocytes. Further in vivo studies show that Apo-NVs and Apo-NVs-miR-124 alleviate synovial inflammation and protect cartilage tissue from degeneration in OA mice. The study highlights the potential of Apo-NVs in treating OA and other inflammation-related diseases.

Decellularized Scaffolds with Double-Layer Aligned Microchannels Induce the Oriented Growth of Bladder Smooth Muscle Cells: Toward Urethral and Ureteral Reconstruction.

There is a great clinical need for regenerating urinary tissue. Native urethras and ureters have bidirectional aligned smooth muscle cells (SMCs) layers, which plays a pivotal role in micturition and transporting urine and inhibiting reflux. Thus far, urinary scaffolds have not been designed to induce the native-mimicking aligned arrangement of SMCs. In this study, a tubular decellularized extracellular matrix (dECM) with an intact internal layer and bidirectional aligned microchannels in the tubular wall, which is realized by the subcutaneous implantation of a template, followed by the removal of the template, and decellularization, is engineered. The dense and intact internal layer effectively increases the leakage pressure of the tubular dECM scaffolds. Rat-derived dECM scaffolds with three different sizes of microchannels are fabricated by tailoring the fiber diameter of the templates. The rat-derived dECM scaffolds exhibiting microchannels of ≈65 µm show suitable mechanical properties, good ability to induce the bidirectional alignment and growth of human bladder SMCs, and elevated higher functional protein expression in vitro. These data indicate that rat-derived tubular dECM scaffolds manifesting double-layer aligned microchannels may be promising candidates to induce the native-mimicking regeneration of SMCs in urethra and ureter reconstruction.

Functionalization of in vivo tissue-engineered living biotubes enhance patency and endothelization without the requirement of systemic anticoagulant administration.

Vascular regeneration and patency maintenance, without anticoagulant administration, represent key developmental trends to enhance small-diameter vascular grafts (SDVG) performance. In vivo engineered autologous biotubes have emerged as SDVG candidates with pro-regenerative properties. However, mechanical failure coupled with thrombus formation hinder translational prospects of biotubes as SDVGs. Previously fabricated poly(ε-caprolactone) skeleton-reinforced biotubes (PBs) circumvented mechanical issues and achieved vascular regeneration, but orally administered anticoagulants were required. Here, highly efficient and biocompatible functional modifications were introduced to living cells on PB lumens. The 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-methoxy (DMPE)-PEG-conjugated anti-coagulant bivalirudin (DPB) and DMPE-PEG-conjugated endothelial progenitor cell (EPC)-binding TPS-peptide (DPT) modifications possessed functionality conducive to promoting vascular graft patency. Co-modification of DPB and DPT swiftly attained luminal saturation without influencing cell viability. DPB repellent of non-specific proteins, DPB inhibition of thrombus formation, and DPB protection against functional masking of DPT's EPC-capture by blood components, which promoted patency and rapid endothelialization in rat and canine artery implantation models without anticoagulant administration. This strategy offers a safe, facile, and fast technical approach to convey additional functionalization to living cells within tissue-engineered constructs.

Electrospun membranes of carboxylated poly(ester urethane)urea/gelatin encapsulating pterostilbene for adaptive and antioxidative purposes.

Oxidative stress caused by the harsh microenvironment after implantation of an artificial graft with mismatching mechanical properties usually triggers inflammation responses, which have adverse impacts on tissue regeneration. For coping with these problems, in this work, bioactive fibrous scaffolds were developed from specially synthesized carboxylated poly(ester urethane)urea (PEUU) and gelatin (Gel) by encapsulating pterostilbene (Pte) for the first time. The prepared electrospun membranes exhibited self-adaptable mechanical properties with high elasticity owing to the bonded electrospun fibers, cross-linking network between PEUU and Gel, and the inherent flexibility of the PEUU polymer in the fibrous matrix. The PEUU/Gel/Pte electrospun membrane containing 7% Pte could promote in vitro proliferation of human umbilical vein endothelial cells, and regulate vascular smooth muscle cells with excellent antioxidant properties via free radical scavenging. In vivo results in a rat subcutaneous implantation model further demonstrated the positive effect of the specially prepared PEUU/Gel/Pte scaffold on both normal cell growth and anti-inflammatory by promoting cellularization and polarizing macrophages toward the M2 phenotype. These PEUU/Gel/Pte electrospun membranes with adaptability benefit to tissue regeneration by modulating inflammation responses, especially applications in vascular regeneration.

Short-wave infrared computed tomography.

We demonstrate a short-wave infrared computed tomography method. It uses a fiber-coupled 1.44µm super-luminescent diode as light source, a PbSe photodiode as infrared detector, and an electronically controlled rotation and translation stage for high-speed Radon scanning. It is a safe and low power nondestructive testing method that can be used for the detection of plastic polymers, biological tissue and other materials that visible light cannot penetrate. We analyze the theoretical resolution of the method and build a short-wave infrared computed tomography system, which realizes the tomography and 3D reconstruction of black plastic bottles and artificial blood vessels. The measured resolution reaches10µm.

Mechanically reinforced biotubes for arterial replacement and arteriovenous grafting inspired by architectural engineering.

There is a lack in clinically-suitable vascular grafts. Biotubes, prepared using in vivo tissue engineering, show potential for vascular regeneration. However, their mechanical strength is typically poor. Inspired by architectural design of steel fiber reinforcement of concrete for tunnel construction, poly(ε-caprolactone) (PCL) fiber skeletons (PSs) were fabricated by melt-spinning and heat treatment. The PSs were subcutaneously embedded to induce the assembly of host cells and extracellular matrix to obtain PS-reinforced biotubes (PBs). Heat-treated medium-fiber-angle PB (hMPB) demonstrated superior performance when evaluated by in vitro mechanical testing and following implantation in rat abdominal artery replacement models. hMPBs were further evaluated in canine peripheral arterial replacement and sheep arteriovenous graft models. Overall, hMPB demonstrated appropriate mechanics, puncture resistance, rapid hemostasis, vascular regeneration, and long-term patency, without incidence of luminal expansion or intimal hyperplasia. These optimized hMPB properties show promise as an alternatives to autologous vessels in clinical applications.

Long-term evaluation of vascular grafts with circumferentially aligned microfibers in a rat abdominal aorta replacement model.

Long-term results of implants in small animal models can be used to optimize the design of grafts to further promote tissue regeneration. In previous study, we fabricated a poly(ɛ-caprolactone) (PCL) bi-layered vascular graft consisting of an internal layer with circumferentially aligned microfibers and an external layer with random nanofibers. The circumferentially oriented vascular smooth muscle cells (VSMCs) were successfully regenerated after the grafts were implanted in rat abdominal aorta for 3 months. Here we investigated the long-term (18 months) performance of the bi-layered grafts in the same model. All the grafts were patent. No thrombosis, aneurysm, or stenosis occurred. The endothelium maintained complete. However, most of circumferentially oriented VSMCs migrated to luminal surface of the grafts to form a neointima with uniform thickness. Accordingly, extracellular matrix including collagen, elastin, and glycosaminoglycan displayed high density in neointima layer while with low density in the grafts wall because of the incomplete degradation of PCL. A small amounts of calcification occurred in the grafts. The contraction and relaxation function of regenerated neoartery almost disappeared. These data indicated that based on the structure design, many other factors of grafts should be considered to achieve the regenerated neoartery similar to the native vessels after long-term implantation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2596-2604, 2018.