Integrated Osteoinductive Factors─Exosome@MicroRNA-26a Hydrogel Enhances Bone Regeneration.

MicroRNAs (miRNAs) are a new therapeutic tool that can target multiple genes by inducing translation repression and target mRNA degradation. Although miRNAs have gained significant attention in oncology and in work on genetic disorders and autoimmune diseases, their application in tissue regeneration remains hindered by several challenges, such as miRNA degradation. Here, we reported Exosome@MicroRNA-26a (Exo@miR-26a), an osteoinductive factor that can be substituted for routinely used growth factors, which was constructed using bone marrow stem cell (BMSC)-derived exosomes and microRNA-26a (miR-26a). Exo@miR-26a-integrated hydrogels significantly promoted bone regeneration when implanted into defect sites; as the exosome stimulated angiogenesis, miR-26a promoted osteogenesis while the hydrogel enabled a site-directed release. Moreover, BMSC-derived exosomes further facilitated healthy bone regeneration by repressing osteoclast differentiation-related genes rather than damaging osteoclasts. Taken together, our findings demonstrate the promising potential of Exo@miR-26a for bone regeneration and provide a new strategy for the application of miRNA therapy in tissue engineering.

A functional PVA aerogel-based membrane obtaining sutureability through modified electrospinning technology and achieving promising anti-adhesion effect after cardiac surgery.

Pericardial barrier destruction, inflammatory cell infiltration, and fibrous tissue hyperplasia, trigger adhesions after cardiac surgery. There are few anti-adhesion materials that are both functional and sutureable for pericardial reconstruction. Besides, a few studies have reported on the mechanism of preventing pericardial adhesion. Herein, a functional barrier membrane with sutureability was developed via a modified electrospinning method. It was composed of poly(l-lactide-co-caprolactone) (PLCL) nanofibers, poly(vinyl alcohol) (PVA) aerogel, and melatonin, named PPMT. The PPMT had a special microstructure manifested as a staggered arrangement of nanofibers on the surface and a layered macroporous aerogel structure in a cross-section. Besides providing the porosity and hydrophilicity obtained from PVA, the structure also had suitable mechanical properties for stitching due to the addition of PLCL nanofibers. Furthermore, it inhibited the proliferation of fibroblasts by suppressing the activation of Fas and P53, and achieved anti-inflammatory effects by affecting the activity of inflammatory cells and reducing the release of pro-inflammatory factors, such as interleukin 8 (IL-8) and tumor necrosis factor α (TNF-α). Finally, in vivo transplantation showed that it up-regulated the expression of matrix metalloproteinase-1 (MMP1) and tissue inhibitor of metalloproteinase-1 (TIMP1), and down-regulated the expression of Vinculin and transforming growth factor ß (TGF-ß) in the myocardium, thereby reducing the formation of adhesions. Collectively, these results demonstrate a great potential of PPMT membrane for practical application to anti-adhesion.

A self-healing neutral aqueous rechargeable Zn/MnO2 battery based on modified carbon nanotubes substrate cathode.

Rechargeable aqueous Zn/MnO2 batteries show great potential for grid-scale storage due to their low cost, high safety, and energy density, yet suffer from continuous capacity decay during operation. Therefore, this work proposes a capacity self-healing aqueous Zn/MnO2 (Zn/cCNTs-MnO2) battery using carboxyl-modified carbon nanotubes (cCNTs) as the cathode substrate, ZnSO4 + MnSO4 mixed aqueous solution as the electrolyte, and Zn foil as the anode. Based on the controllable electrodeposition reaction of MnO2, the specific capacity of Zn/cCNTs-MnO2 batteries can be achieved or recovered by operating several cycles under a low current density (0.1 mA cm-2). Then, the batteries can stably perform under a high current density (1 mA cm-2). By repeating the above steps, a capacity self-healing usage scheme was established, which can significantly improve the cycling performance of Zn/cCNTs-MnO2 batteries. Moreover, the results of the proposed Zn/cCNTs-MnO2 batteries verify the MnO2 electrodeposition mechanism and introduce a novel method for the development of durable aqueous rechargeable Zn/MnO2 batteries.

Rational Design of a High-Performance Quinoxalinone-Based AIE Photosensitizer for Image-Guided Photodynamic Therapy.

Because light exhibits excellent spatiotemporal resolution, photodynamic therapy (PDT) is becoming a promising method for cancer treatment. However, in a single photosensitizer (PS), it remains a big challenge to achieve all key properties including effective singlet oxygen (1O2) production under long-wavelength laser and bright near-infrared (NIR) emission without toxicity in the dark. In addition, clinically used traditional PSs encounter quenched fluorescence and decreased 1O2 production because of molecular aggregation in aqueous solution. To solve the aforementioned issues, quinoxalinone CN (QCN) with effective 1O2 generation under long-wavelength (530 nm) laser irradiation and aggregation-induced NIR emission is rationally designed by precise optimization of the quinoxalinone scaffold. After being encapsulated by an amphiphilic polymer (DSPE-PEG), the yielded nanoparticles exhibit highly efficient 1O2 production and stable NIR fluorescence located at 800 nm without obvious toxicity under the dark. Both in vitro and in vivo evaluation identify that QCN would be a promising PS for image-guided PDT of tumors.

Construction and performance evaluation of Hep/silk-PLCL composite nanofiber small-caliber artificial blood vessel graft.

To meet the growing clinical demand for small-caliber blood vessel grafts to treat cardiovascular diseases, it is necessary to develop safe and long-term unobstructed grafts. In this study, a biodegradable graft made of composite nanofibers is introduced. A composite nanofiber core-shell structure was prepared by a combination of conjugate electrospinning and freeze-dry technology. The core fiber was poly(l-lactide-co-caprolactone) (PLCL)-based and the core fibers were coated with heparin/silk gel, which acted as a shell layer. This special structure in which the core layer was made of synthetic materials and the shell layer was made of natural materials took advantage of these two different materials. The core PLCL nanofibers provided mechanical support during vascular reconstruction, and the shell heparin/silk gel layer enhanced the biocompatibility of the grafts. Moreover, the release of heparin in the early stage after transplantation could regulate the microenvironment and inhibit the proliferation of intima. All of the graft materials were biodegradable and safe biomaterials, and the degradation of the graft provided space for the growth of regenerated tissue in the late stage of transplantation. Animal experiments showed that the graft remained unobstructed for more than eight months in vivo. In addition, the regenerated vascular tissue provided a similar function to that of autogenous vascular tissue when the graft was highly degraded. Thus, the proposed method produced a graft that could maintain long-term patency in vivo and remodel vascular tissue successfully.

Ultrasonication-promoted synthesis of luminescent sulfur nano-dots for cellular imaging applications.

An ultrasonication-promoted strategy was proposed to synthesize luminescent S-dots, which reduced the synthesis time from the commonly used 5 days to several hours. The as-synthesized S-dots show a high photostability and low cytotoxicity, and are then successfully applied for cellular imaging.

Evaluation of a simple off-the-shelf bi-layered vascular scaffold based on poly(L-lactide-co-ε-caprolactone)/silk fibroin in vitro and in vivo.

Purpose: In the field of small-caliber vascular scaffold research, excellent vascular remodeling is the key to ensuring anticoagulant function. We prepared an off-the-shelf bi-layered vascular scaffold with a dense inner layer and a loose outer layer and evaluated its remodeling capabilities by in vivo transplantation. Materials and Methods: Based on poly(L-lactide-co-ε-caprolactone) (PLCL), silk fibroin(SF), and heparin (Hep), PLCL/SF/Hep bi-layered scaffolds and PLCL/Hep bi-layered scaffolds were prepared by electrospinning. The inner layer was a PLCL/SF/Hep or PLCL/Hep nanofiber membrane, and the outer layer was PLCL/SF nano yarn. The in vitro tests included a hydrophilicity test, mechanical properties test, and blood and cell compatibility evaluation. The in vivo evaluation was conducted via single rabbit carotid artery replacement and subsequent examinations, including ultrasound imaging, immunoglobulin assays, and tissue section staining. Results: Compared to the PLCL/Hep nanofiber membrane, the hydrophilicity of the PLCL/SF/Hep nanofiber membrane was significantly improved. The mechanical strength met application requirements. Both the blood and cell compatibility were optimal. Most importantly, the PLCL/SF/Hep scaffolds maintained lumen patency for 3 months after carotid artery transplantation in live rabbits. At the same time, CD31 and α-SMA immunofluorescence staining confirmed bionic endothelial and smooth muscle layers remodeling. Conclusion: Using this hybrid strategy, PLCL and SF were combined to manufacture bi-layered small-caliber vascular scaffolds; these PLCL/SF/Hep scaffolds showed satisfactory vascular remodeling.

Electrospun Nanofibers for Tissue Engineering with Drug Loading and Release.

Electrospinning technologies have been applied in the field of tissue engineering as materials, with nanoscale-structures and high porosity, can be easily prepared via this method to bio-mimic the natural extracellular matrix (ECM). Tissue engineering aims to fabricate functional biomaterials for the repairment and regeneration of defective tissue. In addition to the structural simulation for accelerating the repair process and achieving a high-quality regeneration, the combination of biomaterials and bioactive molecules is required for an ideal tissue-engineering scaffold. Due to the diversity in materials and method selection for electrospinning, a great flexibility in drug delivery systems can be achieved. Various drugs including antibiotic agents, vitamins, peptides, and proteins can be incorporated into electrospun scaffolds using different electrospinning techniques and drug-loading methods. This is a review of recent research on electrospun nanofibrous scaffolds for tissue-engineering applications, the development of preparation methods, and the delivery of various bioactive molecules. These studies are based on the fabrication of electrospun biomaterials for the repair of blood vessels, nerve tissues, cartilage, bone defects, and the treatment of aneurysms and skin wounds, as well as their applications related to oral mucosa and dental fields. In these studies, due to the optimal selection of drugs and loading methods based on electrospinning, in vitro and in vivo experiments demonstrated that these scaffolds exhibited desirable effects for the repair and treatment of damaged tissue and, thus, have excellent potential for clinical application.

Three-dimensional electrospun nanofibrous scaffolds displaying bone morphogenetic protein-2-derived peptides for the promotion of osteogenic differentiation of stem cells and bone regeneration.

Tissue scaffolds with three-dimensional (3D) nanofibrous biomimetic structures have attracted attention in the field of bone regeneration. In recent years, emerging strategies based on electrospinning technologies have facilitated the preparation of 3D nanofibrous scaffolds. Based on these developments, in this study, 3D scaffolds possessing both nanofibrous morphologies and interconnected pores were fabricated for their potential in bone tissue engineering. By combining homogenizing, freeze-drying, and thermal crosslinking techniques, nano-hydroxyapatite/PLLA/gelatin (nHA/PLA/GEL) 3D nanofibrous scaffolds were prepared using pre-fabricated electrospun nanofibers. Then, utilizing a polydopamine (pDA)-assisted coating strategy, bone morphogenetic protein-2 (BMP-2)-derived peptides were further immobilized onto the 3D scaffolds to obtain the resulting nano-hydroxyapatite/PLLA/gelatin-peptide (nHA/PLA/GEL-PEP) 3D nanofibrous scaffolds capable of sustained release. Bone mesenchymal stem cells (BMSCs) were cultured on the 3D nanofibrous scaffolds, then relative cell viability, alkaline phosphatase (ALP) activity, and gene expression assays were performed to study the effects of the scaffolds on cell growth and osteogenic differentiation in vitro. Furthermore, the ability of bone formation in vivo was evaluated using a rat cranial bone defect model. In vitro and in vivo results demonstrated that the 3D nanofibrous scaffolds incorporated with nHA and BMP-2 peptides exhibited favorable biocompatibility and osteoinductivity. Therefore, these nanofibrous scaffolds have excellent potential in bone regenerative medicine.

Electrospun Bilayer Composite Vascular Graft with an Inner Layer Modified by Polyethylene Glycol and Haparin to Regenerate the Blood Vessel.

In this study, we prepared a composite vascular graft with two layers. The inner layer, which was comprised of degradable Poly(lactic-co-glycolic acid) (PLGA)/Collagen (PC) nanofibers modified by mesoporous silica nanoparticles (MSN), was grafted with polyethylene glycol (PEG) and heparin to promote cell proliferation and to improve blood compatibility. The outer layer was comprised of polyurethane (PU) nanofibers in order to provide mechanical support. The growth and proliferation of human umbilical vein endothelial cells (HUVECs) in the inner layer was significant, and blood compatibility testing showed that the inner layer had good blood compatibility. The MSN-PEG-Heparin on the fiber surface was observed in vitro during the degradation of the inner layer. After 60 days, the weight of fiber membrane decreased by 92.4%. The inner layer did not cause an inflammatory reaction during the degradation process in vivo and there was uniform cellular growth on the PC/MSN-PEG-Heparin fiber membrane. Composite grafts implanted into the rabbit carotid artery were evaluated for 8 weeks by H&E and immunohistochemical staining, demonstrating that a monolayer of endothelium (CD31-labeled) and smooth muscle (αSMA-labeled) regenerated on the composite graft. Our results demonstrate that the composite graft, with a functional inner layer, has potential to be used for small-caliber blood vessels with long-term patency.

A Method for Preparation of an Internal Layer of Artificial Vascular Graft Co-Modified with Salvianolic Acid B and Heparin.

Studies have shown that salvianolic acid B (SAB), which is derived from Chinese salvia ( Salvia miltiorrhiza), a plant used in traditional Chinese medicine, can promote the proliferation and migration of endothelial cells. The inner layer of an artificial vascular graft was fabricated using the coaxial electrospinning method and was loaded with the anticoagulant heparin and SAB. The release of heparin and SAB was sustained for almost 30 days and without an initial burst release of SAB. Furthermore, the combined effect of SAB and heparin contributed to promoting human umbilical vein endothelial cell (HUVEC) growth and improved the blood compatibility of the graft. In addition, upregulation of GRP78 by SAB protected human endothelial cells from oxidative stress-induced cellular damage. In vivo evaluation through Masson's trichrome and H&E staining was performed after the graft was subcutaneously embedded in SD rats for 2 weeks and indicated that the graft possessed satisfactory biocompatibility and did not cause a significant immune response. Hence, the functional inner layer is promising for preventing acute thrombosis and promotes rapid endothelialization of artificial vascular grafts.

Preparation and evaluation of poly(ester-urethane) urea/gelatin nanofibers based on different crosslinking strategies for potential applications in vascular tissue engineering.

Due to the brittleness of gelatin, the resulting absence of mechanical performance restricts its applications in vascular tissue engineering. In this research, the fabrication of poly(ester-urethane) urea/gelatin (PU75) nanofibers via an electrospinning technique, followed by different crosslinking methods, resulted in the improvement of its mechanical properties. Poly(ester urethane) urea (PEUU) nanofibrous scaffolds and PU75-based nanofibrous scaffolds were characterized using scanning electron microscopy (SEM), attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, wide-angle X-ray diffraction (WAXRD), a mechanical properties test, a cytocompatibility assay, a hemolysis assay, and a histological analysis. Water contact angle (WCA) tests confirmed that the PU75-GA (PU75 nanofibers crosslinked with glutaraldehyde vapor) nanofibrous scaffold surfaces became more hydrophilic compared with other crosslinked nanofibrous scaffolds. The results show that the PU75-GA nanofibrous scaffold exhibited a combination of excellent mechanical properties, suitable pore diameters, hydrophilic properties, good cytocompatibility, and reliable hemocompatibility. Overall, PU75-GA nanofibers may be a potential scaffold for artificial blood vessel construction.