Integrated oral microgel system ameliorates renal fibrosis by hitchhiking co-delivery and targeted gut flora modulation.

BACKGROUND: Renal fibrosis is a progressive process associated with chronic kidney disease (CKD), contributing to impaired kidney function. Active constituents in traditional Chinese herbs, such as emodin (EMO) and asiatic acid (AA), exhibit potent anti-fibrotic properties. However, the oral administration of EMO and AA results in low bioavailability and limited kidney accumulation. Additionally, while oral probiotics have been accepted for CKD treatment through gut microbiota modulation, a significant challenge lies in ensuring their viability upon administration. Therefore, our study aims to address both renal fibrosis and gut microbiota imbalance through innovative co-delivery strategies. RESULTS: In this study, we developed yeast cell wall particles (YCWPs) encapsulating EMO and AA self-assembled nanoparticles (NPYs) and embedded them, along with Lactobacillus casei Zhang, in chitosan/sodium alginate (CS/SA) microgels. The developed microgels showed significant controlled release properties for the loaded NPYs and prolonged the retention time of Lactobacillus casei Zhang (L. casei Zhang) in the intestine. Furthermore, in vivo biodistribution showed that the microgel-carried NPYs significantly accumulated in the obstructed kidneys of rats, thereby substantially increasing the accumulation of EMO and AA in the impaired kidneys. More importantly, through hitchhiking delivery based on yeast cell wall and positive modulation of gut microbiota, our microgels with this synergistic strategy of therapeutic and modulatory interactions could regulate the TGF-ß/Smad signaling pathway and thus effectively ameliorate renal fibrosis in unilateral ureteral obstruction (UUO) rats. CONCLUSION: In conclusion, our work provides a new strategy for the treatment of renal fibrosis based on hitchhiking co-delivery of nanodrugs and probiotics to achieve synergistic effects of disease treatment and targeted gut flora modulation.

An Intrinsically Magnetic Epicardial Patch for Rapid Vascular Reconstruction and Drug Delivery.

Myocardial infarction (MI) is a major cause of mortality worldwide. The major limitation of regenerative therapy for MI is poor cardiac retention of therapeutics, which results from an inefficient vascular network and poor targeting ability. In this study, a two-layer intrinsically magnetic epicardial patch (MagPatch) prepared by 3D printing with biocompatible materials like poly (glycerol sebacate) (PGS) is designed, poly (ε-caprolactone) (PCL), and NdFeB. The two-layer structure ensured that the MagPatch multifariously utilized the magnetic force for rapid vascular reconstruction and targeted drug delivery. MagPatch accumulates superparamagnetic iron oxide (SPION)-labelled endothelial cells, instantly forming a ready-implanted organization, and rapidly reconstructs a vascular network anastomosed with the host. In addition, the prefabricated vascular network within the MagPatch allowed for the efficient accumulation of SPION-labelled therapeutics, amplifying the therapeutic effects of cardiac repair. This study defined an extendable therapeutic platform for vascularization-based targeted drug delivery that is expected to assist in the progress of regenerative therapies in clinical applications.

Self-healing polyurethane-elastomer with mechanical tunability for multiple biomedical applications in vivo.

The unique properties of self-healing materials hold great potential in the field of biomedical engineering. Although previous studies have focused on the design and synthesis of self-healing materials, their application in in vivo settings remains limited. Here, we design a series of biodegradable and biocompatible self-healing elastomers (SHEs) with tunable mechanical properties, and apply them to various disease models in vivo, in order to test their reparative potential in multiple tissues and at physiological conditions. We validate the effectiveness of SHEs as promising therapies for aortic aneurysm, nerve coaptation and bone immobilization in three animal models. The data presented here support the translation potential of SHEs in diverse settings, and pave the way for the development of self-healing materials in clinical contexts.

A perfusable, multifunctional epicardial device improves cardiac function and tissue repair.

Despite advances in technologies for cardiac repair after myocardial infarction (MI), new integrated therapeutic approaches still need to be developed. In this study, we designed a perfusable, multifunctional epicardial device (PerMed) consisting of a biodegradable elastic patch (BEP), permeable hierarchical microchannel networks (PHMs) and a system to enable delivery of therapeutic agents from a subcutaneously implanted pump. After its implantation into the epicardium, the BEP is designed to provide mechanical cues for ventricular remodeling, and the PHMs are designed to facilitate angiogenesis and allow for infiltration of reparative cells. In a rat model of MI, implantation of the PerMed improved ventricular function. When connected to a pump, the PerMed enabled targeted, sustained and stable release of platelet-derived growth factor-BB, amplifying the efficacy of cardiac repair as compared to the device without a pump. We also demonstrated the feasibility of minimally invasive surgical PerMed implantation in pigs, demonstrating its promise for clinical translation to treat heart disease.

Injectable Citrate-Based Hydrogel as an Angiogenic Biomaterial Improves Cardiac Repair after Myocardial Infarction.

Implanted medical biomaterials are closely in contact with host biological systems via biomaterial-cell/tissue interactions, and these interactions play pivotal roles in regulating cell functions and tissue regeneration. However, many biomaterials degrade over time, and these degradation products also have been shown to interact with host cells/tissue. Therefore, it may prove useful to specifically design implanted biomaterials with degradation products which greatly improve the performance of the implant. Herein, we report an injectable, citrate-containing polyester hydrogel which can release citrate as a cell regulator via hydrogel degradation and simultaneously show sustained release of an encapsulated growth factor Mydgf. By coupling the therapeutic effect of the hydrogel degradation product (citrate) with encapsulated Mydgf, we observed improved postmyocardial infarction (MI) heart repair in a rat MI model. Intramyocardial injection of our Mydgf-loaded citrate-containing hydrogel was shown to significantly reduce scar formation and infarct size, increase wall thickness and neovascularization, and improve heart function. This bioactive injectable hydrogel-mediated combinatorial approach offers myriad advantages including potential adjustment of delivery rate and duration, improved therapeutic effect, and minimally invasive administration. Our rational design combining beneficial degradation product and controlled release of therapeutics provides inspiration toward the next generation of biomaterials aiming to revolutionize regenerative medicine.

Elastic 3D-Printed Hybrid Polymeric Scaffold Improves Cardiac Remodeling after Myocardial Infarction.

Myocardial remodeling, including ventricular dilation and wall thinning, is an important pathological process caused by myocardial infarction (MI). To intervene in this pathological process, a new type of cardiac scaffold composed of a thermoset (poly-[glycerol sebacate], PGS) and a thermoplastic (poly-[ε-caprolactone], PCL) is directly printed by employing fused deposition modeling 3D-printing technology. The PGS-PCL scaffold possesses stacked construction with regular crisscrossed filaments and interconnected micropores and exhibits superior mechanical properties. In vitro studies demonstrate favorable biodegradability and biocompatibility of the PGS-PCL scaffold. When implanted onto the infarcted myocardium, this scaffold improves and preserves heart function. Furthermore, the scaffold improves several vital aspects of myocardial remodeling. On the morphological level, the scaffold reduces ventricular wall thinning and attenuated infarct size, and on the cellular level, it enhances vascular density and increases M2 macrophage infiltration, which might further contribute to the mitigated myocardial apoptosis rate. Moreover, the flexible PGS-PCL scaffold can be tailored to any desired shape, showing promise for annular-shaped restraint device application and meeting the demands for minimal invasive operation. Overall, this study demonstrates the therapeutic effects and versatile applications of a novel 3D-printed, biodegradable and biocompatible cardiac scaffold, which represents a promising strategy for improving myocardial remodeling after MI.

[Experimental study on mechanical properties of decellularized porcine aortic valve and effects of precoating methods of biological scaffold on histocompatibility].

OBJECTIVE: To observe the mechanical properties of decellularized porcine aortic valve, and to explore the effects of precoating methods of biological scaffold on histocompatibility. METHODS: Fresh porcine aortic valves were decellularized using trypsin, TritonX-100 and nuclease. Treated valves were evaluated by light microscopy, scanning electron microscopy (SEM) and mechanical test. Three groups of scaffold were precoated with phosphate buffered saline (PBS), poly-L-lysine (PLL) and fetal bovine serum (FBS) respectively. Myofibroblasts were seeded onto each scaffold. Light and electron microscopic observation was performed and MTT test was used to examine efficiency of cell attachment. RESULTS: HE stain and SEM showed that cells were almost absent in the treated leaflet. The wave-like collagen together with the whole three-dimensional structure was maintained. Compared with normal valves, the Max-load, Max-stress and elastic modulus decreased while the Max-strain increased (P<0.05). The result of MTT test showed more cells were attached on the valves treated with FBS compared to the other two groups. Histological investigations also confirm that the high degree of cell attachment on the valves precoated with FBS (F=129.26, P=0.000). CONCLUSIONS: Enzyme combined with detergent and nucleases can remove cells from porcine aortic valves. Meanwhile the mechanical properties of these valves may be altered. Precoating porcine aortic valve with FBS is an effective method to improve cell attachment, growth and increasing.

Spatiotemporal delivery of nanoformulated liraglutide for cardiac regeneration after myocardial infarction.

The local, intramyocardial injection of proteins into the infarcted heart is an attractive option to initiate cardiac regeneration after myocardial infarction (MI). Liraglutide, which was developed as a treatment for type 2 diabetes, has been implicated as one of the most promising protein candidates in cardiac regeneration. A significant challenge to the therapeutic use of this protein is its short half-life in vivo. In this study, we evaluated the therapeutic effects and long-term retention of liraglutide loaded in poly(lactic-co-glycolic acid)-poly(ethylene glycol) (PLGA-PEG) nanoparticles (NP-liraglutide) on experimental MI. PLGA-PEG nanoparticles (NPs) have been shown to efficiently load liraglutide and release bioactive liraglutide in a sustained manner. For in vitro test, the released liraglutide retained bioactivity, as measured by its ability to activate liraglutide signaling pathways. Next, we compared the effects of an intramyocardial injection of saline, empty NPs, free liraglutide and NP-liraglutide in a rat model of MI. NPs were detected in the myocardium for up to 4 weeks. More importantly, an intramyocardial injection of NP-liraglutide was sufficient to improve cardiac function (P<0.05), attenuate the infarct size (P<0.05), preserve wall thickness (P<0.05), promote angiogenesis (P<0.05) and prevent cardiomyocyte apoptosis (P<0.05) at 4 weeks after injection without affecting glucose levels. The local, controlled, intramyocardial delivery of NP-liraglutide represents an effective and promising strategy for the treatment of MI.

Hybrid small-diameter vascular grafts: Anti-expansion effect of electrospun poly ε-caprolactone on heparin-coated decellularized matrices.

Small-diameter vascular grafts (SDVGs) (D < 6 mm) are increasingly needed in clinical settings for cardiovascular disease, including coronary artery and peripheral vascular pathologies. Vessels made from synthetic polymers have shortcomings such as thrombosis, intimal hyperplasia, calcification, chronic inflammation and no growth potential. Decellularized xenografts are commonly used as a tissue-engineering substitute for vascular reconstructive procedures. Although acellular allogeneic vascular grafts have good histocompatibility and antithrombotic properties, the decellularization process may damage the biomechanics and accelerate the elastin deformation and degradation, finally resulting in vascular graft expansion and even aneurysm formation. Here, to address these problems, we combine synthetic polymers with natural decellularized small-diameter vessels to fabricate hybrid tissue-engineered vascular grafts (HTEV). The donor aortic vessels were decellularized with a combination of different detergents and dehydrated under a vacuum freeze-drying process. Polycaprolactone (PCL) nanofibers were electrospun (ES) outside the acellular aortic vascular grafts to strengthen the decellularized matrix. The intimal surfaces of the hybrid small-diameter vascular grafts were coated with heparin before the allograft transplantation. Histopathology and scanning electron microscope revealed that the media of the decellularized vessels were severely injured. Mechanical testing of scaffolds showed that ES-PCL significantly enhanced the biomechanics of decellularized vessels. Vascular ultrasound and micro-CT angiography showed that all grafts after implantation in a rat model were satisfactorily patent for up to 6 weeks. ES-PCL successfully prevented the occurrence of vasodilation and aneurysm formation after transplantation and reduced the cell inflammatory infiltration. In conclusion, the HTEV with perfect histocompatibility and biomechanics provide a facile and useful technique for the development of SDVGs.

Scalable units for building cardiac tissue.

Scalable units for building cardiac tissue are fabricated from biodegradable elastomeric polymers by pairwise stacking of heart-cell scaffolds with sinusoidal internal pore architectures and dedicated perfusable microvessels with rapidly degrading porous interfaces in a parallel flow configuration. This platform supports viable heart cells in vitro and, if validated in vivo, may aid in the regenerative repair of vascularized tissues.

3D structural patterns in scalable, elastomeric scaffolds guide engineered tissue architecture.

Microfabricated elastomeric scaffolds with 3D structural patterns are created by semiautomated layer-by-layer assembly of planar polymer sheets with through-pores. The mesoscale interconnected pore architectures governed by the relative alignment of layers are shown to direct cell and muscle-like fiber orientation in both skeletal and cardiac muscle, enabling scale up of tissue constructs towards clinically relevant dimensions.

Lignocellulosic hydrolysates and extracellular electron shuttles for H2 production using co-culture fermentation with Clostridium beijerinckii and Geobacter metallireducens.

A co-culture of Clostridium beijerinckii and Geobacter metallireducens with AH2QDS produced hydrogen from lignocellulosic hydrolysates (biomass of Miscanthus prepared by hydrothermal treatment with dilute acids). This co-culture system enhanced hydrogen production from lignocellulosic hydrolysates by improving substrate utilization and diminishing acetate accumulation, despite the presence of fermentation inhibitors in the hydrolysates. The improvements were greater for xylose-rich hydrolysates. The increase in maximum cumulative hydrogen production for hydrolysates with glucose:xylose mass ratios of 1:0.2, 1:1 and 1:10 g/g was 0%, 22% and 11%, respectively. Alternative extracellular electron shuttles (EES), including indigo dye, juglone, lawsone, fulvic acids and humic acids, were able to substitute for AH2QDS, improving hydrogen production in the co-culture system using xylose as model substrate. Increased utilization of xylose-rich hydrolysates and substitution of alternative EES make the co-culture with EES system a more attractive strategy for industrial biohydrogen production.

The effect of Heparin-VEGF multilayer on the biocompatibility of decellularized aortic valve with platelet and endothelial progenitor cells.

The application of polyelectrolyte multilayer films is a new, versatile approach to surface modification of decellularized tissue, which has the potential to greatly enhance the functionality of engineered tissue constructs derived from decellularized organs. In the present study, we test the hypothesis that Heparin- vascular endothelial growth factor (VEGF) multilayer film can not only act as an antithrombotic coating reagent, but also induce proliferation of endothelial progenitor cells (EPCs) on the decellularized aortic heart valve. SEM demonstrated the adhesion and geometric deformation of platelets. The quantitative assay of platelet activation was determined by measuring the production of soluble P-selectin. Binding and subsequent release of heparin and VEGF from valve leaflets were assessed qualitatively by laser confocal scanning microscopy and quantitatively by ELISA methods. Human blood derived EPCs were cultured and the adhesion and growth of EPCs on the surface modified valvular scaffolds were assessed. The results showed that Heparin-VEGF multilayer film improved decellularized valve haemocompatibility with respect to a substantial reduction of platelet adhesion. Release of VEGF from the decellularized heart valve leaflets at physiological conditions was sustained over 5 days. In vitro biological tests demonstrated that EPCs achieved better adhesion, proliferation and migration on the coatings with Heparin-VEGF multilayer film. Combined, these results indicate that Heparin-VEGF multilayer film could be used to cover the decellularized porcine aortic valve to decrease platelet adhesion while exhibiting excellent EPCs biocompatibility.

A biodegradable microvessel scaffold as a framework to enable vascular support of engineered tissues.

A biodegradable microvessel scaffold comprised of distinct parenchymal and vascular compartments separated by a permeable membrane interface was conceptualized, fabricated, cellularized, and implanted. The device was designed with perfusable microfluidic channels on the order of 100 µm to mimic small blood vessels, and high interfacial area to an adjacent parenchymal space to enable transport between the compartments. Poly(glycerol sebacate) (PGS) elastomer was used to construct the microvessel framework, and various assembly methods were evaluated to ensure robust mechanical integrity. In vitro studies demonstrated the differentiation of human skeletal muscle cells cultured in the parenchymal space, a 90% reduction in muscle cell viability due to trans-membrane transport of a myotoxic drug from the perfusate, and microvessel seeding with human endothelial cells. In vivo studies of scaffolds implanted subcutaneously and intraperitoneally, without or with exogenous cells, into nude rats demonstrated biodegradation of the membrane interface and host blood cell infiltration of the microvessels. This modular, implantable scaffold could serve as a basis for building tissue constructs of increasing scale and clinical relevance.

Polyelectrolyte multilayer film on decellularized porcine aortic valve can reduce the adhesion of blood cells without affecting the growth of human circulating progenitor cells.

Polyelectrolyte multilayer film modification could be an effective method to reduce the immunological and inflammatory response of the xenogeneic scaffold in vivo, and may also be applied to tissue-engineered heart valve in contact with blood. The objectives of this study are to test heparin-chitosan multilayer film as an antithrombotic coating reagent for decellularized aortic heart valve and the biocompatibility of the modified valvular surface. The adhesion and geometric deformation of platelets were demonstrated by scanning electron microscopy. The quantitative assay of platelet activation was determined by measuring the production of soluble P-selectin. Moreover, the leukocytes' adhesion, erythrocyte hemolysis, and whole blood clotting time studies were performed to gain information on the hemocompatibility of this biomaterial. Human-blood-derived endothelial progenitor cells (EPCs) were cultured and the adhesion and growth of EPCs on the surface-modified PDAV were assessed. The results showed that heparin-chitosan multilayer film could be coated on the decellularized valvular scaffolds, and improved their hemocompatibility with respect to a substantial reduction of platelet adhesion and activation. The modified valve also significantly reduced leukocytes adhesion, erythrocyte hemolysis, and whole blood clotting time. Seeding with EPCs achieved a confluent monolayer on the surface of the decellularized matrix. The in vitro studies performed in this work suggest that it may be reasonable to use heparin-chitosan multilayer film as a means of surface modification to improve the blood compatibility of decellularized valvular scaffold.

Voltage-driven translocation of DNA through a high throughput conical solid-state nanopore.

Nanopores have become an important tool for molecule detection at single molecular level. With the development of fabrication technology, synthesized solid-state membranes are promising candidate substrates in respect of their exceptional robustness and controllable size and shape. Here, a 30-60 (tip-base) nm conical nanopore fabricated in 100 nm thick silicon nitride (Si(3)N(4)) membrane by focused ion beam (FIB) has been employed for the analysis of λ-DNA translocations at different voltage biases from 200 to 450 mV. The distributions of translocation time and current blockage, as well as the events frequencies as a function of voltage are investigated. Similar to previously published work, the presence and configurations of λ-DNA molecules are characterized, also, we find that greater applied voltages markedly increase the events rate, and stretch the coiled λ-DNA molecules into linear form. However, compared to 6-30 nm ultrathin solid-state nanopores, a threshold voltage of 181 mV is found to be necessary to drive DNA molecules through the nanopore due to conical shape and length of the pore. The speed is slowed down ∼5 times, while the capture radius is ∼2 fold larger. The results show that the large nanopore in thick membrane with an improved stability and throughput also has the ability to detect the molecules at a single molecular level, as well as slows down the velocity of molecules passing through the pore. This work will provide more motivations for the development of nanopores as a Multi-functional sensor for a wide range of biopolymers and nano materials.

Enhancement of mesenchymal stem cell attachment to decellularized porcine aortic valve scaffold by in vitro coating with antibody against CD90: a preliminary study on antibody-modified tissue-engineered heart valve.

The importance of cell adhesion to the scaffold in the tissue-engineered heart valve remains to be determined. The current study examined the feasibility of conjugating antibody against CD90 to a decellularized porcine aortic valve scaffold and binding mesenchymal stem cells to that scaffold through interaction with a cell surface antigen. After decellularization, the porcine aortic valve was reacted with biotin, avidin, and biotinylated anti-rat CD90 antibody sequentially and inserted into a laminar flow system used to test the effect of laminar shear stress. Rat bone mesenchymal stem cells (BMSC) were injected and circulated in a flow system to study the ability of anti-CD90 antibody to trap and immobilize cells on the valve surface. The results demonstrated that anti-CD90 antibody on the valve surface remains bound, even under high shear conditions. Compared with the control valve (no antibody), the modified (antibody-coated) valve immobilized significantly more rat BMSC (p < 0.05). Thus, the avidin-biotin system can be used to attach anti-CD90 antibody to these valves, and the bound antibody can immobilize rat BMSC in a flow chamber, suggesting that antibody-modified scaffolds might be used to fabricate shear stress-resistant, tissue-engineered heart valves.