Development of Graphene-Based Materials in Bone Tissue Engineaering.

Bone regeneration-related graphene-based materials (bGBMs) are increasingly attracting attention in tissue engineering due to their special physical and chemical properties. The purpose of this review is to quantitatively analyze mass academic literature in the field of bGBMs through scientometrics software CiteSpace, to demonstrate the rules and trends of bGBMs, thus to analyze and summarize the mechanisms behind the rules, and to provide clues for future research. First, the research status, hotspots, and frontiers of bGBMs are analyzed in an intuitively and vividly visualized way. Next, the extracted important subjects such as fabrication techniques, cytotoxicity, biodegradability, and osteoinductivity of bGBMs are presented, and the different mechanisms, in turn, are also discussed. Finally, photothermal therapy, which is considered an emerging area of application of bGBMs, is also presented. Based on this approach, this work finds that different studies report differing opinions on the biological properties of bGBMS due to the lack of consistency of GBMs preparation. Therefore, it is necessary to establish more standards in fabrication, characterization, and testing for bGBMs to further promote scientific progress and clinical translation.

Coaxial-printed small-diameter polyelectrolyte-based tubes with an electrostatic self-assembly of heparin and YIGSR peptide for antithrombogenicity and endothelialization.

Low patency ratio of small-diameter vascular grafts remains a major challenge due to the occurrence of thrombosis formation and intimal hyperplasia after transplantation. Although developing the functional coating with release of bioactive molecules on the surface of small-diameter vascular grafts are reported as an effective strategy to improve their patency ratios, it is still difficult for current functional coatings cooperating with spatiotemporal control of bioactive molecules release to mimic the sequential requirements for antithrombogenicity and endothelialization. Herein, on basis of 3D-printed polyelectrolyte-based vascular grafts, a biologically inspired release system with sequential release in spatiotemporal coordination of dual molecules through an electrostatic self-assembly was first described. A series of tubes with tunable diameters were initially fabricated by a coaxial extrusion printing method with customized nozzles, in which a polyelectrolyte ink containing of ε-polylysine and sodium alginate was used. Further, dual bioactive molecules, heparin with negative charges and Tyr-Ile-Gly-Ser-Arg (YIGSR) peptide with positive charges were layer-by-layer assembled onto the surface of these 3D-printed tubes. Due to the electrostatic interaction, the sequential release of heparin and YIGSR was demonstrated and could construct a dynamic microenvironment that was thus conducive to the antithrombogenicity and endothelialization. This study opens a new avenue to fabricate a small-diameter vascular graft with a biologically inspired release system based on electrostatic interaction, revealing a huge potential for development of small-diameter artificial vascular grafts with good patency.

Temperature-programmable and enzymatically solidifiable gelatin-based bioinks enable facile extrusion bioprinting.

The development of exceptional bioinks with excellent printability, high fidelity, and excellent cell viability maintenance for extrusion bioprinting remains a major challenge. Gelatin is an ideal candidate bioink due to its biocompatibility, biodegradability, and non-immunogenicity. However, its inherently low viscosity and unstable physical gelation under physiological conditions make it unsuitable for direct extrusion bioprinting of tissue-like gelatin constructs with high fidelity. Herein, sequential chemical modification using reversible quadruple-hydrogen-bonded ureido-pyrimidinone (UPy) and enzyme-responsive tyramine moieties (Tyr) were devloped to endow the gelatin with a temperature-programmable viscosity and enzyme-controlled solidification, thus realizing enhanced printability and superior fidelity. As demonstrated in a proof-of-concept study, various cell-laden constructs were built based on our modified gelatin, including two-dimensional human bone marrow mesenchymal stem cell (hBMSC)-laden patterns, three-dimensional interconnected hBMSC-laden scaffolds, a reversible twisting-tension human-scale hBMSC-laden ear, a bicellular tibia-like construct containing hBMSCs and endothelial cells and a hexagonal prism-shaped hepatocyte-laden scaffold. The loaded cells in the construct have high viability of over 90% at 24 h, and show proliferation and protein secretion over one week, suggesting that Gel-UPy-Tyr-based constructs under physiological temperature not only can keep high fidelity, but also can support the growth and functions of the loaded cells.

Simulated digestion and fermentation in vitro by human gut microbiota of polysaccharides from Helicteres angustifolia L.

Helicteres angustifolia L. is a familiar herbal plant, which exhibits various bioactivities with potential benefits for human health. In this study, the simulated digestion (saliva, simulated gastric and small intestinal conditions) and fermentation in vitro of polysaccharides from H. angustifolia L. (HaLPs) were evaluated, and the effects of HaLPs on gut microbiota were examined using high-throughput sequencing technology. The results indicated that the simulated gastrointestinal digestion had no effect on HaLPs, so HaLPs could reach the large intestine safely. However, the molecular weight of HaLPs and the reducing sugar decreased significantly after fermentation under anaerobic conditions. It was found that HaLPs had the significantly alternation effect on the composition of the gut microbiota. Furthermore, the total short-chain fatty acids content increased significantly after fermentation (from 2.25 ±â€¯0.13 mM to 22.45 ±â€¯4.56 mM). These results provided an understanding of the digestive characteristics of HaLPs and afforded a viable basis for their development.

3D-Bioprinted Osteoblast-Laden Nanocomposite Hydrogel Constructs with Induced Microenvironments Promote Cell Viability, Differentiation, and Osteogenesis both In Vitro and In Vivo.

An osteoblast-laden nanocomposite hydrogel construct, based on polyethylene glycol diacrylate (PEGDA)/laponite XLG nanoclay ([Mg5.34Li0.66Si8O20(OH)4]Na0.66, clay)/hyaluronic acid sodium salt (HA) bio-inks, is developed by a two-channel 3D bioprinting method. The novel biodegradable bio-ink A, comprised of a poly(ethylene glycol) (PEG)-clay nanocomposite crosslinked hydrogel, is used to facilitate 3D-bioprinting and enables the efficient delivery of oxygen and nutrients to growing cells. HA with encapsulated primary rat osteoblasts (ROBs) is applied as bio-ink B with a view to improving cell viability, distribution uniformity, and deposition efficiency. The cell-laden PEG-clay constructs not only encapsulated osteoblasts with more than 95% viability in the short term but also exhibited excellent osteogenic ability in the long term, due to the release of bioactive ions (magnesium ions, Mg2+ and silicon ions, Si4+), which induces the suitable microenvironment to promote the differentiation of the loaded exogenous ROBs, both in vitro and in vivo. This 3D-bioprinting method holds much promise for bone tissue regeneration in terms of cell engraftment, survival, and ultimately long-term function.

Purification and analysis of the composition and antioxidant activity of polysaccharides from Helicteres angustifolia L.

The antioxidant activity of polysaccharides has attracted tremendous research interest in recent years. In this study, we aimed to investigate the antioxidant properties of Helicteres angustifolia L. polysaccharides (HALP) that was acquired by water extraction and alcohol precipitation using ultrasound-assisted method with decolorization and protein removal. The polysaccharides were then successively purified using chromatography on DEAE-cellulose and Sephadex S-300 columns with two major fractions: HALPs1-1 and HALPs2-1. The physicochemical properties, structural characterization, and antioxidant activities of these fractions were investigated. The results indicated that HALPs1-1 was a glucan with an average molecular weight of 151.70kDa, and HALPs2-1 was composed of glucuronic acid with an average molecular weight of 114.81kDa. Infrared spectroscopies and iodine potassium iodide tests showed that the HALP were acid α-pyranoses. The antioxidant activities of the polysaccharide fractions (HALP, HALPs1-1, and HALPs2-1) were evaluated in vitro. The results suggested that HALPs2-1 had the highest scavenging activities for ABTS, hydroxyl and DPPH radicals than other polysaccharides. Taken together, the results of this study suggested that polysaccharides from Helicteres angustifolia L. could be used as novel potential antioxidants.

Strontium incorporation improves the bone-forming ability of scaffolds derived from porcine bone.

Although heterogeneous bone scaffolds have shown potential in bone defect repair, their capability of aiding bone regeneration need to be further enhanced. Strontium, one important trace element in bone, has a well-known favorable effect on bone repair. Here a strontium containing scaffold (CPB/PCL/Sr) based on superficially porous calcined porcine bone (CPB) was obtained straightforwardly by sequential coating of SrCl2 and polycaprolactone (PCL). The basic characterization revealed that PCL coating could simultaneously improve the mechanical properties and, more importantly, restrain strontium release. Moreover, in vitro behaviors of human MSCs on CPB, CPB/PCL, and CPB/PCL/Sr were studied in detail. The comprehensive results of proliferation, osteogenic gene expression, ALP staining, and ALP activity demonstrated that PCL coating slightly impaired the bone repair potential of CPB. In contrast, CPB/PCL/Sr better supported the osteogenic differentiation of MSCs than CPB，highlighting the role of strontium. The in vivo test confirmed a better new bone formation of CPB/PCL/Sr than CPB and CPB/PCL. These results verified the superiority of incorporating strontium to improve the bone-forming ability of CPB, offering a promising alternative for bone defect repair.

Antidiabetic activity of Helicteres angustifolia root.

Context The root of Helicteres angustifolia L. (Sterculiaceae) has been used as folk herbal drug to treat cancer, bacterial infections, inflammatory, and flu in China. However, there is no report on its antidiabetic activity. Objective This study evaluates the antidiabetic activity of ethanol extract from H. angustifolia root. Materials and methods The promoting effect of H. angustifolia root ethanol extract (25, 50, and 100 µg/mL) on glucose uptake was evaluated using HepG2 cell, differentiated C2C12 myotubes, and differentiated 3T3-L1 adipocytes. The antidiabetic activity of the extract was assessed in vivo using STZ-induced diabetic rats by orally administration of the extract (200 and 400 mg/kg b.w.) once per day for 28 d. Blood glucose, TG, TC, TP, HDL-C, UA, BUN, AST, ALT, insulin, and HOMA-IR were analyzed. Results The results showed that the extract increased glucose uptake in C2C12 myotubes and 3T3-L1 adipocytes with an IC50 value of 79.95 and 135.96 µg/mL, respectively. And about 12%, 19%, and 10% (p < 0.05) in HepG2 cells when compared with the control at the concentration of 25, 50, and 100 µg/mL, respectively. After 28 days' treatment with the extract, significant reduction was observed in blood glucose, HOMA-IR, TC, TG, UA, BUN, AST, and ALT levels, while the levels of TP and HDL cholesterol increased. Discussion and conclusion These results suggest that H. angustifolia root ethanol extract possess potent antidiabetic activity, which is the first report on antidiabetic activity of this plant.

Bottom-up topography assembly into 3D porous scaffold to mediate cell activities.

Native cells live in a three-dimensional (3D) extracellular matrix (ECM) capable of regulating cell activities through various physical and chemical factors. Designed topographies have been well proven to trigger significant difference in cell behaviours. However, present topographies are almost all constructed on two-dimensional (2D) substrates like discs and films, which are far from features like 3D and porosity required in application like bone repair. Here we bottom-up assembled poly(lactic-co-glycolic acid)/calcium carbonate (PLGA/CC) microspheres with superficial porous topography intactly into a 3D porous scaffold. Because the scaffold was obtained through a mild technique, the bioactivity of released BMP-2 was well retained. Mouse bone marrow mesenchymal stem cells (mMSCs) were cultured on produced scaffolds having different 3D topographies. It turned out that osteogenic differentiation of mMSCs did respond to the 3D topographies, while proliferation didn't. Gene expression of αv and ß1 integrins revealed that adhesion was supposed to be the underlying mechanism for osteogenic response. The study provides insight into enhancing function of practical scaffolds by elaborate topography design. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1056-1063, 2016.

Evaluation of anti-hyperglycemic effect of Actinidia kolomikta (Maxim. etRur.) Maxim. root extract.

This study aimed to evaluate the anti-hyperglycemic effect of ethanol extract from Actinidia kolomikta (Maxim. etRur.) Maxim. root (AKE).An in vitro evaluation was performed by using rat intestinal α-glucosidase (maltase and sucrase), the key enzymes linked with type 2 diabetes. And an in vivo evaluation was also performed by loading maltose, sucrose, glucose to normal rats. As a result, AKE showed concentration-dependent inhibition effects on rat intestinal maltase and rat intestinal sucrase with IC(50) values of 1.83 and 1.03mg/mL, respectively. In normal rats, after loaded with maltose, sucrose and glucose, administration of AKE significantly reduced postprandial hyperglycemia, which is similar to acarbose used as an anti-diabetic drug. High contents of total phenolics (80.49 ± 0.05mg GAE/g extract) and total flavonoids (430.69 ± 0.91mg RE/g extract) were detected in AKE. In conclusion, AKE possessed anti-hyperglycemic effects and the possible mechanisms were associated with its inhibition on α-glucosidase and the improvement on insulin release and/or insulin sensitivity as well. The anti-hyperglycemic activity possessed by AKE maybe attributable to its high contents of phenolic and flavonoid compounds.

Engineering poly(lactic-co-glycolic acid)/calcium carbonate microspheres with controllable topography and their cell response.

Polymeric porous microspheres can be used as functional vehicles in drug delivery and cell culture. However, the conventional porous microspheres are somewhat complicated with respect to preparation, and limited in composition and controllability. Here we report the preparation of poly(lactic-co-glycolic acid)/calcium carbonate (PLGA/CC) composite microspheres with uniform superficial macropores through a facile single-step method, where CC acts simultaneously as in situ pore-forming agent and reinforcing phase. The SEM images and quantified results from mercury intrusion porosimetry indicated that the size and density of the superficial macropores were highly controllable via changing the starting parameters such as size and content of CC particles and concentration of PLGA. Mouse bone mesenchymal stem cells were cultured on microspheres of different topographies to investigate the cell-substrate interaction. The results showed that cells adhered and grew well on all microspheres, while the topography with smaller and more discrete macropores exhibited the highest proliferation, indicating that cells responded to the topography of the microsphere. This work provides a novel approach to obtain diverse porous composite microspheres designed for tissue repair and study of cell-substrate interaction.