Utilization of PVPA and its effect on the material properties and biocompatibility of PVA electrospun membrane

This study aims to investigate on the effect of the addition of polyvinylphosphonic acid (PVPA) on the material properties and biocompatibility of a polyvinyl alcohol (PVA) polymer tissue engineering scaffold fabricated by electrospinning process. Stabilization of the membranes was carried out by heat application. Fourier transform infrared spectroscopy confirmed the presence of P?O bonds and P–OH bonds and increase in the atomic percentage of phosphorus in Energy Dispersive Spectroscopy (EDS) indicated successful incorporation of PVPA to the PVA. Scanning electron microscope fiber morphology and microstructure showed that addition of PVPA reduced the average fiber diameter of the scaffold. After 1 hr of phosphate buffered saline immersion, scaffolds were observed to swell to almost 200% of their original weight. Increased tensile strength was observed at 0.1% PVPA addition but reduced values were observed when the concentration was greater than 0.1%. Cytocompatibility was examined with M3CT3‐E1 preosteoblast cells through cell viability after exposure to extract solutions and Live/Dead cell staining. Cell proliferation was quantified by MTT assay; cell adhesion was visualized by scanning electron microscope and confocal microscopy images. Bone regeneration was also observed and quantified using histomorphometric analysis and histological staining methods after implantation in rat calvarium for 4 weeks. Copyright © 2013 John Wiley & Sons, Ltd.     

Reversibly Controlling Preferential Protein Adsorption on Bone Implants by Using an Applied Weak Potential as a Switch

A facile method is needed to control the protein adsorption onto biomaterials, such as, bone implants. Herein we doped taurocholic acid (TCA), an amphiphilic biomolecule, into an array of 1D nano‐architectured polypyrrole (NAPPy) on the implants. Doping TCA enabled the implant surface to show reversible wettability between 152° (superhydrophobic, switch‐on state) and 55° (hydrophilic, switch‐off state) in response to periodically switching two weak electrical potentials (+0.50 and ?0.80 V as a switch‐on and switch‐off potential, respectively). The potential‐switchable reversible wettability, arising from the potential‐tunable orientation of the hydrophobic and hydrophilic face of TCA, led to potential‐switchable preferential adsorption of proteins as well as cell adhesion and spreading. This potential‐switchable strategy may open up a new avenue to control the biological activities on the implant surface.     

ROS Scavenging Graphene-Based Hydrogel Enhances Type H Vessel Formation and Vascularized Bone Regeneration via ZEB1/Notch1 Mediation

The regeneration strategy for bone defects is greatly limited by the bone microenvironment, and excessive reactive oxygen species (ROS) seriously hinder the formation of new bone. Reduced graphene oxide (rGO) is expected to meet the requirements because of its ability to scavenge free radicals through electron transfer. Antioxidant hydrogels based on gelatine methacrylate (GM), acrylyl-β-cyclodextrin (Ac-CD), and rGO functionalized with β-cyclodextrin (β-CD) are developed for skull defect regeneration, but the mechanism of how rGO-based hydrogels enhance bone repair remains unclear. In this work, it is confirmed that the GM/Ac-CD/rGO hydrogel has good antioxidant capacity, and promotes osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and angiogenesis of human umbilical vein endothelial cells (HUVECs). The rGO-based hydrogel affects ZEB1/Notch1 to promote tube formation. Furthermore, two-photon laser scanning microscopy is used to observe the ROS in a skull defect. The rGO-based hydrogel promotes type H vessel formation in a skull defect. In conclusion, the hydrogel neutralizes ROS in the vicinity of a skull defect and stimulates ZEB1/Notch1 to promote the coupling of osteogenesis and angiogenesis, which may be a possible approach for bone regeneration.     

Modified-Hydroxyapatite-Chitosan Hybrid Composite Interfacial Coating on 3D Polymeric Scaffolds for Bone Tissue Engineering

Three dimensional (3D) scaffolds have huge limitations due to their low porosity, mechanical strength, and lack of direct cell-bioactive drug contact. Whereas bisphosphonate drug has the ability to stimulate osteogenesis in osteoblasts and bone marrow mesenchymal stem cells (hMSC) which attracted its therapeutic use. However it is hard administration low bioavailability, and lack of site-specificity, limiting its usage. The proposed scaffold architecture allows cells to access the bioactive surface at their apex by interacting at the scaffold's interfacial layer. The interface of 3D polycaprolactone (PCL) scaffolds has been coated with alendronate-modified hydroxyapatite (MALD) enclosed in a chitosan matrix, to mimic the native environment and stupulate the through interaction of cells to bioactive layer. Where the mechanical strength will be provided by the skeleton of PCL. In the MALD composite's hydroxyapatite (HAP) component will govern alendronate (ALD) release behavior, and HAP presence will drive the increase in local calcium ion concentration increases hMSC proliferation and differentiation. In results, MALD show release of 86.28 ± 0.22. XPS and SEM investigation of the scaffold structure, shows inspiring particle deposition with chitosan over the interface. All scaffolds enhanced cell adhesion, proliferation, and osteocyte differentiation for over a week without in vitro cell toxicity with 3.03 ± 0.2 kPa mechanical strength.     

Enhanced Protein Adsorption,Cell Attachment,and Neural Differentiation with the Help of Amine Functionalized Polycaprolactone Scaffolds

Today, neurodegenerative diseases are very common among people. As a result, researchers are investigating methods for treatment of these diseases. One therapeutic approach is differentiating stem cells into neural cells to replace damaged areas of the brain. Cell attachment is the first, necessary step for the process of differentiation. Hence, we tried to enhance cell adhesion and proliferation of bone marrow stem cells on poly(?-caprolactone) (PCL) scaffolds through modifying this substrate with amine functional groups. The presence of amine groups was confirmed by Fourier transform infrared spectrometry (FTIR). Protein adsorption was measured at 280 nm via UV-spectrometry. The proliferation of differentiated neurons was assessed by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (a dye) and cresyl violet staining. Finally, the morphology of differentiated neurons was shown by scanning electron microscopy (SEM). Results showed that amine modification of PCL scaffolds enhanced protein absorption and, consequently, cell adhesion and proliferation.     

Heterobifunctional Poly(ethylene glycol) Derivatives for the Surface Modification of Gold Nanoparticles Toward Bone Mineral Targeting

Heterobifunctional poly(ethylene glycol)s can be used for many biomedical applications ranging from solubility enhancement of hardly soluble compounds to surface modification of medical devices. In order to modify gold nanoparticles as model particles for drug targeting applications, PEG derivatives are synthesized that possess a high affinity for gold surfaces, namely a thioalkyl function, known to form stable monolayers on gold. Additionally a bisphosphonate function is introduced in the PEG molecule to allow targeting of hydroxyapatite rich tissues, like bone. Gold nanoparticles are modified using the synthesized bifunctional PEG and investigated for their stability in biological fluids and their ability to bind to hydroxyapatite granules in these fluids.

     

IL-17-deficient allogeneic bone marrow transplantation prevents the induction of collagen-induced arthritis in DBA/1J mice

IL-17-producing CD4⁺ T cells (Th17) play important functions in autoimmune diseases and allograft rejection of solid organs. We examined the effects of IL 17 and its mechanism of action on arthritis in a murine collagen-induced arthritis (CIA) model using bone marrow transplantation (BMT) system. DBA/1J mice were administered a lethal radiation dose and then rescued with bone marrow derived from either wild-type (WT) or IL-17^-/- mice on C57BL/6 background mice. CIA was induced after the bone marrow transplant, and disease progression was characterized. DBA/1J mice with CIA that received IL-17^-/- donor bone marrow showed potently inhibited development and severity of clinical arthritis as compared with CIA mice that received WT bone marrow. Reduced secretion of the pro-inflammatory cytokines tumor necrosis factor-α, IL-1β, and IL-6, and collagen-specific T cell responses were observed in mice that received IL-17^-/- bone marrow. IL-17 blockade also inhibited effector T cell proliferation by reciprocally regulating the Treg/Th17 ratio. IL-17 blockade prevented joint destruction in mice with CIA. These findings suggest that CIA with BMT is a viable method of immunological manipulation and that IL-17 deficiency suppresses severe joint destruction and inflammation in CIA mice. There may be clinical benefits in blocking IL-17 and BMT in the treatment of rheumatoid arthritis.     

环境及遗传因素对温州地区绝经女性骨质疏松影响的研究

为探讨温州地区绝经后女性骨质疏松症（Postmenopausal osteoporosis,PMOP）与雌激素受体基因（Estrogen receptor,ERa）多态性及相关因素的关联性,将203例观察对象按骨质疏松症诊断标准分为骨量正常组、骨量减少组与PMOP组,分别检测各组观察对象的腰椎L1~4正位骨密度、PvuⅡ、XbaⅠ基因型并调查其膳食行为与运动习惯等项目。结果显示：（1）年龄越大、文化程度越低、身材矮小及BMI低者、妊娠次数越多、产次越多者易罹患PMOP;经常饮用牛奶、豆浆、茶,经常食用水产品、豆类、肉类、蛋类、水果、坚果及每月运动次数较多者与高骨密度有关;在校正年龄、文化程度、BMI等干扰因素之后,饮茶、食用水果、坚果类的次数与BMD呈正相关。（2）该研究未发现PvuⅡ与XbaⅠ基因型与PMOP间有关联。提示该地区膳食行为、环境因素与PMOP关系密切,但雌激素受体α多态性与骨密度间关系尚不明确,其对骨质疏松的预测价值尚待深入研究。     

Promotion of cell affinity of porous PLLA scaffolds by immobilization of RGD peptides via plasma treatment

In the present work, RGDS (Arg-Gly-Asp-Ser) was immobilized on PLLA scaffolds with plasma treatment. The amount of immobilization, determined by HPLC, was confirmed to be in the effective order. Results from the culture of rat osteosarcoma (ROS), osteoblastic-like cells, demonstrate that the immobilization of RGDS could effectively enhance the attachment of ROS cells on PLLA and increase the cell density in PLLA scaffolds. In addition, experiments of in vitro mineralization indicate that there were more cells and mineralization focci in the RGDS-immobilized scaffolds, suggesting a tendency to form bone-like tissues, compared with the unmodified PLLA scaffold. On the other hand, the PLLA scaffolds immobilized with RGES (Arg-Gly-Glu-Ser) were much less effective in promotion of ROS attachment, suggesting that the enhancement on cell attachment was mainly due to the recognition of RGDS by the adhesion receptors on the cell membrane. The results presented in this work demonstrate that RGDS could be successfully immobilized on PLLA scaffolds with plasma treatment and such modification can make PLLA scaffolds more suitable for culture of osteoblast-like cells and for generation of bone-like tissues.     

Effective PCR detection of animal species in highly processed animal byproducts and compound feeds

In this paper we present a polymerase chain reaction (PCR)-based method for detecting meat and bone meal (MBM) in compound feedingstuffs. By choosing adequate DNA targets from an appropriate localisation in the genome, the real-time PCR method developed here proved to be robust to severe heat treatment of the MBM, showing high sensitivity in the detection of MBM. The method developed here permits the specific detection of processed pig and cattle materials treated at 134 °C in various feed matrices down to a limit of detection of about 0.1%. This technique has also been successfully applied to well-characterised MBM samples heated to as high as 141 °C, as well as to various blind feed samples with very low MBM contents. Finally, the method also passed several official European ring trials.