Nanofibrous gelatine scaffolds integrated with nerve growth factor‐loaded alginate microspheres for brain tissue engineering

Neural regeneration research is designed in part to develop strategies for therapy after nerve damage due to injury or disease. In this study, a new gelatine‐based biomimetic scaffold was fabricated for brain tissue engineering applications. A technique combining thermally induced phase separation and porogen leaching was used to create interconnected macropores and nanofibrous structure. To promote tissue regeneration processes, the scaffolds were integrated with nerve growth factor (NGF)‐loaded alginate microspheres. The results showed that nanofibrous matrix could only be obtained when gelatine concentration was at least 7.5% (w/v). The scaffold with a modulus value (1.2 kPa) similar to that of brain tissue (0.5–1 kPa) was obtained by optimizing the heat treatment time, macropore size and gelatine concentration. The encapsulation efficiencies of NGF into 0.1% and 1% alginate microspheres were 85% and 100%, respectively. The release rate of NGF from the microspheres was controlled by the alginate concentration and the poly(L‐lysine) coating. The immobilization of the microspheres in the scaffold reduced burst release and significantly extended the release period. The nanofibrous architecture and controlled release of NGF from the microspheres induced neurite extension of PC12 cells, demonstrating that the released NGF was in an active form. The results suggest that the scaffolds prepared in this study may have potential applications in brain tissue engineering due to topologic and mechanical properties similar to brain tissue and pore structure suitable for cell growth and differentiation. Copyright © 2016 John Wiley & Sons, Ltd.     

Bioactive injectable aggregates with nanofibrous microspheres and human dental pulp stem cells: A translational strategy in dental endodontics

Regeneration of the pulp–dentin complex with stem cells is a potential alternative to conventional root canal treatments. Human dental pulp stem cells (hDPSCs) have been extensively studied because of their ability to proliferate and differentiate into mineralized dental and non‐dental tissues. Here we combined hDPSCs with two types of injectable poly‐l ‐lactic acid (PLLA) microsphere with a nanofibrous or smooth surface to form bioactive injectable aggregates, and examined their ability to promote pulp regeneration in the root canal in an in vivo model. We investigated the biocompatibility, biosafety and odontogenic potential of fibrous (F‐BIM) and smooth bioactive injectable microspheres (S‐BIM) in vitro and in vivo. Our results demonstrated that PLLA microspheres and hDPSCs were able to form bioactive injectable aggregates that promoted dentin regeneration in both in vitro and in vivo models. Our results suggest that F‐BIM and S‐BIM may induce dentinogenesis upon in vivo grafting, and propose that the potential usefulness of the microsphere–hDPSC aggregates described here should be evaluated in clinical settings. Copyright © 2017 John Wiley & Sons, Ltd.     

克百威分子印迹固相萃取微球的制备及性能

以偶氮二异丁腈为引发剂,2~3 μm的聚苯乙烯微球为种球,克百威为模板分子,甲基丙烯酸为功能单体,乙二醇二甲基丙烯酸酯为交联剂,采用单步溶胀法制备粒径均一的克百威分子印迹聚合物微球（MIPMs）。通过扫描电镜（SEM）、吸附平衡实验和竞争吸附实验分析了克百威MIPMs的形貌及其对克百威的结合特性及吸附选择性,并比较了克百威分子印迹固相萃取柱（MISPE）与C18固相萃取柱（C18 SPE）富集水中克百威的效果。结果表明：合成的MIPMs粒径约10 μm,表面呈蜂窝状;在90 min内可达到饱和吸附,最大吸附量为25.94 mg/g;在克百威、灭多威和三羟基克百威共存的条件下, 克百威MIPMs可实现对克百威的专一性吸附;与C18 SPE相比,克百威MISPE重复使用6次后加标回收率仍在85%以上,可用于水体中痕量克百威的检测。     

在CPS微球表面实现N-羟基邻苯二甲酰亚胺的同步合成与固载及固体催化剂催化氧化性能初探

通过分子设计的构思,仅通过两步大分子反应,便实现了N-羟基邻苯二甲酰亚胺(NHPI)在交联聚苯乙烯(CPS)微球表面的同步合成与固载,并制得了非均相催化剂微球CPS-NHPI。以氯化偏苯三酸酐(TMAC)为试剂、Lewis酸为催化剂,通过Friedel-Crafts酰基化反应,先将邻苯二甲酸酐(PA)基团键合在CPS微球表面,得到改性微球CPS-PA;再与盐酸羟胺进行酰亚胺反应,制备出固载有NHPI基团的非均相催化剂微球CPS-NHPI。重点研究了CPS微球表面发生Friedel-Crafts酰基化反应的影响因素。采用红外光谱(FT-IR)及扫描电子显微镜(SEM)等对微球CPS-NHPI进行表征,将微球CPS-NHPI分别用于分子氧氧化乙苯及环己烷两种烃类物质的氧化过程,初步考察了该微球的催化活性。研究结果表明,对于微球CPS与TMAC之间的Friedel-Crafts酰基化反应,适宜的溶剂为氯仿与N,N-二甲基乙酰胺(DMAC)混合溶剂(氯仿与DMAC的体积比为7:3),适宜的Lewis酸催化剂为SnCl₄。初步探索实验表明,催化剂微球CPS-NHPI与Co(OAc)₂ 所构成的共催化体系,在分子氧氧化乙苯及环己烷的催化氧化过程中,都表现出了良好的催化活性,温和条件下,反应35 h时乙苯氧化为苯乙酮的转化率可达37%,反应30 h时环己烷氧化为环己酮的转化率可达21%。     

Lineage-related and particle size-dependent cytotoxicity of chitosan nanoparticles on mouse bone marrow-derived hematopoietic stem and progenitor cells

Chitosan nanoparticles (CSNPs) have potential applications in stem cell research. In this study, ex vivo cytotoxicity of CSNPs on mouse bone marrow-derived (MBMCs) hematopoietic stem and progenitor cells (HSPCs) was determined. MBMCs were exposed to CSNPs of different particle sizes at various concentrations for up to 72 h. Cytotoxicity effect of CSNPs on MBMCs was determined using MTT, Live/Dead Viability/Cytotoxicity assays and flow cytometry analysis of surface antigens on HSCs (Sca-1⁺), myeloid-committed progenitors (CD11b⁺, Gr-1⁺), and lymphoid-committed progenitors (CD45⁺, CD3e⁺). At 24 h incubation, MBMCs' viability was not affected by CSNPs. At 48 and 72 h, significant reduction was detected at higher CSNPs concentrations. Small CSNPs (200 nm) significantly reduced MBMCs' viability while medium-sized particle (∼400 nm) selectively promoted MBMCs growth. Surface antigen assessment demonstrated lineage-dependent effect. Significant decrease in Sca-1⁺ cells percentage was observed for medium-sized particle at the lowest CSNPs concentration. Meanwhile, reduction of CD11b⁺ and Gr-1⁺ cells percentage was detected at high and intermediate concentrations of medium-sized and large CSNPs. Percentage of CD45⁺ and CD3e⁺ cells along with ROS levels were not significantly affected by CSNPs. In conclusion, medium-sized and large CSNPs were relatively non-toxic at lower concentrations. However, further investigations are necessary for therapeutic applications.     

Chitosan based micro- and nanoparticles for colon-targeted delivery of vancomycin prepared by alternative processing methods

The aim of this work was to prepare chitosan (CH) based particulate formulations for colon delivery of vancomycin (VM). Chitosan microparticles (MPs) and nanoparticles (NPs) loaded with VM were prepared using different CH/tripolyphosphate (TPP) molar ratios and different technological processes. In particular, nanoparticles were prepared by ionic gelation and freeze-drying to recover these particles, or, alternatively, by spray-drying method. Microparticles were prepared using a different spray-dryer. Micro- and nanoparticles were characterized in terms of size distributions by photon correlation spectroscopy (PCS), while encapsulation and drug loading efficiencies were studied using a dialysis method. Fourier Transform Infrared Spectroscopy (FT-IR) was employed to determine the surface composition of the micro- and nanoparticles respectively, and the morphologies of the developed systems were studied by scanning electron microscopy (SEM). Water uptake as well as drug release profiles were also measured. Antibacterial activity against Staphylococcus aureus, a Gram-positive model strain, was evaluated. FT-IR results suggested an electrostatic interaction between VM and CH/TPP particles. Moreover, the particles were found to hold a positive zeta-potential, indicating the presence of CH on the particle surfaces. Particle size and encapsulation efficiency were mainly influenced by the different manufacturing processes employed. Nanoparticles obtained by spray-drying showed the best results in terms of water uptake and drug release rate. Moreover, they showed a good bactericidal activity against S. aureus.     

Efficacy study of two novel hyaluronic acid-based formulations for viscosupplementation therapy in an early osteoarthrosic rabbit model

Viscosupplementation (VS) is a therapy for osteoarthrosis (OA) consisting of repetitive intra-articular injections of hyaluronic acid (HA). It is known to be clinically effective in relieving pain and increasing joint mobility by restoring joint homeostasis. In this study, the effects of two novel HA-based VS hydrogel formulations were assessed and challenged against a pure HA commercial formulation for the first time and this in a rabbit model of early OA induced by anterior cruciate ligament transection (ACLT). The first formulation tested was a hybrid hydrogel composed of HA and reacetylated chitosan, a biopolymer considered to be chondroprotective, assembled thanks to an ionic shielding. The second formulation consisted of a novel HA polymer grafted with antioxidant molecules (HA-4AR) aiming at decreasing OA oxidative stress and increasing HA retention time in the articulation.ACLT was performed on rabbits in order to cause structural changes comparable to traumatic osteoarthrosis. The protective effects of the different formulations were observed on the early phase of the pathology in a full randomized and blinded manner. The cartilage, synovial membrane, and subchondral bone were evaluated by complementary investigation techniques such as gross morphological scoring, scanning electron microscopy, histological scoring, and micro-computed tomography were used.In this study, ACLT was proven to successfully reproduce early OA articular characteristics found in humans. HA and HA-4AR hydrogels were found to be moderately protective for cartilage as highlighted by μCT. The HA-4AR was the only formulation able to decrease synovial membrane hypertrophy occurring in OA. Finally, the hybrid HA-reacetylated chitosan hydrogel surprisingly led to increased subchondral bone remodeling and cartilage defect formation. This study shows significant effects of two innovative HA modification strategies in an OA rabbit model, which warrant further studies toward more effective viscosupplementation formulations.     

90Y树脂微球治疗患者尿液中90Y放射性活度检测和分析

目的研究患者接受90Y树脂微球选择性内放射治疗(SIRT)后48 h内所排泄尿液中90Y的放射性活度, 为术后患者排泄物的管理提供建议。方法收集3名患者在术后0～24 h和24～48 h两个时间段内排泄的尿液, 并对尿液中的90Y放射性活度进行检测和分析。结果 3名患者术后0～24 h和24～48 h尿液中的90Y放射性活度排泄量分别为(1 266±258)kBq/GBq和(140±106)kBq/GBq, 90Y放射性活度浓度分别为(640±113)kBq/L和(53±12)kBq/L。结论 90Y树脂微球治疗术后肝癌患者0～24 h排泄尿液中的90Y放射性活度比24～48 h高。术后患者可通过增加排泄尿量的方式来加速排出体内游离的90Y;患者住院期间的排泄物应按照HJ 1188-2020《核医学辐射防护与安全要求》的要求处理。     

90Y树脂微球选择性内放射治疗放射防护检测与剂量评估

目的对90Y树脂微球选择性内放射治疗过程进行放射防护检测和剂量评估, 为放射防护工作提供参考。方法对90Y树脂微球介入手术治疗各操作环节和患者体表的外照射水平进行检测, 估算相关人员的受照剂量水平。结果 90Y树脂微球分装及转运过程的剂量率水平为1.12～454 μSv/h, 手术操作过程为2.06～58.2 μSv/h;3名患者术后0.5 h, 体表5 cm和1 m处的剂量率分别为22.7～64.1和0.82～2.55 μSv/h。按照每年200例患者的工作量, 90Y树脂微球药物操作对工作人员年个人有效剂量贡献为0.12～1.03 mSv/年, 术后患者对公众、家属及陪护志愿者的个人有效剂量贡献为0.02～0.24 mSv/年。结论在患者治疗、护理和出院过程中, 工作人员、陪护志愿者和公众的照射剂量均低于(GB 18871-2002《电离辐射防护与辐射源安全基本标准》)中的剂量限值和医疗机构设定的管理目标值。     

Thiocarbohydrazide-crosslinked chitosan as a bioinspired corrosion inhibitor for protection of stainless steel in 3.5% NaCl

A facile single-step synthesis was performed to cross-link chitosan with thiocarbohydrazide to yield thiocarbohydrazide-chitosan (TC-Cht) which was for the first time evaluated as an inhibitor for corrosion of stainless steel in 3.5% NaCl solution. A comprehensive electrochemical analysis employing electrochemical impedance spectroscopy (EIS), potentiodynamic polarization (PDP), and cyclic voltammetry (CV) was undertaken and showed that the TC-Cht acts by adsorption on the steel surface and exhibits mixed type behavior with predominantly cathodic nature. The adsorption of TC-Cht molecules on the surface of stainless steel followed the Langmuir isotherm. The TC-Cht showed a high inhibition efficiency of >94% at 500 mg L^?1 concentration. Surface investigation using SEM and EDX supported the inhibitor adsorption on the steel surface.