Cell Sheet Comprised of Mesenchymal Stromal Cells Overexpressing Stem Cell Factor Promotes Epicardium Activation and Heart Function Improvement in a Rat Model of Myocardium Infarction

Cell therapy of the post-infarcted myocardium is still far from clinical use. Poor survival of transplanted cells, insufficient regeneration, and replacement of the damaged tissue limit the potential of currently available cell-based techniques. In this study, we generated a multilayered construct from adipose-derived mesenchymal stromal cells (MSCs) modified to secrete stem cell factor, SCF. In a rat model of myocardium infarction, we show that transplantation of SCF producing cell sheet induced activation of the epicardium and promoted the accumulation of c-kit positive cells in ischemic muscle. Morphometry showed the reduction of infarct size (16%) and a left ventricle expansion index (0.12) in the treatment group compared to controls (24–28%; 0.17–0.32). The ratio of viable myocardium was more than 1.5-fold higher, reaching 49% compared to the control (28%) or unmodified cell sheet group (30%). Finally, by day 30 after myocardium infarction, SCF-producing cell sheet transplantation increased left ventricle ejection fraction from 37% in the control sham-operated group to 53%. Our results suggest that, combining the genetic modification of MSCs and their assembly into a multilayered construct, we can provide prolonged pleiotropic effects to the damaged heart, induce endogenous regenerative processes, and improve cardiac function.     

聚乙烯醇/牛I型胶原支架材料的制备及评价

研究了一种聚乙烯醇(PVA)和胶原(COL)复合支架材料的制备方法。采用氨基硅烷对PVA海绵表面进行了氨基化修饰后,通过戊二醛溶液交联牛Ⅰ型胶原(COL),最后通过赖氨酸溶液封闭,获得一种PVA/COL复合支架材料。采用扫描电镜(SEM)、X光电子能谱仪(XPS)、傅里叶红外光谱(FT-IR)等手段对支架材料的理化性能进行表征,并通过细胞实验对支架材料的生物学性能进行评价。结果表明,经过COL修饰的PVA孔隙率为21.33%,平均孔径为168.68 ?m且均匀分布,支架材料接触角为20.03°。对支架材料的生物学评价结果表明C3A细胞在复合材料上黏附良好,优于PVA组;CCK-8增殖检测结果表明细胞在复合材料上呈增殖生长趋势,与对照组PVA相比差异显著(P?0.01)。将PVA和COL复合制备得到的支架材料具有良好的理化及生物学特性,具有广阔的应用前景。     

森林脑炎病毒不同感染途径对小鼠模型的感染性分析

目的分析森林脑炎病毒不同感染途径对小鼠模型的致病性、病毒分布及增殖动态。方法将适应原代地鼠肾(primary hamster kidney,PHK)细胞的森林脑炎病毒进行10倍系列稀释,取4个连续稀释度的病毒液,分别通过脑腔注射、灌胃、滴鼻、肌肉注射、腹腔注射的感染方式,对昆明小鼠进行攻毒,逐日连续观察14 d,计算半数致死量(median lethal dose,LD₅₀);并分别取攻毒后第3、5、7天发病典型的腹腔感染组小鼠脑、肺、肾脏、肝脏、心脏、小肠、血液7种组织器官,采用蚀斑法检测病毒滴度。结果脑腔、灌胃、滴鼻、肌肉注射和腹腔注射攻毒方式的LD₅₀分别为10、10^5.1、10⁵、10^1.2和10²PFU/mL。所有攻毒途径小鼠的脑腔中病毒滴度最高,达10⁹PFU/mL,肺组织为10^6.5PFU/mL,肾脏为10⁵PFU/mL,心脏和肝脏中病毒滴度较低,分别为10²     

MDCK细胞库的建立、检定及其生物学特性

目的建立MDCK细胞的主细胞库及工作细胞库,并进行检定,同时评价其生物学特性。方法从欧洲细胞保藏中心(European Collection of Authenticated Cell Culture,ECACC)引进1株MDCK细胞,经传代扩大培养后,分别建立主库及工作细胞库。按照《中国药典》三部(2015版)规定的方法对细胞库进行细胞活力、细胞形态、无菌、支原体及细胞内外源病毒因子检查,并对细胞生长状况、传代稳定性、流感病毒敏感性进行检测。结果建立的MDCK细胞主种子库及工作种子库细胞形态呈不规则的上皮样细胞状,生长状态良好,细胞活率分别为96. 0%和95. 5%,无菌、支原体及细胞内外源病毒因子检查结果均合格。MDCK细胞生长曲线均呈S型,培养48～96 h进入对数生长期,随后进入平台期。连续传10代48 h细胞活率在93. 0%～95. 5%之间,较稳定。MDCK细胞对流感病毒较敏感,对B型流感病毒的敏感性高于A型流感病毒。结论建立的主细胞库及工作细胞库各项检测结果均符合《中国药典》规定,具有良好的传代稳定性及流感病毒敏感性。     

重组人淋巴细胞活化基因-3蛋白胞外段的真核表达及鉴定

目的真核表达重组人淋巴细胞活化基因-3(lymphocyte activation gene-3,LAG-3)蛋白胞外段,并进行鉴定。方法用植物血球凝集素(phytohaemagg lutinin,PHA)刺激Jurkat细胞,流式细胞术检测Jurkat细胞中LAG-3蛋白的表达;提取Jurkat细胞总mRNA,RT-PCR法扩增人LAG-3蛋白胞外段基因片段,同时在蛋白C-末端引入His标签,将其克隆入载体pcDNA3. 1+,构建重组质粒,转染Expi293F真核细胞,当细胞活率低于50%时收获细胞上清,经镍柱亲合层析纯化。纯化产物进行4%～20%SDS-PAGE、HPLC及Western blot分析,BCA法测定浓度。结果经菌液PCR、双酶切及测序鉴定,表明质粒构建正确。重组表达蛋白的相对分子质量约60 000,纯化后纯度达95%以上,与鼠抗LAG-3单克隆抗体可发生特异性结合,浓度为2. 4 mg/mL。结论成功构建了重组真核表达质粒LAG-3/pcDNA3. 1+,并于Expi293F细胞中表达,纯化获得了纯度较高的LAG-3蛋白,为后期LAG-3蛋白的相关研究及其单抗的制...     

Investigation of LSM-8YSZ cathode within an all ceramic SOFC. Part I: Chemical interactions

This work focuses on a novel, co-sintered, all-ceramic solid oxide fuel cell (SOFC) concept. The objective is the understanding of interaction and degradation mechanisms of the cathode and current collector layers within the design during co-sintering. Half cells consisting of silicate mechanical support, lanthanum strontium manganite (LSM) current collector, LSM mixed with 8 mol% yttria-stabilized zirconia (8YSZ) composite cathode and 8YSZ electrolyte were co-sintered at 1150 °C < T < 1250 °C. Crystallographically stable LSM compositions within the design were identified. However, the cathode and silicate/electrolyte interacted by interdiffusion of Zn (gas diffusion) and Mn (solid diffusion), and by the formation of several reaction phases (between silicate and cathode only). Introducing silicate poisoning decreased the electrochemical performance of the cell by around 40%. This is likely due to the formation of the Zn- and Mn-rich phase in the cathode, but may also be caused by a higher ohmic resistance of the current collector.     

Investigation of LSM/8YSZ cathode within an all-ceramic SOFC,Part II: Optimization of performance and co-sinterability

This paper focuses on the cathode and current collector layers of a co-sintered, all-ceramic solid oxide fuel cell (SOFC) concept. Challenges to reach good electrochemical performance have to be overcome, due to more demanding manufacturing conditions, including a relatively high co-sintering temperature. Master sintering curves show that the sintering activity of lanthanum strontium manganite (LSM) is significantly higher than that of 8-mol% yttria stabilized zirconia (8YSZ). By applying a double-layered cathode and a current collector with optimized microstructures the best electrochemical performance of the cathode is 0.26 $\mathrm{\Omega }$cm${}^2$ at 800 $°$C, evaluated from polarization resistances of 8YSZ electrolyte-supported symmetric cells post-sintered at 1150 $°$C $<T<1250$ $°$C. The cathode and current collector materials are adapted to fit the co-sintering process by adjustment of the paste compositions. Half-cells consisting of silicate mechanical support, LSM current collector, LSM mixed with 8YSZ composite cathode and 8YSZ electrolyte are co-sintered porous and defect-free at 1150 $°$C $<T<1250$ $°$C.     

A novel flow field with controllable pressure gradient to enhance mass transport and water removal of PEM fuel cells

An easily machined novel flow field with controllable pressure gradient across adjacent channels was designed and a two dimensional, across-the-channel, two-phase model was developed to study the gas transport and water removal of the novel configuration. The effect of channel-rib width ratio, GDL thickness and pressure gradient on the profiles of oxygen concentration and water saturation within the GDL were investigated. Special attention was paid to the mechanisms of the promoted mass transport and water removal rates under a pressure gradient. The model was validated by experiments with various channel-rib ratios and GDL thicknesses at different operating pressure. The results revealed that, oxygen concentration was increased, and the water saturation was reduced under the rib with a pressure gradient generated across the adjacent channels. The optimal pressure gradient is between 0.1 to 0.2 atm for the studied channel geometry and configuration. The mechanisms of the improved cell performance were elucidated.     

Branched poly(biphenylene-co-sulfone)ether ion exchange membranes containing perfluorocyclobutane groups for fuel cell applications

A series of branched poly(biphenylene-co-sulfone) ion exchange membranes containing perfluorocyclobutane groups were prepared for fuel cells. Two bifunctional trifluorovinyloxy-terminated monomers (sulfonable 4,4′-bis(trifluorovinyloxy)biphenyl and insulfonable 4,4′-sulfonyl-bis(trifluorovinyloxy)biphenyl) and a trifunctional trifluorovinyloxy-terminated branching agent (1,1,1-tris(4′-trifluorovinyloxyphenyl)ethane) were synthesized and terpolymerized via thermal [2π + 2π] cyclodimerization to obtain partially fluorinated and branched polymers containing 0–5 mol% of the branching agent. They were then postsulfonated by chlorosulfonic acid at room temperature, cast as membranes, and characterized to evaluate their electrochemical properties for fuel cell applications. As the branching agent content was increased, their polydispersity values highly increased, indicating they became highly branched. It was confirmed that higher branching agent content also increased the ion exchange capacity, water uptake, and proton conductivity of the branched ion exchange membranes containing perfluorocyclobutane groups. This indicates that their electrochemical properties can be easily controlled by the degree of branching. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48373.     

Finite element optimization of sample geometry for measuring the torsional shear strength of glass/metal joints

Assessment of mechanical properties of glass/metal joints is a challenging process, especially when the application relevant conditions of the joints have to be considered in the test design. In this study, a finite element method (FEM) is implemented to analyze a torsional shear strength test designed for glass-ceramic/steel joints aiming towards solid oxide fuel/electrolysis cells application. Deviations from axial symmetry of the square flanges (ends) of respective hourglass-shaped specimens and also supporting and loading sockets of the test set-up are included in the model to simulate conditions close to reality. Undesirable tensile stress and also shear stress concentration appear at the outer edge of glass-ceramic layers, which are less for the hollow-full specimen. The simulation results show that for a specimen with either 9 mm thick square- or 6 mm thick triangular-flanges, locally enhanced tensile stresses almost disappear, resulting in a symmetric shear stress distribution. The difference between the analytically derived nominal shear strength and the real critical shear stress derived via simulation reduces with decreasing the fracture torque.