A three-dimensional in vitro culture model for primary neonatal rat ventricular myocytes

To overcome bad prognosis of patients with heart failure and the lack of organ donors, cardiac tissue engineering has developed as a biomimetic approach to repair, replace, and regenerate the damaged cardiac tissue. During the past decade years, researchers are devoted to find different natural and/or synthetic materials that can build appropriate physical structures to contain and organize implanted cells. In this study, we present a new method for primary neonatal rat cardiomyocytes culture in vitro using alginate/collagen/chitosan hydrogel. To investigate the feasibility of this material as scaffold for cardiac myocytes, neonatal rat ventricular myocytes were isolated and encapsulated in alginate-based beads cross-linked with calcium ion. The growth of cells was evaluated by staining with α-Sarcomeric actin (α-SCA) and Troponin T type 2 (TNNT2), and the viability of cardiomyocytes was studied in vitro by assessing the expression levels of several cardiac ion channels, including CACNL1A1, Connexin 43 and SCN5A. The results showed a significant increase in cardiac myocytes number, and the expression levels of CACNL1A1, Connexin 43 (Cx43) were up-regulated significantly except SCN5A, as compared with two-dimensional cultures. Moreover, extracellular matrix produced by the seeded cells themselves was observed by staining with fibronectin. Taken together, these findings indicate that this alginate/collagen/chitosan hydrogel bead is suitable for supporting the growth and retaining the morphologic and electrophysiologic characteristics of primary cultured rat cardiac muscle cells.     

明胶改性聚醚醚酮微载体的制备、表征及性能

微载体因其具有较高的表面积/体积比等优点可以大大提高哺乳动物细胞培养效率,被广泛应用于生物制药和组织工程等领域。 但微载体多为一次性使用,不耐高温,且主要依赖进口,价格昂贵,因而限制了其国内的应用和推广。 聚醚醚酮(PEEK)材料具有良好的生物相容性、化学稳定性及耐高温等特性,是一种优异的微载体材料,但存在熔点高,加工方法单一和生物惰性等缺陷。 本文以浓硫酸为溶剂,乙醇溶液为萃取剂,采用气流辅助滴注/相分离法,将PEEK制备成448 μm左右,尺寸均匀的微球;经氨丙基三乙氧基硅烷(APTES)处理,获得表面氨基化修饰的PEEK微球(PEEK-N);进一步,以N,N'-羰基二咪唑(CDI)为活性中间体,将明胶分子接枝到PEEK-N微球表面,获得表面明胶修饰的PEEK微载体(PEEK-G)。 对材料的物理化学性质、表面接枝量进行表征;并通过体外细胞实验评估其细胞毒性、细胞粘附效率和细胞增殖能力。 结果显示,通过该方法制备成功了3种不同明胶接枝含量的PEEK细胞微载体(PEEK-G1,PEEK-G2,PEEK-G3),其中明胶含量较高的PEEK-G3毒性最低,细胞粘附和增殖效果最理想。     

Modelling toxin effects on protein biosynthesis in eukaryotic cells

We present a rather generic model for toxin (ricin) inhibition of protein biosynthesis in eukaryotic cells. We also study reduction of the ricin toxic effects with application of antibodies against the RTB subunit of ricin molecules. Both species initially are delivered extracellularly. The model accounts for the pinocytotic and receptor-mediated toxin endocytosis and the intact toxin exocytotic removal out of the cell. The model also includes the lysosomal toxin destruction, the intact toxin motion to the endoplasmic reticulum (ER) for separation of its molecules into the RTA and RTB subunits, and the RTA chain translocation into the cytosol. In the cytosol, one portion of the RTA undergoes degradation via the ERAD. The other its portion can inactivate ribosomes at a large rate. The model is based on a system of deterministic ODEs. The influence of the kinetic parameters on the protein concentration and antibody protection factor is studied in detail.     

Fabrication of the First Disposable Three‐dimensional Paper‐based Concentration Cell as Ammonia Sensor with a New Method for Paper Hydrophobization by Laser Patterned Parafilm®

A new, simple and low‐cost method for patterning hydrophobic barriers in porous support such as paper by Parafilm® has been introduced. This method is then used for electrochemical paper‐based ammonia sensor construction. Ammonia sensor is based on electrochemical concentration cell which ammonia reaction with electrolyte in halves cell caused in concentration gradient and therefore potential difference dependent on ammonia concentration. The effect of concentrations of the involved chemicals, time periods of the required processes, the presence of Faraday cage as well as the effects of different salts used in the salt bridge on the response of the sensor, were investigated in order to find the optimized conditions.     

Preparation of 17β‐estradiol surface molecularly imprinted polymers and their application to the analysis of biological samples

17β‐Estradiol (E2) surface molecularly imprinted polymers have been prepared using functionalized monodispersed poly(glycidyl methacrylate‐co‐ethylene dimethacrylate) beads as a support. The resulting polymers were found to be uniform in size (5 μm), and the surfaces of the microspheres possessed large pore‐like structures. A chromatographic experiment demonstrated that the resulting microspheres exhibited high levels of recognition and selectivity toward the target molecule. The particles were employed as a novel sorbent in a molecularly imprinted SPE protocol. A method was then developed involving the combination of the pretreatment with HPLC to determine the levels of estrogen secreted from Michigan Cancer Foundation‐7 cells. The obtained results revealed that the extraction recoveries of E2 from real samples were in the range of 73.0–97.5% with RSDs of < 7.5% (n = 3). Calibration curves were established with R values > 0.9996 for concentrations in the range of 0.50–100.00 ng/mL. The LOD of this new method was 0.14 ng/mL. Compared with traditional C₁₈ SPE agents, the particles showed high selectivity and extraction efficiency for E2 in the pretreatment process. The particles could therefore be used to determine trace estrogen in biological samples with a UV detector only.     

SYNTHESIS OF A NOVEL POLYSACCHARIDE HYDROGEL

ABSTRACT

A modified hyaluronic acid (HA) biopolymer was synthesized that can be photocrosslinked to form a stable hydrogel. The chemical and physical properties including the amount of modification of the polymer with methacrylate anhydride, the viscosity of the modified biopolymer, and the solute diffusion characteristics of the polymer have been determined.     

Synthesis and in vitro biological evaluation of novel 2-aminoimidazolone derivatives as anti-tumor agents

Novel 2-aminoimidazolone derivatives were synthesized.Most compounds displayed strong anticancer activities against human carcinoma cells in vitro.Compounds 8a,8b and 8j exhibited optimal activity superior to 5-FU in most cancer cells tested.Especially,the lC₅₀s of 8b（12.6-21.5μmol/L） against five tumor cells were 1 -4 fold less than those of 5-FU（18.4-56.1μmol/L） in vitro.Furthermore,comp以ound 8b could induce SMMC-7721 cell apoptosis in a dose-dependent manner.Therefore,our novel findings may provide a new framework for the design of new 2-aminoimidazolone derivatives for the treatment of cancer.     

Construction of microscale structures in enclosed microfluidic networks by using a magnetic beads based method

A large number of microscale structures have been used to elaborate flowing control or complex biological and chemical reaction on microfluidic chips. However, it is still inconvenient to fabricate microstructures with different heights (or depths) on the same substrate. These kinds of microstructures can be fabricated by using the photolithography and wet-etching method step by step, but involves time-consuming design and fabrication process, as well as complicated alignment of different masters. In addition, few existing methods can be used to perform fabrication within enclosed microfluidic networks. It is also difficult to change or remove existing microstructures within these networks. In this study, a magnetic-beads-based approach is presented to build microstructures in enclosed microfluidic networks. Electromagnetic field generated by microfabricated conducting wires (coils) is used to manipulate and trap magnetic beads on the bottom surface of a microchannel. These trapped beads are accumulated to form a microscale pile with desired shape, which can adjust liquid flow, dock cells, modify surface, and do some other things as those fabricated microstructures. Once the electromagnetic field is changed, trapped beads may form new shapes or be removed by a liquid flow. Besides being used in microfabrication, this magnetic-beads-based method can be used for novel microfluidic manipulation. It has been validated by forming microscale dam structure for cell docking and modified surface for cell patterning, as well as guiding the growth of neurons.     

Synthesis and characterization of new selective Zn fluorescent probes for functionalization: in vitro Cell imaging applications

Herein the synthesis and characterization of new, lipophilic highly Zn²⁺-selective fluorescent probes are reported. High affinity for zinc (K_d 1.1–8.0 nM) over other biologically relevant metals and mixtures of metals was observed. Excitation at 360 nm afforded an emission spectrum with maximum at 530 nm for the zinc bound complex. The linear relationship between fluorescence intensity and zinc concentration indicates that FZnA-probes can be used for quantification. The probes have been synthesized in 28–45% overall yield and the feasibility for further functionalization with biologically relevant side chains has been demonstrated. In vitro studies using PC12 cells and 10 μM of one of the novel probes (FZnA-Ada) visualized endogenous labile Zn²⁺ after 45 min incubation time.