电纺直写制备聚己内酯支架形貌及精度

作为一种新型的微纳制造技术，熔体直写电纺被广泛用于组织工程支架的可控制备,有序的纤维沉积是该领域应用的前提条件。对于支架成型精度的探究，本文使用生物可降解材料聚己内酯（PCL），采用自行设计的熔体电纺三维可控成型设备进行实验，考察了纤维间距对二维并行纤维沉积形貌及成型精度的影响，以及纺丝电压和网格大小对三维网格结构形貌及精度影响。结果表明，随着并行纤维设定距离的增大，纤维的沉积误差减小，并最终趋于平稳。对于三维网格结构，随电压的增加，最大沉积层数量先增大后减小，当纺丝电压为6kV时达到最大沉积层数15层。成型精度误差先减小后增大，当纺丝电压为7kV时，精度最高误差小于5%。随设定网格边长的增大，沉积层数不断增大。成型精度逐渐提高，当网格边长大于等于1.5mm时，沉积误差趋于稳定，并维持在5%左右。     

Biomimetic strain hardening in interpenetrating polymer network hydrogels

In this paper, we present the systematic development of mechanically enhanced interpenetrating polymer network (IPN) hydrogels with Young's moduli rivaling those of natural load-bearing tissues. The IPNs were formed by synthesis of a crosslinked poly(acrylic acid) (PAA) network within an end-linked poly(ethylene glycol) (PEG) macromonomer network. The strain-hardening behavior of these PEG/PAA IPNs was studied through uniaxial tensile testing and swelling measurements. The interaction between the independently crosslinked networks within the IPN was varied by (1) changing the molecular weight of the PEG macromonomer, (2) controlling the degree of PAA ionization by changing pH, and (3) increasing the polymer content in the PAA network. Young's moduli and the maximum stress-at-break of the swollen hydrogels were normalized on the basis of their polymer content. Strain hardening in the IPNs exhibited a strong dependence on the molecular weight of the first network macromonomer, the pH of the swelling buffer, as well as the polymer content of the second network. The results indicate that the mechanical enhancement of these IPNs is mediated by the strain-induced intensity of physical entanglements between the two networks. The strain can be applied either by mechanical deformation or by changing the pH to modulate the swelling of the PAA network. At pHs below the pK_a of PAA (4.7), entanglements between PEG and PAA are reinforced by interpolymer hydrogen bonds, yielding IPNs with high fracture strength. At pHs above 4.7, a “pre-stressed” IPN with dramatically enhanced modulus is formed due to ionization-induced swelling of the PAA network within a static PEG network. The modulus enhancement ranged from two-fold to over 10-fold depending on the synthesis conditions used. Variation of the network parameters and swelling conditions enabled “tuning” of the hydrogels' physical properties, yielding materials with water content between 58% and 90% water, tensile strength between 2.0 MPa and 12.0 MPa, and initial Young's modulus between 1.0 MPa and 19.0 MPa. Under physiologic pH and salt concentration, these materials attain “biomimetic” values for initial Young's modulus in addition to high tensile strength and water content. As such, they are promising new candidates for artificial replacement of natural tissues such as the cornea, cartilage, and other load-bearing structures.     

Tissue-type plasminogen activator variants with domain duplications and rearrangements

The interactions between tPA domains that are important forcatalysis are poorly understood. We have probed the functionof interdomain interactions by generating tPA variants in whichdomains are duplicated or rearranged. The proteins were expressedin a transient mammalian expression system and tested in vitrofor their ability to activate plasminogen, induce fibrinolysisand bind to a forming fibrin clot. Duplication of the heavychain domains of tPA produced enzymatically active tPA variants,many of which demonstrated similar in vitro amidolytic and fibrinolyticactivity and similar fibrin affinity to the parent molecule.Zymographic analysis of the domain duplication tPA variantsshowed one major active species for each variant. Selectionof the residues duplicated and the interdomain spacing werefound to be critical considerations in the design of tPA variantswith duplicated domains. We also rearranged the domains of tPAsuch that kringle 1 replaced the second kringle domain and viceversa. An analysis of these variants indicates that the firstkringle domain can confer fibrin affinity to a tPA variant andfunction in place of kringle 2. Therefore, in wild-type tPA,the functions of kringle 1 and kringle 2 must be dependent partiallyon their orientation within the heavy chain of the protein.The functional autonomy of the heavy and light chains of tPAis demonstrated by the activity of a tPA variant in which theorder of the heavy and light chains was reversed.     

弓形虫复合黏膜疫苗滴鼻免疫小鼠的黏膜相关淋巴细胞及其亚群的动态变化

目的探讨弓形虫复合黏膜疫苗滴鼻免疫小鼠的黏膜相关淋巴细胞及其亚群的动态变化。方法取BALB/c小鼠96只,随机分为实验组和对照组,实验组以弓形虫复合黏膜疫苗20μl/只滴鼻免疫,对照组以PBS滴鼻。滴鼻2次(间隔2周)后,分别于第1、2、3、4、6、8、10、12周处死小鼠,分离鼻相关淋巴组织(NALT)、Peyer结(PP)和上皮内淋巴细胞(IEL),制备淋巴细胞悬液,计数并涂片。免疫细胞化学法检测CD4+、CD8+T细胞亚群水平。结果实验组NALT、PP和IEL的T淋巴细胞均显著增生,PP和IEL的T淋巴细胞第2周达高峰,之后逐渐降低。与对照组相比,NALT的T淋巴细胞于第1、2、6、8、12周显著增生,PP的T淋巴细胞数于第1、2、3周显著增生,IEL于第1～4周显著增生。NALT和PP中主要以CD4+T细胞增生为主,肠IEL以CD8+T细胞增生为主。结论弓形虫复合黏膜疫苗滴鼻免疫BALB/c小鼠,可诱导鼻相关淋巴组织和肠相关淋巴组织T淋巴细胞明显增生,同时诱导其向不同亚群分化。     

组织工程多孔支架制备技术进展

综述了天然或合成生物降解材料为原料的组织工程多孔支架的几种制备方法，包括无纺织物／纤维粘结法、溶液浇铸／粒子洗出法、相分离／冻干法、气体成孔法和三维“印刷”等方法。特别评述了近期组织工程多孔支架研究进展及其在骨、软骨和皮肤等组织修复中的应用。     

肾缺血再灌注细胞内钙水平与氧化应激损伤研究

目的 观察肾细胞中游离钙([Ca~(2+)]i)及氧自由基在缺血再灌注损伤过程中的改变情况,探讨二者在再灌注肾损伤中发生作用机制。方法 摘除Wistar大鼠左肾,钳夹右侧肾蒂,建立急性缺血再灌注肾损伤模型,应用Fura-2/AM荧光指示剂测定缺血再灌注大鼠肾细胞内[Ca~(2+)]i浓度的变化,同时测定谷胱甘肽过氧化物酶(GSH-Px)活性、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果 缺血再灌注不同时期肾细胞内[Ca~(2+)]i浓度均有不同程度增高,与对照组相比差异有显著意义,各实验组SOD活力水平降低,与对照组相比差异有显著意义,MDA生成高于对照组,GSH-Px在再灌注早期活力减弱,明显低于对照组,晚期活力基本恢复。结论 1.缺血再灌注不同时期大鼠肾细胞内[Ca~(2+)]i超载和不同程度的氧化侵袭,在缺血再灌注肾损伤病理过程中起重要作用;2.再灌注时间与肾细胞内[Ca~(2+)]i超载呈正相关,提示再灌注损伤通过不同机制加重细胞钙超载。     

基于C-V模型和MRF的脑部MRI分割

为准确分割脑部磁共振图像(MRI)的灰质、白质和背景,提出一种基于C-V模型和马尔可夫随机场的全自动分割方法。采用C-V模型与形态学相结合的方法对脑MRI进行预处理,去除多余脑组织,获得待分割图像。引入灰度场局部熵的思想对惩罚因子进行估计,利用马尔可夫随机场模型建模实现脑灰白质的分割,并运用形态学方法获得最终分割结果。对96幅IBSR图像和46幅临床图像进行实验,结果表明,该方法能够实现脑部MRI灰白质的全自动分割,且具有较好的分割精度和较快的处理速度。     

Simulation of the Impact Between Head and Seat Head Restraint During REC Under Consideration of the Multiphase and Nonlinear Features

The impact process between car drivers head and seat head restraint during a car rear end collision is simulated, where the multiphase and nonlinear features of the human brain tissues and the foam material of the seat head restraint are considered. Using the FEM-software LS-DYNA, the brain tissue deformation caused by a car REC is calculated. The purpose of this work is to supply references for improving the design of the head protective devices. The results show that the maximum shear strain appears near the boundaries of different phases and there is a great shear strain gradient in the brain tissues.     

Evaluation of PLGA/chitosan/HA conduits for nerve tissue reconstruction

A micro-envioment for nerve cells and tissue growth were designed and constructed via surface modification of poly(L-lactide-co-glycolide)(PLGA) with chitosan and hydroxyapatite(HA). The poly(L-lactide-co-glycolide)/chitosan/hydroxyapatite (PLGA/chitosan/HA) conduits were manufactured by a combined solvent casting and particulate leaching technique. The conduits were highly porous with an interconnected pore structure and 76.5% porosity. Micropores with 50–100 micrometer diameter were formed in the conduits. In vivo application of PLGA/chitosan/HA conduits for reconstruction of 10 mm sciatic nerve defect was assessed by the walking track analysis, the quantifying of the wet weight of tibialis anterior muscle and the histological assessment. The conduits in host rats in vivo can not only be an effective in promoting regenerating of nerves but can also lead to favorable nerve functional recovery. Funded by the National Natural Science Foundation of China(No.50774096)     

Design and preparation of bone tissue engineering scaffolds with porous controllable structure

A novel method of designing and preparing bone tissue engineering scaffolds with controllable porous structure of both macro channels and micro pores was proposed. The CAD software UG NX3.0 was used to design the macro channels’ shape, size and distribution. By integrating rapid prototyping and traditional porogen technique, the macro channels and micro pores were formed respectively. The size, shape and quantity of micro pores were controlled by porogen particulates. The sintered β-TCP porous scaffolds possessed connective macro channels of approximately 500 μm and micro pores of 200–400 μm. The porosity and connectivity of micro pores became higher with the increase of porogen ratio, while the mechanical properties weakened. The average porosity and compressive strength of β-TCP scaffolds prepared with porogen ratio of 60wt% were 78.12% and 0.2983 MPa, respectively. The cells’ adhesion ratio of scaffolds was 67.43%. The ALP activity, OCN content and cells micro morphology indicated that cells grew and proliferated well on the scaffolds. Funded by the Postdoctor Science Fund of China (No. 20070410715) and Shanghai Excellent Youth Special Fund (No. 17014)