Fiber-Based Chitosan Tubular Scaffolds for Soft Tissue Engineering： Fabrication and in Vitro Evaluation

Porous, two-ply tubular chitosan conduits for guided tissue regeneration were fabricated by combining the textile technique （inner layer） with the thermally induced phase separation process （outer layer）. A hollow chitosan tube was prepared using an industrial warp knitting process with chitosan yarns. Then, an appropriate diameter mandrel was inserted into the pre-fabricated tube. The tube and the mandrel were dipped into the chitosan solution together, taken out, and freeze-dried. After being neutralized in alkaline solution and dried at room temperature, the mandrel was removed to create the chitosan tubular scaffold. Scanning electron micrographs show that the resulting tubes have a biphasic wall structure, with a fibrous inner layer and a semipermeable outer layer. The swelling properties and the mechanical strength before and after in vitro degradation were investigated. The biocompatibility of the scaffolds was also investigated by co-culturing neuroblastoma cells （N2A, mouse） with the scaffolds. The results suggest that these chitosan tubular scaffolds are useful for the regeneration of tissues requiring a tubular scaffold.     

Electrospun Nanofibers of Hydroxyapatite/Collagen/Chitosan Promote Osteogenic Differentiation of BMSCs

Bone tissue engineering, aiming at developing bone substitutes for repair and regeneration of bone defects instead of using autologous bone grafts, has attracted wide attention in the field of tissue engineering and regenerative medicine. Developing biomimetic biomaterial scaffolds able to regulate osteogenic differentiation of stem cells could be a promising strategy to improve the therapeutic efficacy. In this study, clectrospun composite nanofibers of hydroxyapatite/collagen/chitosan （ HAp/Col/CTS ） resembling the fibrous nanostructure and constituents of the hierarchically organized natural bone, were prepared to investigate their capacity for promoting bone mesenchymal stem cells （BMSCs） to differentiate into the osteogenic lineage in the absence and presence of the osteogenlc supplementation, respectively. Call morphology, proliferation and quantified specific osteogenic protein expression on the electrospun HAp/Coi/CTS scaffolds were evaluated in comparison with different controls including dectrospun nanofibrous CTS, HAp/CTS and tissue culture plate. Our remits showed that the nanofibrous HAp/Col/CTS scaffolds supported better spreading and proliferation of the BMSCs than other substrates （ P 〈 0.01 ）. Expressions of osteogenesis protein markers, alkaline phosphatase （ALP） and Col, were significantly upregulated on the HAp/Col/CTS than those on the CTS （P 〈0.01） and HAp/ CTS （P 〈 0. 05 ） scaffolds in the absence of the osteogeulc supplementation. Moreover, presence of osteogeulc supplementation also proved to enhance osteogeule differentiation of BMSCs on HAp/ Col/CTS scaffolds, indicative of a synergistic effect. This study highlights the potential of BMSCs/HAp/Col/CTS cell-scaffold system for functional bone repair and regeneration applications.     

组装法制备壳聚糖/羟基磷灰石/海藻酸复合膜及其性能测定

利用壳聚糖和海藻酸能通过正负电荷吸引形成聚电解质膜的特点,采用层层组装的方法制备了壳聚糖/羟基磷灰石/海藻酸复合膜.用电子扫描电镜观察了复合膜的断裂面和膜内羟基磷灰石(HA)的形貌.研究了复合膜的透光率、溶胀率及热稳定性.结果表明复合膜内HA为贝壳状和花瓣状的纳米晶体,复合膜的断裂面为紧密的连续结构.随着组装次数增加,复合膜的溶胀率变化不大,复合膜的分解温度为301℃.这种具有片状纳米HA的复合膜可能成为一种潜在的骨细胞培养支架材料.     

Dexamethasone-Loaded PLGA Microspheres Incorporated PLLA/PLGA/PCL Composite Scaffold for Bone Tissue Engineering

The combination of micro-carriers and polymer scaffolds as promising bone grafts have attracted considerable interest in recent decades.The poly(L-lactic acid)/poly(lactic-co-glycolic acid)/polycaprolactone(PLLA/PLGA/PCL)composite scaffold with porous structure was fabricated by thermally induced phase separation(TIPS).Dexamethasone(DEX)was incorporated into PLGA microspheres and then loaded on the PLLA/PLGA/PCL scaffoldtopreparethedesiredcompositescaffold.The physicochemical properties of the prepared composite scaffold were characterized.The morphology of rat bone marrow mesenchymal stem cells(BMSCs)grown on scaffolds was observed using scanning electron microscope(SEM)and fluorescence microscope.The resultsshowedthatthePLLA/PLGA/PCLscaffoldhad interconnected macropores and biomimetic nanofibrous structure.In addition,DEX can be released from scaffold in a sustained manner.More importantly,DEX loaded composite scaffold can effectively support the proliferation of BMSCs as indicated by fluorescence observation and cell proliferation assay.The results suggested that the prepared PLLA/PLGA/PCL composite scaffold incorporating drug-loaded PLGA microspheres could hold great potential for bone tissue engineering applications.     

Green Electrospun Silk Fibroin/Galactose Chitosan Composite Nanofibrous Scaffolds for Hepatic Tissue Engineering

The electrospun nanofibrous scaffolds made of proteins and polysaccharides were thought to be able to simulate the structure of natural extracellular matrix well.Silk fibroin(SF)and chitosan(CS)are probably the most widely used natural materials in biomedical fields including liver tissue engineering for their good properties and wide variety of sources.The asialoglycoprotein receptors of hepatocyte were reported to specifically recognize and interact with galactose.In this work,a green electrospun SF/galactosylated chitosan(GC)composite nanofibrous scaffold was fabricated and characterized.The data indicated that the addition of GC greatly influenced the spinning effect of SF aqueous solution,and the average diameter of the composite nanofibers was about 520nm.Moreover,the green electrospun SF/GC nanofibrous scaffolds were demonstrated significantly enhancing the adhesion and proliferation of hepatocyte(RH35)according to our data.The present study did a useful exploration on constructing scaffolds for liver regeneration by green electrospinning,and also laid a good foundation for the further applicative research of this green electrospun scaffolds in liver tissue engineering.     

星星草Puccinellia tenuiflora（Turcz.）Scribn.et Merr.解剖研究

利用扫描电镜技术，研究了星星草Ｐｕｃｃｉｎｅｌｌｉａｔｅｎｕｉｆｌｏｒａ（Ｔｕｒｃｚ．）Ｓｃｒｉｂｎ．ｅｔＭｅｒｒ．的营养器官，发现其根、茎、叶各部器官的组织具有显著的旱生结构特征，即根的皮层细胞破裂，形成发达的通气组织￣［１］；茎的表皮细胞排列紧密，角质层很厚，维管束不成两轮排列，而星散分布于基本组织中，髓腔较小；叶的表皮具乳突状表皮毛，泡状细胞数目较少，且深陷。叶脉维管束鞘不为典型的Ｃ＿３植物结构，两层维管束鞘的细胞壁厚度相等，内层维管束鞘壁并不加厚，外层维管束鞘含叶绿体。这些特征都表明星星草受外界生态因素影响，而演化出各种各样的形态结构来适应所生长的客观生态环境￣［２］。     

聚氨酯改性及复合支架应用于血管组织工程

通过查阅近年来将聚氨酯作为血管组织工程支架材料的相关文献,介绍了聚氯酯改性的常用方法在血管组织工程支架中的应用情况,综述了聚氨酯表面改性支架和复合支架在血管组织工程中的应用.     

角膜移植的免疫学研究——实验性角膜移植排斥反应的病理组织学观察

角膜移植排斥反应现已成为角膜移植手术失败的主要原因,研究角膜移植免疫反应具有重要意义。本文报告了用同一供体皮肤移植引起兔同种穿透性角膜移植排斥反应的动物模型,描述了角膜移植术后和皮肤移植术后的经过,排斥反应的临床表现,组织学标本,包括光镜、扫描电镜和透射电镜标本的制作方法,重点报告了排斥反应实验组的角膜和对照组透明角膜的组织学观察结果。发现对照组的透明角膜,除了在供、受体组织交界处形成疤痕外,植片和受主角膜表现与正常角膜相同。而在发生排斥反应的角膜,则有大量新生血管侵入,植片有大量的浸润细胞,这些细胞主要是淋巴细胞,还有巨噬细胞、浆细胞、多形核白细胞、成纤维细胞等。植片组织遭到不同程度的损害,受主角膜除了有新生血管生长和有少量浸润细胞附着以外,没有受损。本文讨论了有关动物模型、排斥反应的临床特征、病理组织学与免疫机理及功能的联系,新生血管在排斥反应中起的作用等问题。我们认为本组实验动物模型的临床表现和组织学改变均符合典型的特异性免疫反应。这种反应以细胞免疫为主,亦有体液免疫参与。血管在免疫排斥反应的传入、传出弧中均超重要作用。     

壳聚糖多孔支架的制备与生物学性质

用一种蛋白改性壳聚糖,通过冷冻干燥方法制备三维多孔支架,对支架材料的细胞毒性、贴壁率和细胞增殖等生物学特性进行评价.结果显示,采用-20,-70,-196 ℃不同温度预冻后冻干,可以制成孔结构不同的支架材料.通过冻干法制备的蛋白改性壳聚糖多孔支架对细胞没有毒性,L929细胞和人成纤维细胞可以在支架表面和内部正常生长和增殖,角质细胞在材料表面可以正常老化、角质化.     

抗血小板剂对内皮化人工血管动静脉瘘通畅率的影响

目的：探索抗血小板剂对人工血管动静脉瘘血栓形成的影响。方法：将人工血管接种内皮细胞后,给动物犬股动静脉造成动静脉瘘（ＡＶＦ）,术后用抗血小板剂,观察对ＡＶＦ血栓形成的影响,结果：使用抗血小板剂动物的动静脉瘘肉眼血栓形成率为２２．２２％,对照组为４１．６７％（Ｐ＜０．０５）。结论：抗血小板剂能改善实验动物内皮细胞作衬里的人工血管移植动静脉瘘的通畅率。     

秦岭滑蜥中脑视叶组织学结构观察

光镜下观察了秦岭滑蜥（Scincella tsinlingensis）中脑视叶的组织学结构.秦岭滑蜥中脑视叶分为背侧的顶盖和腹侧的被盖两部分,两者之间无明显界限.顶盖处灰质和白质交替排列.神经细胞主要在灰质内,白质中有小细胞零星分布.由深层至表层依次为室管膜层、深白质层、深灰质层、中白质层、中灰质层、浅白质层、外灰质层和分子层.被盖处细胞层次不明显.左右视叶与中脑水管在视叶中部相通.视叶前部有横行的纤维将左右视叶联系起来.与其他脊椎动物中脑的比较结果表明秦岭滑蜥在爬行类进化上处于较低等地位.     

青海玉树多目涡虫咽部组织结构的显微观察

利用连续石蜡切片(包括水平切、矢状切和横切)技术,H.E染色和光镜观察的方法,对青海玉树多目涡虫咽部的组织结构进行了初步的研究,结果表明:咽壁包括三层结构,外层由单层柱状上皮构成,似合胞体,上皮细胞游离面可见三层嗜酸性带状结构,其中,外层带呈刷状;咽壁的中层分为内侧发达的环肌和外侧较薄的纵肌两部分;邻接咽腔的内层上皮在游离面呈指状突起,未见上皮细胞间界限.     

Fabrication and Characterization of Chitosan Nerve Conduits with Microtubular Architectures

Porous multi-channel chitosan conduits were fabricated using a novel phase-separation technique with an axial temperature gradient. First, porous chitosan tubes were made with a mold that was composed of two concentric polytetrafiuoroethylene tubes. Then 1%-3% （w/v） chitosan solution was injected into the chitosan tube while the two ends of the tube were closed with steel rods. Then the outside of the tube was wrapped with a layer of thermal insulating material to reduce the heat transfer through the outside, and the tubes were placed in a freezer. The resulting phase separation then occurred in the presence of an axial temperature gradient. The porosity, microtubule diameter, and orientation were controlled by adjusting the polymer concentration and temperature gradient. After the preparation course, no poisonous substances remained on the conduits. The mechanical properties, swelling, and biodegradability of the chitosan conduits were investigated, and a scanning electron microscope was used to observe the tubular morphology and growth of neuroblastoma cells （N2A, mouse） in the conduits. The results demonstrate that the multi-channel chitosan conduits have suitable mechanical strength, swelling, degradation properties, and nerve cell affinity, so they hold promise for use as neural tissue engineering scaffolds.     

华南五针松种皮发育研究

运用石蜡切片法,在透射电子显微镜和光学显微镜下观察华南五针松(Pinus kwangtungensis)种皮的形态结构和发育,用PAS反应显示细胞内淀粉粒分布,研究细胞内淀粉粒的含量.结果表明,华南五针松的种皮的发育开始在大孢子母细胞发育的时期,开始时在雌配子体的周围形成比较薄而且致密,颜色较深的几层;随着种子的不断发育,从种子的横切面可见,种皮可以明显分为外种皮,中种皮和内种皮三部分.这三部分种皮的形态结构各有不同:外种皮是几层扁平状硬化薄壁细胞;中种皮细胞较多,细胞层较厚,细胞较大,形状不规则,壁厚;内种皮细胞受到发育的胚的挤压,比较扁小,排列紧密.另外,PAS反应显示出,中种皮与内种皮所含的淀粉粒比较丰富,特别是在种子发育的初期,形成了一层富含淀粉粒的细胞层,这层细胞所含的淀粉粒随着种子的发育越来越少,这与种子发育过程需要分解淀粉粒为细胞分裂提供能量有关.     

镀锌钢上钼酸盐/硅烷复合膜的结构与耐蚀性

将热镀锌钢板用二步法（先后浸入钼酸盐钝化液和硅烷溶液）和一步法（一次浸入加有钼酸盐钝化剂的硅烷溶液）获得钼酸盐/硅烷复合膜,用XPS和AES对 2种复合膜的化学成分进行表面分析和剥层分析,用Tafel极化曲线测量和盐雾腐蚀试验（NSS）对膜层的耐蚀性能进行了研究,并将它们与单独的钼酸盐转化膜、硅烷膜对比。结果表明,由一步法制备的复合膜与二步法制备的复合膜具有相似的双层结构,内层以钼酸盐转化膜（含O、Mo、Zn、P）为主而外层以硅烷膜（含C、O、Si）为主,内外层之间及膜与锌基体之间的化学成分呈梯度变化;和单独的钼酸盐膜、硅烷膜相比,2种复合膜对腐蚀的阴极过程的抑制明显增加,自腐蚀电流减小至单层膜的1/5以下,耐蚀性显著提高,接近常规铬酸盐钝化膜。     

壳聚糖/氧化石墨烯复合水凝胶的流变学及其pH响应性能研究

利用壳聚糖与氧化石墨烯之间的静电作用力,制备出了具有p H响应的壳聚糖/氧化石墨烯复合水凝胶体系.运用扫描电镜、X射线衍射、紫外可见光谱和红外光谱对复合水凝胶的微观内部结构与形成机理进行了分析探索.结果表明,壳聚糖与氧化石墨烯的最佳用量比为1:6时,能形成十分稳定的凝胶.此外,这种凝胶表现出明显的p H响应特性.当体系p H在4.98时,复合水凝胶具有最高的粘度1060Pa·s,远高于同浓度的单独壳聚糖与氧化石墨烯溶液,但p H改变后,凝胶粘度均会变小.这种复合凝胶体系不仅制备简便,而且具有p H响应性能,可望扩展石墨烯凝胶材料在生物领域中的应用范围.     

VE TPGS-Loaded Silk Fibroin/Hydroxybutyl Chitosan Nanofibrous Scaffolds for Skin Care Application

Vitamin E（ VE） is an ideal antioxidant and a stabilizing agent in biological membranes. In this study,silk fibroin（ SF） /hydroxybutyl chitosan（ HBC） nanofibrous scaffolds are loaded with VE tocopherol polyethylene glycol 1000 succinate（ VE TPGS） via electrospinning. SEM images show that the average nanofibrous diameter has no significant difference when the content of VE TPGS increases to 4. 0%（ SF / HBC）. However,the average nanofibrous diameter decreases largely to 200 nm when the VE TPGS content reaches 6. 0%. Furthermore,VE TPGS presents a sustained release behavior from the nanofibrous scaffolds. Cell viability studies of mouse skin fibroblasts（ L929） demonstrate that VE TPGS loaded SF / HBC nanofibrous scaffolds present good cellular compatibility.Moreover,the incorporation of VE TPGS could strengthen the ability of SF / HBC nanofibrous scaffolds on protecting the cells against oxidation stress using the Tertbutyl hydroperoxide（ t-BHP）-induced oxidative injury model. Therefore,VE TPGS-loaded SF /HBC nanofibrous scaffolds might be potential candidates for personal skin care,wound dressing and skin tissue engineering scaffolds.     

Hippocampal Neuron-Derived Extracellular Matrix Coated Nanofibrous Scaffold for Neural Tissue Engineering

The aim of this study is to prepare poly-L-lactide( PLLA) electrospun nanofibrous scaffolds coated with hippocampal neuron-derived extracellular matrix( N-ECM)and construct a novel neural tissue engineering scaffold.Neonatal rat hippocampal neurons were seeded on PLLA nanofibers,and then decellularized to derive a cell-free extracellular matrix loaded N-ECM/PLLA modified scaffolds. The morphology and ingredients of N-ECM/PLLA were observed by scanning electron microscopy( SEM) and immunofluorescence staining respectively, and the cytocompatibility of the composite scaffolds was characterized by cell count kit-8( CCK-8) assay. The N-ECM was clearly identified loading on scaffolds when being imaged via SEM and immunofluorescence staining results showed that the N-ECM was made up of fibronectin and laminin. Most importantly, compared with tissue culture polystyrene and pure scaffolds, N-ECM/PLLA scaffolds could effectively facilitate the proliferation of rat adrenal neuroma cells( PC12 cells),indicating their better cell compatibilities. Based on the combination of N-ECM and PLLA biomaterials,the present study has fabricated a unique and versatile neural tissue engineering scaffold,offering a new thought for future neural tissue engineering.     

可降解聚醚酯弹性体PTCG的合成及初步生物学评价

用熔融缩聚法合成聚对苯二甲酸丁二醇酯-co-聚对苯二甲酸环己烷二甲醇酯-b-聚乙二醇(PTCG)聚醚酯弹性体.按照ISO 10993标准,采用L929小鼠成纤细胞对其进行体外细胞毒性测试;以狗的血管平滑肌细胞 (SMCs)为模板细胞,测试细胞在材料表面的贴附性能.采用旋转成型/粒子洗出法制备三维血管支架,接种SMCs后在生物反应器上进行动态培养,探索PTCG作为组织工程血管支架材料的可行性.结果表明,合成的PTCG聚醚酯弹性体无细胞毒性;SMCs在PTCG薄膜表面贴附生长良好;所制备支架的孔径、孔隙率和力学性能等满足组织工程血管支架的要求;体外培养3 d后,大量SMCs长入血管支架并开始分化.     

二例肝泡球蚴病的组织病变考察

报道塔城地区二例泡球蚴病肝的病理病变．组织切片显示病肝组织被无数小泡囊所取代．泡囊的角质层较厚，常呈破裂，内层的生发膜产生的小雏囊均不含原头蚴，全系不育囊．肝的基本组织，包括肝小叶、肝细胞、肝窦、胆小管等均遭受严重的破坏．在病灶中心，可见有坏死与钙化的肝组织，泡囊的周围形成肉芽炎症反应，整个病灶形成肉芽结节．肝组织里并见有些泡囊破裂，生发层浸润周围组织，产生新的小泡囊并见于病灶的纤维组织内．