丝素纤维人工韧带材料表面改性及其体外矿化性能

人工韧带移植是临床治疗韧带断裂及缺损性疾病的有效手段,然而现有人工韧带尚存在不能降解及骨愈合不良的问题。该文设计并编织了一种丝素纤维人工韧带材料,并采用多巴胺溶液对材料表面进行涂层改性。对该人工韧带材料的力学性能、体外矿化性能及细胞相容性进行了一系列表征。结果表明,编织法可以快速成型人工韧带材料,通过调节编织角可调控人工韧带材料不同部位的结构。多巴胺涂层可显著提高丝素纤维人工韧带材料的体外矿化性能。矿化层Ca∶P元素比约为1. 57,结构为无定形。细胞试验结果表明,多巴胺涂层人工韧带材料细胞相容性良好。     

可分解食道支架缝合线体外降解性能的研究

采用磷酸盐缓冲溶液(PBS)(pH=7.4)为降解介质,37℃下对可分解食道支架聚(L-丙交酯)(PLLA)和聚(乙交酯-丙交酯)(PGLA)缝合线进行8周的体外降解实验。通过测试降解过程中其质量损失、pH值、特性粘度、抗张强度、取向度、结晶度、熔点及表面形貌等变化情况,对其体外降解行为进行研究。结果表明PLLA缝合线各项性能无明显变化,而PGLA缝合线各项性能发生了显著变化。随着降解进行PGLA缝合线的pH值、特性粘度、抗张强度、取向度和结晶度逐渐下降,质量损失率增加,6周时抗张强度几乎为零,8周时质量损失率近70%。DSC结果表明,随着体外降解PGLA缝合线晶区熔点保持不变,晶区的熔融热焓逐渐增加,而无定形区产生新的有序区.并且含量不断增加。SEM结果表明PGLA缝合线在降解过程中,涂层首先脱落,然后线体以横向断裂方式断裂。因此,PGLA缝合线适用于良性食道狭窄的可分解食道支架,而PLLA缝合线制备的支架分解时间大于2个月,不适于良性狭窄的治疗。     

丝素蛋白/聚己内酯纳米纤维支架生物相容性评价

目的:改善再生丝素蛋白的降解性及力学性能,评价丝素蛋白/聚己内酯纳米纤维支架神经生物材料的生物相容性。方法:采用静电纺丝技术制备丝素蛋白/聚己内酯纳米纤维支架。体外培养雪旺细胞并与支架及其浸提液共培养,通过荧光染色,细胞毒性试验(MTT法)检测其细胞生物相容性。将纤维支架材料在体外置于蛋白酶ⅪV溶液评价其体外降解行为;通过皮下埋植实验观察纤维材料在体内的局部组织反应。结果:丝素蛋白/聚己内酯支架材料,呈现三维网状结构。雪旺细胞具有良好的生长形态;无细胞毒性。随着丝素蛋白比例的降低,能够显著增加混合支架的降解速度。皮下移植实验未引起明显免疫排斥反应,炎症反应轻。结论:丝素蛋白/聚己内酯支架具有良好的生物相容性和生物可降解性,有望用于神经组织工程支架材料修复神经缺损。     

纤维连接蛋白表面修饰对促进新型PET材料人工韧带（LARS）细胞黏附的实验研究

目的通过体外实验探讨采用表面水解法处理纤维连接蛋白表面修饰增进新型PET材料人工韧带生物相容性的有效性。方法采用碱性水解法表面处理后再与纤维连接蛋白反应修饰该新型人工韧带,使用红外透射光谱分析和X线衍射分析技术测定材料表面蛋白结合含量,采用大鼠成纤维细胞与聚脂纤维人工韧带材料体外共培养的方法,观察细胞粘附、增殖以及分化情况,对比采用纤维连接蛋白表面修饰的材料和未处理的材料的生物相容性的差别。结果红外透射光谱分析和X线衍射分析技术证实通过纤维连接蛋白表面修饰后该新型聚脂纤维人工韧带表面蛋白含量明显提高,采用纤维连接蛋白预处理的聚脂材料上细胞能够早期粘附,附着生长情况更好,数量更多。结论采用碱性水解表面处理纤维连接蛋白修饰的方法简便、有效,能够增进新型PET材料人工韧带的生物相容性。     

新型聚乳酸与纳米羟基磷灰石复合人工骨的性能测试研究

目的　通过原位聚合法制备新型的聚乳酸与纳米羟基磷灰石复合材料,找到两者复合的最佳配比,从而得到理想的人工骨移植材料。 方法　采用原位聚合法按一定的配比（纳米羟基磷灰石质量分数分别为0,10%,20%,30%,40%)聚乳酸与纳米羟基磷灰石复合人工骨,对这类新型的人工骨进行性能测试,通过抗弯,抗压,弹性模量,电镜扫描,体外降解实验,观测该人工骨的力学性能、微观结构、纳米羟基磷灰石在聚乳酸中的分散情况以及复合材料的降解性能。 结果　（1）力学测试显示：随着n-HA含量的增加,复合材料的拉伸强度逐渐减少。复合材料的弯曲强度在n-HA微粒的质量分数为20%时弯曲强度出现峰值(156.8 MPa)。复合材料的弯曲模量随着n-HA微粒质量分数的增加而增大。（2）SEM扫描显示：纯PDLLA材料断裂表面较平整;在n-HA含量为10%时出现大量的韧窝, 明显的粗糙断裂面;在n-HA含量为20%断裂表面凹凸不平,形成大量的韧窝;在n-HA含量为30%以上时断口又变得越来越平整,尚有许多小的韧窝。（3）体外降解实验显示：随着降解时间的延长,降解液的pH值均逐渐降低,复合材料的力学性能也逐渐的产生一定的衰减。 结论　当n-HA含量为20%时,该人工骨复合材料有着更好的力学性能和降解性能,筛选出该种新型人工骨的最佳配比,制备出性能良好的PDLLA/n-HA复合人工骨材料。     

丝素纤维人工韧带材料的表面改性及细胞相容性

该文研究设计并制备了一种可降解的丝素纤维人工韧带材料,并对其进行了表面改性和细胞相容性研究。以丝素纤维为原材料编织仿生结构的人工韧带,对其材料表面进行低温氧等离子体处理,再通过EDC/NHS接枝I型胶原蛋白。同法接枝人纤维蛋白原和转化因子TGF-β1结合肽到材料表面。对材料的细胞相容性进行评价,结果表明,经过等离子体处理和生物活性分子接枝后,细胞相容性有了进一步提升。成纤维细胞在接枝了人纤维蛋白原的材料表面活性更高。     

甲壳素短纤维增强聚己内酯复合材料体外降解实验研究

将纯聚己内酯（PCL）、甲壳素短纤维增强PCL复合材料和聚DL-乳酸分别置于生理盐水溶液中,进行体外降解实验研究。于2、4、8、12、16、24周分别取材,测试降解液pH值、试样失重率及力学性能的变化,并行扫描电镜观察。结果显示甲壳素短纤维增强PCL初始强度大于纯PCL,在降解过程中,浸泡液pH值呈弱酸性和中性,失重速率快于纯PCL,力学性能先高后低,强度和模量值24周与初始值相差不大。通过以上体外降解实验可以得出结论：PCL树脂复合甲壳素纤维后,加快降解速度,提高力学性能,缓冲材料局部pH值的下降,是一种很有前途的胸壁缺损修补及骨科内固定材料。     

PGLA降解产物对材料降解性能影响的体外研究

为了探讨PGLA材料的降解产物对材料本体降解性能的影响，本研究设计了体外两种浸泡体系——磷酸缓冲液（PBS）和人工血浆，分别在替换和不替换浸泡介质两组不同条件下，分析材料的质量损耗情况，并将不替换组的各时间段降解液进行pH值的测定，以判断环境的酸碱度对材料本体降解的作用。实验降解周期分别为2、3、4、6、8、10周。结果显示：在浸泡初期，人工血浆和PBS降解液中的质量损耗两者间无显著性差异（P〉0．05）；大约从2周到6周，不替液组中材料在两种介质内的降解速度显著快于相对应的同期换液组（P〈0．01）；不替换液组材料在PBS液中的降解快于在人工血浆中的降解，而且在整个降解周期内PBS降解液的pH值基本保持稳定（pH≈7．0～7．4），而人工血浆降解液中的pH值随着降解过程的不断进行，其pH值由7．5逐渐降到5．7左右，两种降解液pH值变化之间的差异具有显著的统计学意义（P〈0．01）。由此提示：在不替换浸泡介质的条件下，降解液中持续存在的降解产物显然影响材料本体的降解速度，PGLA材料的降解产物具有加速材料质量损耗的作用；环境的酸碱度对材料的降解会产生一定的影响，pH值越低，材料的降解速度越缓慢。     

聚乙交酯丙交酯体内外生物降解性能的相关研究

通过对聚乙交酯丙交酯 (PGL A)进行体内和体外生物降解性的实验研究 ,探讨两种降解过程之间的关联性。体外实验是将 PGL A材料 (1cm× 1cm)分别浸泡在人工唾液和 PBS溶液中 ,体内 (动物 )试验是将材料直接植入 Wistar大白鼠的皮下组织 ,浸泡后或植入后 1～ 10周 ,每周计算材料的质量损耗率 ,每 2周进行分子量测定。实验结果显示 :PGL A材料在人工唾液中的降解要快于在 PBS液中的降解 ;材料质量损耗的发生慢于分子量的变化 ;体外的降解周期大约为 9～ 10周 ,体内降解周期为 8周左右 ,体内组的降解速率是体外组的 1.33倍。结论为PGL A体外降解主要是化学降解过程 ,通过酯键的水解来进行。体内降解过程中 ,应力环境和生物因素都会对材料的降解动力学产生影响 ,使降解过程明显加快 ,但体内和外降解都符合脂肪族聚酯降解的动力学模型 ,PGL A的体内外生物降解性之间存在一定的相关性。     

不同降解材料制备的分解食道支架体外降解研究

采用磷酸盐缓冲溶液(PBS)(pH =7.4)和人工胃液(pH=1.5)为降解介质,37℃下对由PLLA和PGLA丝线缝合的分解食道支架进行了体外降解实验.通过测试降解过程中支架的径向支撑力及丝线的特性粘度、熔点、结晶性及表面形貌等变化,对支架体外降解行为进行研究.结果表明,上述降解介质中由PL-LA丝线缝合的支架的径向支撑力及丝线的各性能变化不明显;由PGLA丝线缝合的支架在PBS中6周时分解,丝线的特性粘度保留率随降解时间的延长线性下降,而在人工胃液中9周时分解,特性粘度保留率随降解时间的延长呈“Z”字型变化,即初期和后期变化平缓,中期迅速下降;SEM显示在上述介质中,支架分解时PGLA丝线均以横向方式断裂.因此,PGLA丝线制备的支架可用于治疗良性食道狭窄,而PLLA丝线制备的支架分解时间大于2个月,不适于良性狭窄的治疗.     

Enzymatic degradation behavior of porous silk fibroin sheets

We investigated the degradation behavior of porous silk fibroin sheets by in vitro enzymatic experiments with alpha-chymotrypsin, collagenase IA, and protease XIV. With 1.0 U/ml protease XIV, 70% of a silk fibroin sheet was degraded within 15 days at 37 degrees C. When the fibroin sheet was exposed to collagenase IA, the amount of Silk II crystalline structure in the sheets decreased slightly, and a small amount of Silk I crystalline structure was formed. When protease XIV was used, almost all Silk II disappeared, but the crystallinity increased overall because the amount of Silk I increased. During digestion with protease XIV, the pore size of the fibroin sheets increased with increasing degradation time, until the sheets finally collapsed and became totally shapeless. The average molecular weight of the products after degradation with the three enzymes followed the order protease XIV < collagenase IA < alpha-chymotrypsin. More than 50% of the products resulting from degradation with protease XIV were free amino acids.     

Biodegradation behavior of silk fibroin membranes in repairing tympanic membrane perforations

Silk fibroin (SF) from silkworms has been widely studied as a biomaterial. The degradation behavior of silk biomaterials is important for medical applications, but few studies have examined long-term degradation behavior in vivo. In this study, we investigated the degradation behavior of SF membranes in vitro and in vivo. For the in vitro assay, we observed degradation of silk membranes in phosphate buffered saline, culture media, and an enzyme (proteinase K) solution. In the proteinase K solution, 80% of the silk membranes degraded within 10 days. Silk membranes exhibited no cytotoxicity toward L929 cells and rat tissues. To investigate the degradation of silk membranes in vivo, they were implanted subcutaneously in rats and harvested 19 months after surgery. Scanning electron microscopy imaging and histological analysis of silk membrane explants showed that they broke into several pieces after 16 months. Results show that silk membranes are biocompatible and display excellent long-term degradation behavior when used as biomaterials.     

Degradation behaviour of self-reinforced 80L/20G PLGA devices in vitro

In vitro degradation of self-reinforced PLGA 80L/20G material and bioabsorbable stents was studied in artificial urine and phosphate buffer solution (PBS) to define if the media have an effect on the degradation rate in urological applications. After six weeks, the Mv of the samples immersed in PBS was 40% (16.7 kDa) from the initial value and 57% (24.0 kDa) for the samples immersed in artificial urine. The strength loss of samples that were immersed in PBS was slower when compared with samples in artificial urine. The bending strength of samples immersed 15 weeks in artificial urine was 43% (21.7 MPa) of the bending strength of samples immersed in PBS (50.9 MPa), and the shear strength was 13% (artificial urine 3.7 MPa, PBS 28.8 MPa), respectively. The maximum compression force in PBS was slightly over at the initial level after 2 weeks of immersion. It decreased to half (102.2N) of the initial value (204.1N) in 8 weeks, and after 12 weeks it was 25% (50.8 N) of the initial value. The compression force in artificial urine was 35% (66.8 N) of the initial value (193.9 N) after 8 weeks. In 12 weeks it had lowered to 26 N in artificial urine, which was 14% of the initial value. The degradation rate of self-reinforced L-lactic and glycolic acid stents in vitro tests in artificial urine was coinciding with our clinical test. Based on these results, it is possible to make a sufficiently accurate in vitro model for the degradation rate of bioabsorbable polymers for urological applications.     

聚丁二酸丁二醇酯/聚碳酸亚丙酯生物膜的制备及相关性能评价

背景：聚丁二酸丁二醇酯、聚碳酸亚丙酯是近年来发展的新型医用材料,具有良好的生物相容性、可降解性及价格低廉等优点。 目的：采用静电纺丝法制备聚丁二酸丁二醇酯/聚碳酸亚丙酯生物膜,评价其理化性能、体外降解性能及对细胞增殖的影响。 方法：采用静电纺丝法制备聚丁二酸丁二醇酯/聚碳酸亚丙酯生物膜：常温下称取聚丁二酸丁二醇酯、聚碳酸亚丙酯各0.9 g,溶解于10 mL三氯甲烷中,磁力搅拌,待其充分溶解后,将纺丝液加入纺丝管内,调整纺丝距离约15 cm,最后开启电源将电压调至18 kV即可,扫描电镜观察其表面形态,检测其强度、接触角、吸水率及体外降解过程中的pH值和失重率。将人骨肉瘤细胞MG63与生物膜共培养7 d后,采用CCK-8检测细胞增殖。 结果与结论：扫描电镜可见聚丁二酸丁二醇酯/聚碳酸亚丙酯生物膜呈均匀多孔状,孔孔交通,纤维直径约  0.88 μm,平均孔径约5.68 μm,孔隙率为78.3%,断裂强度平均为2.31 MPa,断裂伸长率为23.48%,接触角约为87°,吸水率为68.54%;生物膜降解过程中pH值呈缓慢降低,第12周时pH值为6.76,同时该生物膜处于均匀缓慢的降解过程中,12周时失重率为6.04%。CCK-8实验结果显示该生物膜对细胞的增殖具有一定的促进作用。表明静电纺丝聚丁二酸丁二醇酯/聚碳酸亚丙酯生物膜具有良好的理化性能,具备良好的空间维持能力、湿润性及降解性,能为骨组织再生提供充足的时间。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

Biocompatibility evaluation of silk fibroin with peripheral nerve tissues and cells in vitro

Silk-based materials have been used in the field of bone or ligament tissue engineering. In order to explore the feasibility of using purified silk fibroin to construct artificial nerve grafts, it is necessary to evaluate the biocompatibility of silk fibroin material with peripheral nerve tissues and cells. We cultured rat dorsal root ganglia (DRG) on the substrate made up of silk fibroin fibers and observed the cell outgrowth from DRG during culture by using light and electron microscopy coupled with immunocytochemistry. On the other hand, we cultured Schwann cells from rat sciatic nerves in the silk fibroin extract fluid and examined the changes of Schwann cells after different times of culture. The results of light microscopy, MTT test and cell cycle analysis showed that Schwann cells cultured in the silk fibroin extract fluid showed no significant difference in their morphology, cell viability and proliferation as compared to that in plain L15 medium. Furthermore, no significant difference was found in expression of the factors secreted by Schwann cells, such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and S-100, between Schwann cells cultured in the silk fibroin extraction fluid and in plain L15 medium by the aid of immunocytochemistry, RT-PCR and Western analysis. Collectively, these data indicate that silk fibroin has good biocompatibility with DRG and is also beneficial to the survival of Schwann cells without exerting any significant cytotoxic effects on their phenotype or functions, thus providing an experimental foundation for the development of silk fibroin as a candidate material for nerve tissue engineering applications.     

丝蛋白应用于牙周组织工程的可行性

背景：丝蛋白是有利于表皮细胞、成纤维细胞、成骨细胞、血管内皮细胞、胶质细胞黏附和生长的一种新型生物材料。 目的：评估丝蛋白作为支架材料应用于牙周组织工程的可行性。 方法：采用组织块法培养人牙周膜细胞,将第5代细胞悬液以2×10⁷ L^-1的浓度接种到丝蛋白支架材料上复合培养,并以1%,10%,50%,100%的丝蛋白支架浸提液培养,观察人牙周膜细胞在丝蛋白上及在丝蛋白浸提液中生长状况,用MTT法测定浸提液培养人牙周膜细胞的活力。 结果与结论：扫描电镜可见人牙周膜细胞在丝蛋白支架上伸展充分,生长旺盛,不同浓度丝蛋白支架浸提液培养对人牙周膜细胞的增殖与碱性磷酸酶活性均无影响。说明丝蛋白材料具有良好的生物相容性、独特的力学性能,可作为人牙周膜细胞黏附生长的理想支架材料较好地应用于牙周组织工程中。     

丝素蛋白作为组织工程材料应用于角膜的研究进展

背景：丝素蛋白纤维材料具有透明性、结构可塑性、成分单一性、力学强韧性及生物相容性等特点。 目的：综述国内外丝素蛋白应用于角膜组织工程的研究进展。 方法：由第一作者在标题和摘要中以“silk fibroin, corneal, ocular”或“丝素,角膜”为检索词,检索1980至2011年PubMed及1990至2011年CNKI数据库中关于丝素蛋白角膜的文章。 结果与结论：从天然蚕丝中提取的高分子丝素蛋白,因其良好的生物相容性、独特的力学性能、光学透明性及降解速率可控性,既可以单独应用于角膜组织结构的重建,又可与其他组织材料联合应用,成为角膜组织工程学应用的理想材料。现已证明多种角膜细胞可在丝素纤维膜上良好生长,但体外培养的细胞应用于动物模型的相关研究较少;此外丝素蛋白材料植入角膜内对其产生何种影响的研究数据较缺乏,这些均是亟待解决的问题。     

In vitro degradation of silk fibroin

A significant need exists for long-term degradable biomaterials which can slowly and predictably transfer a load-bearing burden to developing biological tissue. In this study Bombyx mori silk fibroin yarns were incubated in 1mg/ml Protease XIV at 37 degrees C to create an in vitro model system of proteolytic degradation. Samples were harvested at designated time points up to 12 weeks and (1) prepared for scanning electron microscopy (SEM), (2) lyophilized and weighed, (3) mechanical properties determined using a servohydraulic Instron 8511, (4) dissolved and run on a SDS-PAGE gel, and (5) characterized with Fourier transform infrared spectroscopy. Control samples were incubated in phosphate-buffered saline. Fibroin was shown to proteolytically degrade with predictable rates of change in fibroin diameter, failure strength, cycles to failure, and mass. SEM indicated increasing fragmentation of individual fibroin filaments from protease-digested samples with time of exposure to the enzyme; particulate debris was present within 7 days of incubation. Gel electrophoresis indicated a decreasing amount of the silk 25 kDa light chain and a shift in the molecular weight of the heavy chain with increasing incubation time in protease. Results support that silk is a mechanically robust biomaterial with predictable long-term degradation characteristics.