多孔HA/PU复合支架材料生物相容性的评估

进行了三维多孔立体结构的纳米羟基磷灰石/聚氨酯(HA/PU)复合支架材料体外细胞培养和体内肌肉埋植实验研究,评估材料的生物相容性。实验选用SD大鼠的骨髓基质干细胞(BMSCs)和健康的SD雌性大鼠,进行细胞相容性、形态学观察和组织学切片分析。HA/PU支架材料的多孔性为细胞的生长提供了良好的微环境,细胞在内部贴壁爬行、增殖并分化,细胞毒性为零级,材料与周围组织有良好的结合,降解的空间有结缔组织纤维长入。实验表明,HA/PU复合支架材料具有良好的细胞亲和性和组织学相容性,可作为一类新型组织工程支架材料。     

胶原-纳米羟基磷灰石复合支架的细胞相容性

背景：观察成骨细胞在生物材料上的形态、增殖和分化等项目,可评估生物支架材料的生物相容性。 目的：观察复合支架材料纳米羟基磷灰石/胶原对成骨细胞增殖、分化的影响。 方法：取新生24 h内Wistar大鼠的颅盖骨,采用改良胶原酶消化法进行成骨细胞原代培养,取第3代细胞与纳米羟基磷灰石/胶原支架或普通羟基磷灰石材料体外复合培养。培养3,6,9 d后,观察材料周边的细胞形态及支架材料对细胞分化、增殖的影响。 结果与结论：纳米羟基磷灰石/胶原材料较普通的羟基磷灰石材料更有利于成骨细胞的黏附、生长、分化、增殖,证实其生物相容性更好,有望成为一种新型的骨组织工程支架材料。     

血管组织工程生物支架材料细胞生物相容性实验研究

目的采用新方法研究天然生物支架材料胶原/透明质酸膜,明胶海绵的细胞相容性。方法应用WST-1法测定平滑肌细胞与材料的粘附率,增殖力,^3H-TDR掺入法测定DNA合成率,BrdU细胞标记免疫组化鉴定,分析组织工程生物支架材料的细胞相容性。结果WST-1法测定,^3H-TDR掺入法测定DNA合成率,BrdU细胞标记结果表明：胶原/透明质酸膜与平滑肌细胞的粘附率最高,细胞的增殖和代谢状况较好,明胶海绵较低。结论应用WST-1法测定,^3H-TDR掺入法测定DNA合成率,BrdU细胞标记方法研究细胞相容性方法简便可行;天然复合生物材料胶原/透明质酸膜具有较理想的细胞相容性。     

三维多孔支架材料PLLA/n-HA的体外生物相容性研究

目的：研究三维多孔支架材料聚乳酸/纳米羟基磷灰石(PLLA/n-HA)的体外生物相容性,探讨其作为细胞培养材料和骨组织工程支架的可行性。方法将大鼠成骨细胞接种于PLLA/n-HA复合支架上,体外共同培养后,CCK-8法检测大鼠成骨细胞增殖活性,荧光倒置显微镜、扫描电子显微镜下观察PLLA/n-HA复合支架材料表面和孔隙内细胞粘附情况。结果 CCK-8法检测显示实验组复合支架材料上细胞的增殖与空白对照组的差异无统计学意义（P>0.05）;电镜观察到细胞在复合支架材料表面和孔隙内大量黏附、生长,并且随着共培养时间的增加,材料表面的细胞数量呈几何级增长。结论三维多孔支架材料PLLA/n-HA的生物相容性较好,可望成为一种性能良好的骨组织工程支架材料。     

多孔纳米羟基磷灰石/壳聚糖支架材料复合成骨细胞的异位成骨

目的评价多孔纳米羟基磷灰石/壳聚糖﹙nHA/CS﹚支架与成骨细胞复合植入大鼠股部肌袋模型内的异位成骨能力。方法采用共沉淀和粒子沥滤法制备多孔nHA/CS。分离培养培养SD大鼠成骨细胞,并将其与多孔nHA/CS共同培养来构建组织工程骨。将所构建的组织工程骨和空白nHA/CS支架材料分别植入SD大鼠股部肌袋模型内。分别在植入2、4、6、8周后,将植入的支架材料取出行苏木精-伊红染色观察新骨形成情况,并应用JEDA-801D形态学图象分析系统来计算新骨生成率。用SPSS13.0软件包对测定结果进行单因素方差分析。结果在各观察时间段内成骨细胞复合多孔nHA/CS组新骨生成量均多于nHA/CS。结论多孔nHA/CS复合成骨细胞支架材料具有异位成骨能力。     

磁性多孔磷酸三钙陶瓷的生物相容性分析

一种新型骨移植替代材料,磁性多孔磷酸三钙陶瓷(MPTCP)可能为骨缺损的治疗开辟一条新途径.为检验其生物相容性及毒性,我们用40只大白鼠(雌雄各半)进行了以下实验:将平均重量为15.3mg的该材料颗粒植入大白鼠双侧股骨近端人工钻孔中,一半动物仅钻孔不植入材料作为对照组.术后30天作动物体重、器官重、器官/体重以及血液学、临床生化、X线拍片、普通切片、扫描电镜检查.结果表明,该材料生物相容性好,无异物及毒性反应,且材料周围骨细胞增生活跃,并向陶瓷孔隙中生长.故认为,MPTCP是一种很值得探讨的骨移植替代材料.     

含氟羟基磷灰石涂层体外细胞相容性研究

在羟基磷灰石（HA）中掺入氟可以降低其在体内的溶解性，对提高钛合金植入体表面生物活性改性层（涂层）的长效性有着重要的意义。本研究采用溶胶-凝胶法在钛合金基板上制备了含氟羟基磷灰石（Ca10（PO4）6（OH）2-xFx）（FHA）涂层。X光衍射（XRD）和X光电子能谱（XPS）分析结果显示：本实验中获得的涂层是一个纯磷灰石晶相，含氟涂层中x为1．2。在体外成骨细胞相容性的研究中，以HA涂层为对照，通过定时细胞的计数测定了细胞生长曲线，用流式细胞仪法测定了细胞周期，用MTT比色法分析了涂层浸提液的细胞毒性。实验结果表明：在HA涂层中引入氟后，FHA涂层浸提液对细胞毒性级别为0级，该涂层不但没有对细胞产生毒性，而且会促进成骨细胞的贴壁生长，有着更好的细胞生长速度和增殖活性。     

丝素蛋白/壳聚糖复合支架材料的细胞相容性**

背景：大量研究表明丝素蛋白、壳聚糖为天然高分子材料,具有良好的细胞生物相容性。 目的：探讨丝素蛋白/壳聚糖复合支架材料与诱导的兔骨髓间充质干细胞的生物相容性。 方法：将兔骨髓间充质干细胞分离培养、诱导后,与丝素蛋白/壳聚糖三维支架材料体外共培养,以材料的细胞毒性、细胞增殖活力、材料细胞黏附率及扫描电镜等检测评价材料的细胞相容性。 结果与结论：经诱导后的骨髓间充质干细胞在支架材料上黏附、生长良好,保持正常的分裂增殖速度;随时间的增加,细胞黏附率增加,材料组较对照组黏附率强,差异有显著性意义(P < 0.05)。扫描电镜观察发现细胞接种48 h后细胞生长良好,与支架黏附紧密,增殖分裂活跃。说明丝素蛋白/壳聚糖三维支架材料具有良好的细胞相容性。     

软骨脱细胞细胞外基质多孔支架与山羊髓核细胞生物相容性研究

目的 制备软骨脱细胞细胞外基质多孔支架,并探讨其与山羊髓核细胞的生物相容性.方法 猪关节软骨经研磨、脱细胞、冷冻干燥技术等处理制成三维多孔支架;从山羊腰椎间盘中分离出髓核细胞,培养后获取P1代细胞;四甲基偶氮唑蓝(MTT)检测支架浸提液毒性;将髓核细胞以5×106/ml的密度接种在支架上体外培养48 h,通过倒置显微镜、HE染色、死活细胞染色(LIVE/DEAD染色)、扫描电镜观察细胞在支架上的黏附及活性.结果 软骨脱细胞基质多孔支架在敷水状态下光滑透明,分离的髓核细胞呈典型的软骨细胞样形态;MTT检测各组间增殖,其差异不具有统计学意义(P＞0.05);倒置显微镜、电镜观察髓核细胞呈球状或短梭形均匀地贴附在支架内部,HE染色观察可见髓核细胞均匀分布在支架内部,LIVE/DEAD染色显示全部为绿色荧光(活细胞),未见红色荧光(死细胞).结论 软骨脱细胞基质多孔支架在组成上与髓核组织相似,与山羊髓核细胞具有良好的生物相容性,可以作为髓核组织工程的支架材料.     

新型纳米纤维支架的细胞生物相容性

背景：高孔隙率聚己内酯纳米纤维支架具有适合血管平滑肌细胞黏附、增殖的多级孔径结构,具有良好的细胞生物相容性。 目的：探讨高孔隙率聚己内酯静电纺丝纳米纤维支架的细胞相容性。 方法：根据支架的制作工艺不同分为传统支架组、新型纳米纤维支架组两组,另设单纯细胞组为对照组。采用组织块贴壁法体外原代培养兔主动脉平滑肌细胞并进行传代,用3~6代细胞作为实验用种子细胞。应用WST-1法测定平滑肌细胞黏附率、增殖力,光镜及扫描电镜观察细胞形态,评估支架的细胞生物相容性。 结果与结论：高孔隙率聚己内酯纳米纤维支架对细胞形态无明显影响,新型支架上的种子细胞黏附、增殖及代谢活性情况较传统支架好。提示,高孔隙率聚己内酯静电纺丝纳米纤维支架具有较高的细胞相容性。     

A bioactive titanium foam scaffold for bone repair

While titanium has been clinically successful as an orthopedic or dental implant material, performance problems still persist related to implant-bone interfacial strength and mechanical modulus mismatch between titanium and tissue. We describe here the preparation of a titanium foam as a better mechanical match to tissue with surfaces attractive to bone cells through deposition of an organically-modified apatite layer (organoapatite). In a rotating bioreactor, these organoapatite-coated foams are successfully colonized by preosteoblastic cells. Finite element analyses suggest that ingrown tissue in these systems may improve both implant performance and tissue formation through load-sharing and stress distribution. The novel metal-ceramic-polymer hybrid materials described here hold great promise for bone tissue engineering.     

新型海藻酸钠组织工程支架材料与人成纤维细胞的相容性

目的 利用N-乙基.N'-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)作为催化剂-乙二胺作为交联剂制备新型海藻酸钠组织工程支架材料(简称材料)并测试其细胞相容性.方法 以体外培养的人成纤维细胞作为对象.采用四甲基偶氮唑盐(MTT)法检测材料浸渍液对细胞增殖情况的影响,光镜下观察材料浸渍液中细胞的生长状况.将人成纤维细胞悬液接种于材料表面,制备人成纤维细胞-材料复合物,扫描电镜下观察培养7 d的复合物表面细胞的黏附和生长状态,评价材料的细胞相容性.结果 材料浸渍液作用1、2,4、7 d后人成纤维细胞的相对增殖率(RGR)分别为98.00%、104.10%、110.80%、93.17%,毒性均为0级或1级.倒置显微镜观察人成纤维细胞在材料浸渍液中生长形态良好.扫描电镜下培养7 d的人成纤维细胞-材料复合物表面人成纤维细胞能很好的与材料黏附,材料表面细胞均生长旺盛形成许多伪足状突起,并分泌产生细胞基质.结论 新型海藻酸钠组织工程支架材料与人成纤维细胞的相容性良好,为其作为细胞生长支持物和进一步的医学应用提供了实验依据.     

Porous scaffold of gelatin-starch with nanohydroxyapatite composite processed via novel microwave vacuum drying

Hydroxyapatite (HA) is a fundamental mineral-based biomaterial, used for preparing composites for bone repair and regeneration. Gelatin blended with starch results in scaffold composites with enhanced mechanical properties. A gelatin-starch blend reinforced with HA nanocrystals (nHA) gave biocompatible composites with enhanced mechanical properties. In this study, a porous scaffold of gelatin-starch-nHA composites was fabricated through microwave vacuum drying and crosslinking using trisodium citrate. Three different composite scaffolds were prepared at three different percentages of nHA: 20%, 30% and 40%. The microstructures and compositions of the composites were analyzed. Within the porous structure, the nHA crystals were observed to precipitate. The interaction between the gelatin-starch network film and nHA crystalline material was studied using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction analysis (XRD). XRD reflections showed that there are two different minerals present in the scaffold composite. There were strong reflection peaks close to the 26 degrees and 32 degrees 2theta angles of HA, and close to the 8 degrees and 49 degrees 2theta angles for sodium citrate minerals. The FTIR result suggested that carboxyl groups, C=O and amino groups play crucial roles in HA formation on the surface of a gelatin network.     

神经干细胞在新型复合支架中的生长和分化

BACKGROUND:Neural stem cells with self-proliferation and differentiation potential are the ideal seed cells for central nervous tissue engineering. Although collagen and silk fibroin as biological scaffold materials have been widely used, both of them used alone have certain shortcomings. Is it possible to combine the two materials to build a novel neural tissue-engineered scaffold? What is the effect of this novel scaffold on the growth and differentiation of neural stem cells? OBJECTIVE:To observe the growth and differentiation of neural stem cells seeded onto the novel composite scaffold. METHODS:The rat embryonic neural stem cells were inoculated onto new composite scaffolds, and then, their growth and differentiation were observed by light microscopy and scanning electron microscopy. Neural stem cells were cultured in conventional suspension culture as control group. Cell counting kit-8 assay was used to detect viability of neural stem cells in the two groups. Three-dimensional composite scaffolds carrying neural stem cells were sliced into paraffin sections to observe the growth and differentiation of neural stem cells by hematoxylin-eosin staining and immunofluorescence staining. RESULTS AND CONCLUSION: Neural stem cells cultured on the new composite scaffold grew and differentiated well, and interconnected synapses were observed. Cell counting kit-8 assay showed that neural stem cells on the scaffold grew well, and the cell viability was significantly higher in the composite scaffold group than that in the control group (P < 0.05). Hematoxylin-eosin staining and immunofluorescence staining of paraffin sections further provided evidence for good growth and differentiation of neural stem cells on the scaffold. These results indicate that the novel composite scaffold with good biocompatibility benefits the growth and differentiation of neural stem cells, promising a favorable application prospect.     

纳米晶重组类人胶原基/聚乳酸复合支架材料的表征及细胞相容性研究

目的　制备纳米羟基磷灰石/重组类人胶原基/聚乳酸复合支架材料 (nano-hydroxyapatite/ recombinant human- like collagen/polylactic acid,nHA/RHLC/PLA),观察材料的形貌特征,探讨材料对骨髓基质干细胞(BMSCs)增殖、黏附及分化等生物学行为的影响。 方法　制备nHA/RHLC/PLA复合支架材料,应用X 光衍射分析（XRD）、红外光谱分析（FTIR）、ZWICK Z005 测试机对样品的化学成分、机械性能测试和压缩强度进行测试,通过扫描电镜检查等方法观察材料的表征;将犬骨髓基质细胞（BMSCs）接种在支架材料上培养,检测材料-细胞的黏附情况及材料对细胞生长增殖的影响。 结果　nHA/RHLC/PLA复合支架材料压缩强度均大于1MPa,达到了天然松质骨的最低强度。扫描电镜结果显示：支架材料呈三维多孔结构,孔为不规则多边形,孔的走向多样,纵向和横向孔隙互为交通,孔径在几十微米到300微米不等,孔隙率为75%~83%。nHA/RHLC/PLA复合支架材料表面BMSCs的黏附、生长良好;而BMSCs的增殖能力与对照组相比,差异无显著性意义(P>0.05)。 结论　nHA/RHLC/PLA复合支架材料符合组织工程骨支架的力学要求,具有良好的微观结构,无细胞毒性,细胞与支架生物相容性良好。利用重组类人胶原代替动物源性胶原制备纳米晶骨修复材料,规避了动物胶原交叉感染的风险,有望成为一种理想的骨组织工程支架材料。     

双层蛛丝蛋白血管支架的制备及其生物力学性能与细胞相容性研究

目的 应用静电纺丝法制备一种双层蛛丝蛋白血管支架,观察血管支架的微观结构,并研究其生物力学性能和细胞相容性。方法 配制纺丝液,通过静电纺丝,以旋转接收棒为收集装置,制备(pNSR16/PCL/CS)/(pNSR16/PCL/Gt)双层蛛丝蛋白血管支架。探讨质量分数和管壁厚度对血管支架孔隙率、爆破强度、拉伸性能、缝合强度和水渗透性的影响,并检测血管支架的细胞毒性和细胞黏附性能。结果 血管支架的微观结构为纤维随机分布的三维多孔网状,爆破强度、拉伸强度和缝合强度大小均与支架的质量分数和管壁厚度成正比,孔隙率、水渗透性和断裂伸长率大小与支架的质量分数和管壁厚度成反比。血管支架爆破强度的范围为43~183 kPa,高于生理血压;缝合强度高于0.19 N,符合体内移植要求;拉伸强度高于人体桡动脉血管,满足体内移植的要求;水渗透性为0.3~0.6 mL?min-1?cm-2。血管支架无细胞毒性,并有利于内皮细胞细胞黏附及增殖。结论 使用静电纺丝法制备的双层蛛丝蛋白血管支架是可行的,其优异的生物力学性能和生物相容性能表明其能应用于体外组织工程血管的构建,具有进一步应用于血管移植物研究的前景,为临床应用奠定了一定的基础。     

成人成骨细胞与多孔钛联合培养观察

目的观察成人成骨细胞在多孔钛表面的生长情况，评价多孔钛的生物相容性。方法将成人骨髓来源的成骨细胞与多孔钛联合培养，以多孔羟基磷灰石（hydroxyapatite，HA）为对照，倒置显微镜、扫描电镜观察细胞生长情况，MTT法检测细胞活性。结果成骨细胞在钛微孔表面生长良好，MTT法检测细胞活性，两组吸光度值无显著性差异（P〉0．05）。结论多孔钛具有良好的生物相容性，是比较理想的成骨细胞载体。     

Assessment of tissue scaffold degradation using electrochemical techniques

Degradation of a commercially available collagen–glycosaminoglycan dermal equivalent matrix was studied using electrochemical techniques. Degradation was accelerated by exposure to gamma radiation followed by storage at elevated temperatures or exposure to enzymes. The time-dependent diffusion of a small, electrochemically active, molecular probe, potassium ferrocyanide, through the matrix was monitored via changes in the oxidation peak currents of cyclic voltammograms. These measurements were made using a two-compartment diffusion chamber with the sample positioned well away from the working electrodes and a single-compartment electrode cell where the matrix was in direct contact with the working electrode. The relative merits of these two approaches are considered. Regardless of the approach chosen, amperometry appears well suited to monitoring progressive diffusivity changes through mechanically weak porous structures subject to different solution environments.     

The Effects of Morphology, Confluency, and Phenotype on Whole-Cell Mechanical Behavior

Emerging evidence indicates that cellular mechanical behavior can be altered by disease, drug treatment, and mechanical loading. To effectively investigate how disease and mechanical or biochemical treatments influence cellular mechanical behavior, it is imperative to determine the source of large inter-cell differences in whole-cell mechanical behavior within a single cell line. In this study, we used the atomic force microscope to investigate the effects of cell morphological parameters and confluency on whole-cell mechanical behavior for osteoblastic and fibroblastic cells. For nonconfluent cells, projected nucleus area, cell area, and cell aspect ratio were not correlated with mechanical behavior (p ≥ 0.46), as characterized by a parallel-spring recruitment model. However, measured force-deformation responses were statistically different between osteoblastic and fibroblastic cells (p < 0.001) and between confluent and nonconfluent cells (p < 0.001). Osteoblastic cells were 2.3–2.8 times stiffer than fibroblastic cells, and confluent cells were 1.5–1.8 times stiffer than nonconfluent cells. The results indicate that structural differences related to phenotype and confluency affect whole-cell mechanical behavior, while structural differences related to global morphology do not. This suggests that cytoskeleton structural parameters, such as filament density, filament crosslinking, and cell–cell and cell–matrix attachments, dominate inter-cell variability in whole-cell mechanical behavior.