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1.
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。  相似文献   

2.
背景:目前胶原作为牙周组织工程支架材料仍具有机械强度差、降解速度快等缺点,将其与壳聚糖复合可改善上述问题。 目的:评估新型壳聚糖-胶原支架材料的体外生物相容性。 方法:通过MTT法评估100%,75%,50%,25%壳聚糖-胶原支架材料浸提液对人牙周膜细胞的毒性。选择第4-6代生长状态良好的人牙周膜细胞与壳聚糖-胶原支架共培养,观察细胞在支架上的生长情况,并检测与壳聚糖-胶原支架复合培养前后人牙周膜细胞碱性磷酸酶活性的变化。 结果与结论:新型壳聚糖-胶原支架具有双层结构,一侧表面致密,一侧表面疏松多孔。MTT法检测不同浓度材料浸提液毒性评级为0或1级。扫描电子显微镜及组织学观察可见细胞在壳聚糖-胶原支架上增殖良好,且致密层可起屏障膜作用,阻挡细胞进入支架内部;复合培养24 h后,人牙周膜细胞的碱性磷酸酶活性与复合培养前无明显差异(P > 0.05),复合培养48,72 h后人牙周膜细胞的碱性磷酸酶活性高于复合培养前(P < 0.05)。以上结果提示新型壳聚糖-胶原支架具有良好的生物相容性及屏障功能,可进一步应用于牙周组织工程的研究。  相似文献   

3.
背景:有关纳米含氟羟基磷灰石牙种植体材料生物相容性的报道较少。 目的:检测纳米含氟羟基磷灰石牙种植体材料的体外生物相容性。 方法:采用溶胶凝胶技术分别制备羟基磷灰石与纳米含氟羟基磷灰石。①溶血性实验:在0.2 mL稀释兔抗凝血中分别加入0.01,0.15,0.2 g/L纳米含氟羟基磷灰石溶液、生理盐水及蒸馏水各10 mL,检测各组上清液吸光度值。②体外细胞毒性实验:分别以100%,50%纳米含氟羟基磷灰石浸提液、100%羟基磷灰石浸提液、苯酚溶液及RPMI1640培养液培养传至第2代的L929细胞,MTT法检测培养2,4,7 d的吸光度值。 结果与结论:体外溶血性实验显示,各浓度梯度纳米含氟羟基磷灰石的溶血率均在5%以内,符合医用材料的溶血要求。体外细胞毒性实验显示,随着培养时间的增加,100%,50%纳米含氟羟基磷灰石浸提液组细胞贴壁覆盖率增加,细胞密度增高,细胞为长梭形或多角形,细胞增殖及形态与RPMI1640培养液组、羟基磷灰石组无明显差别,细胞毒性为0级。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;组织工程全文链接:  相似文献   

4.
背景:国内外的研究证实普通碳酸钙陶瓷作为骨替代材料时具有细胞支架作用。 目的:观察多孔碳酸钙陶瓷与成骨细胞的相容性,及作为骨组织工程支架的可能性。 方法:SD大鼠骨髓基质干细胞经矿化诱导培养、扩增并检测证实其已具成骨细胞表型后,分别与多孔碳酸钙陶瓷支架、普通羟基磷灰石陶瓷支架体外复合培养。 结果与结论:骨髓基质干细胞经体外诱导形成成骨细胞,钙结节、Ⅰ型胶原和碱性磷酸酶免疫染色结果阳性。多孔碳酸钙陶瓷支架材料与羟基磷灰石陶瓷材料皆有细胞附着生长,但多孔碳酸钙陶瓷支架材料细胞的黏附能力、增殖活力及成骨活性均强于羟基磷灰石陶瓷材料。提示多孔碳酸钙陶瓷支架材料与SD大鼠骨髓基质干细胞源性成骨细胞有良好相容性。  相似文献   

5.
背景:硼硅酸盐不仅可通过矿化作用形成羟基碳酸盐磷灰石层,而且具有强化学反应活性,可促进骨细胞再生。 目的:通过体外培养实验观察硼硅酸盐生物玻璃对兔成骨细胞生长行为的影响。 方法:根据 ISO10993-12:2007 的要求制备硼硅酸盐生物玻璃初次浸提液与二次浸提液。分离培养兔骨髓间充质干细胞,取第2代细胞诱导生成成骨细胞。取第5-15代成骨细胞,分别以硼硅酸盐生物玻璃初次浸提液、硼硅酸盐生物玻璃二次浸提液与α-MEM培养基培养,观察硼硅酸盐生物活性玻璃对成骨细胞增殖、蛋白合成、碱性磷酸酶活性、细胞凋亡及细胞横向与纵向迁移的影响。 结果与结论:初次浸提液组与二次浸提液组成骨细胞增殖优于α-MEM培养基组(P < 0.05),且初次浸提液组成骨细胞增殖优于二次浸提液组(P < 0.05)。初次浸提液组成骨细胞总蛋白含量高于二次浸提液组与α-MEM培养基组(P < 0.05)。3组间成骨细胞碱性磷酸酶活性、凋亡率、横向迁移距离及Transwell 中穿膜细胞数比较差异均无显著性意义。表明硼硅酸盐生物玻璃具有良好的细胞相容性,对成骨细胞增殖有一定的良性调节作用。  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  相似文献   

6.
背景:前期实验采用仿生学原理制备了可注射性纳米羟基磷灰石/壳聚糖/半水硫酸钙复合材料,但其与骨髓间充质干细胞的生物相容性还不十分清楚。目的:探讨纳米羟基磷灰石/壳聚糖/半水硫酸钙作为注射型骨组织工程支架材料的可行性。方法:将第3代兔骨髓间充质干细胞与可注射纳米羟基磷灰石/壳聚糖/半水硫酸钙支架复合培养,作为实验组;以单纯接种培养的骨髓间充质干细胞为对照组,倒置显微镜下观察细胞生长情况,MTT法检测细胞增殖,扫描电镜观察细胞在材料表面生长与增殖。将纳米羟基磷灰石/壳聚糖/半水硫酸钙支架埋植在家兔背部肌袋内,埋植后2,4,6,8周进行病理学观察。结果与结论:实验组细胞生长、增殖良好,与对照组无明显差异。支架埋植后2周,材料周围有中等量中性粒细胞、淋巴细胞和巨细胞浸润,可见小血管与纤维母细胞增生,材料已被炎性细胞分割、围绕散碎;埋植后4周,可见少量淋巴细胞、纤维母细胞聚集,炎症反应进一步消退,肌纤维排列、形态正常;埋植后6周,材料周围炎症反应轻微,组织水肿不明显;埋植后8周,炎症反应基本消退,材料基本降解完成,肌纤维形态基本正常。表明纳米羟基磷灰石/壳聚糖/半水硫酸钙复合物具有良好的细胞相容性和生物降解性,可作为注射型支架材料。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;组织工程全文链接:  相似文献   

7.
背景:经处理后的珊瑚转化羟基磷灰石具有良好的生物相容性和可降解性,其三维相通的孔隙结构不仅可为种子细胞的黏附、增殖提供足够的内部空间和表面积,而且有利于营养成分的渗透和血管化形成。 目的:对珊瑚转化羟基磷灰石进行细胞相容性和细胞毒性的研究,以初步探讨其应用于牙周组织工程的可行性。 方法:将体外培养的人牙周膜细胞接种到珊瑚转化羟基磷灰石支架上体外三维复合培养,通过细胞计数方法和扫描电镜观察人牙周膜细胞在珊瑚转化羟基磷灰石支架上的附着、生长情况,并设立阴性对照组和阳性对照组,采用MTT测试法和碱性磷酸酶活性检测法评估稀释浓度分别为100%,50%,10%,1%的珊瑚转化羟基磷灰石浸提液对人牙周膜细胞增殖及功能表达的影响。 结果与结论:珊瑚转化羟基磷灰石具有良好的多孔网状结构,人牙周膜细胞在珊瑚转化羟基磷灰石支架上生长旺盛,伸展充分,而不同浓度的材料浸提液对人牙周膜细胞的生长、增殖及碱性磷酸酶活性的影响差异均无显著性意义。提示珊瑚转化羟基磷灰石具有良好的三维空间结构和细胞相容性,且无细胞毒性,有望用作牙周组织工程的支架材料。关键词:珊瑚转化羟基磷灰石;细胞相容性;细胞毒性;支架材料;组织工程 缩略语注释:CHA:coral hydroxyapatite,珊瑚转化羟基磷灰石;HPDLCs:human periodontal ligament cells,人牙周膜细胞 doi:10.3969/j.issn.1673-8225.2012.16.025   相似文献   

8.
背景:低温快速成型技术具有支架成型可控性、保持材料生物学活性和易于实现支架材料的三维多孔立体结构等优势,被迅速用于骨组织工程支架的制备。 目的:采用低温快速成型制备聚乙二醇改性聚乳酸-乙醇酸/纳米羟基磷灰石复合支架,并检测其性能。 方法:采用低温快速成型设备分别制备聚乙二醇改性聚乳酸-乙醇酸/纳米羟基磷灰石与聚乳酸-乙醇酸/纳米羟基磷灰石复合支架,通过电镜观察支架超微结构,以介质(乙醇)浸泡法测定支架孔隙率,采用电子试验机检测支架力学性能;将两种支架材料分别与大鼠成骨细胞共培养,培养12 h采用沉淀法检测细胞黏附率,培养1,3,5,7,9,12 d采用CCK-8法检测细胞增殖。 结果与结论:两组支架孔径均在理想范围内并具有较高孔隙率,但聚乙二醇改性聚乳酸-乙醇酸/纳米羟基磷灰石支架的孔径波动范围大,孔径均值较聚乳酸-乙醇酸/纳米羟基磷灰石支架小且部分有闭塞现象。聚乙二醇改性聚乳酸-乙醇酸/纳米羟基磷灰石支架的细胞黏附率及表面细胞增殖活性高于聚乳酸-乙醇酸/纳米羟基磷灰石支架(P < 0.05),力学性能低于聚乳酸-乙醇酸/纳米羟基磷灰石支架(P < 0.05)。表明聚乙二醇改性聚乳酸-乙醇酸/纳米羟基磷灰石复合支架具有良好的细胞相容性。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;组织工程全文链接:  相似文献   

9.
背景:课题组拟对纳米级二氧化锆增韧的羟基磷灰石生物陶瓷进行一些初步研究,主要集中在生物力学匹配性,化学稳定性,以及生物相容性实验,其中体外细胞培养实验具有可控性,可重复性,能很好地反映材料的生物相容性。 目的:比较纳米级二氧化锆增韧的羟基磷灰石、纯羟基磷灰石两种材料对兔骨髓基质干细胞增殖、分化的影响。 方法:将骨髓基质干细胞置于含体积分数为20%胎牛血清的DMEM培养基中培养,传代后改用含β-甘油磷酸钠,地塞米松和维生素C的条件培养基培养。取传至第3代的成骨细胞,以1.0×108 L-1浓度接种于放有材料块的细胞培养板中,培养第1~10天倒置相差显微镜观察细胞生长情况,绘制细胞生长曲线,并进行碱性磷酸酶活性检测。培养第6天的细胞和材料复合物用多聚甲醛固定进行扫描电镜观察。 结果与结论:MTT法测得两种材料培养的细胞生长曲线无显著差异。复合培养的兔骨髓基质干细胞能够保持正常分泌碱性磷酸酶的功能。电镜照片也同样证实了两种材料表面均有细胞的附着。说明纳米级二氧化锆增韧的羟基磷灰石、纯羟基磷灰石均不影响成骨细胞增长分化,具有优良的成骨细胞相容性。  相似文献   

10.
壳聚糖/纳米羟基磷灰石分层复合支架的生物相容性研究   总被引:2,自引:0,他引:2  
制备壳聚糖/纳米羟基磷灰石(CS/nHA)分层复合支架,对其进行细胞毒性评价.分离培养大鼠软骨细胞接种于支架,相差显微镜和扫描电镜观察细胞的黏附及生长情况.动物皮下埋植试验观察其组织相容性.实验结果证实壳聚糖/纳米羟基磷灰石分层复合支架具有良好的生物相容性,有望成为较好的骨软骨组织工程支架.  相似文献   

11.
Calcium phosphate-chitosan composite scaffolds for bone tissue engineering   总被引:6,自引:0,他引:6  
Macroporous calcium phosphate-chitosan composite scaffolds were fabricated and evaluated for use in bone tissue engineering. Human osteoblast-like MG63 cells were cultured on the composite scaffolds, and their response to the materials was studied. Cell morphology, total protein content, and expression of classic markers for osteoblast differentiation were characterized. MG63 cells on the hydroxyapatite scaffolds nesting chitosan sponges (HC1) showed significantly higher alkaline phosphatase (ALP) level and osteocalcin (OC) production during the 11-day culture period, compared with the control culture on tissue culture plates. Cells on the chitosan scaffolds incorporated with hydroxyapatite powders (HC2) exhibited lower ALP activity during the 11-day culture period and OC secretion during the first 7 days, in comparison with that on HC1. The addition of calcium phosphate glass as in HC3 scaffolds increased the ALP and OC levels of MG63 cells. Our study indicated that the hydroxyapatite-matrix composite scaffolds might enhance the phenotype expression of MG63 cells, in comparison with chitosan-matrix scaffolds. Soluble calcium phosphate glasses should be added to the scaffolds to prevent chitosan from fast degradation that may affect the differentiation of osteoblast cells.  相似文献   

12.
A three-dimensional (3-D) scaffold is one of the major components in many tissue engineering approaches. We developed novel 3-D chitosan/poly(lactic acid-glycolic acid) (PLAGA) composite porous scaffolds by sintering together composite chitosan/PLAGA microspheres for bone tissue engineering applications. Pore sizes, pore volume, and mechanical properties of the scaffolds can be manipulated by controlling fabrication parameters, including sintering temperature and sintering time. The sintered microsphere scaffolds had a total pore volume between 28% and 37% with median pore size in the range 170-200microm. The compressive modulus and compressive strength of the scaffolds are in the range of trabecular bone making them suitable as scaffolds for load-bearing bone tissue engineering. In addition, MC3T3-E1 osteoblast-like cells proliferated well on the composite scaffolds as compared to PLAGA scaffolds. It was also shown that the presence of chitosan on microsphere surfaces increased the alkaline phosphatase activity of the cells cultured on the composite scaffolds and up-regulated gene expression of alkaline phosphatase, osteopontin, and bone sialoprotein.  相似文献   

13.
背景:木通皂苷D具有促进成骨细胞的增殖与分化、提高成骨细胞活性与数量、促进基质钙化与骨痂生长等诸多作用,主要被用于治疗骨质疏松与促进骨折愈合,将其应用于骨缺损修复的研究较少见。目的:以纳米羟基磷灰石/壳聚糖支架为载体,将包裹木通皂苷D的缓释微球负载于其中,观察其骨缺损修复作用。方法:采用W/O/W方法制作包裹木通皂苷D的缓释微球,采用冷冻干燥方法制备负载包裹木通皂苷D缓释微球的纳米羟基磷灰石/壳聚糖支架(以下简称缓释支架)与单纯的纳米羟基磷灰石/壳聚糖支架(以下简称空白支架),检测缓释微球与缓释支架的体外释药能力。将小鼠来源前成骨细胞MC3T3-E1分别接种于两种支架上,以单独培养的细胞为对照,分析细胞的黏附、增殖与分化情况。在24只成年新西兰大白兔双侧桡骨中段制造1.5 cm的骨缺损,分别植入空白支架与缓释支架,术后4,12周时进行大体观察、Micro-CT扫描影像学检查及组织学观察。结果与结论:(1)包裹木通皂苷D的缓释微球与缓释支架均具有缓释作用,其中缓释支架的药物释放速率更加平稳、持久;(2)CCK-8实验显示,缓释支架上的细胞增殖速率快于空白支架、对照组(P <0.05...  相似文献   

14.
Shor L  Güçeri S  Wen X  Gandhi M  Sun W 《Biomaterials》2007,28(35):5291-5297
Computer-aided tissue-engineering approach was used to develop a novel precision extrusion deposition (PED) process to directly fabricate Polycaprolactone (PCL) and composite PCL/hydroxyapatite (PCL-HA) tissue scaffolds. The process optimization was carried out to fabricate both PCL and PCL-HA (25% concentration by weight of HA) with a controlled pore size and internal pore structure of the 0 degrees /90 degrees pattern. Two groups of scaffolds having 60% and 70% porosity and with pore sizes of 450 and 750 microm, respectively, were evaluated for their morphology and compressive properties using scanning electron microscopy (SEM) and mechanical testing. Our results suggested that inclusion of HA significantly increased the compressive modulus from 59 to 84 MPa for 60% porous scaffolds and from 30 to 76 MPa for 70% porous scaffolds. In vitro cell-scaffolds interaction study was carried out using primary fetal bovine osteoblasts to assess the feasibility of scaffolds for bone tissue-engineering application. The cell proliferation and differentiation were calculated by Alamar Blue assay and by determining alkaline phosphatase activity. The osteoblasts were able to migrate and proliferate over the cultured time for both PCL as well as PCL-HA scaffolds. Our study demonstrated the viability of the PED process to the fabricate PCL and PCL-HA composite scaffolds having necessary mechanical property, structural integrity, controlled pore size and pore interconnectivity desired for bone tissue engineering.  相似文献   

15.
Porous scaffolds of biphasic calcium phosphate (BCP)/polyamide 6 (PA6) with weight ratios of 30/70, 45/55, and 55/45 have been fabricated through a modified thermally induced phase separation technique. The chemical structure properties, macrostructure, and mechanical strength of the scaffolds were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, scanning electron microscopy, and mechanical testing. The results indicated that the BCP/PA6 scaffolds had an interconnected porous structure with a pore size mainly ranging from 100 to 900 μm and many micropores on the rough pore walls. The mechanical property of the scaffold was significantly enhanced by the addition of BCP inorganic fillers. The 55/45 BCP/PA6 composite scaffold with 76.5% ± 2.1% porosity attained a compressive strength of 1.86 ± 0.14 MPa. Moreover, the BCP/PA6 porous scaffold was cultured with rat calvarial osteoblasts to investigate the cell proliferation, viability, and differentiation function (alkaline phosphatase). The type I collagen expression was also used to characterize the differentiation of rat calvarial osteoblasts on BCP/PA6 composite scaffold by immunocytochemistry. The in vitro cytocompatibility evaluation demonstrated that the BCP/PA6 scaffold acted as a good template for the cells adhesion, spreading, growth, and differentiation. These results suggest that the BCP/PA6 porous composite could be a candidate as an excellent substitute for damaged or defect bone.  相似文献   

16.
Three-dimensional oriented chitosan (CS)/hydroxyapatite (HA) scaffolds were prepared via in situ precipitation method in this research. Scanning electron microscopy (SEM) images indicated that the scaffolds with acicular nano-HA had the spoke-like, multilayer and porous structure. The SEM of osteoblasts which were polygonal or spindle-shaped on the composite scaffolds after seven-day cell culture showed that the cells grew, adhered, and spread well. The results of X-ray powder diffractometer and Fourier transform infrared spectrometer showed that the mineral particles deposited in the scaffold had phase structure similar to natural bone and confirmed that particles were exactly HA. In vitro biocompatibility evaluation indicated the composite scaffolds showed a higher degree of proliferation of MC3T3-E1 cell compared with the pure CS scaffolds and the CS/HA10 scaffold was the highest one. The CS/HA scaffold also had a higher ratio of adhesion and alkaline phosphate activity value of osteoblasts compared with the pure CS scaffold, and the ratio increased with the increase of HA content. The ALP activity value of composite scaffolds was at least six times of the pure CS scaffolds. The results suggested that the composite scaffolds possessed good biocompatibility. The compressive strength of CS/HA15 increased by 33.07% compared with the pure CS scaffold. This novel porous scaffold with three-dimensional oriented structure might have a potential application in bone tissue engineering.  相似文献   

17.
Gravel M  Gross T  Vago R  Tabrizian M 《Biomaterials》2006,27(9):1899-1906
Macroporous composites made of coralline:chitosan with new microstructural features were studied for their scaffolding potential in in vitro bone regeneration. By using different ratios of natural coralline powder, as in situ gas forming agent and reinforcing phase, followed by freeze-drying, scaffolds with controlled porosity and pore structure were prepared and cultured with mesenchymal stem cells (MSCs). Their supportive activity of cellular attachment, proliferation and differentiation were assessed through cell morphology studies, DNA content, alkaline phosphatase (ALP) activity and osteocalcin (OC) release. The coralline scaffolds showed by far the highest evaluation of cell number and ALP activity over all the other chitosan-based scaffolds. They were the only material on which the OC protein was released throughout the study. When used as a component of the chitosan composite scaffolds, these coralline's favourable properties seemed to improve the overall performance of the chitosan. Distinct cell morphology and osteoblastic phenotype expression were observed depending on the coralline-to-chitosan ratios composing the scaffolds. The coralline-chitosan composite scaffolds containing high coralline ratios generally showed higher total cell number, ALP activity and OC protein expression comparing to chitosan scaffolds. The results of this study strongly suggest that coralline:chitosan composite, especially those having a high coralline content, may enhance adhesion, proliferation and osteogenic differentiation of MSCs in comparison with pure chitosan. Coralline:chitosan composites could therefore be used as attractive scaffolds for developing new strategies for in vitro tissue engineering.  相似文献   

18.
背景:目前骨组织工程常用的支架材料主要有无机材料、有机高分子材料及天然衍生材料等,上述材料各有优缺点,为了充分发挥各类材料的优势,弥补其不足,目前多采用联合材料制备复合支架。 目的:制备新型仿生支架材料骨形态发生蛋白7多肽/壳聚糖/纳米羟基磷灰石/胶原,并观察其对骨髓间充质干细胞增殖、黏附及分化的影响。 方法:制备壳聚糖/纳米羟基磷灰石/胶原复合支架材料,扫描电镜观察支架材料表面微观形貌;采用真空吸附法将骨形态发生蛋白7多肽与支架材料复合,高效液相色谱仪检测骨形态发生蛋白7多肽在体外的释放规律;将骨髓间充质干细胞接种到复合骨形态发生蛋白7多肽的仿生支架材料上,以未复合多肽的支架材料作为对照,检测支架材料表面细胞增殖、黏附率、生长形态及碱性磷酸酶活性。 结果与结论:壳聚糖/纳米羟基磷灰石/胶原支架材料呈多孔状,孔径10~100 µm;骨形态发生蛋白7多肽可以从支架材料中缓慢释出;在复合多肽的仿生支架材料表面,骨髓间充质干细胞的黏附及向成骨细胞方向分化能力均明显强于对照组(P < 0.05),而增殖能力与对照组差异无显著性意义(P > 0.05)。说明新型仿生支架材料骨形态发生蛋白7多肽/壳聚糖/纳米羟基磷灰石/胶原是一种理想的骨组织工程支架材料,具有良好的细胞相容性。  相似文献   

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