蚕丝蛋白液与聚乙烯醇作为骨缺损修复材料的性能比较

目的探索不同材料与丝素蛋白液(SP)及聚乙烯醇(PVA)水凝胶混合后制成材料的理化、生物性能。方法取羊椎骨粉和纳米级羟基磷灰石,分别与丝素蛋白溶液、PVA水凝胶以及丝素蛋白复合PVA溶液混合,通过抗溶解性、圧缩力、剪切力及材料细胞毒性的检测,分析比较各组材料的机械生物性能。结果 (1)抗溶解性检测:丝素蛋白溶液混合材料72 h后均达到Ⅲ级溶解,丝素蛋白复合PVA溶液混合材料力学性能优于单纯丝素蛋白组;(2)力学性能检测:丝素蛋白溶液的力学性能较弱,PVA力学性能优良,丝素蛋白溶液混合PVA后力学性能可增强。各组间差异有统计学意义,抗压缩力F=333.667,P<0.05,抗剪切力F=59.997,P<0.05;(3)材料体外细胞毒性检测:各组毒性均为0级或1级,丝素蛋白溶液/骨粉组、PVA/骨粉组、丝素蛋白复合PVA溶液/骨粉组、丝素蛋白溶液/NHA组、PVA/NHA组、丝素蛋白复合PVA溶液/NHA组均无细胞毒性,单纯丝素蛋白溶液混合粉剂材料细胞相容性更好。结论丝素蛋白具有良好的生物相容性,但力学性能欠佳,可以将蚕丝蛋白混合其他材料作为组织工程骨研究的新方向。     

细胞膜片复合3D打印马鹿角粉/丝素蛋白/聚乙烯醇支架对羊下颌骨缺损的修复效果

目的 探讨细胞膜片复合马鹿角粉/丝素蛋白/聚乙烯醇支架修复下颌骨极限性骨缺损的效果.方法 通过3D打印制备马鹿角粉/丝素蛋白/聚乙烯醇支架及纳米级羟基磷灰石/丝素蛋白/聚乙烯醇支架,采用全骨髓培养法制备阿勒泰大尾羊髂骨骨髓血细胞膜片;按观察时间1、2、3个月分别建立阿勒泰大尾羊双侧下颌骨极限性缺损模型4只,实验组为细胞...     

3D打印聚乙烯醇/纳米羟基磷灰石支架与丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架的性能比较

目的　采用3D打印技术制备3D打印聚乙烯醇 /纳米羟基磷灰石支架与丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架,并对其进行表征。方法　采用3D打印技术制作聚乙烯醇/纳米羟基磷灰石支架以及丝素蛋白/聚乙烯醇/纳米羟基磷灰石复合支架。进行孔隙率、扫描电镜、压缩力学性能及细胞毒性检测。 结果　①扫描电镜观察：丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架结构规则,网状结构清晰,交通支连续,层层之间搭接良好,支架空隙均一。相同倍数下,聚乙烯醇/纳米羟基磷灰石支架网状结构连续性较差。②压缩力学性能：相同应力情况下（10 MPa）,3D打印丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架的应变大于3D打印聚乙烯醇/纳米羟基磷灰石支架。③孔隙率：3D打印丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架的孔隙率大于3D打印聚乙烯醇/纳米羟基磷灰石支架。④细胞毒性检测：不同时间点两组支架的细胞增殖率无明显差别。结论　结果表明：3D打印聚乙烯醇 /纳米羟基磷灰石支架与丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架具有良好的理化性能和细胞相容性。     

聚乳酸-纳米羟基磷灰石-丝素蛋白引导骨再生膜的制备及表面特征

目的制备一种新型的聚乳酸-纳米羟基磷灰石-丝素蛋白(polylactic acid/nano-hydroxyapatite/silk fi-broin,PLLA/n-HA/SF)纳米纤维引导骨组织再生膜,初步探讨其作为引导骨再生屏障膜的可行性。方法采用热致相分离法制备PLLA/n-HA/SF纳米纤维复合膜,通过扫描电镜对其形貌进行研究,利用傅里叶红外光谱仪分析纳米羟基磷灰石和丝素蛋白的加入对所制备的复合膜结构的影响,并计算其孔隙率。结果扫描电镜显示该复合膜具有纳米纤维状三维网络结构。纤维直径约为160～320nm,孔径大小为1～4μm,丝素蛋白和纳米羟基磷灰石在复合膜中分散均匀。红外光谱结果表明PLLA/n-HA/SF复合膜具备聚乳酸、纳米羟基磷灰石及丝素蛋白的特征峰表现,3种组分之间结合良好。该膜的孔隙率为92.600%。结论热效相分离法制备的PLLA/n-HA/SF复合膜具有良好的微观结构和较高的孔隙率。     

3D打印丝素蛋白/聚乙烯醇/纳米羟基磷灰石支架体内降解的研究

目的 使用3D打印技术打印出丝素蛋白/聚乙烯醇/纳米羟基磷灰石(SF/PVA/n?HA)支架与聚乙烯醇/纳米羟基磷灰石(PVA/n?HA)支架并用细胞膜片包裹后回植到动物体内,研究并探讨SF/PVA/n?HA与PVA/n?HA支架的降解性能.方法 将SF/PVA/n?HA支架与PVA/n?HA支架分别植入羊下颌骨内,在术后1、2、3个月处死实验动物,取出支架所在部位下颌骨,行影像学观察、HE染色观察、实时定量PCR(RT?PCR),探讨两种支架在体内降解情况的区别以及降解相关因子之间的关系.结果 ①影像学结果:SF/PVA/n?HA支架组与PVA/n?HA支架组可见骨缺损处低密度影均减小,而二者相比,SF/PVA/n?HA支架组低密度影范围较小,高密度影部位也较PVA/n?HA支架明显.②HE染色结果:SF/PVA/n?HA支架组在前两个月仍可见部分支架残余,支架周围前两个月可见炎性细胞,3个月末炎性逐渐消退,可见明显骨陷窝.③RT?PCR:同一时间不同材料比较,SF/PVA/n?HA支架组与PVA/n?HA支架组相比,IL?1、IL?6、M?CSF、NFATc1四种因子表达趋势相近.在术后1、2个月,SF/PVA/n?HA支架组四种因子的mRNA表达量均高于PVA/n?HA支架组;术后3个月,四种因子mRNA表达量SF/PVA/n?HA支架组均低于PVA/n?HA支架组(P<0.05).结论 SF/PVA/n?HA支架具有一定的降解性能,且在降解过程中,能促进破骨相关因子IL?1、IL?6、M?CSF、NFATc1的表达.     

3D打印复合PVA骨组织工程支架研究现状

3D打印复合多孔性骨组织工程支架已成为研究热点。复合聚乙烯醇(polyvinyl alcohol,PVA)具备良好生物相容性及可降解性能,但因不能抵抗较大机械抗力,不能单独制备支架,复合其他机械性能佳及生物相容性良好的材料,可表现出良好的生物相容性、可降解性及机械性能等优点,因此3D打印复合PVA支架材料可以优化PVA支架的性能,本文从3D打印骨组织工程支架技术、PVA、复合PVA支架在体内外骨形成效果等方面作一综述。     

丝素蛋白/左旋聚乳酸支架材料与软骨细胞体外细胞相容性研究

目的:观察静电纺丝支架材料丝素蛋白/左旋聚乳酸(SF/PLLA)的体外细胞相容性,探索其作为软骨组织工程支架材料的可行性,为进一步的体内及动物实验奠定基础。方法:将兔膝关节软骨细胞与丝素蛋白/左旋聚乳酸(SF/PLLA)支架材料复合培养,在第3、7、14天分别作HE染色和阿利新蓝+核固红染色,扫描电镜检验细胞黏着情况,MTT试验检测细胞在支架上的增殖情况。结果:细胞在支架上可以获得良好的粘附,细胞增殖良好,无细胞表型的变化。结论:丝素蛋白/左旋聚乳酸(SF/PLLA)具有良好的细胞相容性,可作为软骨组织工程支架材料。     

电纺聚己内酯/Ⅰ型胶原蛋白/纳米锆酸钙复合支架的制备及其生物相容性的研究

目的:制备聚己内酯(PCL)/Ⅰ型胶原(COLI)/纳米锆酸钙(nCZ)复合支架用于骨组织再生,评价其性能及对人牙周膜细胞(PDLCs)生物相容性及成骨分化的影响.方法:用静电纺丝法制备PCL/COLI、PCL/COLI/纳米羟基磷灰石(nHA)和PCL/COLI/nCZ复合支架,通过扫描电子显微镜表征支架形貌,能量色...     

PLGA/PCL/nHA生物支架复合兔BMSCs体外培养的实验研究

目的:研究新型复合支架聚乳酸-聚乙醇酸/聚己内酯/纳米羟基磷灰石(PLGA/PCL/nHA)的生物相容性,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:将兔骨髓基质细胞(bone marrow stromal cells,BMSCs)接种于PLGA/PCL/nHA复合支架上,体外共同培养后,MTT法检测BMSC...     

3D打印制备不同重量比例的n-HA/PLA/PVA复合膜性能比较

目的:通过3D打印的方法制备的不同重量比例的纳米羟基磷灰石/聚乳酸/聚乙烯醇(n-HA/PLA/PVA)复合膜,并对其相关性能检测。方法:采用3D打印技术制备不同重量比例的n-HA/PLA/PVA复合膜,分别为PLA/PVA复合膜、15%n-HA/PLA/PVA复合膜、50%n-HA/PLA/PVA复合膜、75%n-HA/PLA/PVA复合膜。扫描电镜下观察各组膜形态,对其力学性能、细胞毒性及动物实验相应指标进行检测。结果:扫描电镜下观察,n-HA/PLA/PVA复合膜呈现三维网状结构,各材料间相互结合,孔隙分布不均,大小不一。随着n-HA质量浓度的提高,电镜下见各材料间孔隙逐渐减小,形成结构均匀的复合膜。力学性能及吸水率检测中,随着nHA含量的增加,n-HA/PLA/PVA复合膜的拉伸强度及吸水率呈下降趋势;细胞毒性检测,不同比例复合膜的细胞增殖率无明显差别,无细胞毒性。动物实验测量牙周菌斑指数及龈沟出血指数未发现不同比例n-HA/PLA/PVA复合膜有统计学差异。结论:3D打印不同比例的n-HA/PLA/PVA复合膜具有良好的物理性能和细胞生物相容性,n-HA比例更高的复合膜可能具有更好的物理性能及良好的生物相容性。     

LbL法制备磁性纳米复合纤维膜生物相容性的研究

目的 将磁性纳米材料嫁接到静电纺丝纤维膜上以达到增强其生物相容性的目的。方法 通过“层-层自组装”(LbL)的方法将Fe3O4磁性纳米颗粒组装到聚乳酸-羟基乙酸共聚物（poly(lactic-co-glycolic acid),PLGA）、ε-聚己内酯（ε-caprolactone,PCL）、明胶（gelatin,Gel）共混静电纺丝纤维膜（PPG）上,使用接触角仪、电子万能试验机、扫描电镜和振动样品磁强计对其表面形貌、亲水性、拉伸性能及磁学性能进行测试。将MC3T3-E1细胞接种到复合膜上,通过CCK-8法测试其细胞增殖,并与对照组进行比较。结果 扫描电镜显示Fe3O4组装到膜表面,膜仍然保持类似细胞外基质的疏松纳米纤维结构。振动样品磁强计结果显示PPG膜在修饰了Fe3O4磁性纳米颗粒后具备超顺磁性性能。接触角及吸水率测量表明PPG-F膜亲水性显著改善,CCK-8检测结果显示PPG-F膜细胞增殖活性显著高于对照组PPG膜(P<0.05)。结论 本方法形成的PPG-F纳米纤维复合膜具有良好生物相容性。     

壳聚糖复合材料在骨组织工程中的研究进展

骨组织工程是利用生物学和工程学方法研制能修复或替代人类骨组织材料的科学领域。支架是组织工程的关键要素,为细胞和组织生长提供结构支持和黏附位点。支架可以由金属、聚合物和陶瓷生物材料制成,其中聚合物支架由于其生物相容性、生物可降解性和机械稳定性而被广泛应用。而壳聚糖（CS）作为甲壳素衍生的天然聚合物,过去20年在骨组织工程中发挥了重要作用。近年来,CS复合材料在骨组织工程领域的应用受到相当大的关注,因为它具有更小的异物反应,优良的抗菌性、可塑性,适合细胞向内生长和骨传导。本文将讨论常见的几种CS复合材料在骨组织工程中的生物相容性及体内外促成骨研究。     

3-dimensional composite scaffolds consisting of apatite-PLGA-atelocollagen for bone tissue engineering

We fabricated 3-dimensional scaffolds consisting of biodegradable poly(D, L-lactide-co-glycolic acid)(PLGA)(75/25) with hydroxyapatite particles containing atelocollagen (aAC). The aim of this study was to evaluate this new type of scaffold in regard to its basic properties and biocompatibility. Characterization of the obtained scaffolds was performed to know the porosity, shrinkage, diametral tensile strength, and biocompatibility. Composite scaffolds made of PLGA with hydroxyapatite particles containing atelocollagen (PL-aAC) showed a greater strength and stability than PLGA scaffolds. PL-aAC also exhibited superior performance in terms of cell attachment and proliferation as compared to PLGA, while histological findings showed that PL-aAC had an excellent response toward soft tissues. Our results strongly suggest that PL-aAC is more useful for cell transplantation as compared to PLGA for bone tissue engineering.     

Segmental bone tissue engineering by seeding osteoblast precursor cells into titanium mesh-coral composite scaffolds

The size and specific shape of bone grafts are important for jaw reconstruction. In this experiment, segmental bone grafts were engineered in a predetermined shape via seeding osteoblast precursor cells into titanium mesh-coral composite scaffolds. Titanium meshes were moulded into the shape of a column with length 12 mm and diameter 8mm. The column was filled with natural coral granules and the complex acted as a cell-seeding scaffold. About 4 x 10(7) osteoblast precursor cells in 200 microl cell-culture medium were seeded into each of six scaffolds and incubated in vitro for 2 days. Then, the composites were implanted subcutaneously into the backs of nude mice and incubated in vivo. Two months after implantation, the animals were killed and new bone formed in the scaffolds was investigated by gross inspection, X-ray examination, histological observation and mechanical testing. The results showed that newly formed tissue was red and presented the gross appearance of bone, and kept the original shape of the column. Titanium mesh was situated on the surface of the bone graft. An X-ray blocking shadow was observed in and around the titanium scaffolds; most of the coral granules had been absorbed. Histological observation demonstrated a large amount of new bone formed and integrated well with titanium mesh. Mechanical testing showed that new bone improved the mechanical property of the graft significantly. In conclusion, a titanium mesh-coral composite scaffold with osteoblast precursor cells is an efficient means to engineer segmental bone, possessing the desired shape and mechanical strength.     

Comparison of human alveolar osteoblasts cultured on polymer-ceramic composite scaffolds and tissue culture plates

The effects of medical grade polycaprolactone-tricalcium phosphate (mPCL-TCP) (80:20) scaffolds on primary human alveolar osteoblasts (AOs) were compared with standard tissue-culture plates. Of the seeded AOs, 70% adhered to and proliferated on the scaffold surface and within open and interconnected pores; they formed multi-layered sheets and collagen fibers with uniform distribution within 28 days. Elevation of alkaline phosphatase activity occurred in scaffold-cell constructs independent of osteogenic induction. AO proliferation rate increased and significant decrease in calcium concentration of the medium for both scaffolds and plates under induction conditions were seen. mPCL-TCP scaffolds significantly influenced the AO expression pattern of osterix and osteocalcin (OCN). Osteogenic induction down-regulated OCN at both RNA and protein level on scaffolds (3D) by day 7, and up-regulated OCN in cell-culture plates (2D) by day 14, but OCN levels on scaffolds were higher than on cell-culture plates. Immunocytochemical signals for type I collagen, osteopontin and osteocalcin were detected at the outer parts of scaffold-cell constructs. More mineral nodules were found in induced than in non-induced constructs. Only induced 2D cultures showed nodule formation. mPCL-TCP scaffolds appear to stimulate osteogenesis in vitro by activating a cellular response in AO's to form mineralized tissue. There is a fundamental difference between culturing AOs on 2D and 3D environments that should be considered when studying osteogenesis in vitro.