HA/GCPU复合多孔支架的可控制备及细胞相容性

以脂肪族异佛尔酮二异氰酸酯(IPDI)作为硬段、蓖麻油甘油酯(GCO)作为软段,通过原位聚合法制备了羟基磷灰石/蓖麻油甘油酯基聚氨酯(HA/GCPU)复合多孔支架.利用红外光谱和扫描电子显微镜对复合支架进行表征,并测试其力学性能及孔隙率.研究结果表明,HA/GCPU复合多孔支架的孔隙率和抗压强度依赖羟基磷灰石的含量,并具有明显的可控性.HA质量分数分别为0,20%和40%时,HA/GCPU复合多孔支架孔隙率分别为(61±3)%,(68±2)%和(57±3)%,抗压强度分别为(605±61),(2125±58)和(4588±260)k Pa,可见HA质量分数为20%时,HA/GCPU复合多孔支架具有与松质骨较为匹配的孔隙率和抗压强度.将MG63细胞与多孔支架共培养,通过倒置显微镜和扫描电子显微镜观察,用噻唑蓝(MTT)法表征HA/GCPU复合多孔支架的细胞相容性,结果表明,HA/GCPU复合多孔支架表面细胞黏附,生长良好,无细胞毒性,在骨组织工程领域具有一定的应用前景.     

纳米羟基磷灰石/丝素蛋白复合支架材料的降解特性及生物相容性研究

应用共混法制备了纳米羟基磷灰石/丝素蛋白复合支架材料, 通过体外降解和细胞培养实验研究了复合支架材料的降解特性和生物相容性. 体外降解实验结果显示, 复合支架材料具有稳定的降解能力; 在降解过程中, 羟基磷灰石由于与降解液发生钙、磷等离子的交换, 使其结晶得到了进一步生长和完善. 利用细胞计数法、四甲基偶氮唑盐(MTT)比色法和碱性磷酸酶(ALP)活性测定等分析了复合支架材料的生物相容性, 结果表明, MG63细胞在复合支架材料上具有良好的粘附、增殖能力, 并可引起早期的骨分化. 因此, 纳米羟基磷灰石/丝素蛋白复合支架作为骨组织工程的支架材料具有良好的应用前景.     

非化学计量羟基磷灰石骨水泥的制备及性能研究

采用磷酸四钙和磷酸氢钙混合粉末制备了nCa/nP比为1.58的非化学计量羟基磷灰石骨水泥(n-HAC)及其多孔支架材料。结果表明:与nCa/nP=1.67的化学计量羟基磷灰石骨水泥(HAC)相比,n-HAC的凝结时间和抗压强度没有明显的区别。XRD和IR显示:n-HAC与HAC都为羟基磷灰石结构,但n-HAC在Tris-HCl缓冲溶液的降解性明显大于HAC。细胞培养结果表明:成骨细胞在n-HAC和HAC两种材料上的粘附和细胞形态没有明显的区别,但细胞在n-HAC上的增殖率明显高于HAC。将多孔n-HAC支架材料植入兔股骨缺损处,观察其修复骨缺损情况,组织学分析结果表明:新生骨在多孔支架的表面形成,并长入其内部;n-HAC在体内的降解比HAC快,能明显地促进新骨生成。     

可注射聚氨酯基抗菌复合骨水泥的制备与表征

兼具生物活性和抗菌性能的可注射材料对骨质疏松症椎体骨折的治疗具有更好的效果。本文通过向在位固化的聚氨酯中添加无机抗菌剂磷酸银和纳米羟基磷灰石制备了可注射聚氨酯基复合材料,考察不同磷酸银含量对其物理化学性能和抗菌性能的影响。傅里叶变换红外光谱分析、溶胀率和凝胶含量实验表明,制得了具有交联聚合结构的可注射聚氨酯复合材料。该材料固化时间小于1小时,适宜临床操作,固化后具有与天然松质骨相似的压缩强度、密度以及适宜细胞生长的表面亲水性。另外,抗细菌粘附实验和抑菌率实验表明,材料抗菌性能随磷酸银含量的增加逐渐提高。这些优异的性能表明可注射聚氨酯复合材料在骨质疏松症临床应用上具有广阔的前景。     

主链含四重氢键基元聚氨酯的合成与性能

合成了一种新型含有UPy(2-ureido-4[1H]-pyrimidone)基团的二羟基化合物,以此二羟基化合物作扩链剂,通过与聚氨酯预聚体进行的扩链反应,制备了一系列主链含UPy的聚氨酯(PU-UPy).傅里叶红外光谱(FTIR)、氢核磁共振(1H-NMR)等测试结果表明,在聚氨酯主链中确实含有UPy链段.同时,热性能及力学性能测试表明,聚氨酯中的UPy二聚体会集聚而形成微晶,熔点在60℃附近.在聚氨酯主链中引入UPy,能大幅提高聚氨酯的力学性能,调整软段的分子量,以及在主链中UPy含量可改变聚氨酯弹性体的断裂伸长率和抗张强度.     

软硬段对聚氨酯弹性体结构性能的影响

采用本体聚合和溶液聚合两种方法,合成了一系列用聚乙二酸丁二醇酯二醇作为软段的聚氨酯弹性体。研究了硬段含量和聚醚添加量对聚氨酯弹性体综合性能的影响。利用红外光谱、热分析、力学性能测试、记忆回弹性能和耐水解老化实验等测试手段对样品进行了表征与分析。结果表明:硬段质量含量为31%~35%时,材料的力学性能较优,形状回复率可以达到75%~85%;软段中聚醚添加量在4%~5%(占软段的质量分数)时,弹性体具有较好的力学性能和耐水解性能。     

聚醚醚酮复合材料表面生物活性涂层的制备与性能

以聚醚醚酮/钡玻璃粉(PEEK-BGF)复合材料为基体, 通过硅烷偶联剂, 在复合材料表面构建具有生物活性的纳米羟基磷灰石(nHA)和甲基丙烯酸酯基的光固化树脂复合涂层. 采用扫描电子显微镜(SEM)和X射线光电子能谱(XPS)分析了材料表面形貌和元素分布, 测试了涂层与复合材料之间的粘接强度. 通过检测大鼠成骨细胞总蛋白含量和碱性磷酸酶表达水平, 评价新型光固化纳米羟基磷灰石/聚甲基丙烯酸酯(nHA/PMMA)复合涂层的生物活性. 研究结果表明, nHA填充的光固化复合材料形成粗糙的表面, 随着nHA的填充量提高, 涂层表面生物学活性得到提高.     

基于聚丙交酯的光固化型可生物降解聚氨酯丙烯酸酯的合成和表征

基于聚醚酯的聚氨酯丙烯酸酯具有应用于组织工程支架材料及相关生物医用材料的潜力,研究其可生物降解性及影响因素非常重要.聚醚酯是由PEG(Mw=400)引发L-丙交酯开环聚合而得到的PLLA-PEGPLLA(PLEL)嵌段共聚物.它与二异氰酸酯(异佛尔酮二异氰酸酯和六亚甲基二异氰酸酯)反应,并用甲基丙烯酸羟乙酯封端,得到聚氨酯丙烯酸酯低聚物,然后通过紫外固化得到聚氨酯丙烯酸酯材料(PUA).用NMR和GPC对PLEL二醇和预聚物进行了组成和分子量表征,用DSC和DMA对PUA进行了结构和物理性能表征以及用接触角、吸水率和质量分析方法对材料的亲水性和降解性能进行了表征.结果发现,随着PLLA疏水链段变长,PLEL软段分子量增大,材料的亲水性降低,交联度和降解速率变小.相同的软段,基于硬段HDIHEMA的PUA材料比IPDI-HEMA的PUA有较低的T_g,较高的亲水性和降解速率.因为IPDI有环状结构,降低了PUA与水的相互作用.在3种不同降解条件下,氧化降解速率最高,酶解的速率高于水解.PUA材料的氧化降解速率取决于软段中PEG的含量,PLEL1000-HDI中PEG含量最高,其氧化降解最快,13周内失重率达到82.6%.     

聚氨酯弹性体/蒙脱土纳米复合材料的合成与性能

采用聚氨酯本体预聚法 ,利用原位插层聚合合成了聚氨酯 蒙脱土纳米复合材料 .通过X 射线衍射(XRD)和Molau实验研究了蒙脱土在复合材料中的分散情况 .红外分析 (IR)表明随着蒙脱土含量的增加 ,复合材料羰基氢键减少 .动态力学分析 (DMA)以及差热分析 (DSC)结果说明随着蒙脱土含量的增加 ,材料的玻璃化温度降低 .聚氨酯纳米复合材料的拉伸强度和断裂伸长率同时提高 ,表现出较好的力学性能 .     

纳米羟基磷灰石复合材料在硬组织工程领域中的应用研究

人体骨骼的晶体成分主要是纳米羟基磷灰石(n-HA),n-HA具有优良的生物相容性、骨传导性和骨结合能力,被广泛应用于硬组织修复材料中。本文综述了纳米羟基磷灰石在构建人体骨修复支架材料、骨替代材料和口腔医用材料方面的应用研究。     

Fabrication of poly(lactide‐co‐glycolide) scaffold embedded spatially with hydroxyapatite particles on pore walls for bone tissue engineering

Poly(lactide‐co‐glycolide) (PLGA) scaffolds embedded spatially with hydroxyapatite (HA) particles on the pore walls (PLGA/HA‐S) were fabricated by using HA‐coated paraffin spheres as porogens, which were prepared by Pickering emulsion. For comparisons, PLGA scaffolds loaded with same amount of HA particles (2%) in the matrix (PLGA/HA‐M) and pure PLGA scaffolds were prepared by using pure paraffin spheres as porogens. Although the three types of scaffolds had same pore size (450–600 µm) and similar porosity (90%–93%), the PLGA/HA‐S showed the highest compression modulus. The embedment of the HA particles on the pore walls endow the PLGA/HA‐S scaffold with a stronger ability of protein adsorption and mineralization as well as a larger mechanical strength against compression. In vitro culture of rat bone marrow stem cells revealed that cell morphology and proliferation ability were similar on all the scaffolds. However, the alkaline phosphatase activity was significantly improved for the cells cultured on the PLGA/HA‐S scaffolds. Therefore, the method for fabricating scaffolds with spatially embedded nanoparticles provides a new way to obtain the bioactive scaffolds for tissue engineering. Copyright © 2011 John Wiley & Sons, Ltd.     

Cell adhesive and growth behavior on electrospun nanofibrous scaffolds by designed multifunctional composites

Nanostructured biocomposite scaffolds of poly(l-lactide) (PLLA) blended with collagen (coll) or hydroxyapatite (HA), or both for tissue engineering application, were fabricated by electrospinning. The electrospun scaffolds were characterized for the morphology, chemical and tensile properties by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), water contact angle (WCA), Fourier transform infrared (FTIR) measurement, and tensile testing. Electrospun biocomposite scaffolds of PLLA and collagen or (and) HA in the diameter range of 200-700 nm mimic the nanoscale structure of the extracellular matrix (ECM) with a well-interconnection pore network structure. The presence of collagen in the scaffolds increased their hydrophility, and enhanced cell attachment and proliferation, while HA improved the tensile properties of the scaffolds. The biocompatibility of the electrospun scaffolds and the viability of contacting cells were evaluated by 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) nuclear staining and by fluorescein diacetate (FDA) and propidium iodide (PI) double staining methods. The results support the conclusion that 293T cells grew well on composite scaffolds. Compared with pure PLLA scaffolds a greater density of viable cells was seen on the composites, especially the PLLA/HA/collagen scaffolds.     

Crosslinked Silk Fibroin/Gelatin/Hyaluronan Blends as Scaffolds for Cell-Based Tissue Engineering

3D porous scaffolds fabricated from binary and ternary blends of silk fibroin (SF), gelatin (G), and hyaluronan (HA) and crosslinked by the carbodiimide coupling reaction were developed. Water-stable scaffolds can be obtained after crosslinking, and the SFG and SFGHA samples were stable in cell culture medium up to 10 days. The presence of HA in the scaffolds with appropriate crosslinking conditions greatly enhanced the swellability. The microarchitecture of the freeze-dried scaffolds showed high porosity and interconnectivity. In particular, the pore size was significantly larger with an addition of HA. Biological activities of NIH/3T3 fibroblasts seeded on SFG and SFGHA scaffolds revealed that both scaffolds were able to support cell adhesion and proliferation of a 7-day culture. Furthermore, cell penetration into the scaffolds can be observed due to the interconnected porous structure of the scaffolds and the presence of bioactive materials which could attract the cells and support cell functions. The higher cell number was noticed in the SFGHA samples, possibly due to the HA component and the larger pore size which could improve the microenvironment for fibroblast adhesion, proliferation, and motility. The developed scaffolds from ternary blends showed potential in their application as 3D cell culture substrates in fibroblast-based tissue engineering.     

Fabrication and Properties of Porous Collagen/Hyaluronic Composite Scaffolds Containing Nano-Bioactive Glass for Tissue Engineering

In this work, nano-structured scaffolds were designed for tissue engineering using collagen, hyaluronic acid (HA) and nano-bioactive glass (NBAG) as their main components. The scaffold was prepared via freeze-drying method and the properties including morphology, porosity, compressive strength, swelling ratio and cytotoxicity in-vitro, were also evaluated. The composite scaffolds showed well interconnected macropores with the pore size of ranging from 100 to 500 μm. The porosity percent and swelling ability were decreased with the introduction of NBAG into the collagen/HA hydrogel; however, the compressive strength was enhanced. The cytotoxicity in-vitro study shows that the collagen-HA/NBAG scaffolds have good biocompatibility with improving effect on fibroblastic cells growth. It could be concluded that this scaffold fulfills the main requirements to be considered as a bone substitute.     

In-Vitro Cytotoxicity Study: Cell Viability and Cell Morphology of Carbon Nanofibrous Scaffold/Hydroxyapatite Nanocomposites

Electrospun carbon nanofibers (CNFs), which were modified with hydroxyapatite, were fabricated to be used as a substrate for bone cell proliferation. The CNFs were derived from electrospun polyacrylonitrile (PAN) nanofibers after two steps of heat treatment: stabilization and carbonization. Carbon nanofibrous (CNF)/hydroxyapatite (HA) nanocomposites were prepared by two different methods; one of them being modification during electrospinning (CNF-8HA) and the second method being hydrothermal modification after carbonization (CNF-8HA; hydrothermally) to be used as a platform for bone tissue engineering. The biological investigations were performed using in-vitro cell counting, WST cell viability and cell morphology after three and seven days. L929 mouse fibroblasts were found to be more viable on the hydrothermally-modified CNF scaffolds than on the unmodified CNF scaffolds. The biological characterizations of the synthesized CNF/HA nanofibrous composites indicated higher capability of bone regeneration.     

Fabrication of chitosan/collagen/hydroxyapatite scaffolds with encapsulated Cissus quadrangularis extract

Here, we demonstrated the fabrication of a composite scaffold (chitosan [CS], collagen [Col], and hydroxyapatite [HA]) with the incorporation of encapsulated Cissus quadrangularis (CQ) extract for tissue engineering applications. First, the crude extract of CQ loaded nanoparticles were synthesized via double emulsion technique using polycaprolactone (PCL) and polyvinyl alcohol (PVA) as oil and aqueous phases, respectively. Both PCL (20, 40, and 80 mg/mL) and PVA (0.5%, 1%, and 3% w/v) concentrations were varied to determine the optimum concentrations for CQ‐loaded nanoparticle preparation. The CQ‐loaded PCL nanoparticles (CQ‐PCL NPs), prepared with 20 mg/mL PCL and 0.5% (w/v) PVA, exhibited the smallest size of 334.22 ± 43.21 nm with 95.54 ± 1.49% encapsulation efficiency. Then, the CQ‐PCL NPs were incorporated into the CS/Col/HA scaffolds. These scaffolds were also studied for their ultrastructure, pore sizes, chemical composition, compressive modulus, water swelling, weight loss, and biocompatibility. The results showed that the addition of CQ‐PCL NPs into the scaffolds did not dramatically alter the ultrastructure and properties of the scaffolds, compared to CS/Col/HA scaffolds alone. However, incorporation of CQ‐PCL NPs in the scaffolds improved the release profile of CQ by preventing the initial burst release and prolonging the release rate of CQ. In addition, the CQ‐PCL NPs‐loaded CS/Col/HA scaffolds supported the attachment and proliferation of MC3T3‐E1 osteoblast cells.     

Improving mechanical and biological properties of macroporous HA scaffolds through composite coatings

Interconnected porous hydroxyapatite (HA) scaffolds are widely used for bone repair and replacement, owing to their ability to support the adhesion, transfer, proliferation and differentiation of cells. In the present study, the polymer impregnation approach was adopted to produce porous HA scaffolds with three-dimensional (3D) porous structures. These scaffolds have an advantage of highly interconnected porosity (≈85%) but a drawback of poor mechanical strength. Therefore, the as-prepared HA scaffolds were lined with composite polymer coatings in order to improve the mechanical properties and retain its good bioactivity and biocompatibility at the same time. The composite coatings were based on poly(d,l-lactide) (PDLLA) polymer solutions, and contained single component or combination of HA, calcium sulfate (CS) and chondroitin sulfate (ChS) powders. The effects of composite coatings on scaffold porosity, microstructure, mechanical property, in vitro mineralizing behavior, and cell attachment of the resultant scaffolds were investigated. The results showed that the scaffolds with composite coatings resulted in significant improvement in both mechanical and biological properties while retaining the 3D interconnected porous structure. The in vitro mineralizing behaviors were mainly related to the compositions of CS and ChS powders in the composite coatings. Excellent cell attachments were observed on the pure HA scaffold as well as the three types of composite scaffolds. These composite scaffolds with improved mechanical properties and bioactivities are promising bone substitutes in tissue engineering fields.     

溶液喷丝法制备组织工程无纺布支架

在高压氮气作用下用溶液喷丝制备了组织工程支架.聚乙二醇/聚对苯二甲酸丁二醇酯(PEGT/PBT)共聚物的氯仿溶液经高压氮气流喷丝,继而沉积成无纺布.为了提高支架的生物相容性,将透明质酸水溶液喷涂到PEGT/PBT丝表面.结果表明,所制备的支架材料的生物相容性良好,细胞外基质透明酸能显著提高无纺布支架的生物相容性.