Incorporation of Sphingosine 1-Phosphate Loaded Mesoporous Silica Nanoparticles into Poly( L-lactic acid ) Nanofibrous Scaffolds for Bone Tissue Engineering

Controlled release of the functional factors is the key to improve clinical therapeutic efficacy during the tissue repair and regeneration.The three-dimensional(3D)scaffold can provide not only physical properties such as high strength and porosity but also an optimal environment to enhance tissue regeneration.Sphingosine1-phosphate(S1P),an angiogenic factor,was loaded into mesoporous silica nanoparticles(MSNs)and then incorporated into poly(L-lactic acid)(PLLA)nanofibrous scaffold,which was fabricated by thermally induced phase separation(TIPS)method.The prepared scaffolds were examined by attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),scanning electron microscopy(SEM),and transmission electron microscopy(TEM)and compressive mechanical test.The ATR-FTIR result demonstrated the existence of MSNs in the PLLA nanofibrous scaffold.The SEM images showed that PLLA scaffold had regular pore channel,interconnected pores and nanofibrous structure.The addition of MSNs at appropriate content had no visible effect on the structure of scaffold.The compressive modulus of scaffold containing MSNs was higher than that of the scaffold without MSNs.Furthermore,fluorescein isothiocyanate(FITC)was used as model molecule to investigate the release behavior of S1P from MSNsincorporated PLLA(MSNs/PLLA)nanofibrous scaffold.The result showed that the composite scaffold largely reduced the initial burst release and exhibited prolonged release of FITC than MSNs.Thus,these results indicated that S1P-loaded composite nanofibrous scaffold has potential applications for bone tissue engineering.     

Dexamethasone-Loaded PLGA Microspheres Incorporated PLLA/PLGA/PCL Composite Scaffold for Bone Tissue Engineering

The combination of micro-carriers and polymer scaffolds as promising bone grafts have attracted considerable interest in recent decades.The poly(L-lactic acid)/poly(lactic-co-glycolic acid)/polycaprolactone(PLLA/PLGA/PCL)composite scaffold with porous structure was fabricated by thermally induced phase separation(TIPS).Dexamethasone(DEX)was incorporated into PLGA microspheres and then loaded on the PLLA/PLGA/PCL scaffoldtopreparethedesiredcompositescaffold.The physicochemical properties of the prepared composite scaffold were characterized.The morphology of rat bone marrow mesenchymal stem cells(BMSCs)grown on scaffolds was observed using scanning electron microscope(SEM)and fluorescence microscope.The resultsshowedthatthePLLA/PLGA/PCLscaffoldhad interconnected macropores and biomimetic nanofibrous structure.In addition,DEX can be released from scaffold in a sustained manner.More importantly,DEX loaded composite scaffold can effectively support the proliferation of BMSCs as indicated by fluorescence observation and cell proliferation assay.The results suggested that the prepared PLLA/PLGA/PCL composite scaffold incorporating drug-loaded PLGA microspheres could hold great potential for bone tissue engineering applications.     

Hippocampal Neuron-Derived Extracellular Matrix Coated Nanofibrous Scaffold for Neural Tissue Engineering

The aim of this study is to prepare poly-L-lactide( PLLA) electrospun nanofibrous scaffolds coated with hippocampal neuron-derived extracellular matrix( N-ECM)and construct a novel neural tissue engineering scaffold.Neonatal rat hippocampal neurons were seeded on PLLA nanofibers,and then decellularized to derive a cell-free extracellular matrix loaded N-ECM/PLLA modified scaffolds. The morphology and ingredients of N-ECM/PLLA were observed by scanning electron microscopy( SEM) and immunofluorescence staining respectively, and the cytocompatibility of the composite scaffolds was characterized by cell count kit-8( CCK-8) assay. The N-ECM was clearly identified loading on scaffolds when being imaged via SEM and immunofluorescence staining results showed that the N-ECM was made up of fibronectin and laminin. Most importantly, compared with tissue culture polystyrene and pure scaffolds, N-ECM/PLLA scaffolds could effectively facilitate the proliferation of rat adrenal neuroma cells( PC12 cells),indicating their better cell compatibilities. Based on the combination of N-ECM and PLLA biomaterials,the present study has fabricated a unique and versatile neural tissue engineering scaffold,offering a new thought for future neural tissue engineering.     

Nanofibrous Scaffold Containing Osteoblast-Derived Extracellular Matrix for the Proliferation of Bone Marrow Mesenchymal Stem Cells

Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by reconstituting osteoblast cell-derived ECM( O-ECM) on the electrospun nanofibrous scaffold,and further to evaluate its subsequent application for promoting the proliferation of bone marrow mesenchymal stem cells( BMSCs). To engineer a biomimetic scaffold, calvarial osteoblasts and electrospun poly-llactic acid( PLLA) nanofibers were prepared and subjected to decellularize for O-ECM deposition. To evaluate and characterize the O-ECM/PLLA scaffold, the morphology was examined and several specific mark proteins of osteoblasts matrix were evaluated.Furthermore,the cell counting kit-8( CCK-8) assay was used to detect the proliferation of the BMSCs cultivated on the O-ECM/PLLA scaffold. The results indicated O-ECM/PLLA scaffold was loaded with Collagen I, Fibronectin, and Laminin, as the composition of the marrow ECM. After decellularization,O-ECM deposition was observed in O-ECM/PLLA scaffold. Moreover,the O-ECM/PLLA scaffold could significantly enhance the proliferation of BMSCs,suggesting better cytocompatibility compared to the other groups tested. Taken together,a biomimetic scaffold based on the joint use of O-ECM and PLLA biomaterials,which represents a promising approach to bone tissue engineering, facilitates the expansion of BMSCs in vitro.     

Fabrication and characterization of electrospun poly-L-lactide/gelatin graded tubular scaffolds: Toward a new design for performance enhancement in vascular tissue engineering

In this study, a new design of graded tubular scaffolds have been developed for the performance enhancement in vascular tissue engineering. The graded poly-L-lactide (PLLA) and gelatin fibrous scaffolds produced by electrospining were then characterized. The morphology, degradability, porosity, pore size and mechanical properties of four tubular scaffolds (graded PLLA/gelatin, layered PLLA/gelatin, PLLA and gelatin scaffolds) have been investigated. The tensile tests demonstrated that the mechanical strength and also the estimated burst pressure of the graded scaffolds were significantly increased in comparison with the layered and gelatin scaffolds. This new design, resulting in an increase in the mechanical properties, suggested the widespread use of these scaffolds in vascular tissue engineering in order to prepare more strengthened vessels.     

Immobilization of biomacromolecules on poly-L-lactide surface via a layer-by-layer method for the improving of its cytocompatibility to bone marrow stromal cells

Hyaluronic acid （HA） and chitosan （CS） were immobilized on the surface of poly-L-lactide （PLLA） by the following procedure： Firstly, PLLA was aminolyzed with 1, 6-hexanediamine, and part of the PLLA surface ester groups were converted to free amino groups. Then negatively charged hyaluronic acid and positively charged chitosan were deposited onto the surface of aminolyzed PLLA film in a layer-by-layer assembly manner. The effect of the layer-by- layer deposition was evaluated by ATRoFTIR spectroscopy, Raman spectroscopy and static contact angle measurements. The cytocompatibility of PLLA sample to bone marrow stromal cells （BMSCs） was improved after modification with chitosan and HA. The cell attachment, activity, and proliferation on CS/HA modified PLLA films were enhanced comparing with the control. The cells cultured on the modified PLLA samples excreted abundant cytoplasm and can differentiate to vascular smooth muscle （SM）-like （SM-like） cells. A macroporous three-dimensional PLLA scaffold was prepared by integrating both the technique of freeze-drying and particle leaching. Layer-by-layer modification by HA/CS and cell culture was also applied on this scaffold. The scaffold cultured with BMSCs for 2 weeks has been tested successfully in vivo as a patch for repairing the artificial incision on canine pulmonary artery.     

纳米HAP/CPP/PLLA骨组织工程支架复合材料降解特性

现有支架材料的降解速率与骨细胞生长、繁殖速率不匹配,在降解过程中支架材料的强度、刚度衰减速率与成骨速率不匹配,支架材料在体内降解的酸性副产物会引起炎症反应.为克服以上困难,采用溶媒浇铸、颗粒滤取与气体发泡相结合的方法制备出纳米HAP/CPP/PLLA骨组织工程支架复合材料;选用生理盐水作为模拟体液进行降解实验,测试该支架复合材料的降解性能,用扫描电子显微镜对其在不同降解时期的微观结构进行观察.结果表明:纳米HAP/CPP/PLLA支架复合材料在降解过程中具有三维、连通、微孔网状结构,并具有良好的降解性能和生物相容性,是比较理想的骨组织工程支架材料.     

聚乳酸多孔支架的制备研究

分别采用粒子滤出法和冷冻干燥粒子滤出复合法制备了聚乳酸多孔支架.在粒子滤出法中,采用氯化钠和碳酸氢铵作为造孔剂制备多孔支架;在冷冻干燥法中,采用碳酸氢铵和冰颗粒作为造孔剂制备多孔支架.实验结果表明:采用氯化钠或碳酸氢铵做造孔剂,生成的多孔支架有造孔剂残留;采用冰颗粒造孔则无残留.研究认为,采取适当的工艺措施,加速造孔剂向环境的传质过程,是降低支架中造孔剂残留的有效途径,并提出了在实际中可采取的改进措施,包括选用易于滤出的材料、提高氯仿和造孔剂含量、提高温度和真空度、采用特殊气氛、提高过滤溶液的流动性.     

新型前交叉韧带支架材料的构建及其生物相容性的实验研究

探讨以聚羟基丁酸己酯/聚左旋乳酸(PHB/PLLA1∶1)胶原杂化支架作为前交叉韧带组织工程载体材料的可行性。制备"三明治"样结构PHB/PLLA共聚物并测量其孔隙率等指标。以I型胶原对制备的PHB/PLLA支架进行杂化,获得PHB/PLLA胶原杂化支架。扫描电镜观察其表面结构。将兔皮肤成纤维细胞(SF)接种于PHB/PLLA胶原杂化支架,共培养5d后,扫描电镜下观察其在材料上生长情况。PHB/PLLA支架杂化后胶原填充于纤维空隙,分布比较均匀。体外培养的皮肤成纤维细胞成功种植在支架材料上,在材料上粘附、生长良好。说明构建的支架材料具有良好的三维构型和生物相容性,有望为前交叉韧带损伤的修复提供了一种新型的支架材料。     

Preparation of fiber-microsphere scaffolds for loading bioactive substances in gradient amounts

Gradient scaffolds are needed for interface tissue regeneration. In this study, a technique combining electrospinning and electrospraying was developed for preparing poly(L-lactide-co-glycolide) (PLGA) fiber-microsphere scaffolds for loading bioactive substances in gradient amounts. The gradient fiber-microsphere scaffolds contain two sheets of electrospun membranes and a sheet of microspheres loaded with bioactive substances in gradient amounts between the electrospun membranes. The morphologies of the gradient scaffolds were characterized and bovine serum albumin (BSA) was loaded as a model bioactive substance. The amount of BSA-loaded microspheres decreased gradually along the length of the gradient scaffold. The addition of poly (ethylene glycol) significantly improved the hydrophilicity of the gradient scaffold and the release behavior of BSA with respect to the gradient became apparent, with differences in the release amounts along the length of the gradient scaffold being observed. The biocompatibility of the gradient scaffold was verified using MC3T3-E1 pre-osteoblastic cells. The study demonstrated that the combination of electrospinning and electrospraying was a feasible method for the preparation of gradient scaffolds for potential applications in interface tissue engineering.     

离心—冷冻法制备梯度孔结构的真皮支架

胶原基真皮支架的结构和性能受多方面因素影响,比如除胶原外的其他主要材料、支架的孔径和孔隙率以及交联度等.采用离心—冷冻法结合冷冻干燥法制备梯度孔结构的真皮支架,表征支架的孔径和孔隙率,并以细胞培养检验其生物相容性,其中采用扫描电镜对上中下3部分支架结构及孔径进行表征,并通过细胞培养生长实验检测该支架材料的细胞粘附性能,以达到表征离心力的影响.结果显示,离心力对支架的微观结构产生了影响,支架的致密程度从下到上有明显的不同,具体表现在孔径上:上层孔径52±27.8μm,中层孔径57±8.7μm,下层孔径49±40.4μm.同时,细胞粘附实验表明,致密的下层更适合细胞的粘附生长.     

聚乳酸/聚环氧乙烷纳米纤维膜的结构与性能表征

采用静电纺丝方法制备了聚乳酸(PLLA)/聚环氧乙烷(PEO)纳米纤维膜,利用扫描电子显微镜(SEM)、差示扫描量热(DSC)、X-射线衍射(XRD)和拉伸测试,对PLLA/PEO纳米纤维膜的形貌、结构、结晶行为和力学性能进行了表征,探究了PEO的质量分数对PLLA/PEO纳米纤维膜的结构、结晶行为和力学性能的影响。结果表明,PEO的加入,降低了PLLA/PEO纳米纤维膜的平均直径和直径分布宽度,提高了纤维的结晶速度、结晶度及断裂伸长率;PEO分子可提高聚乳酸分子链的运动能力和降低纤维的取向度,从而降低因取向引起的中间相含量,提高结晶相含量,进而改善聚乳酸纤维的结晶性能和韧性。     

组织工程用聚乳酸梯度支架的制备及分形研究

采用改进的溶液浇铸/粒子沥滤工艺,将聚乳酸（PLLA）与致孔剂粒子粉碎后,制备了双层和多层孔径梯度变化的PLLA泡沫支架。扫描电镜观察表明,材料内部界面处存在一定的扩散现象,且界面处孔的连通性好,通过调整致孔剂粒子的粒径以及PLLA与致孔剂粒子的比例,可有效地调控支架的各项参数（材料大小、孔径、孔隙率）。对材料进行分形研究,表明梯度多孔支架内部无明显界面,孔径过渡平缓。     

皮芯结构的纤维基组织工程肌腱支架的制备及性能研究

以聚乙交酯(polyglycolide,PGA)纤维和聚丙交酯(polylactide,PLA)纤维为原料,采用小口径针织技术制备了一种具有皮芯结构的新型纤维基组织工程肌腱支架.对支架的体外降解行为及细胞在支架上的黏附情况进行了研究.研究表明,支架的整个降解过程分为“强力下降期”、“质量损失期”、“准稳定期”三个阶段,细胞在支架的皮层和芯层上均黏附较好,而且分泌了大量的细胞外基质.     

新型明胶纳米纤维毡的制备及其细胞培养

以水为溶剂通过静电纺丝法制备出了新型明胶纳米纤维。结果表明，当明胶的质量分数为33%和体系温度为40℃时，所得明胶纳米纤维毡具有均匀多孔的微观形貌，纤维直径分布在120～210nm。用质量分数为1.5%的1-(3-二甲氨基丙基)-3-乙基碳二亚胺（EDC）/N-羟基琥珀酰亚胺（NHS）的乙醇/水溶液对纤维毡进行交联，所得明胶纳米纤维毡中纤维的形态得到较好的保持，且其焓值降低，耐热性和力学性能均有所提高。将制备的明胶纳米纤维毡进行牙周基质细胞（PDLCs）培养实验，初步的结果表明PDLCs能够在该纳米纤维毡上粘附、伸展和繁殖。     

低分子量聚乳酸对聚乳酸膜结构及性能的影响

在不引入除溶剂和溶质以外的第三组分情况下,用溶液浇铸法制备表面呈孔聚乳酸膜。通过在高分子量的聚乳酸中混入低分子量的聚乳酸,借助电子扫描电镜、差热分析、力学试验机考察低分子量聚乳酸对聚乳酸膜结构和性能的影响。通过在高分子聚乳酸中混入低分子量的聚乳酸可以制备出表面呈孔膜,该膜可满足力学性能和生物相容性好的临床要求,该方法减少了影响膜性能的因素。     

Preparation and mineralization of PLGA/Gt electrospun fiber mats

Poly(lactic-co-glycolic acid)(PLGA)/gelatin(Gt) ultrafine composite fibers were fabricated via electro-spinning.The effect of gelatin on the morphology and tensile property of the electrospun fiber mats was investigated.Mineralization was carried out in 10×simulated body fluid(10SBF).The deposited calcium phosphate(CaP) was identified by scanning electron microscopy(SEM),energy dispersive spectroscopy(EDS),X-ray diffraction(XRD) and Fourier transform infrared spectroscopy(FTIR).Results indicated that the av...     

新型明胶-无定形磷酸钙3D打印骨修复材料的制备

首先采用共沉淀法制备明胶改性的无定形磷酸钙(ACP)粉末材料,再以纯水作为粘接溶液,通过3D打印机制造骨移植修复材料,并应用红外光谱仪、X射线衍射仪以及扫描电子电镜进行表征.对于明胶稳定无定形磷酸钙化学结构的相关作用机制给出了初步的解释,重点考察了明胶添加量对打印骨成品的微观形貌、抗压强度以及孔隙率的影响规律,确定了明...     

聚乙烯醇及改性聚乙烯醇/明胶多孔支架的体外生物相容性研究

以吐温80为发泡剂,采用乳化发泡/冷冻—解冻法制备多孔的聚乙烯醇(PVA)和改性聚乙烯醇/明胶(PVA/Gel)支架材料,用扫描电子显微镜(SEM)对其形貌进行观察,并通过体外细胞培养对支架的生物相容性进行评价.实验结果表明,用该方法制备的多孔支架具有良好的贯通性,且PVA/Gel支架贯通性高于PVA支架;细胞结果显示,乳化发泡法制备的多孔支架对细胞的黏附、增殖和形貌无不良影响;荧光染色结果表明,PVA/Gel支架由于引入了大量氨基,使得细胞的黏附力优于PVA支架,具有更好的生物相容性.     

Preparation,characterization, and antibacterial activity of γ-irradiated silver nanoparticles in aqueous gelatin

Colloidal silver nanoparticles (Ag-NPs) were obtained through γ-irradiation of aqueous solutions containing AgNO₃ and gelatin as a silver source and stabilizer, respectively. The absorbed dose of γ-irradiation influences the particle diameter of the Ag-NPs, as evidenced from surface plasmon resonance (SPR) and transmission electron microscopy (TEM) images. When the γ-irradiation dose was increased (from 2 to 50 kGy), the mean particle size was decreased continuously as a result of γ-induced Ag-NPs fragmentation. The antibacterial properties of the Ag-NPs were tested against Methicillinresistant Staphylococcus aureus (MRSA) (Gram-positive) and Pseudomonas aeruginosa (P.a) (Gram-negative) bacteria. This approach reveals that the γ-irradiation-mediated method is a promising simple route for synthesizing highly stable Ag-NPs in aqueous solutions with good antibacterial properties for different applications.