超声增强藻酸钙凝胶支架材料孔隙率的研究

藻酸钙凝胶具有三维立体多孔结构, 能为细胞生长提供充分的附着空间, 且具有良好的生物相容性和一定的机械强度, 是一种理想的细胞支架材料. 本文研究了藻酸钙三维支架材料的力学特性与氯化钙/藻酸钠的配比的关系, 并提出采用低强度脉冲超声处理藻酸钙凝胶、基于超声空化效应增强藻酸钙凝胶孔隙率的新方法. 实验采用交联合成方法制备藻酸钙凝胶支架材料, 测量力学特性、孔洞的联通性与孔隙率, 并利用绿色荧光蛋白的表达评价细胞的增殖能力. 结果表明, 当氯化钙/藻酸钠的配比为3:5时, 凝胶的机械强度和弹性较好, 力学性能稳定, 为最佳配比参数. 采用声压0.055 MPa的脉冲超声作用20 min, 可以有效提高凝胶支架的孔隙率; 且细胞在该支架中生长状态良好, 呈现团簇状生长趋势.     

荧光偏振法研究脉冲电场对酿酒酵母细胞膜流动性影响

以DPH(1,6-二苯基-1,3,5-己三烯)为荧光探剂,采用荧光偏振法探讨了脉冲电场(0～25 kV·cm-1,0～266 ms)对酿酒酵母细胞膜流动性影响。经5 kV·cm-1电场处理后,酿酒酵母细胞膜的流动性显著减小,并且随电场强度和处理时间的增加而减小;通过平板计数法和紫外分光光度计法分别检测了脉冲电场对酿酒酵母细胞存活对数及膜通透性影响。结果显示,5 kV·cm-1虽然只能使少量的酵母致死,却能使酵母细胞膜的通透性显著增加,膜流动性显著降低。并且细胞的存活率随电场强度增大而减小,细胞膜的通透性随电场强度增大而增大。这表明细胞膜的流动性降低与细胞膜的通透性升高成正相关,与细胞的存活率成负相关。由此推测脉冲电场在对酿酒酵母灭菌过程中,细胞膜是其作用的一个关键位点,膜流动性减小,细胞膜通透性增强,是细胞死亡的主要原因。     

梯度穿缝型双孔隙率多孔材料的吸声性能

在传统单一孔隙率多孔材料中引入宏观尺度的周期性梯度穿缝结构设计,构造出梯度穿缝型双孔隙率多孔材料,其包含多孔材料基体微孔尺度与穿缝尺度两个尺度。采用分层等效的理论建模方法,将复杂梯度渐变问题变为多层均匀等效层叠加问题。针对不同特征尺寸的多孔材料薄层,分别采用低、高两种渗透率对比度双孔隙率理论,给出了其等效密度和动态压缩系数,再应用传递矩阵方法得到了相邻薄层之间的声压和质点速度传递关系并求得其表面声阻抗,从而建立了梯度穿缝型双孔隙率多孔材料的吸声理论模型。发展了多尺度材料声学有限元数值模型,在所考虑的100~3000 Hz频段范围内数值模拟结果完全吻合理论模型结果。理论与模拟分析了多尺度结构参数对双孔隙率多孔材料吸声性能的影响,结果表明引入多尺度梯度结构设计能够显著提高单一孔隙率多孔材料的吸声性能,且穿缝尺度比穿缝梯度影响更为显著;精细数值模拟获得的声压和能量密度分布云图揭示了多尺度结构设计的吸声增强机制。该工作可用于指导双孔隙率多孔材料的多尺度结构设计,从而提高多孔材料的中低频吸声性能。      

激光参数对纳米金靶向细胞膜通透性的影响

激光参数,比如脉宽、照射能量、脉冲数目以及照射方式等在各个方而尤其是在生物医学应用中对生物组织的不同效应都起着重要作用.采用纳米金颗粒靶向细胞,激光照射后细胞膜的通透性会发生改变.采用不同的激光光源和不同的照射方式(直接照射和扫描照射)、调整不同的激光参数照射细胞微粒结合体,通过流式细胞仪测量细胞对异硫氰酸荧光素葡聚糖(fluorescein isothiocyanate-Dextran,FITC-D)和碘化丙啶(propidium iodide PI)的吸收量来判断细胞膜的暂时通透性和细胞的死亡率,用以研究激光参数变化对细胞膜通透性的影响.采用传热模型对其机理进行了分析.结果表明通过调整激光参数,采用这种方法有可能进行基因转染.     

声场中双空化泡的运动特性

空化泡的运动特性是声场作用下的动力学行为,受空化泡初始半径,声压幅值,驱动声压频率,液体特性等众多因素的影响,是个复杂工程。本文从双空化泡运动方程出发,考虑到液体粘滞系数、空化泡辐射阻尼项的影响,研究了不同初始半径、驱动声压频率、驱动声压幅值、液体粘滞系数下空化泡泡壁的运动情况,研究结果表明不同初始半径、外界驱动声压频率、驱动声压幅值、液体粘滞系数均会对空化泡的膨胀比和空化泡的溃灭时间有一定影响。     

超声空化增强川芎嗪对脑缺血再灌注损伤保护作用机制的研究

川芎的生物活性成分(川芎嗪)已广泛应用于治疗心脑血管疾病。基于谷氨酸诱导PC12细胞损伤建立脑缺血再灌注损伤的细胞模型,探讨超声增强川芎嗪对谷氨酸损伤PC12细胞的保护作用机制。研究结果表明,超声能进一步加强川芎嗪对细胞的保护,其主要作用机制为:(1)抑制氧化应激和细胞凋亡相关的Bcl-2蛋白和Bax蛋白的变化从而达到抗凋亡的效果;(2)降低炎症因子(TNF-α和IL-8)的表达,减轻炎症反应损伤;(3)适当的声压可以增强川芎嗪对谷氨酸损伤PC12细胞的保护作用,但过高的声压会引起细胞损伤,导致细胞凋亡。本文的工作表明超声能够增强川芎嗪对脑缺血再灌注损伤的保护作用,为临床脑缺血再灌注损伤的治疗提供了新治疗策略。      

基于三维分数阶微分增强的边缘曲面重构算法

提出一种基于三维分数阶微分增强的三维边缘曲面重构算法,可重构出更多的三维不规则目标的细节结构信息,克服了原边缘曲面重构算法重构细节信息不充分的缺点.根据分数阶微分傅里叶变换的可分离性原理,将二维分数阶微分算子推广至三维并推导出三维分数阶离散滤波模板,利用三维分数阶微分对图像具有非线性增强作用的特性,对三维切片数据场进行三维分数阶微分增强.与传统三维边缘曲面重构算法相比,经过三维分数阶微分增强且采用分数阶梯度追踪细节改进的重构算法,能够重构出更丰富的三维目标细节结构信息.算法运用于神经元细胞的共焦显微图像中三维不规则目标的边缘曲面重构,实验结果验证了该算法的正确性和高效性,可推广应用至生物医学领域的三维可视化研究.     

超声在液体中的非线性传播及反常衰减

本文从广义的Navier-Stokes流体方程出发,考虑到流体介质的黏滞性和存在的热传导,导出了更接近实际流体的三维非线性声波动方程.鉴于声传播所涉及的空间和时间尺度的复杂性和多样性,文中针对一维情形下的非线性波动方程进行了求解和分析.由方程的二级近似解可以看出,声压振幅的衰减遵循几何级数规律,而且驱动声波的频率越高声压的衰减就越快.在满足条件ωb《ρ0c_0~2时,基波的衰减系数与驱动频率的平方及耗散系数的乘积成正比;二次谐波的衰减规律更加复杂,与频率的更高次幂相关.对声衰减系数及声压的分布进行数值计算发现,声压的分布还与初始的声压幅值及频率有关,初始的声压与频率越高衰减得越快.另外,当声压高于液体的空化阈值时,液体中就会出现大量的空化泡,文中模拟了单个空化泡的运动,发现随着声压的增大空化泡的振动越剧烈、空化泡所受的黏滞力变大,随着声波作用时间的增大黏滞力的幅值迅速增大并与驱动声压值同阶,因而空化泡的非线性径向运动引起的声衰减不容忽视.结果表明,驱动声压越高在空化区域附近引起的声衰减越快、输出的声压越低.     

液体材料超声处理过程中声场和流场的分布规律研究

针对合金熔体等液体材料的超声处理过程,选取水作为透明模型材料,采用数值模拟计算和示踪粒子实验方法,研究了20和490 kHz两种频率超声作用下水中的声场和流场分布.结果表明,增大变幅杆半径能够提高水中声压水平,扩大空化效应的发生区域.当超声频率为20 kHz时,水中声压最大值出现在超声变幅杆下端面处,且声压沿传播距离的增大而显著减小.如果超声频率增加至490 kHz,水中的声压级相比于20 kHz时明显提高,且声压沿着超声传播方向呈现出周期性振荡特征.两种频率超声作用下水中的流场呈现相似的分布特征,且平均流速均随着变幅杆半径增大表现出先升高后降低的趋势.变幅杆半径相同时,20 kHz频率超声作用下水中的平均流速高于490 kHz频率超声.采用示踪粒子图像测速技术实时观察和测定了水中的流速分布,发现其与计算结果基本一致.     

双层生物膜的动力学性质

采用傅里叶空间中的布朗运动方程研究了具有双层结构的生物膜的动力学性质,给出了生物膜在空间热库作用下随机运动的三维图像.研究表明,双层生物膜之间的相对滑动是一个非常重要的动力学过程,对高度-高度相关函数有非常显著的影响. 关键词： 双层细胞膜 相关函数     

Functionalized alginate/chitosan biocomposites consisted of cylindrical struts and biologically designed for chitosan release

A novel alginate/chitosan composite scaffold was developed. The composite scaffolds were fabricated at low temperature using a three-axis robot system connected to a micro-dispenser and a core/shell nozzle. The structure of the composite scaffolds included hollow struts; deposited chitosan on the inner walls (core region) of the struts reacted electrostatically with the alginate layer (shell region). The fabricated, highly porous composite scaffolds exhibited excellent mechanical properties and controllable chitosan release, which was closely dependent on the weight fraction of the alginate in the shell region. The tensile strength in the dry state was ∼1.8-fold greater than that of pure alginate scaffold due to the ionic interaction between alginate and chitosan. To determine the feasibility of using the developed scaffold in tissue regeneration applications, in vitro cellular responses were evaluated using osteoblast-like-cells (MG63). The cell proliferation on the composite scaffold was ∼3.4-fold greater than that on the pure alginate scaffold. Alkaline phosphate activity and calcium deposition of the composite scaffold after 14 and 21 days of cell culture were significantly enhanced (1.6- and 1.8-fold greater, respectively) compared with those of the pure alginate scaffold. These results suggested that the alginate/chitosan composite scaffolds with a controlled chitosan release have great potential for use in regenerating various tissues.     

Influence of Nano-Bioactive Glass (NBG) Content on Properties of Gelatin-Hyaluronic Acid/NBG Composite Scaffolds

The development of three-dimensional (3-D) scaffolds with highly open porous structure is one of the most important issues in tissue engineering. A novel nanocomposite scaffold of gelatin (Gel), hyaluronic acid (HA), and nano-bioactive glass (NBG) was prepared by blending NBG with a Gel and HA solution followed by lyophilization. The effects of NBG content on the properties of the Gel-HA/NBG composite scaffolds, including the morphologies, porosity, compressive strength, swelling behavior, cell viability and alkaline phosphatase (ALP) activity, were investigated. Porous composite scaffolds with interconnected pores were obtained and the pores became cylindrical with increasing NBG content. The porosity percent and swelling ability decreased with increasing NBG content; however, the compressive strength, cell viability and ALP activity were enhanced. All the results showed the addition of NBG particles can improve the physicochemical and biological properties and the Gel-HA/NBG composite scaffolds exhibited good potential for tissue engineering applications.     

Preparation and Characterization of Nanocomposite Scaffolds Based on Polycaprolactone-Polyethylene Glycol/Methylene Diphenyl Diisocyanate/Diethylene Glycol and Nano-Bioactive Glass

Designing and fabricating nanocomposite scaffolds based on biodegradable polymers and bioactive materials are an important topic in the area of bone regeneration. A novel nanocomposite scaffold composed of polyurethane (BPU) and nano-bioactive glass (NBAG) was prepared. The effects of the NBAG content on the properties of the BPU/NBAG composite scaffolds, including the morphologies, porosity and compressive strength, were investigated. The BPU/NBAG composite scaffolds showed an interconnected pore structure with the pore size ranging from 50 to 500?μm for all samples. The porosity percent and swelling ability decreased with increasing NBAG content; however, the compressive strength was enhanced.     

3D polycarprolactone (PCL) scaffold with hierarchical structure fabricated by a piezoelectric transducer (PZT)-assisted bioplotter

The 3D bioplotter, which is one of the rapid-prototyping systems, enables us to produce the design-based scaffolds which could control good mechanical properties and pore structures for mimicking human organs. Although the plotting system has several advantages to fabricate a variety of designed scaffolds, the main disadvantage of scaffolds fabricated by the system is that the strand surfaces are too smooth and tend to discourage initial cell attachment within the scaffolds. To overcome the problem, we suggest a new 3D plotting method supplemented by piezoelectric vibration system for fabricating scaffolds that have hierarchical surface structures, which increase the surface roughness of the scaffold without any additional chemical process. The surface-modified 3D scaffold exhibited various positive qualities including enhanced compressive modulus and improved initial cell attachment and proliferation. Cell culturing results demonstrated that the interactions between chondrocytes and the scaffold were much more favorable than those between the cells and conventionally plotted 3D scaffolds. This process provides a feasible new technique for fabricating high-quality 3D scaffolds for tissue engineering applications.     

Biomimetic formation of apatite on the surface of porous gelatin/bioactive glass nanocomposite scaffolds

There have been several attempts to combine bioactive glasses (BaGs) with biodegradable polymers to create a scaffold material with excellent biocompatibility, bioactivity, biodegradability and toughness. In the present study, the nanocomposite scaffolds with compositions based on gelatin (Gel) and BaG nanoparticles in the ternary SiO₂-CaO-P₂O₅ system were prepared. In vitro evaluations of the nanocomposite scaffolds were performed, and for investigating their bioactive capacity these scaffolds were soaked in a simulated body fluid (SBF) at different time intervals. The scaffolds showed significant enhancement in bioactivity within few days of immersion in SBF solution. The apatite formation at the surface of the nanocomposite samples confirmed by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX) and X-ray powder diffraction (XRD) analyses. In vitro experiments with osteoblast cells indicated an appropriate penetration of the cells into the scaffold's pores, and also the continuous increase in cell aggregation on the bioactive scaffolds with increase in the incubation time demonstrated the ability of the scaffolds to support cell growth. The SEM observations revealed that the prepared scaffolds were porous with three dimensional (3D) and interconnected microstructure, pore size was 200-500 μm and the porosity was 72-86%. The nanocomposite scaffold made from Gel and BaG nanoparticles could be considered as a highly bioactive and potential bone tissue engineering implant.     

Applications of low-intensity pulsed ultrasound to increase monoclonal antibody production in CHO cells using shake flasks or wavebags

Many technologies, such as cell line screening and host cell engineering, culture media optimization and bioprocess optimization, have been proposed to increase monoclonal antibody (mAb) production in Chinese Hamster Ovary (CHO) cells. Unlike the existing biochemical approaches, we investigated stimulation using low-intensity pulsed ultrasound (LIPUS) as a purely physical approach, offering enhanced scalability, contamination control and cost-efficiency, while demonstrating significantly increased cell growth and antibody production. It was found that daily ultrasound treatments at 40 mW/cm² for 5 min during cell culture increased the production of human anti-IL-8 antibody by more than 30% using 10 or 30 mL shake flasks. Further increasing the ultrasound dosage (either intensities or the treatment duration) did not appreciably increase cell growth or antibody production, however feeding the culture with additional highly-concentrated nutrients, glucose and amino acids (glutamine in this case), did further increase cell growth and antibody titer to 35%. Similar ultrasound treatments (40 mW/cm², 5 min per day) when scaled up to larger volume wavebags, resulted in a 25% increase in antibody production. Increased antibody production can be attributed to both elevated cell count and the ultrasound stimulation. Theoretical study of underlying mechanisms was performed through the simulations of molecular dynamics using the AMBER software package, with results showing that LIPUS increases cell permeability. The significance of this study is that LIPUS, as a physical-based stimulation approach, can be externally applied to the cell culture without worrying about contamination. By combining with the existing technologies in antibody production, LIPUS can achieve additional mAb yields. Because it can be easily integrated with existing cell culture apparatuses, the technology is expected to be more acceptable by the end users.     

Effect of the internal microstructure in rapid-prototyped polycaprolactone scaffolds on physical and cellular properties for bone tissue regeneration

Biomedical scaffolds should be designed to optimize their inter-microstructure to enable cell infiltration and nutrient/waste transport. To acquire these properties, several structural parameters, such as pore size, pore shape, porosity, pore interconnectivity, permeability, and tortuosity are required. In this study, we explored the effect of tortuosity on the viable cell proliferation and mineralization of osteoblast-like-cells (MG63) in polycaprolactone scaffolds. For analysis, we designed four different scaffolds of various tortuosities ranging from 1.0 to 1.3 under the same porosity (56?%) and 100?% pore interconnectivity. The pore size of the scaffolds was set as 150 and 300?μm, and a mixture of these sizes. We found that despite the porosity being same, the elastic modulus was dependent on the pore size of the scaffolds due to the distributed stress concentration. In addition, the relative water movement within scaffolds was also related to the internal microstructure. Cell viability and Ca²⁺ deposition of the cell-seeded scaffolds showed that the proliferation of viable cells and mineralization in the scaffolds with appropriate tortuosity (1.2) was relatively high compared to those of the scaffolds displaying low (1.05 and 1.1) or high (1.3) tortuosity. Our findings indicated that the internal microstructure of the scaffolds may influence not only the physical properties, but in addition the cellular behavior.     

Influence of low-intensity pulsed ultrasound on osteogenic tissue regeneration in a periodontal injury model: X-ray image alterations assessed by micro-computed tomography

Objective

This study was conducted to evaluate, with micro-computed tomography, the influence of low-intensity pulsed ultrasound on wound-healing in periodontal tissues.

Methods

Periodontal disease with Class II furcation involvement was surgically produced at the bilateral mandibular premolars in 8 adult male beagle dogs. Twenty-four teeth were randomly assigned among 4 groups (G): G1, periodontal flap surgery; G2, periodontal flap surgery + low-intensity pulsed ultrasound (LIPUS); G3, guided tissue regeneration (GTR) surgery; G4, GTR surgery plus LIPUS. The affected area in the experimental group was exposed to LIPUS. At 6 and 8 weeks, the X-ray images of regenerated teeth were referred to micro-CT scanning for 3-D measurement.

Results

Bone volume (BV), bone surface (BS), and number of trabeculae (Tb) in G2 and G4 were higher than in G1 and G3 (p < 0.05). BV, BS, and Tb.N of the GTR + LIPUS group were higher than in the GTR group. BV, BS, and Tb.N of the LIPUS group were higher than in the periodontal flap surgery group.

Conclusion

LIPUS irradiation increased the number, volume, and area of new alveolar bone trabeculae. LIPUS has the potential to promote the repair of periodontal tissue, and may work effectively if combined with GTR.     

Preparation and Characterization of Poly(L-lactide-co-glycolide-co-ε-caprolactone)/Nano-biaoactive Glass-Nano-β-tricalcium Phosphate Composite Scaffolds

Abstract

Polymeric/ceramic composite scaffolds that are biocompatible and biodegradable are widely used for tissue engineering applications. In this work a series of poly(L-lactide-co-glycolide-co-ε-caprolactone)/nano-biaoactive glass-nano-β-tricalcium phosphate composite scaffolds were successfully fabricated and the influences of the inorganic content and freezing temperature on the physical properties were studied. The composite scaffolds with various inorganic contents showed an interconnected pore structure with irregular shapes. The composite scaffolds had a porosity that was reduced with increasing inorganic content and decreasing freezing temperature. The incorporation of inorganic fillers and decreasing freezing temperature improved the mechanical properties of the hybrid scaffolds. By appropriate control of these two factors (10.0?wt% content of NBAG and β-TCP with freezing at ?30?°C) a suitable composite scaffold was prepared as a potential bone tissue engineering implant.