基于天然水凝胶的生物材料在骨组织工程中的应用*

天然水凝胶是指原材料来自于天然生物材料的水凝胶。由于这种天然的聚合物含有构成生物体的天然成分,与天然组织具有生物学和化学相似性,而受到特别关注。天然水凝胶由于其与细胞外基质高度的相似性被认为是骨组织工程中优良的仿生基质材料。而针对天然水凝胶机械性能差、成骨诱导性能弱等缺陷,通常需要对天然水凝胶进行改性、引入其他材料或生物活性因子,以此来获得更适用于骨组织工程支架材料。对近年来基于天然水凝胶的生物材料在骨组织工程的应用,与其不同的应用形式(可注射水凝胶、多孔水凝胶支架、3D生物打印水凝胶支架等)进行了概述,以期对这类基于天然水凝胶的生物材料在未来骨组织工程中的应用提供参考。     

光交联水凝胶在组织工程中的研究进展

由于生物相容性、可降解性、与天然细胞外基质结构的相似性,水凝胶成为组织工程的研究热点与重点。基于原位形成和可注射性、与现有加工技术(3D打印、静电纺丝)的兼容性,光交联水凝胶在组织工程领域广泛应用。综述了近年来光交联水凝胶在组织工程领域的研究进展,包括其在软骨组织、骨组织、脂肪组织、牙周组织和皮肤组织方面的研究思路及应用进展,以期为后续光交联水凝胶作为组织工程支架的研究提供参考。     

细胞梯度力学微环境体外构建的研究

细胞微环境与细胞的相互作用日益成为细胞生物学领域研究热点。微环境中物理信号(如基底的力学性能、形貌和牵张力)在控制细胞命运中的作用更不容忽视。其中力学刺激常以不均一的梯度形式参与调节发育、炎症、伤口愈合以及癌症过程中不同细胞的增殖、迁移和分化等行为。水凝胶是模拟细胞外基质(extracellular matrix, ECM)二维/三维组织支架的理想材料。先进的微纳制造技术已被广泛应用于支撑或包裹细胞的仿生水凝胶的合成和微环境的个性化定制研究中。本文阐述了体内细胞力学微环境中刚度和拉压应力刺激的构建方法与表征手段的研究现状,并着重综述了近年来水凝胶在细胞梯度力学微环境体外构建中的应用研究,同时也对未来研究中所面临的挑战提出了新的展望。这些工作对于组织工程及再生医学具有重要意义。     

仿生黏弹性高分子水凝胶及其生物医学应用

天然细胞外基质和生物体软组织固有的黏弹性是调控细胞行为和组织修复与再生过程的关键因素.基于动态建构化学反应交联得到的动态高分子水凝胶材料可有效模拟在体细胞或组织的黏弹性力学微环境,为体外调控细胞命运、揭示其力学生物学响应机制提供了重要工具,也为组织修复与再生提供了仿生支架材料.本综述在介绍天然细胞外基质及生物体软组织黏弹性的基础上,重点对仿生黏弹性水凝胶材料的设计思路、性能表征及影响因素等进行了概括和总结,并揭示了黏弹性水凝胶调控细胞、组织行为的规律及机制,最后,分析了目前该领域研究中所存在的问题并对未来发展方向进行了展望.本综述将有助于启发高分子水凝胶的仿生功能化设计思路及材料生物学效应研究,进一步拓展高分子水凝胶材料的生物医学应用.     

琼脂糖在组织工程中的研究进展

琼脂糖是一种来源丰富、成本低廉的天然高分子材料,具有生物安全性和可降解性,利用其凝胶化现象可制成形状可塑的具有三维网络结构的凝胶。与其他天然材料相比,琼脂糖凝胶在机械性能上具有一定优势,如抗拉伸/压缩性、粘弹性、流变性等。其特殊的防粘连作用,使其必须与其他材料复合或者选用适宜的定型方式制成组织工程支架,以提高支架的组织相容性,用于填充、修复或者再生机体缺损组织。琼脂糖在组织工程领域的研究历史虽然不长,但在软骨、神经、骨、角膜和口腔黏膜等方面已经取得一些研究成果,其独特的微观结构和力学性能使其在软骨组织工程方面的研究最为广泛。目前,琼脂糖的组织工程研究离临床应用还有一段距离,材料制备和作用机理探索将是未来研究的重点。     

壳聚糖基温敏水凝胶的研究进展

壳聚糖是一种由甲壳素脱乙酰化得到的氨基多糖,具有生物相容性、低细胞毒性和可生物降解性等特点。壳聚糖/β-甘油磷酸钠溶液温敏水凝胶在组织工程、药物缓释等领域多有报道,其成胶性能取决于凝胶的组分和浓度。针对单纯壳聚糖水凝胶强度较低、降解较快、药物突释等缺陷,通常对壳聚糖进行改性或引入新材料共混,获得更符合实际需要的壳聚糖基温敏水凝胶。对近年来壳聚糖基水凝胶的研究进展进行综述,包括改性壳聚糖、共混体系等,概述了其在组织工程(软骨、血管、神经修复)、药物缓释(癌症药物缓释、糖尿病治疗)领域中研究和应用的新进展,以期为后续温敏水凝胶的进一步研究提供参考。     

丝素蛋白/壳聚糖复合材料在组织工程中应用的研究进展

丝素蛋白(silk fibroin,SF)和壳聚糖(chitosan,CS)具有良好的生物相容性和可降解性,然而单一组分的SF和CS支架材料的诸多缺点限制了其在组织工程研究中的应用。SF/CS复合材料克服单一组分SF和CS支架的缺点,具有力学性能优良、可塑性好、孔隙率及孔径可调和组分优势互补等特点。多种方法制备的SF/CS复合材料(微米/纳米颗粒、膜、纳米纤维、水凝胶和三维多孔支架)已用于骨、软骨、皮肤、神经、脂肪、心脏和角膜等组织工程或组织损伤修复的研究中。目前,国内外对于SF/CS复合材料在组织工程中应用的研究尚处于起步阶段。主要对SF/CS复合材料的特点、制备方法以及在多种组织工程中应用的研究现状进行了简要介绍。     

用于软骨修复的组织工程水凝胶

软骨损伤的发病率逐年增高,治疗要求也从过去的缓解疼痛,到现在的恢复运动功能,达到或接近伤前生活质量水平。传统的治疗方式受限于软骨组织特殊的生理病理条件,难以达到良好的远期治疗效果,所以,开发新型有效的治疗方式是软骨组织工程领域的研究热点。要达到良好的软骨修复效果,除了满足必需的生物活性因子与种子细胞要求外,作为载体的水凝胶也至关重要,只有与软骨本身的生物力学特征相似,才能够为软骨再生提供仿生的微环境。为了达到这一要求,已有多种设计方式与结构被应用于软骨组织工程水凝胶的构建,该文从水凝胶结构设计的角度对其在软骨修复组织工程中的应用进行了系统的综述。     

RADA-16的研究进展

自组装多肽RADA-16(RADARADARADARADA)是化学合成的一种纳米材料,最早是麻省理工学院的张曙光教授合成。研究显示,1%的RADA-16溶液在盐溶液的激发下能迅速自组装成纳米纤维水凝胶;在不同的多肽浓度下,RADA-16形成不同的结构形态,经超声波破碎打断的RADA-16纤维会随着时间的延长而重新组装成较长的纳米纤维;p H值的变化也会影响RADA-16水凝胶的形成,在酸性、碱性、中性条件下分别以3种离子状态存在;RADA-16纳米纤维水凝胶能够提供一个利于细胞生长的三维环境,将不同的细胞种植在水凝胶中能够促进细胞的增殖和迁移;已有科学家将RADA-16应用在组织工程中皮肤缺损、软骨修复、神经修复、止血等方面的研究并取得很好的研究成果。本研究主要对RADA-16的研究进展进行阐述。     

冷冻干燥及DMEM培养液浸泡对海藻酸水凝胶性能的影响

目的：考察冷冻干燥及DMEM培养液浸泡对海藻酸水凝胶性能的影响。方法：制备得到海藻酸水凝胶,对其结构和性能进 行了表征,重点模拟了细胞的培养过程,考察水凝胶- 冻干-DMEM培养液浸泡后水凝胶的力学性能、流变性能的变化。使用 CCK-8 法研究海藻酸水凝胶的细胞毒性。结果：制备了结构和性能可控的海藻酸水凝胶,冻干再浸泡会降低凝胶的拉伸强度和流 变屈服强度。海藻酸水凝胶对上皮细胞和平滑肌细胞没有明显的细胞毒性。结论：冷冻干燥及DMEM培养液浸泡后,凝胶拉伸强 度和屈服强度出现了下降,其原因是浸泡过程中钙离子会从中溶出。     

Dextran and hyaluronan methacrylate based hydrogels as matrices for soft tissue reconstruction

Polysaccharide hydrogels have become increasingly studied as matrices in soft tissue engineering because of their known cytocompatibility. In this work cross-linkable dextran methacrylates and hyaluronan methacrylate were synthesized and their transformation into stable hydrogels was studied. The in vitro degradation behaviour of the formed hydrogels could be controlled by the polysaccharide structure and the cross-linking density. Under in vitro conditions, the formed gels had no cytotoxic effects against fibroblasts, but cells could adhere only inefficiently in long term experiments. The use of composite gels improved the adherence of cells. Different scaffold architectures were studied including porous structures and perforated gel layers. Selected hydrogels were examined in an in vivo pilot study using a rabbit model to evaluate their biocompatibility, stability and degradation. No signs of inflammation were seen and with prolonged duration the material was degraded and lacunas were formed by immigrating or ingrowing cells. Optimizing their mechanical properties, the formed hydrogels represent promising candidates as matrices for soft tissue reconstruction.     

PEGDA hydrogels with patterned elasticity: Novel tools for the study of cell response to substrate rigidity

The ability of cells to migrate in response to mechanical gradients (durotaxis) and differential cell behavior in adhesion, spreading, and proliferation in response to substrate rigidity are key factors both in tissue engineering, in which materials must be selected to provide the appropriate mechanical signals, and in studies of mechanisms of diseases such as cancer and atherosclerosis, in which changes in tissue stiffness may inform cell behavior. Using poly(ethylene glycol) diacrylate hydrogels with varying polymer chain length and photolithographic patterning techniques, we are able to provide substrates with spatially patterned, tunable mechanical properties in both gradients and distinct patterns. The hydrogels can be patterned to produce anisotropic structures and exhibit patterned strain under mechanical loading. These hydrogels may be used to study cell response to substrate rigidity in both two and three dimensions and can also be used as a scaffold in tissue‐engineering applications. Biotechnol. Bioeng. 2010; 105: 636–644. © 2009 Wiley Periodicals, Inc.     

Tunable Collagen I Hydrogels for Engineered Physiological Tissue Micro-Environments

Collagen I hydrogels are commonly used to mimic the extracellular matrix (ECM) for tissue engineering applications. However, the ability to design collagen I hydrogels similar to the properties of physiological tissues has been elusive. This is primarily due to the lack of quantitative correlations between multiple fabrication parameters and resulting material properties. This study aims to enable informed design and fabrication of collagen hydrogels in order to reliably and reproducibly mimic a variety of soft tissues. We developed empirical predictive models relating fabrication parameters with material and transport properties. These models were obtained through extensive experimental characterization of these properties, which include compression modulus, pore and fiber diameter, and diffusivity. Fabrication parameters were varied within biologically relevant ranges and included collagen concentration, polymerization pH, and polymerization temperature. The data obtained from this study elucidates previously unknown fabrication-property relationships, while the resulting equations facilitate informed a priori design of collagen hydrogels with prescribed properties. By enabling hydrogel fabrication by design, this study has the potential to greatly enhance the utility and relevance of collagen hydrogels in order to develop physiological tissue microenvironments for a wide range of tissue engineering applications.     

Reductive alkylation of hyaluronic acid for the synthesis of biocompatible hydrogels by click chemistry

Hyaluronan (HA) based hydrogels have been synthesized combining chemical modification of the polysaccharide by partial oxidation, reductive amination and 'click chemistry'. HA was oxidized by 4-acetamido-TEMPO-mediated reaction, using sodium hypochlorite as primary oxidant and NaBr in buffered pH, so that the produced aldehyde moieties (hemiacetals) were trapped in situ by adding primary amines containing azide or alkyne-terminal groups. The structure of the reaction products, oxidized-HA and primary amines bonded to HA, was elucidated using 2D NMR spectroscopy. SEC-MALLS analysis of the modified substrates showed a negligible degradation of the polysaccharide using this procedure. Furthermore, azido- and alkynyl derivatives underwent cross-linking by click chemistry into hydrogels, which were characterized by NMR, FT-IR, swelling degree and mechanical properties. Possible application of the material as scaffold for tissue engineering was tested by seeding and proliferation of chondrocytes for up to 15 days.     

Evaluation of Physical and Mechanical Properties of Porous Poly (Ethylene Glycol)-co-(L-Lactic Acid) Hydrogels during Degradation

Porous hydrogels of poly(ethylene glycol) (PEG) have been shown to facilitate vascularized tissue formation. However, PEG hydrogels exhibit limited degradation under physiological conditions which hinders their ultimate applicability for tissue engineering therapies. Introduction of poly(_L-lactic acid) (PLLA) chains into the PEG backbone results in copolymers that exhibit degradation via hydrolysis that can be controlled, in part, by the copolymer conditions. In this study, porous, PEG-PLLA hydrogels were generated by solvent casting/particulate leaching and photopolymerization. The influence of polymer conditions on hydrogel architecture, degradation and mechanical properties was investigated. Autofluorescence exhibited by the hydrogels allowed for three-dimensional, non-destructive monitoring of hydrogel structure under fully swelled conditions. The initial pore size depended on particulate size but not polymer concentration, while degradation time was dependent on polymer concentration. Compressive modulus was a function of polymer concentration and decreased as the hydrogels degraded. Interestingly, pore size did not vary during degradation contrary to what has been observed in other polymer systems. These results provide a technique for generating porous, degradable PEG-PLLA hydrogels and insight into how the degradation, structure, and mechanical properties depend on synthesis conditions.     

A homogenization model of the annulus fibrosus

The objective of this study was to use a homogenization model of the anisotropic mechanical behavior of annulus fibrosus (AF) to address some of the issues raised in structural finite element and fiber-reinforced strain energy models. Homogenization theory describes the effect of microstructure on macroscopic material properties by assuming the material is composed of repeating representative volume elements. We first developed the general homogenization model and then specifically prescribed the model to in-plane single lamella and multi-lamellae AF properties. We compared model predictions to experimentally measured AF properties and performed parametric studies. The predicted tensile moduli (E theta and E z) and their dependence on fiber volume fraction and fiber angle were consistent with measured values. However, the model prediction for shear modulus (G thetaz) was two orders of magnitude larger than directly measured values. The values of E theta and E z were strongly dependent on the model input for matrix modulus, much more so than the fiber modulus. These parametric analyses demonstrated the contribution of the matrix in AF load support, which may play a role when protoeglycans are decreased in disc degeneration, and will also be an important design factor in tissue engineering. We next compared the homogenization model to a 3-D structural finite element model and fiber-reinforced energy models. Similarities between the three model types provided confidence in the ability of these models to predict AF tissue mechanics. This study provides a direct comparison between the several types of AF models and will be useful for interpreting previous studies and elucidating AF structure-function relationships in disc degeneration and for functional tissue engineering.     

Facile synthesis of degradable and electrically conductive polysaccharide hydrogels

Degradable and electrically conductive polysaccharide hydrogels (DECPHs) have been synthesized by functionalizing polysaccharide with conductive aniline oligomers. DECPHs based on chitosan (CS), aniline tetramer (AT), and glutaraldehyde were obtained by a facile one-pot reaction by using the amine group of CS and AT under mild conditions, which avoids the multistep reactions and tedious purification involved in the synthesis of degradable conductive hydrogels in our previous work. Interestingly, these one-pot hydrogels possess good film-forming properties, electrical conductivity, and a pH-sensitive swelling behavior. The chemical structure and morphology before and after swelling of the hydrogels were verified by FT-IR, NMR, and SEM. The conductivity of the hydrogels was tuned by adjusting the content of AT. The swelling ratio of the hydrogels was altered by the content of tetraaniline and cross-linker. The hydrogels underwent slow degradation in a buffer solution. The hydrogels obtained by this facile approach provide new possibilities in biomedical applications, for example, biodegradable conductive hydrogels, films, and scaffolds for cardiovascular tissue engineering and controlled drug delivery.