新型骨-软骨一体化修复支架材料的制备

利用可降解聚合物微球的相互粘结制备了一种新型的组织工程支架材料, 可用于软骨和软骨下骨损伤的修复。采用光学显微镜、 扫描电镜对支架的表面形貌、 内部结构进行了表征, 同时研究了支架材料的力学性能, 此外还研究了微球的粒径对支架材料孔隙率的影响。结果显示, 该材料在结构上分为乳酸-羟基乙酸共聚物(PLGA)层和PLGA/生物活性玻璃(BG)层; 材料的孔隙三维连通、 分布均匀; 采用粒径为150~200μm微球所制备的支架孔隙率为(53.37±4.39)%, 在10%的应变下材料压缩强度便已达到了0.9MPa, 显示了较强的力学性能; 随着微球粒径的变小, 材料孔隙率逐渐增大。这种微球支架在骨-软骨组织缺损修复方面有着很大的研究价值和应用价值。      

可注射型天然高分子软骨组织支架材料

作为理想的组织工程支架材料，可注射水凝胶材料受到医学和材料学领域科学家的广泛关注，而天然高分子材料由于具有良好的生物相容性和安全性，在这一领域备受青昧。综述了胶原、纤维蛋白和海藻酸盐在用于可注射型软骨组织支架材料方面的特征和研究现状，展望了这一领域的发展。     

ADSCs-壳聚糖/明胶水凝胶工程化软骨的三维动态构建

组织工程软骨的体外构建被认为是一种有希望治疗关节软骨缺损的有效途径。为评估载脂肪干细胞(Adipose-derived stem cells,ADSCs)壳聚糖/明胶水凝胶支架,在体外动态构建组织工程软骨相对传统静态培养的优势,本研究用壳聚糖/明胶制备了软骨仿生支架,并检测其物理性质。在制备的水凝胶支架上以1×107cells/m L密度接种ADSCs后,分别置于转瓶及T-瓶的软骨诱导基中培养两周,通过试剂染色、代谢检测和电镜观察,考察了细胞的软骨分化能力、活性、生长分布、渗透深度、增殖及胞外基质分泌情况。结果表明,壳聚糖/明胶支架的平均孔径为118.25±19.51μm,孔隙率为82.60±2.34%,吸水率为361.28±0.47%,弹性模量为61.2±0.16 k Pa,具有良好生物相容性。ADSCs生长状态良好,可向软骨细胞分化,适于作为组织工程软骨构建的种子细胞。表征结果显示,转瓶内水凝胶支架中细胞蛋白多糖的表达更显著,细胞生长分布更加均匀,细胞外基质分泌基本填满整个支架。因此,转瓶载壳聚糖/明胶支架所提供的三维动态环境,是体外构建组织工程软骨的良好方法。     

骨软骨组织工程仿生梯度支架研究进展

骨软骨缺损是导致关节发病和残疾的重要原因,骨软骨组织工程是修复骨软骨缺损的方法之一。骨软骨组织工程方法涉及仿生梯度支架的制造,该支架需模仿天然骨软骨组织的生理特性（例如从软骨表面到软骨下骨之间的梯度过渡）。在许多研究中骨软骨仿生梯度支架表现为离散梯度或连续梯度,用于模仿骨软骨组织的特性,例如生物化学组成、结构和力学性能。连续型骨软骨梯度支架的优点是其每层之间没有明显的界面,因此更相似地模拟天然骨软骨组织。到目前为止,骨软骨仿生梯度支架在骨软骨缺损修复研究中已经取得了良好的实验结果,但是骨软骨仿生梯度支架与天然骨软骨组织之间仍然存在差异,其临床应用还需要进一步研究。本文首先从骨软骨缺损的背景、微尺度结构与力学性能、骨软骨仿生梯度支架制造相关的材料与方法等方面概述了离散和连续梯度支架的研究进展。其次,由于3D打印骨软骨仿生梯度支架的方法能够精确控制支架孔的几何形状和力学性能,因此进一步介绍了计算仿真模型在骨软骨组织工程中的应用,例如采用仿真模型优化支架结构和力学性能以预测组织再生。最后,提出了骨软骨缺损修复相关的挑战以及骨软骨组织再生未来研究的展望。例如,连续型骨软骨仿生梯度支架需要更相似地模拟天然骨软骨组织单元的结构,即力学性能和生化性能的过渡更加自然地平滑。同时,虽然大多数骨软骨仿生梯度支架在体内外实验中均取得了良好的效果,但临床研究和应用仍然需要进行进一步深入研究。     

聚氨酯软骨-骨双层复合材料的构建及性能研究

关节软骨损伤是临床上的常见病,由于其组织再生能力差,可能导致骨性关节炎的发生,因此,研究开发骨-软骨移植替代材料非常重要。目的就是设计一体化软骨-骨双层复合材料,以解决软骨与骨的整合问题。该双层复合体上层软骨材料为聚氨酯,软骨下骨为羟基磷灰石/聚氨酯复合支架材料,两层结构中引用了同一种材料——聚氨酯,将双层结构有机黏合在一起,使黏合更牢固。下层多孔HA/PU复合支架材料的孔与孔之间相互贯通,孔隙率约为83%,孔径范围分布在200～600μm。体外细胞相容性实验表明,该一体化双层复合材料为细胞的黏附、增殖以及生存活力的维持提供了有利环境。上述结果表明该双层复合材料有望用于软骨组织工程修复。     

人工软骨支架材料、结构设计与制备技术研究进展

骨软骨是一种半透明状组织,主要功能是传递、吸收应力和减少摩擦。由于结构和功能复杂性,软骨一旦受损很难修复和再生,软骨缺损治疗仍是一大临床难题。随着再生医学蓬勃发展,组织工程人工软骨技术有望在软骨修复和治疗领域发挥重要作用。首先介绍了天然关节软骨不同分层的解剖结构和功能特征,然后重点从人工软骨支架构建材料、结构设计和制备技术等方面系统地综述了人工软骨组织工程技术的最新进展,最后讨论了人工软骨支架当前面临主要问题和未来发展方向,以期为相关研究提供参考。     

可注射海藻酸钙水凝胶的制备研究

海藻酸钙水凝胶因其良好的生物相容性广泛应用于组织工程支架材料的研究。以海藻酸钠(SA),碳酸钙,葡萄糖酸内酯(GDL)为原料,通过原位相转变制备可注射凝胶,用于软骨组织微创修复材料的研究。测定了单一变量条件下不同海藻酸钠浓度、f值(钙离子与羧基的摩尔比)及n值(葡萄糖酸内酯与钙离子的摩尔比)对海藻酸凝胶力学强度、溶胀率、浸提液pH值等的影响,从而获得各组分最适的配比;另外,通过原位接种软骨细胞,研究了软骨细胞在凝胶中的生长行为。综合海藻酸钙凝胶性能,最终确定海藻酸钠浓度为2.5%、f=0.5及n=0.6为最佳配比;细胞培养结果表明软骨细胞在凝胶中具有较高的活性且维持了其软骨细胞形态,证实了研究制得的海藻酸钙水凝胶是一种优良的可注射软骨组织工程支架材料。     

胶原作为组织工程软骨支架材料的研究

本文报导了用胶原作为软骨组织的载体材料在组织工程化软骨中的研究情况.本研究采用冷冻干燥方法制备出了Ⅰ-型胶原(sigma)、Ⅱ-型胶原(sigma)和混合型胶原(本室提取)的胶原海绵,并将其用作软骨组织工程的载体支架,比较研究了这三种胶原材料支架在软骨组织工程应用中的效果,筛选出了较理想的软骨组织工程载体材料.     

CPP/PLLA软骨组织工程支架复合材料初步研究

采用溶媒投放、颗粒滤取技术制备出CPP/PLLA软骨组织工程支架复合材料,测试了该复合材料的物理力学性能和降解性能。研究结果表明,CPP/PLLA软骨组织工程支架复合材料具有高的孔隙率(90%)、良好的生物降解性能和物理力学性能,以及三维连通、微孔、网状微观结构,故该复合材料有希望成为软骨组织工程支架材料之一。     

高润滑水凝胶的结构与吸附-排斥作用

水凝胶因具有与天然软骨材料相近的吸水性、高弹性及低摩擦性能,从而成为关节软骨的理想替代材料.介绍吸附-排斥模型及水凝胶的润滑行为,并论述水凝胶的本体结构与水凝胶润滑性能之间的关系.     

Hydrogels of collagen/chondroitin sulfate/hyaluronan interpenetrating polymer network for cartilage tissue engineering

The network structure of a three-dimensional hydrogel scaffold dominates its performance such as mechanical strength, mass transport capacity, degradation rate and subsequent cellular behavior. The hydrogels scaffolds with interpenetrating polymeric network (IPN) structure have an advantage over the individual component gels and could simulate partly the structure of native extracellular matrix of cartilage tissue. In this study, to develop perfect cartilage tissue engineering scaffolds, IPN hydrogels of collagen/chondroitin sulfate/hyaluronan were prepared via two simultaneous processes of collagen self-assembly and cross linking polymerization of chondroitin sulfate-methacrylate (CSMA) and hyaluronic acid-methacrylate. The degradation rate, swelling performance and compressive modulus of IPN hydrogels could be adjusted by varying the degree of methacrylation of CSMA. The results of proliferation and fluorescence staining of rabbit articular chondrocytes in vitro culture demonstrated that the IPN hydrogels possessed good cytocompatibility. Furthermore, the IPN hydrogels could upregulate cartilage-specific gene expression and promote the chondrocytes secreting glycosaminoglycan and collagen II. These results suggested that IPN hydrogels might serve as promising hydrogel scaffolds for cartilage tissue engineering.     

A comparison study of different physical treatments on cartilage matrix derived porous scaffolds for tissue engineering applications

Native cartilage matrix derived (CMD) scaffolds from various animal and human sources have drawn attention in cartilage tissue engineering due to the demonstrable presence of bioactive components. Different chemical and physical treatments have been employed to enhance the micro-architecture of CMD scaffolds. In this study we have assessed the typical effects of physical cross-linking methods, namely ultraviolet (UV) light, dehydrothermal (DHT) treatment, and combinations of them on bovine articular CMD porous scaffolds with three different matrix concentrations (5%, 15% and 30%) to assess the relative strengths of each treatment. Our findings suggest that UV and UV–DHT treatments on 15% CMD scaffolds can yield architecturally optimal scaffolds for cartilage tissue engineering.     

Formulation of PEG-based hydrogels affects tissue-engineered cartilage construct characteristics

The limited supply of cartilage tissue with appropriate sizes and shapes needed for reconstruction and repair has stimulated research in the area of hydrogels as scaffolds for cartilage tissue engineering. In this study we demonstrate that poly(ethylene glycol) (PEG)-based semi-interpenetrating (sIPN) network hydrogels, made with a crosslinkable poly(ethylene glycol)-dimethacrylate (PEGDM) component and a non-crosslinkable interpenetration poly(ethylene oxide) (PEO) component, and seeded with chondrocytes support cartilage construct growth having nominal thicknesses of 6 mm and relatively uniform safranin-O stained matrix when cultured statically, unlike constructs grown with prefabricated macroporous scaffolds. Even though changing the molecular weight of the PEO from 100 to 20 kDa reduces the viscosity of the precursor polymer solution, we have demonstrated that it does not appear to affect the histological or biochemical characteristics of cartilaginous constructs. Extracellular matrix (ECM) accumulation and the spatial uniformity of the ECM deposited by the embedded chondrocytes decreased, and hydrogel compressive properties increased, as the ratio of the PEGDM:PEO in the hydrogel formulation increased (from 30:70 to 100:0 PEGDM:PEO). Total collagen and glycosaminoglycan contents per dry weight were highest using the 30:70 PEGDM:PEO formulation (24.4+/-3.5% and 7.1+/-0.9%, respectively). The highest equilibrium compressive modulus was obtained using the 100:0 PEGDM:PEO formulation (0.32+/-0.07 MPa), which is similar to the compressive modulus of native articular cartilage. These results suggest that the versatility of PEG-based sIPN hydrogels makes them an attractive scaffold for tissue engineering of cartilage.     

A comparative study of the chondrogenic potential between synthetic and natural scaffolds in an in vivo bioreactor

Abstract

The clinical demand for cartilage tissue engineering is potentially large for reconstruction defects resulting from congenital deformities or degenerative disease due to limited donor sites for autologous tissue and donor site morbidities. Cartilage tissue engineering has been successfully applied to the medical field: a scaffold pre-cultured with chondrocytes was used prior to implantation in an animal model. We have developed a surgical approach in which tissues are engineered by implantation with a vascular pedicle as an in vivo bioreactor in bone and adipose tissue engineering. Collagen type II, chitosan, poly(lactic-co-glycolic acid) (PLGA) and polycaprolactone (PCL) were four commonly applied scaffolds in cartilage tissue engineering. To expand the application of the same animal model in cartilage tissue engineering, these four scaffolds were selected and compared for their ability to generate cartilage with chondrocytes in the same model with an in vivo bioreactor. Gene expression and immunohistochemistry staining methods were used to evaluate the chondrogenesis and osteogenesis of specimens. The result showed that the PLGA and PCL scaffolds exhibited better chondrogenesis than chitosan and type II collagen in the in vivo bioreactor. Among these four scaffolds, the PCL scaffold presented the most significant result of chondrogenesis embedded around the vascular pedicle in the long-term culture incubation phase.     

A biomimetic three-dimensional woven composite scaffold for functional tissue engineering of cartilage

Tissue engineering seeks to repair or regenerate tissues through combinations of implanted cells, biomaterial scaffolds and biologically active molecules. The rapid restoration of tissue biomechanical function remains an important challenge, emphasizing the need to replicate structural and mechanical properties using novel scaffold designs. Here we present a microscale 3D weaving technique to generate anisotropic 3D woven structures as the basis for novel composite scaffolds that are consolidated with a chondrocyte-hydrogel mixture into cartilage tissue constructs. Composite scaffolds show mechanical properties of the same order of magnitude as values for native articular cartilage, as measured by compressive, tensile and shear testing. Moreover, our findings showed that porous composite scaffolds could be engineered with initial properties that reproduce the anisotropy, viscoelasticity and tension-compression nonlinearity of native articular cartilage. Such scaffolds uniquely combine the potential for load-bearing immediately after implantation in vivo with biological support for cell-based tissue regeneration without requiring cultivation in vitro.     

Scaffold design and manufacturing: from concept to clinic

Since Robert Langer and colleagues pioneered the concept of reconstructing tissue using cells transplanted on synthetic polymer matrices in the early 1990s, research in the field of tissue engineering and regenerative medicine has exploded. This is especially true in the development of new materials and structures that serve as scaffolds for tissue reconstruction. The basic tenet of the last two decades holds scaffolds as degradable materials providing temporary function while enhancing tissue regeneration through the delivery of biologics. Although a number of new scaffolding materials and structures have been developed in research laboratories, the application of such materials practice even has been extremely limited. This paper argues that better integration of all these factors is needed to bring scaffolds from "concept to clinic". It reviews current work in all these areas and suggests where future work and funding is needed.     

A critical review on polymer-based bio-engineered materials for scaffold development

Since the last decade, tissue engineering has shown a sensational promise in providing more viable alternatives to surgical procedures for harvested tissues, implants and prostheses. Due to the fast development on biomaterial technologies, it is now possible for doctors to use patients’ cells to repair orthopedic defects such as focal articular cartilage lesions. In order to support the three-dimensional tissue formation, scaffolds made by biocompatible and bioresorbable polymers and composite materials, for providing temporary support of damaged body and cell structures have been developed recently. Although ceramic and metallic materials have been widely accepted for the development of implants, its non-resorbability and necessity of second surgical operation, which induces extra for the patients, limit their wide applications. This review article aims at introducing (i) concept of cartilage tissue engineering, (ii) common types of bio-engineered materials and (iii) future development of biomaterial scaffolds.     

Preparation and characterization of collagen/PLA,chitosan/PLA,and collagen/chitosan/PLA hybrid scaffolds for cartilage tissue engineering

In this study, three-dimensional (3D) porous scaffolds were developed for the repair of articular cartilage defects. Novel collagen/polylactide (PLA), chitosan/PLA, and collagen/chitosan/PLA hybrid scaffolds were fabricated by combining freeze-dried natural components and synthetic PLA mesh, where the 3D PLA mesh gives mechanical strength, and the natural polymers, collagen and/or chitosan, mimic the natural cartilage tissue environment of chondrocytes. In total, eight scaffold types were studied: four hybrid structures containing collagen and/or chitosan with PLA, and four parallel plain scaffolds with only collagen and/or chitosan. The potential of these types of scaffolds for cartilage tissue engineering applications were determined by the analysis of the microstructure, water uptake, mechanical strength, and the viability and attachment of adult bovine chondrocytes to the scaffolds. The manufacturing method used was found to be applicable for the manufacturing of hybrid scaffolds with highly porous 3D structures. All the hybrid scaffolds showed a highly porous structure with open pores throughout the scaffold. Collagen was found to bind water inside the structure in all collagen-containing scaffolds better than the chitosan-containing scaffolds, and the plain collagen scaffolds had the highest water absorption. The stiffness of the scaffold was improved by the hybrid structure compared to plain scaffolds. The cell viability and attachment was good in all scaffolds, however, the collagen hybrid scaffolds showed the best penetration of cells into the scaffold. Our results show that from the studied scaffolds the collagen/PLA hybrids are the most promising scaffolds from this group for cartilage tissue engineering.     

Fabrication and cell affinity of biomimetic structured PLGA/articular cartilage ECM composite scaffold

An ideal scaffold for cartilage tissue engineering should be biomimetic in not only mechanical property and biochemical composition, but also the morphological structure. In this research, we fabricated a composite scaffold with oriented structure to mimic cartilage physiological morphology, where natural nanofibrous articular cartilage extracellular matrix (ACECM) was used to mimic the biochemical composition, and synthetic PLGA was used to enhance the mechanical strength of ACECM. The composite scaffold has well oriented structure and more than 89% of porosity as well as about 107 μm of average pore diameter. The composite scaffold was compared with ACECM and PLGA scaffolds. Cell proliferation test showed that the number of MSCs in ACECM and composite scaffolds was noticeably bigger than that in PLGA scaffold, which was coincident with results of SEM observation and cell viability staining. The water absorption of ACECM and composite scaffolds were 22.1 and 10.2 times respectively, which was much higher than that of PLGA scaffolds (3.8 times). The compressive modulus of composite scaffold in hydrous status was 1.03 MPa, which was near 10 times higher than that of hydrous ACECM scaffold. The aforementioned results suggested that the composite scaffold has the potential for application in cartilage tissue engineering.