Facile fabrication of the glutaraldehyde cross-linked collagen/chitosan porous scaffold for skin tissue engineering

Porous scaffold is one of the key factors in skin tissue engineering. In this study, a facile method was developed to prepare the glutaraldehyde (GA) cross-linked collagen/chitosan porous scaffold (S2). The properties of S2 were compared with the scaffolds prepared by the traditional method (S1). Compared to the rough surface and collapsed inner structure of S1, S2 showed a smooth surface and controlled size. After treated by GA with same concentration, S1 and S2 showed the similar swelling ratios, which are big enough to ensure the nutrient supply in the early stage of wound healing. The effects of the fabrication methods as well as the GA concentration on the cross-linking degree and in vitro degradation degree of the scaffolds were studied. It was found that the cross-linking degree of S2-0.25% was much higher than that of S1. Investigation of the tensile and compression properties of the scaffolds found that the mechanical property of S2-0.04% is closest to that of S1. High performance liquid chromatography (HPLC) was applied to determine the residual GA. The results proved that, compared to water rinse, oven drying is a feasible and effective method to remove the residual GA. Finally, the cytocompatibility of S2 was evaluated by in vitro culture of fibroblasts. The results of cell morphology and cell viability proved that S2-0.04% could retain the original good cytocompatibility of S1 to accelerate cell infiltration and proliferation effectively. All these results indicate that it is a feasible method to prepare the GA cross-linked collagen/chitosan scaffold.     

New paradigms in hierarchical porous scaffold design for tissue engineering

This paper presented an effective method for the three-dimensional (3D) hierarchical porous scaffold design for tissue engineering. To achieve such a hierarchical porous structure with accurately controlled internal pore architectures, the recursive intersection Boolean operation (RIBO) was proposed in order to satisfy computational efficiency and biological function requirements of a porous scaffold. After generating the distance field (DF) for the given anatomic model and required pore architectures, the recursive DF modifications enable us to design hierarchical porous scaffolds with complex combinations of pore morphologies. A variety of experimental results showed that the proposed hierarchical porous scaffold design method has the potential benefits for accurately controlling both the porosity and the pore architecture gradients while preserving the advantages of triply periodic minimal surface pore geometries.     

Morphology-controllable modeling approach for a porous scaffold structure in tissue engineering

Heterogeneous structures represent an important new frontier for twenty-first-century engineering. In this paper, based on the shape function in the finite element method, a morphology-controllable modelling approach for constructing tissue engineering (TE) bone scaffold with various irregular pores is presented. The modelling approach consists of both irregular element modelling and the whole bone scaffold modelling. Accepting the elements’ information after all-hex mesh generation as inputs, the basic pore-making element can be mapped into various irregular elements based on the shape function. In the bone scaffold modelling, the Boolean difference between the contour model of the solid entity and the pore model which can be constructed by the Boolean operation union would generate a porous bone scaffold model. Compared to the stochastic geometry method and the discrete element packing method, the bone scaffold model obtained in this paper has a continuous, smooth contour and various irregular pores. Moreover, a decrease in computational complexity is achieved in this paper.     

A novel surface modification on calcium polyphosphate scaffold for articular cartilage tissue engineering

The surface of porous three-dimensional (3D) calcium polyphosphate (CPP) scaffold was modified by treatment of quenching-after-sintering in the fabrication process. Scanning electron microscopic examination and degradation tests confirmed a new type of surface modification. A rotary-shaking culture was compared to that of a stationary culture and the results showed that rotary shaking led to enhanced extracellular matrices (ECM) secretion of both proteoglycans and collagen. Rotary-shaking cultured results showed that the quenching-treated CPP scaffold produced a better cartilage tissue, with both proteoglycans and collagen secretions enhanced, than the air-cooled-after-sintering scaffolds. Moreover, β-CPP scaffolds were better for the ECM secretion of both proteoglycans and collagen than the β-CPP + γ-CPP multiphase scaffold. However, the multiphase scaffold led to higher growth rate than that of β-CPP scaffold; the quenching-after-sintering treatment reversed this. In addition, the ECM secretions of both proteoglycans and collagen in the quenching-treated β-CPP scaffold were higher than those in the air-cooled one. Thus, the novel treatment of quenching-after-sintering has shown merits to the porous 3D CPP scaffolds for articular cartilage tissue engineering.     

一种新型的骨组织工程用支架材料的制备与性能表征

以硼硅酸盐玻璃粉为原料,采用有机泡沫浸渍工艺,制备了高孔隙率的网眼多孔支架.应用XRD、SEM及ICP-AES等对硼酸盐生物玻璃粉末在生理模拟液中的降解性能、生物活性等进行了测试分析.结果表明,硼硅酸盐生物玻璃的降解性和生物活性与材料的组成配比有关,因此,可以通过调整玻璃的组成有效控制材料的降解性和表面形成的羟基磷灰石晶体的形态.硼硅酸盐生物活性玻璃作为硬组织工程支架材料的研究具有重要的意义和广泛的应用前景.     

Multifunctional nanofibrous scaffold for tissue engineering

In tissue engineering, scaffolds with multiscale functionality, especially with the ability to release locally multiple or specific bioactive molecules to targeted cell types, are highly desired in regulating appropriate cell phenotypes. In this study, poly (epsilon-caprolactone) (PCL) solutions (8% w/v) containing different amounts of bovine serum albumin (BSA) with or without collagen were electrospun into nanofibres. As verified by protein release assay and fluorescent labelling, BSA and collagen were successfully incorporated into electrospun nanofibres. The biological activity of functionalised fibres was proven in the cell culture experiments using human dermal fibroblasts. By controlling the sequential deposition and fibre alignment, 3D scaffolds with spatial distribution of collagen or BSA were assembled using fluorescently labelled nanofibres. Human dermal fibroblasts showed preferential adhesion to PCL nanofibres containing collagen than PCL alone. Taken together, multiscale scaffolds with diverse functionality and tunable distribution of biomolecules across the nanofibrous scaffold can be fabricated using electrospun nanofibres.     

Porous scaffold design for tissue engineering

A paradigm shift is taking place in medicine from using synthetic implants and tissue grafts to a tissue engineering approach that uses degradable porous material scaffolds integrated with biological cells or molecules to regenerate tissues. This new paradigm requires scaffolds that balance temporary mechanical function with mass transport to aid biological delivery and tissue regeneration. Little is known quantitatively about this balance as early scaffolds were not fabricated with precise porous architecture. Recent advances in both computational topology design (CTD) and solid free-form fabrication (SFF) have made it possible to create scaffolds with controlled architecture. This paper reviews the integration of CTD with SFF to build designer tissue-engineering scaffolds. It also details the mechanical properties and tissue regeneration achieved using designer scaffolds. Finally, future directions are suggested for using designer scaffolds with in vivo experimentation to optimize tissue-engineering treatments, and coupling designer scaffolds with cell printing to create designer material/biofactor hybrids.     

Fabrication and properties of porous scaffold of zein/PCL biocomposite for bone tissue engineering

The aim of this study was to fabricate porous scaffolds of zein/poly(ε-caprolactone) (PCL) biocomposite by solvent casting–particulate leaching method using sodium chloride particles as the porogen. Porous biocomposite scaffolds with porosity around 70% and well-interconnected network were obtained. The incorporation of zein into PCL led to the improvement of hydrophilicity as indicated by the results of water contact angle measurement. After immersion in phosphate buffered saline (PBS) in vitro for 28 days, it was observed that the degradation rate of the zein/PCL biocomposite scaffold was faster than the PCL scaffold and that the rate could be tailored by adjusting the amount of zein in the composite. The results demonstrate the potential of the zein/PCL biocomposite scaffolds to be used in tissue engineering strategies to regenerate bone defects.     

溶剂自扩散法制备聚L-乳酸/β-磷酸三钙复合多孔支架材料

采用溶剂自扩散原理从聚L-乳酸(PLLA)/β-磷酸三钙(β-TCP)氯仿液中沉积得到PLLA/β-TCP复合颗粒,研究了不同扩散介质对该过程的影响.研究表明制备复合颗粒以丙酮/无水乙醇混合液为扩散介质效果最佳,以其为扩散介质沉积速率快、沉积充分,且得到的复合颗粒可以经模压成型、粒子沥滤工艺制备PLLA/β-TCP多孔复合支架.对多孔支架进行了SEM、孔隙率、力学性能及有机溶剂残留量测试,结果表明制备的多孔支架孔结构三维贯通,孔隙率60.3%,抗压强度4.40MPa,氯仿、丙酮、无水乙醇残留量分别为3.630×10-5、2.07 × 10-6、2.517×10-5,满足组织工程支架材料要求.     

A novel technique for fabrication of near-net-shape CMCs

A sol-gel vacuum infiltration technique has been developed for the fabrication of near-net-shape ceramic matrix composites (CMCs) using discontinuous mullite fibre preform with 15 vol.% of fibre content and ZrO₂.10 wt.% Y₂O₃ sol as the infiltrant. Effect of sol viscosity, number of infiltration and calcination temperature on physico-mechanical properties of fabricated CMCs were examined. Characterization of the fibre preform, matrix material (in the form of ceramic specimen without fibre) and the developed CMCs were performed by X-ray diffraction (XRD) and scanning electron microscopy (SEM). XRD indicated the presence of cubic (c) and tetragonal (t) zirconia in both the CMCs and the ceramic specimens calcined even at 1400°C. Flexural strength of the CMCs and the ceramic specimens (calcined at 1400°C), determined by the three-point bending test, was found to be about 14 mPa and 1.40 mPa, respectively. SEM indicated multiple fracture of the matrix which gave rise to pseudo elasticity. This is also evident from the load-displacement curve of the three-point bend test. SEM studies also indicated fibre pull-out in the fracture surface of the CMCs.     

Degradation and cytotoxicity of lotus-type porous pure magnesium as potential tissue engineering scaffold material

The lotus-type porous pure magnesium was prepared using a metal/gas eutectic unidirectional solidification method (GASAR process). The corrosion behavior, decay of mechanical property and the cytocompatibility were evaluated with the compact pure Mg as control. The porous pure Mg indicates better corrosion resistance than that of compact pure Mg in SBF at 37 °C. The compressive yield strength of compact and porous pure Mg is (110.3 ± 8.5) MPa and (23.9 ± 4.9) MPa before immersion test, and porous pure Mg exhibits slower decay in compressive yield strength with the extension of immersion period than that of compact pure Mg. With larger exposed surface area, porous pure Mg shows higher Mg concentration in the extract than that of compact pure Mg, which leads to a higher osmotic pressure to cells and might affect its indirect cytotoxicity assay result, but is still within the Grade I RGR value (no toxicity), implying the feasibility as potential tissue engineering scaffold.     

Hierarchical porous materials for tissue engineering

Biological organisms have evolved to produce hierarchical three-dimensional structures with dimensions ranging from nanometres to metres. Replicating these complex living hierarchical structures for the purpose of repair or replacement of degenerating tissues is one of the great challenges of chemistry, physics, biology and materials science. This paper describes how the use of hierarchical porous materials in tissue engineering applications has the potential to shift treatments from tissue replacement to tissue regeneration. The criteria that a porous material must fulfil to be considered ideal for bone tissue engineering applications are listed. Bioactive glass foam scaffolds have the potential to fulfil all the criteria, as they have a hierarchical porous structure similar to that of trabecular bone, they can bond to bone and soft tissue and they release silicon and calcium ions that have been found to up-regulate seven families of genes in osteogenic cells. Their hierarchical structure can be tailored for the required rate of tissue bonding, resorption and delivery of dissolution products. This paper describes how the structure and properties of the scaffolds are being optimized with respect to cell response and that tissue culture techniques must be optimized to enable growth of new bone in vitro.     

Designing porous scaffolds for tissue engineering

Biomaterials are either modified natural or synthetic materials, with an appropriate response in the host tissue, which find application in a wide spectrum of implants and prostheses used in reconstructive medicine. The subsequent integration and longevity of the implanted device depends on the effectiveness of the associated biological repair. Hence, there has been considerable interest in the development of novel, second generation, biomaterials, which are favourably bioactive in terms of promoting the desired cellular response in vivo. Such biomaterials in a porous form can also act as cellular scaffolds and allow in vitro, as well as in vivo incorporation of the appropriate tissue cells, with potential control of the sequence of cell attachment, proliferation and the production of extra-cellular matrix. Such generic tissue engineering depends critically on the porous architecture of the biomaterial scaffold so as to allow both the cellular ingress and vascularization required to create a living tissue. The particular requirements of tissue-engineering scaffolds with respect to macro- and micro-porosity, as well as chemistry, are reviewed.     

骨组织工程用纳米羟基磷灰石/ 壳聚糖多孔支架材料的制备及性能表征

以16.7%(质量分数)的柠檬酸水溶液作溶剂,通过粒子沥滤法制备了 n HA/CS多孔材料,并对其进行了IR、XRD、SEM、孔隙率及力学性能测试。结果表明n HA/CS复合材料中羟基磷灰石呈弱结晶状态,复合前后两组分的化学组成未发生显著变化,但两相间发生了相互作用。多孔材料呈高度多孔结构,孔壁上富含微孔,孔间贯通性高;复合材料/致孔剂质量比为1时,多孔材料的孔隙率为 53%,其抗压强度可达17 MPa左右,可以满足组织工程支架材料的要求。     

Chitosan-collagen/organomontmorillonite scaffold for bone tissue engineering

A novel composite scaffold based on chitosan-collagen/organomontmo-rillonite (CS-COL/OMMT) was prepared to improve swelling ratio, biodegradation ratio, biomineralization and mechanical properties for use in tissue engineering applications. In order to expend the basal spacing, montmorillonite (MMT) was modified with sodium dodecyl sulfate (SDS) and was characterized by XRD, TGA and FTIR. The results indicated that the anionic surfactants entered into interlayer of MMT and the basal spacing of MMT was expanded to 3.85 nm. The prepared composite scaffolds were characterized by FTIR, XRD and SEM. The swelling ratio, biodegradation ratio and mechanical properties of composite scaffolds were also studied. The results demonstrated that the scaffold decreased swelling ratio, degradation ratio and improved mechanical and biomineralization properties because of OMMT.     

Ultrasonic monitoring of foamed polymeric tissue scaffold fabrication

Polymeric tissue scaffolds are central to many regenerative medicine therapies offering a new approach to medicine. As the number of these regenerative therapies increases there is a pressing need for an improved understanding of the methods of scaffold fabrication. Of the many approaches to processing scaffolds, supercritical fluid fabrication methods have a distinct advantage over other techniques as they do not require the use of organic solvents, elevated processing temperatures or leaching processes. The work presented here is centred on the development of a new approach to monitoring supercritical scaffold fabrication based on determination of the scaffold acoustic impedance to inform protocols for scaffold fabrication. The approach taken uses an ultrasonic pulse-echo reflectometer enabling non-invasive monitoring of the supercritical environment on-line. The feasibility of this approach was investigated for two scaffolds of different molecular weight. Acoustic results demonstrate that differences in the physical properties of the two scaffolds could be resolved, particularly during the foaming process which correlated with findings from time-lapsed imaging and micro X-ray computed tomography (micro X-ray CT) images. Thus, this work demonstrates the feasibility of ultrasonic pulse-echo reflectometry to non-invasively study supercritical scaffold fabrication on-line providing a greater understanding of the scaffold fabrication process.     

A smart bilayer scaffold of elastin-like recombinamer and collagen for soft tissue engineering

Elastin-like recombinamers (ELRs) are smart, protein-based polymers designed with desired peptide sequences using recombinant DNA technology. The aim of the present study was to produce improved tissue engineering scaffolds from collagen and an elastin-like protein tailored to contain the cell adhesion peptide RGD, and to investigate the structural and mechanical capacities of the resulting scaffolds (foams, fibers and foam-fiber bilayer scaffolds). The results of the scanning electron microscopy, mercury porosimetry and mechanical testing indicated that incorporation of ELR into the scaffolds improved the uniformity and continuity of the pore network, decreased the pore size (from 200 to 20 μm) and the fiber diameter (from 1.179 μm to 306 nm), broadened the pore size distribution (from 70–200 to 4–200 μm) and increased their flexibility (from 0.007 to 0.011 kPa⁻¹). Culture of human fibroblasts and epithelial cells in ELR-collagen scaffolds showed the positive contribution of ELR on proliferation of both types of cells.     

一种制备纳米羟基磷灰石/聚乙烯醇多孔支架材料的新方法

采用乳化发泡-冷冻法制备了聚乙烯醇和纳米羟基磷灰石/聚乙烯醇两种多孔材料.用SEM对材料进行了表征,结果表明所得样品孔径均匀,孔隙率高,且孔内外贯通,是一种新型的制备多孔材料的方法.文中还讨论了乳化剂op、聚乙烯醇、纳米羟基磷灰石、搅拌速度等因素对孔的大小和孔隙率的影响.     

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.     

Preparation of a biphasic scaffold for osteochondral tissue engineering

Tissue engineering has been developed as a prospective approach for the repair of articular cartilage defects. Engineered osteochondral implants can facilitate the fixation and integration with host tissue, and therefore promote the regeneration of osteochondral defects. A biphasic scaffold with a stratified two-layer structure for osteochondral tissue engineering was developed from biodegradable synthetic and naturally derived polymers. The upper layer of the scaffold for cartilage engineering was collagen sponge; the lower layer for bone engineering was a composite sponge of poly(DL-lactic-co-glycolic acid) (PLGA) and naturally derived collagen. The PLGA–collagen composite sponge layer had a composite structure with collagen microsponge formed in the pores of a skeleton PLGA sponge. The collagen sponge in the two respective layers was connected. Observation of the collagen/PLGA–collagen biphasic scaffold by scanning electron microscopy (SEM) demonstrated the connected stratified structure. The biphasic scaffold was used for culture of canine bone-marrow-derived mesenchymal stem cells. The cell/scaffold construct was implanted in an osteochondral defect in the knee of a one-year old beagle. Osteochondral tissue was regenerated four months after implantation. Cartilage- and bone-like tissues were formed in the respective layers. The collagen/PLGA–collagen biphasic scaffold will be useful for osteochondral tissue engineering.