Structures Of Scaffolds Research Articles

Production of ethyl cellulose scaffolds by supercritical co2 phase separation

Ethyl Cellulose scaffolds were prepared using a supercritical fluid phase inversion process in which carbon dioxide acts as the non-solvent. Ethyl Cellulose is a derivative of cellulose in which some of the hydroxyl groups on the repeating glucose units are converted into ethyl ether groups; it is a biocompatible but non-biodegradable polymer. It is an extensively studied encapsulating material for the controlled release of pharmaceuticals in tissue engineering applications. Series of experiments were performed at various polymer concentrations, temperatures, pressures, and kind of organic solvent. The structures of the resulting scaffolds were analysed using scanning electron microscopy. We operated with polymer concentrations ranging between 5 and 20 % w/w, using N-methyl pyrrolidone and chloroform as organic solvents and we obtained scaffolds characterized by homogeneous cellular structures with different pore mean size. The mean diameter of the pores ranged from 3 μm to 15 μm, decreasing the polymer concentration from 20 to 5 % w/w. Additional experiments were performed varying the operating pressure from 100 and 200 bar, and varying the operating temperature from 35 to 55 °C. Increasing pressure and/or decreasing temperature, the scaffolds pore size largely decreases; both the effects can be related to carbon dioxide density modification. Ethyl Cellulose scaffolds formation mechanisms were also discussed.

Ultrafine fibrous gelatin scaffolds with deep cell infiltration mimicking 3D ECMs for soft tissue repair.

In this research, ultrafine fibrous scaffolds with deep cell infiltration and sufficient water stability have been developed from gelatin, aiming to mimic the extracellular matrices (ECMs) as three dimensional (3D) stromas for soft tissue repair. The ultrafine fibrous scaffolds produced from the current technologies of electrospinning and phase separation are either lack of 3D oriented fibrous structure or too compact to be penetrated by cells. Whilst electrospun scaffolds are able to emulate two dimensional (2D) ECMs, they cannot mimic the 3D ECM stroma. In this work, ultralow concentration phase separation (ULCPS) has been developed to fabricate gelatin scaffolds with 3D randomly oriented ultrafine fibers and loose structures. Besides, a non-toxic citric acid crosslinking system has been established for the ULCPS method. This system could endow the scaffolds with sufficient water stability, while maintain the fibrous structures of scaffolds. Comparing with electrospun scaffolds, the ULCPS scaffolds showed improved cytocompatibility and more importantly, cell infiltration. This research has proved the possibility of using gelatin ULCPS scaffolds as the substitutes of 3D ECMs.

Effects of Forming Temperature on Structure and Chemical Properties of Calcium Phosphate Cement

With similar components to human bone, hydroxyapatite (HAp) has been commonly used in dental and orthopedics practice. Calcium phosphate cement (CPC) can be made into porous HAp scaffolds through reaction; therefore, this study aims to examine how reaction temperatures influence CPC. CPC powder and NaCl are blended, and after diammonium phosphate (DAP) is added, they are infused into a mold for a hydration reaction under a saturated vapor pressure to form CPC scaffolds. XRD and FTIR are used to determine the influence of reaction temperatures on crystallization and functional groups. A scanning electron microscope (SEM) observes the structures of the CPC scaffolds. Test results show that when the temperature of a saturated vapor pressure is 70 °C, the CPC scaffolds have significant crystalline conversion and functional group peaks. SEM observation indicates that the temperature of a saturated vapor pressure does not affect the formation of the CPC scaffolds structure.

Nanoscale Control of Silks for Nanofibrous Scaffold Formation with Improved Porous Structure.

Silk-based porous scaffolds have been used extensively in tissue engineering because of their excellent biocompatibility, tunable biodegradability and robust mechanical properties. Although many silk-based scaffolds have been prepared through freeze-drying, a challenge remains to effectively control porous structures during this process. In the present study silk fibroin with different nanostructures were self-assembled in aqueous solution by repeated drying-dissolving process and then used to improve porous structure formation in lyophilization process. Viscosity, secondary structures and water interactions were also studied to exclude their influence on the formation and control of porous structures. Following nanofiber formation in aqueous solution, silk scaffolds with improved porous structure were directly formed after lyophilization and then stabilized with water or methanol annealing treatments. Compared to silk scaffolds derived from fresh solution, the nanofibrous scaffolds showed significantly better cell compatibility in vitro. Therefore, this nanoscale control of silk offers feasible way to regulate the matrix features including porous structure and nanostructure, which are important in regulating cell and tissue outcomes in tissue engineering and regeneration, and then achieve silk-based scaffolds with improved properties.

Improving the Gene-Regulation Ability of Small RNAs by Scaffold Engineering in Escherichia coli

Noncoding small RNAs are involved in transcriptional and post-transcriptional gene regulation of target mRNAs by modulating mRNA elongation, stability, or translational efficiency. Many natural trans-encoded small RNAs contain a scaffold that allows binding of the RNA chaperone protein Hfq for conditional gene regulation. Here, we improved the gene regulation abilities of small RNAs by directly fusing the natural Escherichia coli trans-encoded small RNA-derived scaffolds, including Hfq-binding and rho-independent transcription terminator sequences, to the 3' end of the small RNAs that mediate RNA-based gene regulation. As target small RNAs to improve their gene regulation abilities, we selected small RNAs of artificial post-transcriptional riboregulators and transcriptional attenuators. Four different small RNA scaffolds were fused to the riboregulator and attenuator-derived small RNAs. Mutations were introduced into the best small RNA scaffold to improve its gene-regulation ability further. As a result, mutations predicted to stabilize the secondary structures of the small RNA scaffolds dramatically increased its ability to regulate gene expression of both the post-transcriptional riboregulator and transcriptional attenuator systems. We believe our engineered small RNA scaffolds are applicable to other RNA regulators for improving regulatory activity, and engineered small RNA scaffolds may present a valuable strategy to regulate target gene expression strongly.

Comparative studies on ectopic bone formation in porous hydroxyapatite scaffolds with complementary pore structures

Vascularized bone grafts were constructed by implanting hydroxyapatite (HA) scaffolds with complementary macro-pore structures into the dorsal muscle of dogs. The relationship between pore structures and ectopic bone formation properties was investigated. Two types of scaffolds with complementary porous structures were fabricated by spherulite-accumulating and porogen-preparing methods, and were named spherulite HA-positive and porogen HA-negative, respectively. After implantation for 1month, histological observation showed that all the scaffolds were encapsulated by normal muscle tissue and multiple vascular net with cells, indicating excellent biocompatibility and pore interconnectivity of the scaffolds. In the spherulite HA-positive scaffolds, a number of osteoclasts and osteoblasts coupled with new bone tissues were found after 3 and 6months’ implantations, which was better than those in the porogen HA-negative scaffolds. Similarly, the improvement of mechanical properties and the reconstruction of materials in the spherulite HA-positive scaffolds were superior to those in the porogen HA-negative scaffolds. The different ectopic bone formation induced by different macro-pore structures after intramuscular implantation demonstrated the significant effect of macro-pore structures of scaffolds on osteoinduction and vascularization.

Well-ordered biphasic calcium phosphate-alginate scaffolds fabricated by multi-channel 3D plotting under mild conditions.

Herein, we present a new type of biphasic organic-inorganic scaffold, which can be fabricated by multi-channel 3D plotting under mild conditions based on a highly concentrated alginate paste and a ready-to-use calcium phosphate cement (CPC) for bone and osteochondral tissue engineering. The structures of scaffolds were characterised by light and scanning electron microscopy (SEM). Results indicated that the concentrated alginate and CPC pastes had comparable plotting consistency, and therefore could be combined in one (biphasic) scaffold by applying predesigned plotting parameters. After crosslinking of alginate and setting of CPC, the biphasic scaffold obtained mechanical and structural stability. Mechanical test data revealed that biphasic CPC-alginate scaffolds had significantly increased compressive strength and modulus compared to pure alginate as well as mixed calcium phosphate (CaP)-alginate scaffolds in a wet state and improved strength and toughness compared to pure CPC scaffolds in both dry and wet conditions. Culture of human mesenchymal stem cells (hMSCs) on these scaffolds over 3 weeks demonstrated the good cytocompatibility of the selected materials. Because of the mild preparation conditions, bovine serum albumin (BSA) as a model protein was loaded in alginate and CPC pastes prior to plotting with high loading efficiency. Release studies in vitro showed that BSA released much faster from alginate strands than from CPC strands, which might allow amount-controlled protein release from biphasic CPC-alginate scaffolds. Furthermore, an upgraded bipartite osteochondral scaffold consisting of an alginate part for chondral and a biphasic CPC-alginate part for bony repair was fabricated based on this technique. This scaffold showed a strong organic-inorganic interface binding due to interlocking and crosslinking of the alginate strands.

Cross-linked Electrospun Fibrous Scaffolds for Tissue Engineering

The structures of scaffolds for tissue engineering have significant effects on the seeding, migration, and proliferation of the tissue cells. Fibrous scaffolds consisting of microfibers or nanofibers have attracted increasing attention and applications in tissue engineering because of their high porosity and surface area and their ability to direct cell growth and alignment. However, water-soluble and hydrophilic polymeric microfibers and nanofibers lose their fibrous morphology in aqueous conditions such as body fluid and culture media. Cross-linking has been proven an effective method for stabilizing fibrous structure and improving mechanical properties of the fibrous scaffolds. In this article, we will review the current technologies for fabricating cross-linked fibrous scaffolds by the combination of electrospinning and different cross-linking methods including chemical, physical and photo-cross-linking. Their effects on solvent resistance, biocompatibility, mechanical properties, and cell viability will be discussed. Chemical cross-linking with nontoxic cross-linkers such as genipin and the reactive electrospinning with in-situ photo-cross-linking are promising new technologies to fabricate novel nanofibrous scaffolds with enhanced solvent resistance and mechanical strength as well as tunable biodegradation property. Keywords: Electrospinning, nanofibers, chemical cross-linking agent, physical cross-linking, photo-cross-linking, solvent resistance, biocompatibility, mechanical properties, hydrophilic, biochemistry.

Synthesis and evaluation of in vitro bioactivity for vesicular acetylcholine transporter inhibitors containing two carbonyl groups

To identify selective high-affinity ligands for the vesicular acetylcholine transporter (VAChT), we have incorporated a carbonyl group into the structures of trozamicol and prezamicol scaffolds, and also converted the secondary amines of the piperidines of trozamicols and prezamicols into amides. Of 18 new racemic compounds, 4 compounds displayed high affinity for VAChT (Ki=10–20nM) and greater than 300-fold selectivity for VAChT over σ1 and σ2 receptors, namely (4-(4-fluorobenzoyl)-4′-hydroxy-[1,3′-bipiperidin]-1′-yl)(3-methylthiophen-2-yl)methanone oxalate (9g) (Ki-VAChT=11.4nM, VAChT/σ1=1063, VAChT/σ2=370), (1′-benzoyl-4′-hydroxy-[1,3′-bipiperidin]-4-yl)(4-methoxyphenyl)methanone oxalate (10c) (Ki-VAChT=15.4nM, VAChT/σ1=374, VAChT/σ2=315), (4′-hydroxy-1′-(thiophene-2-carbonyl)-[1,3′-bipiperidin]-4-yl)(4-methoxyphenyl)methanone oxalate (10e) (Ki-VAChT=19.0nM, VAChT/σ1=1787, VAChT/σ2=335), and (4′-hydroxy-1′-(3-methylthiophene-2-carbonyl)-[1,3′-bipiperidin]-4-yl)(4-methoxyphenyl)methanone oxalate (10g) (Ki-VAChT=10.2nM, VAChT/σ1=1500, VAChT/σ2=2030). These four compounds can be radiosynthesized with C-11 or F-18 to validate their possibilities of serving as PET probes for quantifying the levels of VAChT in vivo.

Micro-patterned atmospheric pressure plasma‐deposited cell non-adhesive coatings

Low temperature atmospheric pressure plasma deposited thin films in micro patterning technique for cell culturing represent a new field of plasma surface engineering for biological applications. It is necessary because of a trend toward the miniaturization of the detailed structures of tissue culture scaffolds. Plasma deposition at atmospheric pressure allows the cell to be micro-patterned onto a cell-adhesive surface. The adhesion-resistance of the thin films deposited at atmospheric pressure was tested with 3T3 mouse fibroblasts. The 3T3 mouse fibroblasts were successfully confined into circular, 100μm2 cell-adhesive areas that were patterned inside a cell-resistant layer. This atmospheric pressure plasma technique can be used to coat heat-sensitive bio-polymeric materials. The experimentally measured gas phase temperature was approximately 30°C to 90°C, which indicates that this atmospheric pressure plasma can treat polymers without unfavorable thermal effects. The simplicity of plasma process at atmospheric pressure and the versatility of the plasma deposited thin films render this technique highly suitable for the fabrication of biomedical and biological devices, where patterned, high contrast surfaces that are resistant to cell-adhesion are needed.

Preparation of <i>Antheraea pernyi</i> Silk Fibroin/Chitosan Blend Scaffolds

In this work, Antheraea pernyi silk fibroin (ASF) /chitosan (CS) blend scaffolds were prepared by freeze-drying. The relationship between preparation conditions and morphological structures of blend scaffolds was also studied. The results indicated that the porosity and pore diameter of the ASF /CS scaffolds decreased with increasing of the proportion of chitosan and decreasing of the freezing temperature. By changing the process conditions, the average pore diameter and porosity of ASF/CS scaffolds could be adjusted in the range of 134 - 527 μm and 71 - 91% respectively. Compared with the pure ASF or CS materials, the ASF / CS (60/40) blend materials were more favor of cell adhesion.

Modulation of immunological properties of allogeneic mesenchymal stem cells by collagen scaffolds in cartilage tissue engineering

Influence of the structures of some collagen scaffolds on immunological properties of the seeded allogeneic mesenchymal stem cells (MSCs) was studied in this article. Hydrogels, sponge, and membrane were prepared from type-I collagen. These scaffolds were seeded with neonatal rabbit MSCs and cultured for different periods. Changes of the immunological properties associated with different scaffolds were analyzed and compared. It was found that the expression of major histocompatibility complex (MHC) class I and II molecules on MSCs increased gradually in all scaffolds, but the least increment was recorded in hydrogels. Mixed lymphocyte reactions (MLR) showed that the MSC-hydrogel constructs invoked considerably low allogeneic lymphocytes proliferation. Even in presence of interferon-γ (IFN-γ), the hydrogels with higher concentration gave comparatively lower increment of MHC-II expression and allogeneic lymphocytes proliferation. These results suggest that different scaffold structures may provide different microenvironments and extents of isolation from the host immune system for the seed cells, thereby affecting their immunological properties. Therefore, scaffold structures may modulate the immunological properties of tissue-engineered cartilage with allogeneic cells. Hydrogels, especially which were prepared from higher collagen concentrations, were found to be a promising scaffold structure, from the perspective of avoiding severe immune rejection.

Engineering porous scaffolds using gas-based techniques

Scaffolds are used in tissue engineering as a matrix for the seeding and attachment of human cells. The creation of porosity in three-dimensional (3D) structures of scaffolds plays a critical role in cell proliferation, migration, and differentiation into the specific tissue while secreting extracellular matrix components. These pores are used to transfer nutrients and oxygen and remove wastes produced from the cells. The lack of oxygen and nutrient supply impedes the cell migration more than 500μm from the surface. The physical properties of scaffolds such as porosity and pore interconnectivity can improve mass transfer and have a great impact on the cell adhesion and penetration into the scaffolds to form a new tissue. Various techniques such as electrospinning, freeze-drying, and solvent casting/salt leaching have been used to create porosity in scaffolds. The major issues in these methods include lack of 3D structure, control on pore size, and pore interconnectivity. In this review, we provide a brief overview of gas-based techniques that have been developed for creating porosity in scaffolds.

Porous hydroxyapatite ceramics by ice templating: Freezing characteristics and mechanical properties

Ice templating produces porous hydroxyapatite (HA) scaffolds with a lamellar morphology and aligned channels when using aqueous HA slurries. We investigated the freezing characteristics of HA slurries with regard to the pore structures of the porous HA scaffolds. We found that by increasing the cooling rate, the lamellar spacing decreased. The average lamellar spacing is about 785.7 μm at a cooling rate of 1.3 °C/min. The porous geometry changes from lamella and well aligned channels to a partial dendrite and partially aligned cavities with a decrease in the initial nucleation temperature and an increase in the degree of supercooling. Additionally, we determined the relationship between compressive strength and porosity. The compressive strength of the porous HA scaffolds reach 6.7 MPa at a porosity of 64% and the lamellar spacing is about 124 μm.

Cryogenic prototyping of chitosan scaffolds with controlled micro and macro architecture and their effect on in vivo neo‐vascularization and cellular infiltration

A major challenge in tissue engineering has been to develop scaffolds with controlled complex geometries, on both the macro- and micro-scale. One group of techniques, using rapid prototyping (RP) processes, has the capability to produce complex three-dimensional structures with good control over the size, geometry, and connectivity of the pores. In this article, a novel technique based on RP technology, that is, cryogenic prototyping (CP), that has the capability to fabricate scaffolds with controlled macro- and micro-structures, is presented. Our in vivo studies showed that the micro architecture (i.e., both pore size and pore orientation) and macro structures of the CP scaffolds affect both cellular infiltration and neo-vascularization. Full cellular infiltration and neo-vascularization were observed after 28 days in scaffolds with micropore sizes of 90 microm. In addition, it was observed that channels (300 microm) created in scaffolds were effective at enhancing cellular infiltration and vascularization. Our results have demonstrated that CP is a viable method for fabricating scaffolds for a wide range of tissue engineering applications.

High-Resolution 1.5-Tesla Magnetic Resonance Imaging for Tissue-Engineered Constructs: A Noninvasive Tool to Assess Three-Dimensional Scaffold Architecture and Cell Seeding

Tissue-engineered scaffolds are made of biocompatible polymers with various structures, allowing cell seeding, growth, and differentiation. Noninvasive imaging methods are needed to study tissue-engineered constructs before and after implantation. Here, we show that high-resolution magnetic resonance imaging (MRI) performed on a clinical 1.5-T device is a reliable technique to assess three-dimensional structures of porous scaffolds and to validate cell-seeding procedures. A high-temperature superconducting detection coil was used to achieve a resolution of 30 x 30 x 30 microm(3) when imaging the scaffolds. Three types of structures with tuneable architectures were prepared from naturally derived polysaccharides and evaluated as scaffolds for mesenchymal stem cell (MSC) culture. To monitor cell seeding, MSCs were magnetically labeled using simple incubation with anionic citrate-coated iron-oxide nanoparticles for 30 min. Iron uptake was quantified using single-cell magnetophoresis, and cell proliferation was checked for 7 days after labeling. Three-dimensional (3D) microstructures of scaffolds were assessed using MRI, revealing lamellar or globular porous organization according to the scaffold preparation process. MSCs with different iron load (5, 12 and 31 pg of iron per cell) were seeded on scaffolds at low density (132 cells/mm(3)) and detected on 3D gradient-echo MR images according to phase distortions and areas of intensely low signal, whose size increased with cell iron load and echo time. Overall signal loss in the scaffold correlated with the number of seeded cells and their iron load. Different organizations of cells were observed depending on the scaffold architecture. After subcutaneous implantation in mice, scaffolds seeded with labeled cells could be distinguished in vivo from scaffold with nonlabeled cells by observation of signal and phase heterogeneities and by measuring the global signal loss. High-resolution 1.5-T MRI combined with efficient intracellular contrast agents shows promise for noninvasive 3D visualization of tissue-engineered constructs before and after in vivo implantation.

Physicochemical properties and cytotoxicities of Sr-containing biphasic calcium phosphate bone scaffolds

This study demonstrates a new biomaterial system composed of Sr-containing hydroxyapatite (Sr-HA) and Sr-containing tricalcium phosphate (Sr-TCP), termed herein Sr-containing biphasic calcium phosphate (Sr-BCP). Furthermore, a series of new Sr-BCP porous scaffolds with tunable structure and properties has also been developed. These Sr-BCP scaffolds were obtained by in situ sintering of a series of composites formed by casting various Sr-containing calcium phosphate cement (Sr-CPC) into different rapid prototyping (RP) porous phenol formaldehyde resins, which acted as the negative moulds for controlling pore structures of the final scaffolds. Results show that the porous Sr-BCP scaffolds are composed of Sr-HA and Sr-TCP. The phase composition and the macro-structure of the Sr-BCP scaffold could be adjusted by controlling the processing parameters of the Sr-CPC pastes and the structure parameters of the RP negative mould, respectively. It is also found that both the compressive strength (CS) and the dissolving rate of the Sr-BCP scaffold significantly vary with their phase composition and macropore percentage. In particular, the compressive strength achieves a maximum CS level of 9.20 +/- 1.30 MPa for the Sr-BCP scaffold with a Sr-HA/Sr-TCP weight ratio of 78:22, a macropore percentage of 30% (400-550 microm in size) and a total-porosity of 63.70%, significantly higher than that of the Sr-free BCP scaffold with similar porosity. All the extracts of the Sr-BCP scaffold exhibit no cytotoxicity. The current study shows that the incorporation of Sr plays an important role in positively improving the physicochemical properties of the BCP scaffold without introducing obvious cytotoxicity. It also reveals a potential clinical application for this material system as bone tissue engineering (BTE) scaffold.

Fabrication and Characterization of Waterborne Biodegradable Polyurethanes 3-Dimensional Porous Scaffolds for Vascular Tissue Engineering

In this study, a series of 3-D interconnected porous scaffolds with various pore diameters and porosities was fabricated by freeze-drying with non-toxic biodegradable waterborne polyurethane (WBPU) emulsions of different concentration. The structures of these porous scaffolds were characterized by scanning electron microscopy (SEM), and the pore diameters were calculated using CIAS 3.0 software. The pores obtained were 3-D interconnected in the scaffolds. The scaffolds obtained at different pre-freeze temperatures showed a pore diameter ranging from 2.8 to 99.9 μm with a pre-freezing temperature of −60°C and from 13.1 to 229.1 μm with a pre-freezing temperature of −25°C. The scaffolds fabricated with WBPU emulsions of different concentration at the same pre-freezing temperature (−25°C) had pores with mean pore diameter between 90.8 and 39.6 μm and porosity between 92.0 and 80.0%, depending on the emulsion concentration. The effect of porous structure of the scaffolds on adhesion and proliferation of human umbilical vein endothelial cells (HUVECs) cultured in vitro was evaluated using the MTT assay and environmental scanning electron microscopy (ESEM). It was found that the better adhesion and proliferation of HUVECs on 3-D scaffolds of WBPU with relative smaller pore diameter and lower porosity than those on scaffolds with larger pore and higher porosity and film. Our work suggests that fabricating a scaffold with controllable pore diameter and porosity could be a good method to be used in tissue-engineering applications to obtain carriers for cell culture in vitro.

The influence of structural design of PLGA/collagen hybrid scaffolds in cartilage tissue engineering

3D biodegradable porous scaffold plays a very important role in articular cartilage tissue engineering. We developed hybrid structures of 3D scaffolds that combined the advantages of natural type I collagen and synthetic PLGA knitted mesh. The mechanically strong PLGA mesh served as a skeleton while the collagen microsponges facilitated cell seeding and tissue formation. The scaffolds were divided into 3 groups: (1) THIN: collagen microsponge formed in interstices of PLGA mesh; (2) SEMI: collagen microsponge formed on one side of PLGA mesh; (3) SANDWICH: collagen sponge formed on both sides of PLGA mesh. Bovine chondrocytes were cultured in these scaffolds and transplanted subcutaneously into nude mice for 2, 4, and 8 weeks. All three groups of transplants showed homogeneous cell distribution, natural chondrocyte morphology, and abundant cartilaginous ECM deposition. Production of GAGs per DNA and the expression of type II collagen and aggrecan mRNA were much higher in the SEMI and SANDWICH groups than in the THIN group. When compared to native articular cartilage, the mechanical strength of the engineered cartilage reached 54.8%, 49.3% in Young's modulus and 68.8%, 62.7% in stiffness, respectively, in SEMI and SANDWICH. These scaffolds could be used for the tissue engineering of articular cartilage with adjustable thickness. The design of the hybrid structures provides a strategy for the preparation of 3D porous scaffolds.

Tomography Observations of Osteoblast Seeding on 3-D Collagen Scaffold by Synchrotron Radiation Hard X-Ray

In recent years, research into three-dimensional (3-D) scaffolds for tissue engineering has become an important topic. To avoid biological rejection, there are many biocompatible materials being developed for in vitro cultivation of autologous cells for implantation into the human body. The 3-D structures of scaffolds are not easily observed using traditional optical or electron microscopes, which only allow examination of the surfaces of samples or ultra thin films, and often require complicated pre-treating processes. The high-brilliance synchrotron radiation (SR) hard X-ray is an emerging technique to acquire tomography. This study involves the successful reconstruction of 3-D images and the animation of the structure of a collagen scaffold. The images were obtained using synchrotron radiation hard X-ray images, which eliminates the need for embedding and microtoming. SR hard X-ray imaging is a non-destructive technique, which has demonstrated a high spatial resolution and transmission, enabling the morphology of osteoblasts adhered on the inner surfaces of the collagen scaffolds to be captured. It is a convenient facility to obtain 3-D tomography images for biomaterial studies. This is the first study to observe the 3-D structures of collagen scaffolds and aquire detailed images of osteoblast attachment on the scaffold, by utilization of third-generation SR hard X-ray radiography.