[show abstract][hide abstract] ABSTRACT: This study was conducted to investigate the effects of an anodized titanium surface with RGD-peptides. Silane was used to immobilize the RGD peptides to the titanium surfaces. The surfaces were examined by scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS). Crystal violet assay, XTT assay and Alkaline phosphatase (ALP) assay were performed to evaluate response of human mesenchymal stem cell. For in vivo study, twenty-four implants and four beagle dogs were used and made the dehiscence defect model (n=12 in each group). Implant stability quotient (ISQ) value was measured and evaluated by histometric analysis. Histological examination revealed an increase of bone height in the test group. There were significant differences in intra-thread bone density (ITBD) between the groups. The application of RGD peptide to the implant surface may contribute to the improvement of osseointegration quality and bone growth in a small defect area around the implant.
Fetal ovine model for in-situ esophagus tissue engineering. 08/2013; 9(4). · 3.16 Impact Factor
[show abstract][hide abstract] ABSTRACT: Highly biocompatible polycaprolactone (PCL)/poly(lactic-co-glycolic acid) (PLGA)/collagen scaffolds in which the PCL/PLGA collagen solution was selectively dispensed into every other space between the struts were fabricated using solid freeform fabrication (SFF) technology, as we described previously. The objective of this study was to evaluate and compare the PCL/PLGA/collagen scaffolds (group 3) with PCL/PLGA-only scaffolds (group 1) and PCL/PLGA scaffolds with collagen by the dip-coating method (group 2) using human adipose-derived stem cells (hASCs) and rat primary hepatocytes. The selectively dispensed collagen formed a three-dimensional (3D) network of nanofibers in group 3, as observed by scanning electron microscopy. The compressive strength and modulus of group 3 were approximately 140 and 510 times higher, respectively, than those of a sponge-type collagen scaffold whose weak mechanical properties were regarded as a critical drawback. Proliferation and osteogenic differentiation of hASCs were promoted significantly in group 3 compared to groups 1 and 2. In addition, we found that the viability and albumin secretion ability of rat primary hepatocytes were highly retained for 10 days in group 3 but not group 1. Interestingly, hepatocyte aggregation, which enhances hepatic function through cell-cell interactions, was observed particularly in group 3. In conclusion, group 3, in which the collagen was selectively dispensed in the 3D space of the porous PCL/PLGA framework, will be a promising 3D scaffold for culturing various cell types.
Journal of Materials Science Materials in Medicine 02/2013; · 2.14 Impact Factor
[show abstract][hide abstract] ABSTRACT: The purpose of this study was to investigate the healing capacity within an 8-mm rabbit calvarial defect using a polycaprolactone (PCL)/poly(lactic-co-glycolic acid) (PLGA) scaffold blended with tri-calcium phosphate (TCP) that was constructed using solid freeform fabrication (SFF) technology. The PCL/PLGA/TCP scaffold showed a 37 % higher compressive strength and rougher surface than the PCL/PLGA scaffold. In animal experiments, new bone formation was analyzed using microcomputed tomography (micro-CT) and histological and histometric analyses. The PCL/PLGA/TCP groups had significantly greater neo-tissue areas as compared with the control groups at 4 and 8 weeks (P < 0.05). The PCL/PLGA/TCP group had significantly greater bone density as compared with the control and PCL/PLGA groups at 4 and 8 weeks (P < 0.005). The results of this study suggest that the PCL/PLGA/TCP scaffold fabricated using SFF technology is useful for recovering and enhancing new bone formation in bony defects in rabbits.
Journal of Materials Science Materials in Medicine 09/2012; · 2.14 Impact Factor
[show abstract][hide abstract] ABSTRACT: This study developed a bioabsorbable-guided bone regeneration membrane made of blended polycaprolactone (PCL), poly(lactic-co-glycolic acid) (PLGA), and beta-tricalcium phosphate (β-TCP) using solid freeform fabrication (SFF) technology. The chemical and physical properties of the membrane were evaluated using field emission scanning electron microscopy, energy dispersive spectroscopy, and a tensile test. In vitro cell activity assays revealed that the adhesion, proliferation, and osteogenic differentiation of seeded adipose-derived stem cells (ADSCs) were significantly promoted by the PCL/PLGA/β-TCP membranes compared with PCL/PLGA membranes. When the PCL/PLGA and PCL/PLGA/β-TCP membranes were implanted on rabbit calvaria bone defects without ADSCs, microcomputed tomography and histological analyses confirmed that the SFF-based PCL/PLGA/β-TCP membranes greatly increased bone formation without the need for bone substitute materials. Moreover, tight integration, which helps to prevent exposure of the membrane, between both membranes and the soft tissues was clearly observed histologically. The SFF-based PCL/PLGA and PCL/PLGA/β-TCP membranes retained their mechanical stability for up to 8 weeks without significant collapse. Furthermore, PCL/PLGA/β-TCP underwent adequate degradation without a significant immune response at 8 weeks.
Tissue Engineering Part A 08/2012; · 4.64 Impact Factor
[show abstract][hide abstract] ABSTRACT: Fabrication of a three-dimensional (3D) scaffold with increased mechanical strength may be an essential requirement for more advanced bone tissue engineering scaffolds. Various material- and chemical-based approaches have been explored to enhance the mechanical properties of engineered bone tissue scaffolds. In this study, the effects of pore architecture and stacking direction on the mechanical and cell proliferation properties of a scaffold were investigated. The 3D scaffold was prepared using solid freeform fabrication technology with a multihead deposition system. Various types of scaffolds with different pore architectures (lattice, stagger, and triangle types) and stacking directions (horizontal and vertical directions) were fabricated with a blend of polycaprolactone and poly lactic-co-glycolic acid. In compression tests, the triangle-type scaffold was the strongest among the experimental groups. Stacking direction affected the mechanical properties of scaffolds. An in vitro cell counting kit-8 assay showed no significant differences in optical density depending on the different pore architectures and stacking directions. In conclusion, mechanical properties of scaffolds can be enhanced by controlling pore architecture and stacking direction.
Journal of Biomedical Materials Research Part A 04/2012; 100(7):1846-53. · 2.83 Impact Factor
[show abstract][hide abstract] ABSTRACT: Solid freeform fabrication (SFF) is recognized as a promising tool for creating tissue engineering scaffolds due to advantages such as superior interconnectivity and highly porous structure. Despite structural support for SFF-based three-dimensional (3-D) scaffolds that can lead to tissue regeneration, lack of cell recognition motifs and/or biochemical factors has been considered a limitation. Previously, recombinant mussel adhesive proteins (MAPs) were successfully demonstrated to be functional cell adhesion materials on various surfaces due to their peculiar adhesive properties. Herein, MAPs were applied as surface functionalization materials to SFF-based 3-D polycaprolactone/poly(lactic-co-glycolic acid) scaffolds. We successfully coated MAPs onto scaffold surfaces by simply dipping the scaffolds into the MAP solution, which was confirmed through X-ray photoelectron spectroscopy and scanning electron microscopy analyses. Through in vitro study using human adipose tissue-derived stem cells (hADSCs), significant enhancement of cellular activities such as attachment, proliferation, and osteogenic differentiation was observed on MAP-coated 3-D scaffolds, especially on which fused arginine-glycine-aspartic acid peptides were efficiently exposed. In addition, we found that in vivo hADSC implantation with MAP-coated scaffolds enhanced bone regeneration in a rat calvarial defect model. These results collectively demonstrate that facile surface functionalization of 3-D scaffolds using MAP would be a promising strategy for successful tissue engineering applications.
[show abstract][hide abstract] ABSTRACT: The aim of this study was to build a mechanically enhanced three-dimensional (3D) bioprinted construct containing two different cell types for osteochondral tissue regeneration. Recently, the production of 3D cell-laden structures using various scaffold-free cell printing technologies has opened up new possibilities. However, ideal 3D complex tissues or organs have not yet been printed because gel-state hydrogels have been used as the principal material and are unable to maintain the desired D structure due to their poor mechanical strength. In this study, thermoplastic biomaterial polycaprolactone (PCL), which shows relatively high mechanical properties as compared with hydrogel, was used as a framework for enhancing the mechanical stability of the bioprinted construct. Two different alginate solutions were then infused into the previously prepared framework consisting of PCL to create the 3D construct for osteochondral printing. For this work, a multi-head tissue/organ building system (MtoBS), which was particularly designed to dispense thermoplastic biomaterial and hydrogel having completely different rheology properties, was newly developed and used to bioprint osteochondral tissue. It was confirmed that the line width, position and volume control of PCL and alginate solutions were adjustable in the MtoBS. Most importantly, dual cell-laden 3D constructs consisting of osteoblasts and chondrocytes were successfully fabricated. Further, the separately dispensed osteoblasts and chondrocytes not only retained their initial position and viability, but also proliferated up to 7 days after being dispensed.
Journal of Micromechanics and Microengineering 01/2012; 22(8). · 1.79 Impact Factor
[show abstract][hide abstract] ABSTRACT: Natural biomaterials such as hyaluronic acid, gelatin and collagen provide excellent environments for tissue regeneration. Furthermore, gel-state natural biomaterials are advantageous for encapsulating cells and growth factors. In cell printing technology, hydrogel which contains cells was printed directly to form three-dimensional (3D) structures for tissue or organ regeneration using various types of printers. However, maintaining the 3D shape of the printed structure, which is made only of the hydrogel, is very difficult due to its weak mechanical properties. In this study, we developed a hybrid scaffold consisting of synthetic biomaterials and natural hydrogel using a multi-head deposition system, which is useful in solid freeform fabrication technology. The hydrogel was intentionally infused into the space between the lines of a synthetic biomaterial-based scaffold. The cellular efficacy of the hybrid scaffold was validated using rat primary hepatocytes and a mouse pre-osteoblast MC3T3-E1 cell line. In addition, the collagen hydrogel, which encapsulates cells, was dispensed and the viability of the cells observed. We demonstrated superior effects of the hybrid scaffold on cell adhesion and proliferation and showed the high viability of dispensed cells.
[show abstract][hide abstract] ABSTRACT: Bone tissue engineering often requires a well-defined scaffold that is highly porous. The multi-head deposition system (MHDS), a form of solid freeform fabrication, has raised great interest as a method for fabricating scaffolds, since it yields a highly porous inter-connective structure without the use of cytotoxic solvents, and permits the diffusion of nutrients and oxygen. However, this method is not suitable for introducing proteins, as it includes a heating process. Hydrogels incorporated with protein coating of the scaffold surface could overcome this MHDS limitation. In the present study, the surface of a scaffold fabricated using MHDS was coated with a mixture of fibrin and hyaluronic acid (HA) and used as a vehicle for delivery of both bone morphogenetic protein-2 (BMP-2) and adipose-derived stromal cells (ASCs). Fibrin/HA coating of the scaffold significantly enhanced initial cell attachment. Furthermore, the in vitro release of BMP-2 from fibrin/HA-coated scaffolds was sustained for 3 days and it stimulated the alkaline phosphatase activity of ASCs seeded on the scaffold for 10 days more actively and continuously than did the soluble BMP-2 that was added to the culture media, not the scaffold itself. Importantly, the transplantation of undifferentiated ASCs inoculated on BMP-2-loaded, fibrin/HA-coated scaffolds resulted in more improved bone formation and mineralization than did the transplantation of undifferentiated ASCs seeded on uncoated scaffolds or on fibrin/HA-coated scaffolds without BMP-2, but containing BMP-2 in the cell suspension medium. These results show that BMP-2-loaded, fibrin/HA-coated scaffolds fabricated using MHDS may be useful in stimulating bone regeneration from undifferentiated ASCs in vivo.
[show abstract][hide abstract] ABSTRACT: A MHDS has been employed to fabricate 3D scaffolds from PLGA with acetyl endgroups to achieve in vivo regeneration of cartilage tissue. The fabricated acetylated-PLGA scaffold showed open pores and interconnected structures. Rabbit chondrocytes were seeded on the PLGA scaffolds and transplanted immediately into subcutaneous sites of athymic mice. Chondrocytes transplantation with untreated PLGA scaffolds served as a control. Histological analysis of the implants at 4 weeks with H&E staining and alcian blue staining revealed higher extracellular matrix and GAG expression at the neocartilage in the PLGA-6Ac scaffolds than that of the PLGA-6OH scaffold group. This endgroup-modified scaffold may be useful for successful cartilage tissue engineering in orthopedic applications.
[show abstract][hide abstract] ABSTRACT: Synchrotron X-ray bioimaging was successfully carried out to observe bone regeneration by a novel artificial bone substitute of bioactive MegaGen Synthetic Bone (MGSB) and hyaluronate (HA) hydrogels. A biphasic calcium phosphate of MGSB was prepared by chemical precipitation method, with a porous spherical morphology. On the basis of the fact that HA plays important roles in bone regeneration and promotes the differentiation, vascularization, and migration of stem cells, HA-cystamine (CYS) hydrogels with cleavable disulfide linkages were prepared to supply HA continuously for effective bone regeneration by their controlled degradation in vivo. Among seven different samples using Bio-OSS®, MGSB, and/or several kinds of HA hydrogels, MGSB/HA-CYS hydrogels resulted in the most significant bone regeneration in the calvarial critical bone defect of New Zealand white rabbits. Histological and histomorphometric analyses revealed that the bone regeneration by MGSB/HA-CYS hydrogels was as high as 43%, occupying 71% of the bone defect area with MGSB in the form of a calvarial bone plate in 4 weeks. After that, MGSB was bioabsorbed and replaced gradually with regenerated bones as observed in 8 weeks. Synchrotron X-ray imaging clearly confirmed the effective bone regeneration by MGSB/HA-CYS hydrogels, showing three-dimensional micron-scale morphologies of regenerated bones interconnected with MGSB. In addition, sequential nondestructive synchrotron X-ray tomographic analysis results from anterior to posterior of the samples were well matched with the histomorphometric analysis results. The clinically feasible artificial bone substitutes of MGSB/HA-CYS hydrogels will be investigated further for various bone tissue engineering applications using the synchrotron X-ray bioimaging systems.
Tissue Engineering Part C Methods 10/2010; 16(5):1059-68. · 4.64 Impact Factor
[show abstract][hide abstract] ABSTRACT: Solid free-form fabrication (SFF) technology is used to fabricate scaffolds with controllable characteristics including well-defined pore size and porosity. The multi-head deposition system (MHDS), one form of SFF technology, may be more advantageous than others for fabricating scaffolds because a MHDS does not require the use of a cytotoxic solvent. This method, however, may induce the thermal degradation of raw materials and a subsequent decrease in the material's molecular weight, whereby hydrolytic degradation, resulting in acidic by-products, might be accelerated. This study investigated whether fabrication of poly(lactic-co-glycolic acid) (PLGA) scaffolds using a MHDS with various residence times in the heating step induces thermal degradation and affects the proliferation of cells seeded on the scaffold in vitro. To answer this question, we fabricated porous three-dimensional PLGA scaffolds using residence times of 1, 3, 5 and 7 days for groups 1 through 4, respectively. Degradation behavior of the scaffolds was observed for 7 weeks in phosphate-buffered saline solution (pH 7.4) at 37 degrees C. The molecular weight, glass transition temperature and mechanical properties were compared for PLGA scaffolds fabricated with each of the four residence times at 120 degrees C. The proliferation rate of MC3T3-E1 cells grown on each group of scaffolds was compared to investigate the effect of acidic by-products on the growth of seeded cells in vitro. The heat process applied in fabrication of SFF-based PLGA scaffolds induced considerable thermal degradation followed by a decrease in molecular weight and mechanical compressive strength of the scaffolds in groups 3 and 4, which had more than 3 days residence time. Moreover, the cell proliferation rate was significantly higher for group 1 than for groups 3 and 4.