Won-Soo Yun

Evaluation of the effective diffusivity of a freeform fabricated scaffold using computational simulation.

In scaffold-based tissue engineering, sufficient oxygen and nutrient supply into cells within a scaffold is essential to increase cell viability and the proliferation rate. Generally, oxygen and nutrients reach the cells through the media by diffusion in vitro or in vivo, assuming there is no convection flow through a scaffold with small-sized pores. The scaffold diffusion rate depends mainly on the scaffold pore architecture. Thus, understanding the effect of scaffold pore architecture on the diffusion mechanism is necessary to design an efficient scaffold model. This study proposes a computational method to estimate diffusivity using the finite element analysis (FEA). This method can be applied to evaluate and analyze the effective diffusivity of a freeform fabricated 3D scaffold. The diffusion application module of commercial FEA software was used to calculate the spatial oxygen concentration gradient in a scaffold model medium. The effective diffusivities of each scaffold could be calculated from the oxygen concentration data, which revealed that the scaffold pore architecture influences its effective diffusivity. The proposed method has been verified experimentally and can be applied to design pore architectures with efficient diffusion by increasing our understanding of how the diffusion rate within a scaffold is affected by its pore architecture.

Precise stacking of decellularized extracellular matrix based 3D cell-laden constructs by a 3D cell printing system equipped with heating modules

Three-dimensional (3D) cell printing systems allow the controlled and precise deposition of multiple cells in 3D constructs. Hydrogel materials have been used extensively as printable bioinks owing to their ability to safely encapsulate living cells. However, hydrogel-based bioinks have drawbacks for cell printing, e.g. inappropriate crosslinking and liquid-like rheological properties, which hinder precise 3D shaping. Therefore, in this study, we investigated the influence of various factors (e.g. bioink concentration, viscosity, and extent of crosslinking) on cell printing and established a new 3D cell printing system equipped with heating modules for the precise stacking of decellularized extracellular matrix (dECM)-based 3D cell-laden constructs. Because the pH-adjusted bioink isolated from native tissue is safely gelled at 37 °C, our heating system facilitated the precise stacking of dECM bioinks by enabling simultaneous gelation during printing. We observed greater printability compared with that of a non-heating system. These results were confirmed by mechanical testing and 3D construct stacking analyses. We also confirmed that our heating system did not elicit negative effects, such as cell death, in the printed cells. Conclusively, these results hold promise for the application of 3D bioprinting to tissue engineering and drug development.

Evaluation of 3D printed PCL/PLGA/β-TCP versus collagen membranes for guided bone regeneration in a beagle implant model.

Here, we compared 3D-printed polycaprolactone/poly(lactic-co-glycolic acid)/β-tricalcium phosphate (PCL/PLGA/β-TCP) membranes with the widely used collagen membranes for guided bone regeneration (GBR) in beagle implant models. For mechanical property comparison in dry and wet conditions and cytocompatibility determination, we analyzed the rate and pattern of cell proliferation of seeded fibroblasts and preosteoblasts using the cell counting kit-8 assay and scanning electron microscopy. Osteogenic differentiation was verified using alizarin red S staining. At 8 weeks following implantation in vivo using beagle dogs, computed tomography and histological analyses were performed after sacrifice. Cell proliferation rates in vitro indicated that early cell attachment was higher in collagen than in PCL/PLGA/β-TCP membranes; however, the difference subsided by day 7. Similar outcomes were found for osteogenic differentiation, with approximately 2.5 times greater staining in collagen than PCL/PLGA/β-TCP, but without significant difference by day 14. In vivo, bone regeneration in the defect area, represented by new bone formation and bone-to-implant contact, paralleled those associated with collagen membranes. However, tensile testing revealed that whereas the PCL/PLGA/β-TCP membrane mechanical properties were conserved in both wet and dry states, the tensile property of collagen was reduced by 99% under wet conditions. Our results demonstrate in vitro and in vivo that PCL/PLGA/β-TCP membranes have similar levels of biocompatibility and bone regeneration as collagen membranes. In particular, considering that GBR is always applied to a wet environment (e.g. blood, saliva), we demonstrated that PCL/PLGA/β-TCP membranes maintained their form more reliably than collagen membranes in a wet setting, confirming their appropriateness as a GBR membrane.

Using lateral vibration for thermosonic flip-chip interconnection with anisotropic conductive film

In this paper, thermosonic flip-chip bonding with anisotropic conductive film (ACF) is considered. To shorten the ACF curing time, an ultrasonic vibration of 40 kHz in the lateral direction is applied, in addition to the thermocompression process. By measuring the internal temperature of CF, the fast curing of ACF due to ultrasonic vibration is verified. The experiments with a commercialized ACF for the interconnection of LCD driver chips exhibit a notable reduction of bonding time by 77% as well as a reduction of tool-tip temperature while ensuring the bonding reliability. Also, as a typical feature of the proposed method, the sliding phenomenon between the contact surface of the specimen and the horn tool-tip is reported, which significantly enhances the reliability of thermosonic flip-chip bonding. Through the observation of pressured conductive particle marks, the water absorption test and the shear test, the quality of interconnection is proven.

Comparative Efficacies of Collagen-Based 3D Printed PCL/PLGA/β-TCP Composite Block Bone Grafts and Biphasic Calcium Phosphate Bone Substitute for Bone Regeneration

The purpose of this study was to compare bone regeneration and space maintaining ability of three-dimensional (3D) printed bone grafts with conventional biphasic calcium phosphate (BCP). After mixing polycaprolactone (PCL), poly (lactic-co-glycolic acid) (PLGA), and β-tricalcium phosphate (β-TCP) in a 4:4:2 ratio, PCL/PLGA/β-TCP particulate bone grafts were fabricated using 3D printing technology. Fabricated particulate bone grafts were mixed with atelocollagen to produce collagen-based PCL/PLGA/β-TCP composite block bone grafts. After formation of calvarial defects 8 mm in diameter, PCL/PLGA/β-TCP composite block bone grafts and BCP were implanted into bone defects of 32 rats. Although PCL/PLGA/β-TCP composite block bone grafts were not superior in bone regeneration ability compared to BCP, the results showed relatively similar performance. Furthermore, PCL/PLGA/β-TCP composite block bone grafts showed better ability to maintain bone defects and to support barrier membranes than BCP. Therefore, within the limitations of this study, PCL/PLGA/β-TCP composite block bone grafts could be considered as an alternative to synthetic bone grafts available for clinical use.

Effects of 3D-Printed Polycaprolactone/β-Tricalcium Phosphate Membranes on Guided Bone Regeneration

This study was conducted to compare 3D-printed polycaprolactone (PCL) and polycaprolactone/β-tricalcium phosphate (PCL/β-TCP) membranes with a conventional commercial collagen membrane in terms of their abilities to facilitate guided bone regeneration (GBR). Fabricated membranes were tested for dry and wet mechanical properties. Fibroblasts and preosteoblasts were seeded into the membranes and rates and patterns of proliferation were analyzed using a kit-8 assay and by scanning electron microscopy. Osteogenic differentiation was verified by alizarin red S and alkaline phosphatase (ALP) staining. An in vivo experiment was performed using an alveolar bone defect beagle model, in which defects in three dogs were covered with different membranes. CT and histological analyses at eight weeks after surgery revealed that 3D-printed PCL/β-TCP membranes were more effective than 3D-printed PCL, and substantially better than conventional collagen membranes in terms of biocompatibility and bone regeneration and, thus, at facilitating GBR.

Development of a compact micro-stereolithography (MSTL) system using a Blu-ray optical pickup unit

In this paper, a compact micro-stereolithography (MSTL) apparatus of tabletop size was designed and constructed using a Blu-ray optical pickup unit (OPU), which came from a commercial Blu-ray optical disk driver. This is the very first application of a Blu-ray OPU to a light source of a MSTL system. A three-axis stage was adopted to obtain 3D motions, and a simple optic system was added to extend the focal length of a Blu-ray OPU for avoiding the physical interference. A liquid photo-curable resin suitable for the system was recomposed by modifying previously reported one. Then, the fabrication characteristics were investigated by constructing single-line structures at varying parameters such as laser power, scan speed and aperture diameter. The cured depth and width of the single-line structures decreased as the irradiated energy, which is determined by laser power and scan speed, decreased. Interestingly, the result also showed that the aperture diameter can effectively control the aspect ratio of cured depth and width. The increase of aperture diameter brings about the decrease of cured depth and increase of cured width. Based on the results of single-line exposure experiment, various 3D micro-structures were successfully fabricated. From this study, it can be shown that the Blu-ray-based MSTL system and the resin are suitable for fabricating complicated 3D micro-structures with reproducibility.

Elastic region of an ACF joint for thermosonic flip-chip bonding

In this paper, a thermosonic flip-chip bonding method using vibration in the lateral direction is proposed. To enhance the reliability and alignment of the specimen after the thermosonic process, an elastic region of anisotropic conductive film (ACF) joints is investigated, within which ultrasonic vibration is applied. By creep-recovery tests, it is shown that the elastic region gradually develops up to 7 µm as the degree of cure increases to 90%. Based on this result, a two-step lateral thermosonic process composed of a pure thermal curing process at the viscoelastic stage of the ACF epoxy and a thermal and ultrasonic curing process within an established elastic region is proposed. To demonstrate the feasibility of the proposed approach in practice, experiments with a commercialized ACF and LCD driver chips are conducted. The experiment shows a notable enhancement in the alignment of the specimen after the thermosonic process, with a bonding time reduction of 33.3% compared with the conventional thermocompression process.

Ultrasonic ACF Bonding Technique for Mounting LCD Driver ICs

In the paper, we develop the ultrasonic bonding technique for LCD driver chips having small size and high pin-density. In general, the mounting technology for LCD driver ICs is a thermo-compression method utilizing the ACF (An-isotropic Conductive Film). The major drawback of the conventional approach is the long process time. It will be shown that the conventional ACF method based on thermo-compression can be remarkably enhanced by employing the ultrasonic bonding technique in terms of bonding time. The proposed approach is to apply the ultrasonic energy together with the thermo-compression methodology for the ACF bonding process. To this end, we design a bonding head that enables pre-heating, pressure and ultrasonic excitation. Through the bonding experiments mainly with LCD driver ICs, we present the procedures to select the best combination of process parameters with analysis. We investigate the effects of bonding pressure, bonding time, pre-heating temperature before bonding, and the power level of ultrasonic energy. The addition of ultrasonic excitation to the thermo-compression method reduces the pre-heating temperature and the bonding process time while keeping the quality bonding between the LCD pad and the driver IC. The proposed concept will be verified and demonstrated with experimental results.

Osteogenesis of Adipose-Derived and Bone Marrow Stem Cells with Polycaprolactone/Tricalcium Phosphate and Three-Dimensional Printing Technology in a Dog Model of Maxillary Bone Defects

Bone graft material should possess sufficient porosity and permeability to allow integration with native tissue and vascular invasion, and must satisfy oxygen and nutrient transport demands. In this study, we have examined the use of three-dimensional (3D)-printed polycaprolactone/tricalcium phosphate (PCL/TCP) composite material in bone grafting, to estimate the scope of its potential application in bone surgery. Adipose-derived stem cells (ADSCs) and bone marrow stem cells (BMSCs) are known to enhance osteointegration. We hypothesized that a patient-specific 3D-printed solid scaffold could help preserve seeded ADSCs and BMSCs and enhance osteointegration. Diffuse osteogenic tissue formation was observed by micro-computed tomography with both stem cell types, and the ADSC group displayed similar osteogenesis compared to the BMSC group. In histological assessment, the scaffold pores showed abundant ossification in both groups. Reverse transcription polymerase chain reaction (RT-PCR) showed that the BMSC group had higher expression of genes associated with ossification, and this was confirmed by Western blot analysis. The ADSC- and BMSC-seeded 3D-printed PCL/TCP scaffolds displayed promising enhancement of osteogenesis in a dog model of maxillary bone defects.