Ching-Shiow Tseng

Synthesis and 3D Printing of Biodegradable Polyurethane Elastomer by a Water‐Based Process for Cartilage Tissue Engineering Applications

Biodegradable materials that can undergo degradation in vivo are commonly employed to manufacture tissue engineering scaffolds, by techniques including the customized 3D printing. Traditional 3D printing methods involve the use of heat, toxic organic solvents, or toxic photoinitiators for fabrication of synthetic scaffolds. So far, there is no investigation on water-based 3D printing for synthetic materials. In this study, the water dispersion of elastic and biodegradable polyurethane (PU) nanoparticles is synthesized, which is further employed to fabricate scaffolds by 3D printing using polyethylene oxide (PEO) as a viscosity enhancer. The surface morphology, degradation rate, and mechanical properties of the water-based 3D-printed PU scaffolds are evaluated and compared with those of polylactic-co-glycolic acid (PLGA) scaffolds made from the solution in organic solvent. These scaffolds are seeded with chondrocytes for evaluation of their potential as cartilage scaffolds. Chondrocytes in 3D-printed PU scaffolds have excellent seeding efficiency, proliferation, and matrix production. Since PU is a category of versatile materials, the aqueous 3D printing process developed in this study is a platform technology that can be used to fabricate devices for biomedical applications.

Water-based polyurethane 3D printed scaffolds with controlled release function for customized cartilage tissue engineering

Conventional 3D printing may not readily incorporate bioactive ingredients for controlled release because the process often involves the use of heat, organic solvent, or crosslinkers that reduce the bioactivity of the ingredients. Water-based 3D printing materials with controlled bioactivity for customized cartilage tissue engineering is developed in this study. The printing ink contains the water dispersion of synthetic biodegradable polyurethane (PU) elastic nanoparticles, hyaluronan, and bioactive ingredients TGFβ3 or a small molecule drug Y27632 to replace TGFβ3. Compliant scaffolds are printed from the ink at low temperature. These scaffolds promote the self-aggregation of mesenchymal stem cells (MSCs) and, with timely release of the bioactive ingredients, induce the chondrogenic differentiation of MSCs and produce matrix for cartilage repair. Moreover, the growth factor-free controlled release design may prevent cartilage hypertrophy. Rabbit knee implantation supports the potential of the novel 3D printing scaffolds in cartilage regeneration. We consider that the 3D printing composite scaffolds with controlled release bioactivity may have potential in customized tissue engineering.

Fabrication of Precision Scaffolds Using Liquid-Frozen Deposition Manufacturing for Cartilage Tissue Engineering

The fused deposition manufacturing (FDM) system has been used to fabricate tissue-engineered scaffolds with highly interconnecting and controllable pore structure, although the system is limited to a few materials. For this reason, the liquid-frozen deposition manufacturing (LFDM) system based on an improvement of the FDM process was developed. Poly(D,L-lactide-co-glycolide) (PLGA) precision scaffolds were fabricated using LFDM from PLGA solutions of different concentrations. A greater concentration of PLGA solution resulted in greater mechanical strength but also resulted in less water content and smaller pore size on the surface of the scaffolds. LFDM scaffolds in general had mechanical strength closer to that of native articular cartilage than did FDM scaffolds. Neocartilage formation was observed in LFDM scaffolds seeded with porcine articular chondrocytes after 28 days of culture. Chondrocytes in LFDM scaffolds made from low concentrations (15-20%) of PLGA solution maintained a round shape, proliferated well, and secreted abundant extracellular matrix. In contrast, the FDM PLGA scaffolds had low cell numbers and poor matrix production because of heavy swelling. The LFDM system offered a useful way to fabricate scaffolds for cartilage tissue-engineering applications.

The substrate-dependent regeneration capacity of mesenchymal stem cell spheroids derived on various biomaterial surfaces

Mesenchymal stem cells (MSCs) are widely used for their self-renewal and multipotent abilities, which can be further enhanced by growing MSCs as three-dimensional (3D) cellular spheroids on certain substrates. Although various surfaces have been used to generate 3D MSC spheroids, the answer to whether all these spheroids have similar in vitro and in vivo properties remains unclear. In this study, adipose-derived adult stem cells (ADSCs) were cultured on a non-adherent Petri dish, polyvinyl alcohol, chitosan (CS), or chitosan-hyaluronan (CS-HA) to form 3D spheroids. The expression of the cell adhesion molecule, N-cadherin, was analyzed by qRT-PCR and Western blotting. The functional migration ability was tested using the transwell assay. The capacity for chondral regeneration of various ADSC spheroids was further evaluated in a rabbit model. We demonstrated that ADSC spheroids derived on the CS or CS-HA surface had the greater expression of N-cadherin and better migration ability. The latter was consistent with the higher expression levels of chemokine/receptor SDF-1/CXCR4 for the spheroids derived on CS or CS-HA. Animal studies also revealed significantly better cartilage repair in defects loaded with CS- or CS-HA-derived spheroids. In particular, CS-HA-derived spheroids gave rise to the best regeneration when combined with a 3D printed scaffold. This study suggested that MSC spheroids derived on different surfaces may have distinct in vitro and in vivo properties, which appeared to be associated with the surface-bound calcium as well as the calcium-dependent N-cadherin and CXCR4 signaling. The substrate-dependent properties may eventually lead to different regeneration capacities of various MSC spheroids in vivo.

Air plasma treated chitosan fibers-stacked scaffolds.

Chitosan is a nontoxic, biodegradable and biocompatible polymer. Rapid prototyped chitosan scaffolds were manufactured by liquid-frozen deposition of chitosan fibers in this study. To investigate if the air plasma (AP) treatment could be used to improve the surface properties of these scaffolds for cell attachment, chitosan films were first prepared and treated with AP under different conditions. Under the optimized condition, the water contact angle of chitosan films was significantly reduced from 90 ± 1° to 19 ± 1° after AP treatment. On the other hand, the surface charge and nanometric roughness of chitosan films increased after AP treatment. X-ray photoelectron spectroscopy measurement on AP-treated three-dimensional chitosan scaffolds showed that nitrogen and oxygen increased at each location inside the scaffolds as compared to the untreated ones, which indicated that AP could permeate through the fibrous stacks of the scaffolds and effectively modify the interior (visible) surface of the scaffolds. Moreover, AP treatment enabled the migration of MC3T3-E1 cells into the scaffolds, facilitated their proliferation and promoted the bone mineral deposition. These results suggested that fibers-stacked chitosan scaffolds may be produced by liquid-frozen deposition and treated with AP for bone tissue engineering applications.

Preparation and characterization of a biodegradable polyurethane hydrogel and the hybrid gel with soy protein for 3D cell-laden bioprinting

3D printing shows great potential for fabricating customized scaffolds for tissue regeneration. Using hydrogel as a bioink for cell printing provides a biological platform for basic research and potential medical treatments. In this study, a waterborne poly(ε-caprolactone) (PCL)-based biodegradable polyurethane (PU) with a soft segment replaced with 20 mol% of poly(l-lactide) (PLLA) diol or poly (d,l-lactide) (PDLLA) diol was prepared. These two PUs formed compact packing structures at temperatures ≥37 °C. They responded differently to temperature changes and the presence of electrolytes because of the difference in the free volume. With their thermal-responsive properties, both PU dispersions could form a gel in 3 min with the gel modulus reaching about 6-8 kPa after 30 min. To enhance the structural integrity during layer-by-layer deposition, the hybrid hydrogel of PU and soy protein isolate (PU/SPI hybrid) was further developed. The PU/SPI hybrid dispersion could undergo rapid gelation at 37 °C with the modulus reaching 130 Pa in 1 min. Moreover, the PU/SPI hybrid gel was readily blended with cells and printed at 37 °C without preheating. Neural stem cells (NSCs) were embedded in the hydrogels and analyzed for cell viability, metabolism, proliferation, and gene expression of neural-related markers. Cells cultured in the PU/SPI hybrid construct had better survival and proliferation than those in the PU gel. The PU/SPI hybrid ink may provide unique rheological properties for direct cell/tissue printing at 37 °C and a biomimetic microenvironment for cell survival, growth, and differentiation.

Registration of 2D C-Arm and 3D CT Images for a C-Arm Image-Assisted Navigation System for Spinal Surgery.

C-Arm image-assisted surgical navigation system has been broadly applied to spinal surgery. However, accurate path planning on the C-Arm AP-view image is difficult. This research studies 2D-3D image registration methods to obtain the optimum transformation matrix between C-Arm and CT image frames. Through the transformation matrix, the surgical path planned on preoperative CT images can be transformed and displayed on the C-Arm images for surgical guidance. The positions of surgical instruments will also be displayed on both CT and C-Arm in the real time. Five similarity measure methods of 2D-3D image registration including Normalized Cross-Correlation, Gradient Correlation, Pattern Intensity, Gradient Difference Correlation, and Mutual Information combined with three optimization methods including Powells method, Downhill simplex algorithm, and genetic algorithm are applied to evaluate their performance in converge range, efficiency, and accuracy. Experimental results show that the combination of Normalized Cross-Correlation measure method with Downhill simplex algorithm obtains maximum correlation and similarity in C-Arm and Digital Reconstructed Radiograph (DRR) images. Spine saw bones are used in the experiment to evaluate 2D-3D image registration accuracy. The average error in displacement is 0.22 mm. The success rate is approximately 90% and average registration time takes 16 seconds.

3D printing of polyurethane biomaterials

Polyurethanes (PUs) have evolved rapidly from traditional polymers to materials with high performance physical properties in recent years. From a chemical point of view, the urethane group (–NH–COO–) in PUs does not exist in nature but appears to be a combination of amide (–NH–CO–) and ester (–COO–) groups that are present in the structure of peptides and lipids, respectively. In addition, the “segmented” structure of PU resembles the “domain” structure of a protein from a physical point of view. The unique chemical and physical characteristics of PUs also make them suitable for various applications including three-dimensional (3D) printing for biomedical applications. We introduce PUs as materials useful for biomaterial applications and then describe their potential to be processed using 3D-printing technology.

An Ultrasound Imaging-Guided Robotic HIFU Ablation Experimental System and Accuracy Evaluations

In recent years, noninvasive thermal treatment by using high-intensity focused ultrasound (HIFU) has high potential in tumor treatment. The goal of this research is to develop an ultrasound imaging-guided robotic HIFU ablation system for tumor treatment. The system integrates the technologies of ultrasound image-assisted guidance, robotic positioning control, and HIFU treatment planning. With the assistance of ultrasound image guidance technology, the tumor size and location can be determined from ultrasound images as well as the robotic arm can be controlled to position the HIFU transducer to focus on the target tumor. After the development of the system, several experiments were conducted to measure the positioning accuracy of this system. The results show that the average positioning error is 1.01 mm with a standard deviation 0.34, and HIFU ablation accuracy is 1.32 mm with a standard deviation 0.58, which means this system is confirmed with its possibility and accuracy.