Morphometric analysis of the proximal ulna using three-dimensional computed tomography and computer-aided design: varus, dorsal, and torsion angulation.

PURPOSE: The proximal ulna, particularly the course of the posterior border, has a complex three-dimensional (3D) morphology which has been highlighted recently due to its clinical relevance in relation to surgical treatments. 3D computed tomography (CT) reconstruction and computer-aided design (CAD) based software can help to visualize the complex anatomy and thus aid the investigation of the more detailed morphology of the proximal ulna. METHODS: In our current study, 3D CT reconstruction images of 20 cadavers were imported into the 3D CAD program. Three morphologic angle parameters of the proximal ulna were measured including the dorsal, varus and torsion angulation. The torsion angulation was measured using the flat spot of olecranon dorsal aspect. We measured the total length of the ulna and the distance between the olecranon tip and the apex of dorsal and varus angulation. Furthermore, the thickness of olecranon was also measured for all the specimens. RESULTS: The results showed that the mean dorsal, varus, and torsion angulation was 4.3° (range 2.6°-5.9°), 12.1° (range 7.9°-17.6°), and 22.5° (range 16.6°-30.5°), respectively. The average length ratio of the dorsal and varus angulation apex to the total ulnar length was 26.4 % (range 19.8-30.7 %) and 32.7 % (range 27.5-37.5 %), respectively. The average of olecranon thickness at the proximal tip, mid-olecranon fossa, and at coronoid tip level was 17.8 mm (range 14.1-22.8 mm), 19.7 mm (range 15.8-23.1 mm), and 35.1 mm (range 27.9-41.8 mm), respectively. CONCLUSION: In conclusion, variations in the proximal ulna have to be considered when anatomically contoured dorsal plates are applied. Knowledge of the 3D morphologic anatomy of the proximal ulna would provide important information on fracture reductions, and the design of a precontoured dorsal plate or a prosthetic ulnar stem.

In Vivo Biophysical-stimulation Platform Applied to Rodent Calvarial Bone-defect Models.

Biophysical strain has been applied widely for bone regeneration. However, application of low-magnitude strains to cells on small-thickness scaffolds is problematic, especially in rodent calvarial defect models, because general translation systems have limitations in terms of generating low-magnitude smooth signals. To overcome these limitations, we developed an in vitro biophysical-stimulation platform for stimulation of cells on small-thickness scaffolds for rodent calvarial bone defects. The customized flexure-based translational nanoactuator enables generation of low-magnitude smooth signals at the subnano- to micrometer-scale. This nanoactuator, which is equipped with a piezoelectric actuator, is suitable for biological applications because it can generate friction-free motion with a high resolution. Moreover, its operation without wear or deterioration eliminates contamination factors in cell culture environments. The developed in vitro biophysical-stimulation platform using these nanoactuators showed predictable operational characteristics. Also, a few-micrometer sinusoidal signal was generated successfully without any distortion. Three-dimensional scaffolds fitting the critical-size rat calvarial defect model were fabricated using poly(caprolactone), poly(lactic-co-glycolic acid), and tricalcium phosphate. Runt-related transcription factor 2 expression was increased upon stimulation of human adipose-derived stem cells (ASCs) on these scaffolds were stimulated in the in vitro biophysical-stimulation platform. Additionally, the use of this platform resulted in up-regulation of alkaline phosphate, osteopontin, and osterix expression compared to the non-stimulated group. These preliminary in vitro results suggest that the biophysical environment provided by the in vitro biophysical-stimulation platform influences the osteogenic differentiation of ASCs.

Flexure-based device for cyclic strain-mediated osteogenic differentiation.

Application of low-magnitude strains to cells on small-thickness scaffolds, such as those for rodent calvarial defect models, is problematic, because general translation systems have limitations in terms of generating low-magnitude smooth signals. To overcome this limitation, we developed a cyclic strain generator using a customized, flexure-based, translational nanoactuator that enabled generation of low-magnitude smooth strains at the subnano- to micrometer scale to cells on small-thickness scaffolds. The cyclic strain generator we developed showed predictable operational characteristics by generating a sinusoidal signal of a few micrometers (4.5 µm) without any distortion. Three-dimensional scaffolds fitting the critical-size rat calvarial defect model were fabricated using poly(caprolactone), poly(lactic-co-glycolic acid), and tricalcium phosphate. Stimulation of human adipose-derived stem cells (ASCs) on these fabricated scaffolds using the cyclic strain generator we developed resulted in upregulated osteogenic marker expression compared to the nonstimulated group. These preliminary in vitro results suggest that the cyclic strain generator successfully provided mechanical stimulation to cells on small-thickness scaffolds, which influenced the osteogenic differentiation of ASCs.

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.

Development of a 3D cell printed construct considering angiogenesis for liver tissue engineering.

Several studies have focused on the regeneration of liver tissue in a two-dimensional (2D) planar environment, whereas actual liver tissue is three-dimensional (3D). Cell printing technology has been successfully utilized for building 3D structures; however, the poor mechanical properties of cell-laden hydrogels are a major concern. Here, we demonstrate the printing of a 3D cell-laden construct and its application to liver tissue engineering using 3D cell printing technology through a multi-head tissue/organ building system. Polycaprolactone (PCL) was used as a framework material because of its excellent mechanical properties. Collagen bioink containing three different types of cells-hepatocytes (HCs), human umbilical vein endothelial cells , and human lung fibroblasts--was infused into the canals of a PCL framework to induce the formation of capillary--like networks and liver cell growth. A co-cultured 3D microenvironment of the three types of cells was successfully established and maintained. The vascular formation and functional abilities of HCs (i.e., albumin secretion and urea synthesis) demonstrated that the heterotypic interaction among HCs and nonparenchymal cells increased the survivability and functionality of HCs within the collagen gel. Therefore, our results demonstrate the prospect of using cell printing technology for the creation of heterotypic cellular interaction within a structure for liver tissue engineering.

Combined effect of three types of biophysical stimuli for bone regeneration.

Pretreatment using various types of biophysical stimuli could provide appropriate potential to cells during construction of the engineered tissue in vitro. We hypothesized that multiple combinations of these biophysical stimuli could enhance osteogenic differentiation in vitro and bone formation in vivo. Cyclic strain, an electromagnetic field, and ultrasound were selected and combined as effective stimuli for osteogenic differentiation using a developed bioreactor. Here we report the experimental evaluation of the osteogenic effects of various combinations of three different biophysical stimuli in vitro and in vivo using human adipose-derived stem cells (ASCs). Osteogenic differentiation of ASCs was accelerated by multiple-combination biophysical stimulation in vitro. However, both single stimulation and double-combination stimulation were sufficient to accelerate bone regeneration in vivo, while the osteogenic marker expression of those groups was not as high as that of triple-combination stimulation in vitro. We inferred from these data that ASCs appropriately differentiated into the osteogenic lineage by biophysical stimulation could be a better option for accelerating bone formation in vivo than relatively undifferentiated or completely differentiated ASCs. Although many questions remain about the mechanisms of combined effects of various biophysical stimuli, this approach could be a more powerful tool for bone tissue regeneration.