Yoshio Shirasaki

Promotion of bone formation using highly pure porous β-TCP combined with bone marrow-derived osteoprogenitor cells

Beta-tricalcium phosphate (TCP) exhibits rapid degradation and weak mechanical properties, which has limited its application as bone graft substitutes, though it has good biocompatibility and osteoconductivity. We hypothesized that a composite of highly pure porous beta-TCP and bone marrow-derived osteoprogenitor cells (BMO) could improve bone formation, and slow down the degradation of beta-TCP. A highly pure porous beta-TCP with 75% porosity was fabricated. The pores averaged 200-400 microm in diameter, with interconnecting paths 100-200 microm. Blocks of beta-TCP 5 mm3 were combined with BMO, and incubated 2 weeks with (+) or without (-) osteogenic medium. They were then implanted into subcutaneous sites of syngeneic rats for 24 weeks. These composites were harvested at different time points. The alkaline phosphatase activity and bone osteocalcin content of the composites (+) were much higher than corresponding values in the composites (-) of the control group (p<0.01). Light microscopy revealed mature bone and lots of blood vessels only in the TCP/BMO composite (+). The amount of newly formed bone increased until week 24. Slow resorptive activity could be found. The mechanical parameters of the composites were much improved over those of dry beta-TCP blocks. These results showed that tissue engineering treatment on incubating the composites of beta-TCP and BMO cells in osteogenic medium results in a good osteogenic activity.

Paradoxical Adaptation of Mature Radius to Unilateral Use in Tennis Playing

The positive effects of physical activity on human bone mass have been well documented in many cross-sectional studies comparing athletes with sedentary controls as well as in longitudinal follow-up. By applying peripheral quantitative computed tomography (pQCT), which has the advantage of measuring volumetric bone mineral density (BMD) and the ability to distinguish among trabecular and cortical components, it was demonstrated that cortical BMD of the dominant arm was not greater than that of the nondominant arm. Cortical drift toward the periosteal direction and an increase in cortical thickness resulted in an improvement of mechanical characteristics of the playing arms midradius. An improvement in the mechanical properties of young adult bone in response to long-term exercise was therefore related to geometric adaptation, but not to an increase in BMD. The manner in which the recruitment and function of bone cells are coordinated differs between the growing and the nongrowing skeleton. In the former, modeling is the dominant mode, and in the latter it is remodeling. In the present study, the side-to-side difference of 92 middle-aged female tennis players who initiated training after bone had matured was analyzed by pQCT. The side-to-side difference detected suggested a paradoxical adaptation of the mature radius to unilateral use during tennis playing, and that tennis playing after bone had matured did not stimulate cortical drift in the periosteal direction, unlike that seen in young subjects. Unexpectedly, the cross-sectional areas (periosteal and endocortical area) of the radius were smaller in the dominant arm than in the nondominant arm in the middle-aged female players. The findings suggest that unilateral use of the arm after the third decade of life suppresses age-related changes in bone geometry.

T2 and Apparent Diffusion Coefficient of MRI Reflect Maturation of Tissue-Engineered Auricular Cartilage Subcutaneously Transplanted in Rats.

In cartilage regenerative medicine, autologous chondrocyte implantation (ACI) has been applied clinically for partial defects of joint cartilage or nasal augmentation. To make treatment with ACI more effective and prevalent, modalities to evaluate the quality of transplanted constructs noninvasively are necessary. In this study, we compared the efficacy of several noninvasive modalities for evaluating the maturation of tissue-engineered auricular cartilage containing a biodegradable polymer scaffold. We first transplanted tissue-engineered cartilage consisting of human auricular chondrocytes, atelocollagen gel, and a poly-l-lactic acid (PLLA) porous scaffold subcutaneously into the back of athymic nude rats. Eight weeks after transplantation, the rats were examined by magnetic resonance imaging (MRI), X-ray, and ultrasound as noninvasive modalities. Then, the excised constructs were examined by histological and biochemical analysis including toluidine blue (TB) staining, glycosaminoglycans content, and enzyme-linked immunosorbent assay of type II collagen. Among the modalities examined, transverse relaxation time (T2) and apparent diffusion coefficient of MRI showed quite a high correlation with histological and biochemical results, suggesting that these can effectively detect the maturation of tissue-engineered auricular cartilage. Since these noninvasive modalities would realize time-course analysis of the maturation of tissue-engineered auricular cartilage, this study provides a substantial insight for improving the quality of tissue-engineered cartilage, leading to improvement of the quality and technique in cartilage regenerative medicine.

Relation between speed of sound measured by using ultrasound and magnetic resonance images and elasticity in tissue-engineered cartilage

It is important to evaluate the elasticity of tissue-engineered cartilage, for evaluating its structural strength. However, in vivo evaluation of tissue-engineered cartilage elasticity has not been established. On the other hand, since the speed of sound (SOS) is available for elasticity evaluation, we have proposed in vivo measurement method of SOS using ultrasound and MR images. This method determines the SOS based on the thickness measurement using the MR image and the time-of-flight (TOF) measurement using the ultrasound. In this study, this method was applied to the SOS measurement in tissue-engineered cartilage, and then relation between in vivo SOS and elasticity was investigated. As the result, in vivo SOS had a high correlation with Youngs modulus. This result reveals that in vivo evaluation of tissue-engineered cartilage elasticity is available through in vivo SOS measurement.

Calibration Method in Elasticity Evaluation of Regenerating Cartilage Based on Ultrasonic Particle Velocity

It is important in regenerative medicine to evaluate the maturity of regenerating tissue. In the maturity evaluation of regenerating cartilage, it is useful to measure the temporal change in elasticity because the maturity of regenerating tissue is closely related to its elasticity. In this study, a quantitative elasticity evaluation of extracted regenerating cartilage samples, which is based on the laser Doppler measurement of ultrasonic particle velocity and calibration, was experimentally investigated using agar-based phantoms with different Youngs moduli and regenerating cartilage samples extracted from beagles in animal experiments. The experimental results verified the feasibility of the proposed method for the elasticity evaluation of regenerating cartilage samples.

Characteristic X-ray absorptiometry applied to the assessment of tissue-engineered cartilage development.

BACKGROUND Transmission and tomographic X-ray measurements are useful in assessing bone structures, but only a few studies have examined cartilage growth because of the poor contrast in conventional X-ray imaging. OBJECTIVE In this study, we attempted to use the linear attenuation coefficient (LAC) as a metric of tissue-engineered cartilage development, which would be useful in high-throughput screening of cartilage products. METHODS Assuming that the LAC is related to the amount of extracellular matrix (ECM) in terms of the density and its atomic components, we measured X-ray absorption through tissue-engineered cartilage constructs. Characteristic X-ray beams from a molybdenum microfocus X-ray tube were employed to avoid beam hardening. The correlation of the LAC with mechanical properties was analyzed for verification. RESULTS The LAC was higher for chondrocyte constructs and lower for fibroblast-dominant constructs and was consistent with the quantification of toluidine blue staining, which is a proof of ECM production. The LAC was positively correlated with the bending modulus but negatively correlated with the dynamic elastic modulus and stiffness, possibly because of the remaining scaffold. CONCLUSIONS The LAC has the potential to be used as a metric of development of tissue-engineered cartilage. However, the calcified regions should be excluded from analysis to avoid decreasing the correlation between the LAC and the amount of ECM.

Biomechanics of Hydrocephalus: Part I. Mechanical properties of the brain

In considering the pathophysiology of hydrocephalus in children, it is necessary to consider properties of brain tissue itself in addition to circulation of cerebrospinal fluid. The first step of biomechanical approach to hydrocephalus is to clarify the mechanical properties of brain tissue.