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Correct assessment of the bone healing process is required for the management of limb immobilization during the treatment of bone injuries, including fractures and defects. Although the monitoring of bone healing using ultrasound poses several advantages regarding cost and ionizing radiation exposure compared with other dominant imaging methods, such as radiography and computed tomography (CT), traditional ultrasound B-mode imaging lacks reliability and objectivity. However, the body structures can be quantitatively observed by ultrasound frequency-based methods, and therefore, the disadvantages of B-mode imaging can be overcome. In this study, we created a femoral bone hole model of a rat and observed the bone healing process using the quantitative ultrasound method and micro-CT, which provides a reliable assessment of the tissue microstructure of the bone. This study analyzed the correlation between these two assessments. The results revealed that the quantitative ultrasound measurements correlated with the CT measurements for rat bone healing. This ultrasound frequency-based method could have the potential to serve as a novel modality for quantitative monitoring of bone healing with the advantages of being less invasive and easily accessible. Impact statement Bone healing monitoring with ultrasound is advantageous as it is less invasive and easily accessible; however, the traditional B-mode method lacks reliability and objectivity. This study demonstrated that the proposed ultrasound frequency-based monitoring method can quantitatively observe bone healing and strongly correlates with the computed tomography measurements for rat bone healing. This method has the potential to become a reliable modality for monitoring bone healing.

Most spine fusion procedures involve the use of prosthetic fixation devices combined with autologous bone grafts rather than biological treatment. We had shown that spine fusion could be achieved by injection of bone morphogenetic protein-2 (BMP-2)-expressing mesenchymal stem cells (MSCs) into the paraspinal muscle. In this study, we hypothesized that posterior spinal fusion achieved using genetically modified MSCs would be mechanically comparable to that realized using a mechanical fixation. BMP-2-expressing MSCs were injected bilaterally into paravertebral muscles of the mouse lumbar spine. In one control group BMP-2 expression was inhibited. Microcomputed tomography and histological analyses were used to evaluate bone formation. For comparison, a group of mouse spines were bilaterally fused with stainless steel pins. The harvested spines were later tested using a custom four-point bending apparatus and structural bending stiffness was estimated. To assess the degree to which MSC vertebral fusion was targeted and to quantify the effects of fusion on adjacent spinal segments, images of the loaded spine curvature were analyzed to extract rigidity of the individual spinal segments. Bone bridging of the targeted vertebrae was observed in the BMP-2-expressing MSC group, whereas no bone formation was noted in any control group. The biomechanical tests showed that MSC-mediated spinal fusion was as effective as stainless steel pin-based fusion and significantly more rigid than the control groups. Local analysis showed that the distribution of stiffness in the MSC-based fusion group was similar to that in the steel pin fusion group, with the majority of spinal stiffness contributed by the targeted fusion at L3-L5. Our findings demonstrate that MSC-induced spinal fusion can convey biomechanical rigidity to a targeted segment that is comparable to that achieved using an instrumental fixation.

The biological effect of the perfluorocarbon-based artificial oxygen carrier (Oxygent) was investigated in tissue-engineered trachea (TET) construction. Media supplemented with and without 10% Oxygent were compared in all assessments. Partial tissue oxygen tension (PtO(2)) was measured with polarographic microprobes; epithelial metabolism was monitored by microdialysis inside the TET epithelium perfused with the medium underneath. Chondrocyte-DegraPol constructs were cultured for 1 month with the medium before glycosaminoglycan assessment and histology. Tissue reaction of TET epithelial scaffolds immersed with the medium was evaluated on the chick embryo chorioallantoic membrane. Oxygent perfusion medium increased the TET epithelial PtO(2) (51.2 +/- 0.3 mm Hg vs. 33.4 +/- 0.3 mm Hg at 200 microm thickness; 12.5 +/- 0.1 mm Hg vs. 3.1 +/- 0.1 mm Hg at 400 microm thickness, p < 0.01) and decreased the lactate concentration (0.63 +/- 0.08 vs. 0.80 +/- 0.06 mmol/L, p < 0.05), lactate/pyruvate (1.87 +/- 0.26 vs. 3.36 +/- 10.13, p < 0.05), and lactate/glucose ratios (0.10 +/- 0.00 vs. 0.29 +/- 0.14, p < 0.05). Chondrocyte-DegraPol in Oxygent group presented lower glycosaminoglycan value (0.03 +/- 0.00 vs. 0.13 +/- 0.00, p < 0.05); histology slides showed poor acid mucopolysaccharides formation. Orthogonal polarization spectral imaging showed no difference in functional capillary density between the scaffolds cultured on chorioallantoic membranes. The foreign body reaction was similar in both groups. We conclude that Oxygent increases TET epithelial PtO(2), improves epithelial metabolism, does not impair angiogenesis, and tends to slow cartilage tissue formation.

Because cardiomyocytes lose their ability to divide after birth, any subsequent cell loss or dysfunction results in pathologic cardiac rhythm initiation or impulse conduction. Strategies to restore and control the electrophysiological activity of the heart may, therefore, greatly affect the regeneration of cardiac tissue functionality. Using lentivirus-derived particles to regulate the bone morphogenetic protein-2 (BMP-2) gene expression in a pristinamycin- or gaseous acetaldehyde-inducible manner, we demonstrated the adjustment of cardiomyocyte electrophysiological characteristics. Complementary metal oxide semiconductor-based high-density microelectrode arrays (HD-MEAs) were used to monitor the electrophysiological activity of neonatal rat cardiomyocytes (NRCs) cultured as monolayers (NRCml) or as microtissues (NRCmt). NRCmt more closely resembled heart tissue physiology than did NRCml and could be conveniently monitored using HD-MEAs because of their ability to detect low-signal events and to sub-select the region of interest, namely, areas where the microtissues were placed. Cardiomyocyte-forming microtissues, transduced using lentiviral vectors encoding BMP-2, were capable of restoring myocardial microtissue electrical activity. We also engineered NRCmt to functionally couple within a cardiomyocyte monolayer, thus showing pacemaker-like activity upon local regulation of transgenic BMP-2 expression. The controlled expression of therapeutic transgenes represents a crucial advance for clinical interventions and gene-function analysis.

We have previously shown that penile corporal structures engineered using autologous cells are able to achieve erection, penetration, and ejaculation. However, fully functional corpora could not be engineered because of the limited cellular content present within the corporal tissue construct. In this study, we investigated whether a dynamic seeding approach would improve cellularity within the corporal tissue construct and thereby restore normal erectile function. Corporal cells were either statically or dynamically seeded on acellular corporal tissue matrices and maintained in a bioreactor system. After 48 h, the cell-matrix complexes were implanted subcutaneously in athymic mice and analyzed for cell attachment, survival, and distribution using histological and molecular techniques. Native tissues and matrices without cells served as controls. The seeded cells attached and proliferated within the sinusoidal walls of the matrices. After completing the seeding, the DNA and cellular content of the dynamically seeded matrices reached 71% of normal corpora, whereas the statically seeded matrices reached 39% of normal corpora. These findings were confirmed histologically, biochemically, and using scanning electron microscopy. This study demonstrates that dynamic cell attachment, using a bioreactor system, leads to the formation of morphologically and biochemically improved corporal tissue, which may be useful for penile reconstruction.