Distraction osteogenesis(DO) is defined as a gradual mechanical process of mechanical stretching two vascularized bone surface apart with a critical rate and rhythm such that new bone forms within the expanding gap, reliably bridges the gap, and ultimately remodels to normal structure. DO has become a mainstay in bone tissue engineering and has significantly improved our armamentarium for reconstructive craniomaxillofacial procedures. But the molecular and biological mechanisms that regulate the formation of new bone during distraction osteogenesis are not completely understood. BMPs are potent osteoinductive agents. Our hypothesis was that BMPs, especially BMP-2 and BMP-4, might play an importent role in the signaling pathways that link the mechanical forces created by distraction to biological responses and in promting new bone formation. Using a rabbit's mandible, we investigated the expression of BMP-2, -4 at different time points during distraction osteogenesis. The purpose of this study is to research the pattern of expression of BMP-2, -4 in new bone formation during distraction osteogenesis of the rabbit mandible. The experimental group was applied gradual distraction (0.7mm a day by twice a day, 4.9mm in total, for 7 days) and the control group was carried out osteotomy alone. They were examined clinically, histologically, and by RT-PCR analysis. On 3 days after osteotomy, the high level of expression of BMP-2, -4 was detected. But, the expression of BMP-4 was decreased during latency period. As distraction was started, its expression was increased and maintained till postoperative 28days. In control group, the expression of BMP-4was remarkably decreased till postoperative 14 days. On the other hand, the expression of BMP-2 was no difference between experimental group and control group. The expression of BMP-4 was maintanined at high level during the entire experimental period in both group. These findings suggested that excellent bone formation during distraction osteogenesis is associated with enhanced expression of BMP-4 genes by mechanical tension stress.
Bone and Bones ; Hand ; Latency Period (Psychology) ; Mandible* ; Mechanical Processes ; Osteogenesis ; Osteogenesis, Distraction* ; Osteotomy

BACKGROUND: Despite the importance of soft tissue balancing during total knee arthroplasty (TKA), all estimating techniques are dependent on a surgeon's manual distraction force or subjective feeling based on experience. We developed a new device for dynamic gap balancing, which can offer constant load to the gap between the femur and tibia, using pneumatic pressure during range of motion. METHODS: To determine the amount of distraction force for the new device, 3 experienced surgeons' manual distraction force was measured using a conventional spreader. A new device called the consistent load pneumatic tensor was developed on the basis of the biomechanical tests. Reliability testing for the new device was performed using 5 cadaveric knees by the same surgeons. Intraclass correlation coefficients (ICCs) were calculated. RESULTS: The distraction force applied to the new pneumatic tensioning device was determined to be 150 N. The interobserver reliability was very good for the newly tested spreader device with ICCs between 0.828 and 0.881. CONCLUSIONS: The new pneumatic tensioning device can enable us to properly evaluate the soft tissue balance throughout the range of motion during TKA with acceptable reproducibility.
Arthroplasty, Replacement, Knee/*instrumentation/methods ; Biomechanics ; Equipment Design ; Femur/surgery ; Humans ; Knee Joint/physiology/*surgery ; Mechanical Processes ; Range of Motion, Articular ; Reproducibility of Results ; Tibia/surgery

BACKGROUND: Various decellularization methods have been studied in order to develop tissue graft which is less immunogenic and more durable. This study was performed to investigate the physico-dynamic and histological effect of trypsin pretreatment on decellularization protocols. MATERIAL AND METHOD: Two groups of bovine pericardium specimen each underwent decellularization process based on SDS and Triton X-100 or N-lauroylsarcosinate and Triton X-100. Two more groups additionally underwent pretreatment with 0.1% Trypsin/0.1% EDTA. After decellularization process, mechanical tensile strength was tested, then iomechanical test of permeability and compliance was tested before and after fatigue test. Light microscopy and electron microscopy was performed to observe histological findings. RESULT: There was no difference in mechanical tensile strength between groups, but permeability and compliance was decreased in trypsin pretreated groups. Light microscopic and electron microscopic findings revealed damage of the extracellular matrix in trypsin pretreated groups and in groups which underwent the fatigue test also. CONCLUSION: Trypsin pretreatment in decellularizing process of bovine pericardium damages extracellular matrix and increases permeability and compliance of the bovine pericardium, but did not decrease tensile strength. Further studies are needed to use enzymatic treatments in decellularization protocols.
Biomedical Engineering ; Bioprosthesis ; Compliance ; Edetic Acid ; Electrons ; Extracellular Matrix ; Fatigue ; Light ; Mechanical Processes ; Microscopy ; Microscopy, Electron ; Octoxynol ; Pericardium ; Permeability ; Sarcosine ; Tensile Strength ; Transplants ; Trypsin

We observed how combined mechanical stimuli affect the proliferation and differentiation of pre-osteoblasts. For this research, a bioreactor system was developed that can simultaneously stimulate cells with cyclic strain and ultrasound, each of which is known to effectively stimulate bone tissue regeneration. MC3T3-E1 pre-osteoblasts were chosen for bone tissue engineering due to their osteoblast-like characteristics. 3-D scaffolds were fabricated with polycaprolactone and poly-L-lactic acid using the salt leaching method. The cells were stimulated by the bioreactor with cyclic strain and ultrasound. The bioreactor was set at a frequency of 1.0 Hz and 10% strain for cyclic strain and 1.0 MHz and 30 mW/cm2 for ultrasound. Three experimental groups (ultrasound, cyclic strain, and combined stimulation) and a control group were examined. Each group was stimulated for 20 min/day. Mechanical stimuli did not affect MC3T3-E1 cell proliferation significantly up to 10 days when measured with the cell counting kit-8. However, gene expression analysis of collagen type-I, osteocalcin, RUNX2, and osterix revealed that the combined mechanical stimulation accelerated the matrix maturation of MC3T3-E1 cells. These results indicate that the combined mechanical stimulation can enhance the differentiation of pre-osteoblasts more efficiently than simple stimuli, in spite of no effect on cell proliferation.
Animals ; Bioreactors ; *Bone Regeneration ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Lactic Acid/chemistry ; *Mechanical Processes ; Mechanotransduction, Cellular/physiology ; Mice ; Osteoblasts/cytology/*metabolism ; Polyesters/chemistry ; Polymers/chemistry ; Tissue Engineering/methods ; Tissue Scaffolds/chemistry/utilization