Three-dimensional reconstruction technique in gastrocnemius flap surgery: initial clinical application / 南方医科大学学报

<p><b>OBJECTIVE</b>To discuss the experience with three-dimensional reconstruction technique in initial clinical application in gastrocnemius muscle flap surgery.</p><p><b>METHOD</b>From 2007 to 2008, 7 patients received gastrocnemius muscle flap surgeries to repair the wounds. Preoperative CT angiography or magnetic resonance imaging (MRI) was performed after injection of the contrast media for individualized three-dimensional gastrocnemius muscle flap reconstruction using Amira4.1 software. According to the size of the defect in the wound, individualized three-dimensional gastrocnemius muscle flap was designed and harvested from the posterior leg.</p><p><b>RESULTS</b>Individualized three-dimensional reconstruction of the gastrocnemius flap was performed in 7 cases, and the reconstructed flaps clearly displayed the blood vessels, skin and the adjacent three-dimensional structures. In 6 cases the main perforating branched and trunk of the blood vessels in the designed flap were consistent with the surgical findings; in 1 case, the perforating branches failed to be clearly displayed in the designed flap, and surgical examination identified perforating branches with an average diameter of 0.5 mm (minimally 0.3 mm). The flaps survived in all the 7 cases.</p><p><b>CONCLUSIONS</b>Three-dimensional reconstruction of the gastrocnemius flap based on the lower limb CT angiography or MRI allows three-dimensional observation of the anatomy of the flap and accurate marking of the extent of the flap to be harvested, therefore avoiding intraoperative injuries to the blood vessels to better survival of the flaps.</p>

Green fluorescent protein as a tracer of bone marrow stromal cells in bone tissue engineering in rhesus / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the role of green fluorescent protein (GFP) in tracing rhesus bone marrow stromal cells (rBMSCs) during tissue-engineered bone formation in vivo.</p><p><b>METHODS</b>Ad5.CMV-GFP was amplified by infecting QBI-293A cells, and the bone marrow was harvested from the ilium of adult male rhesus to obtain rBMSCs, which were cultured and passaged in vitro. GFP was transfected into the third-passage rBMSCs via adenovirus vector and the labeled cells were inoculated into absorbable HA scaffold and cultured for 3 days, with untransfected rBMSCs as control, before the cell-matrix compounds were implanted into the latissimus dorsi muscles of rhesus. Samples were harvested at 6 week and embedded in paraform, and ground sections of the bone tissue were prepared to observe green fluorescence under laser scanning confocal microscope. Propidium iodide staining of the sections was also performed for observation.</p><p><b>RESULTS</b>The rBMSCs grew well after GFP transfection, and green fluorescence could be seen 24 h after the transfection and became stronger till 48 h, with a positive transfection rate beyond 80%. Six weeks after cell implantation, the rBMSCs labeled by GFP-emitted green fluorescence were detected in the bone tissue under laser scanning confocal microscope.</p><p><b>CONCLUSION</b>GFP can effectively trace BMSCs during bone tissue engineering, and the transplanted BMSCs constitute the main source of bone-forming cells in bone tissue engineering.</p>

Distribution and property of nerve fibers in human long bone tissue / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To observe the distribution of the nerve fibers in the bone tissue and the entry points of these fibers into the bone.</p><p><b>METHODS</b>The adult tibia was used for the ground sections which were afterwards made into the slice sections by decalcification in ethylenediamine tetraacetic acid (EDTA). The ground sections were stained in silver and the slice sections were stained in silver and haematoxylin and eosin (HE) respectively. Then, the samples of the transmission electron microscope and the atomic force microscope were made and observed.</p><p><b>RESULTS</b>In the human long bone tissue, many nerve fibers were distributed in the membrane, cortical bone, cancellous bone and marrow. The nerve fibers entered the bone from the nutrient foramen, and passed through the nutrient canal, Haversian's canal and Volkmann's canal, and finally into the bone marrow. In the nutrient canal, the nerve fibers, mainly the medullary nerve fibers, followed the blood vessel into the bone. In the cortical bone, the nerve fibers also followed the blood vessels and were mainly distributed along Haversian's canal and Volkmann's canal. In the bone trabecular and bone marrow, there were many nerve fiber endings arranged around the blood vessels, mainly around the tunica media of medium-size arteries in the marrow and around capillary blood vessels, and a few scattered in the bone marrow. There were sporadic nerve endings in epiphyseal plate and no nerve fibers permeated epiphysis to diaphysis. No distribution of nerve fibers could be found in cartilaginous part.</p><p><b>CONCLUSIONS</b>There are many nerve fibers in bone and the nerve passageway is nutrient foramen, Volkman's canal, Haversian's canal and bone marrow.</p>

Perfusion-weighted magnetic resonance imaging for monitoring vascularization in tissue-engineered bone in rhesuses / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of perfusion-weighted magnetic resonance (MR) imaging (PWMRI) in monitoring vascularization in tissue-engineered bone graft.</p><p><b>METHODS</b>Tibial diaphyseal defect of 20 mm was induced in 25 lower limbs of 13 rhesuses and fixed with an AO reconstruction plate with 7 holes. The monkeys were randomized into 5 groups according to the materials used for defect filling: group A, with beta-tricalcium phosphate (beta-TCP), bone marrow stromal cells (BMSCs) and blood vessel bundles; group B, with beta-TCP and blood vessel bundles; group C, with beta-TCP and BMSCs; group D, with beta-TCP, and group E without filling. PWMRI, X-ray, and radionuclide imaging were carried out at weeks 4, 8, 12 postoperatively. The maximum slope rates of the single intensity-time curve (SS(max)) and the baseline values (SI(baseline)) on the same time points were calculated. Transmittances on the X-ray films and isotope counts in the region of interest (ROI) were assessed and calculated.</p><p><b>RESULTS</b>Compared with other groups, group A showed the highest SS(max) at weeks 4, 8, and 12 postoperatively, and its SS(max) at week 8 was significantly higher than that at week 4 (P=0.003). The SS(max) was positively related to isotope counts in ROI at week 8 after operation (r(s)=0.899, P=0.038), and inversely related to transmittance on X-ray films at week 12 (r(s)=-0.892, P=0.042).</p><p><b>CONCLUSION</b>The SS(max) of the single intensity-time curve can accurately reflect the vascularization of the tissue-engineered bone graft, and PWMRI allows sensitive, quantitative, noninvasive and radiation-free vascularization monitoring.</p>

The regulatory effect of human bone morphogenetic protein 7 gene transfer on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To detect the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSc) transferred by retroviral vector carrying human bone morphogenetic protein 7 (hBMP-7) gene.</p><p><b>METHODS</b>hBMP-7-expressing replication-deficient retroviral vector(PT-PLNCX2-hBMP7) was reconstructed using clone technique and recombinant DNA technique. BMSc were infected with the virus granules. The protein of BMP-7 gene in transferred cells were determined by immunohistochemistry. The proliferativity of the transferred cell were assayed by methabenzthiazuron (MTT) method and flow cytometer. Alkaline phosphatase (ALP) were also detected using enzyme kinetics.</p><p><b>RESULTS</b>Cells transferred by PT-PLNCX2-hBMP7 expressed abundant hBMP7 protein in the cytoplasm. Positive findings were not found in those cells that were not transferred. After infected with virus there were not significant difference of cell proliferation and cell cycle between the cells transferred by hBMP-7 or not (P > 0.05). ALP activity in transferred cells were increased significantly (P < 0.01).</p><p><b>CONCLUSIONS</b>hBMP-7 can be transferred and stably expressed in the cultured rabbit bone marrow stem cells. Proliferation and cell cycle of the transferred cell were not affected. hBMP7 gene transfer can be used to induce differentiation of BMSc into osteoblast-like cells.</p>

The method of accelerating osteanagenesis and revascularization of tissue engineered bone in big animal in vivo / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study whether tissue engineered bone can repair the large segment bone defect of large animal or not. To observe what character the fascia flap played during the osteanagenesis and revascularization process of tissue engineered bone.</p><p><b>METHODS</b>9 Chinese goats were made 2 cm left tibia diaphyseal defect. The repairing effect of the defects was evaluated by ECT, X-ray and histology. 27 goats were divided into three groups: group of CHAP, the defect was filled with coral hydroxyapatite (CHAP); group of tissue engineered bone, the defect was filled with CHAP + bone marrow stroma cells (BMSc); group of fascia flap, the defect was filled with CHAP + BMSc + fascia flap. After finished culturing and inducing the BMSc, CHAP of group of tissue engineered bone and of fascia flap was combined with it. Making fascia flap, different materials as described above were then implanted separately into the defects. Radionuclide bone imaging was used to monitor the revascularization of the implants at 2, 4, 8 weeks after operation. X-ray examination, optical density index of X-ray film, V-G staining of tissue slice of the implants were used at 4, 8, 12 weeks after operation, and the biomechanical character of the specimens were tested at 12 weeks post operation.</p><p><b>RESULTS</b>In the first study, the defect showed no bone regeneration phenomenon. 2 cm tibia defect was an ideal animal model. In the second study, group of CHAP manifested a little trace of bone regeneration, as to group of tissue engineered bone, the defect was almost repaired totally. In group of fascia flap, with the assistance of fascia flap which gave more chance to making implants to get more nutrient, the repair was quite complete.</p><p><b>CONCLUSIONS</b>The model of 2 cm caprine tibia diaphyseal defect cannot be repaired by goat itself and can satisfy the tissue engineering's demands. Tissue engineered bone had good ability to repair large segment tibia defect of goat. Fascia flap can accelerate the revascularization process of tissue engineered bone. And by this way, it augment the ability of tissue engineered bone to repair the large bone defect of goat.</p>