Objective To make a comparison for the change of maximum tensile intensity and stiffness of a whole implant that is placed into bone tunnel with various lengths tendon, by using beagle dog’s autogenous flexor tendons to reconstruct anterior cruciate l igament (ACL). Methods Sixty male beagle dogs were included in the experiment (weighting 13-16 kg). Three dogs were used for intact flexor tendon of both knees (normal control group), 3 dogs for the intact ACL andfemur-graft-tibia complex (auto control group) and 54 dogs (108 knees) for models of reconstructed ACL (6 experimentalgroups according to different lengths of tendon: 5, 9, 13, 17, 21 and 25 mm in the bone tunnel). The tensile intensity and stiffness were measured after 45, 90 and 180 days separately after operation. Results In the normal control group, the maximum tensile intensity of the intact flexor tendon was (564.15 ± 36.18) N, the stiffness was (59.89 ± 4.28) N/ mm. In the auto control group, the maximum tensile intensity of the intact ACL was (684.75 ± 48.10) N, the stiffness was (74.34 ± 6.99) N/ mm, all ruptured through the intra-articular portion of the graft. The maximum tensile intensity of femur-graft-tibia complex in the auto control group was (301.92 ± 15.04) N, the stiffness was (31.35 ± 1.97) N/mm. After 45 days of operation, all failure occurred at the tibial or femoral insertion site. After 90 days of operation, 24 of the breakpoints were scattered in tendon-bone junction, 12 (3 in 17 mm group, 5 in 21 mm group, 4 in 25 mm group) ruptured through the intra-articular portion. After 180 days of the operation, all breakpoints were distributed inside joint of the implant. The maximum tensile intensity and the stiffness were ber in 17, 21 and 25 mm groups than in 5, 9 and 13 mm groups after operation (P lt; 0.05). Conclusion Tendon with 17 mm length, which will be implanted into bone tunnel, is an appl icable index, in reconstruction of ACL by autogenous tendons.
Objective-To apply self-pulmonary tissue flap to reconstruct esophagus directly or with alloy stent in this research. Methods Twenty-four dogs were divided into two groups, middle bronchus was ligated to prepare pulmonaryflap and incised, a 4 to 6 cm long and 1/2 to 2/3 perimeter defect was made in esophageal wall. Esophagus defect was repaired only with pulmonary flap (experimental group) and with pulmonary flap having self-expanded stent inside (control group). The gross appearance, histological apearance and barium X-ray films were observed at 2,4,6,8,10 and 12 weeks after operation. Results Two dogs died of anatomotic leak in experimental group, three dogs died of anatomotic leak and two dogs died of perforation of ulcer in control group. The growth of esophagus epithelium was observed from periphery area to central area after 8 to 10 weeks of operation. In pulmonary flap mass fibrous tissue proliferated and fibroblasts were active, but no necrosis occurred. Barium X-ray ofregenerated esophagus showed that mild stenosis and weakened peristalisis were observed in the middle of resophagus replacement, and that no obstruction, leakage, and dilation above anastomotic stoma occurred. Conclusion Pulmonary tissue flap can well support the mucosa crawl in the defect of esophagus. It is necessary to find a more suitable and satisfied stent for repairing segmental defect.
Objective To investigate an improved large vascular reconstruction method in the canine liver transplantation and see whether it can shorten the anheptic time and thus reduce the harmful effects during the anhepatic phase. Methods Thirty-two mongrel dogs were enrolled and divided into two groups randomly:the donor group (n=16) and the acceptorgroup(n=16). The dogs in the acceptor group were divided into two groups, according to the different reconstruction methods: Group A using the magnetic rings for a large vein reconstruction in the canine liver transplantation (n=10), and Group B using a handsewing large vein reconstruction in the canine liver transplantation (n=6). The operation time, hemodymics change, anastomosis site, and survival were observed. Results The operation time was as follows: In Group A, the total operation time, the inferior vena cava anastomosistime, and the anheptic phase time were significantly shorter than those in Group B (3.24±0.49 h vs 4.12±0.51 h，5.89±2.27 min vs 28.33±6.04 min，3.89±0.73 min vs 12.16±3.72 min),with a significant difference between the two groups (Plt;0.01). The haemodymics changes were as follows: In Group A, MAP dropped during the anhepatic phase, but it soon recovered after reperfusion，and there was only 730.56±150.56 ml of fluid including the donor blood that needed to be transfused, with no pressor agent required. In Group B, blood pressure dropped during the anhepatic phase，but it slowly recovered，and there was 2241.67±390.78 ml of fluid. In Group A, all the stomas had no errhysis, twistor thrombus. The twisted stomas could be corrected by the revolving of the magnetic rings. The endangium at the site of anastomosis was smooth. In Group B, most of the stomas had errthysis. In Group A, 3 dogs survived for more than 7 days, 6dogs survived for 3-6 days, and 1 dog survived for only 12 hours. In Group B, 2 dogs survived for 3-6 days, 3 dogs survived for 1-2 days, and 1 dog survivedfor only 12 hours. Conclusion Using the magnetic rings for a large vascular reconstruction in the canine liver transplantation is an improvedmethod, which can simplify the anastomosis procedures and significantly shortenthe anheptic phase time. However, the magnetic rings have to be placed in the abdomen, so this method remains to be further improved.
The model of acute hemorrhagic necrotizing pancreatitis (AHNP) was produced by retrograde forced injection of autogenous bile into the main pancreatic duct. The result showed that urgent surgical (transduodenal) intra-pancreatic duct drainage reduced the mortality rate of the experimental animals. Keeping the drainage patent and clear can both abate the pathological changes and promote the renovation of the dogs pancreas. So it can guide the clinical treatment in some way.
Objective To explore an effective method of culturing the canine bladder smooth muscle cells, observe the morphological characteristics of the bladder smooth muscle cells growing on acellular small intestinal submucosa（SIS） and offer an experimental basis for reconstruction of the bladder smooth muscle structure by the tissue engineering techniques. Methods The enzymetreatment method and the explant method were respectively used to isolate and harvest the canine bladder smooth muscle cells, and then a primary culture of these cells was performed. The canine bladder smooth musclecells were seeded on the SIS scaffold, and the composite of the bladder smooth muscle cells and the SIS scaffold were co cultured for a further observation. At 5,7 and 9 days of the co culture, the specimens were taken; the bladder smooth muscle cells growing on the SIS scaffold were observed by the hematoxylin staining, the HE staining, and the scanning electron microscopy. The composite of the bladder smooth muscle cells on the SIS scaffold was used as the experimental group, and the bladder smooth muscle cells with no SIS were used as the control group. In each group, 9 holes were chosen for the seeded bladder smooth muscle cells, and then the cells were collected at 3, 5 and 7 days for the cell counting after the enzyme treatment. Morphological characteristics of the cells were observed under the phase contrast microscope and the transmission electron microscope. Expression of the cell specific marker protein was assessed by the immunohistochemical examinaiton. The proliferation of the cells was assessed by the cell counting after the seeding on the SIS scaffold. Results The primary bladder smooth muscle cells that had been harvested by the enzyme treatment method were rapidly proliferated, and the cells had good morphological characteristics. After the primary culture in vitrofor 5 days, the bladder smooth muscle cells grew in confluence. When the bladder smooth muscle cells were seeded by the explant method, a small amount of the spindleshaped bladder smooth muscle cells emigrated from the explant at 3 days. The cells were characterized by the welldeveloped actin filaments inthe cytoplasm and the dense patches in the cell membrane under the transmissionelectron microscope. The immunohistochemical staining showed the canine bladdersmooth muscle cells with positive reacting α actin antibodies. The bladder smooth muscle cells adhered to the surface of the SIS scaffold, growing and proliferating there. After the culture in vitro for 5 days, the smooth muscle cells covered all the surface of the scaffold, showing a singlelayer cellular structure. The cell counts at 3, 5 and 7 days in the experimental group were（16.85±0.79）×105，（39.74±2.16）×105 and （37.15±2.02）×105, respectively. Thecell counts in the control group were（19.43±0.54）×105，（34.50±1.85）×105 and （33.07±1.31）×105, respectively. There was a significant difference between the two groups at 5 days （P＜0.05）. ConclusionWith the enzyme treatment method, the primarily cultured canine bladder smooth muscle cells can produce a great amount of good and active cells in vitro. The acellular SIS can offer an excellent bio scaffold to support the bladder smooth muscle cells to adhere and grow, which has provided the technical foundation for a further experiment on the tissue engineered bladder reconstruction.
To evaluatae the role of cathepsin B in the pathogenesis of acute pancreatitis. The experimental dogs were divided into three groups based on the severity. An acute edematous pancreatitis group (n＝11), an acute hemorrhagic necrotizing pancreatitis group (n＝12), and a control group (n＝7). Distribution of cathepsin B in pancreatic aciner cell was studied. Results: there was a redistribution of cathepsin B from the lysosomal fraction to the zymogen fraction. The results indicated that cathepsin B play a important role in the pathoagenesis of acute pancreatitis.
Abstract: Objective To observe the changes in morphology, structure, and ventricular function of infarct heart after bone marrow mononuclear cells (BMMNC) implantation. Methods Twenty-four dogs were divided into four groups with random number table, acute myocardial infarction (AM I) control group , AM I-BMMNC group , old myocardial infarct ion (OMI) control group and OM I-BMMNC group , 6 dogs each group. Autologous BMMNC were injected into infarct and peri-infarct myocardium fo r transplantation in AM I-BMMNC group and OM I-BMMNC group. The same volume of no-cells phosphate buffered solution (PBS) was injected into the myocardium in AM Icontrol group and OM I-control group. Before and at six weeks of cell t ransplantation, ult rasonic cardiography (UCG) were performed to observe the change of heart morphology and function, then the heart was harvested for morphological and histological study. Results U CG showed that left ventricular end diastolic dimension (LV EDD) , left ventricular end diastolic volume (LVEDV ) , the thickness of left ventricular postwall (LVPW ) in AM I-BMMNC group were significantly less than those in AM I-control group (32. 5±5. 1mm vs. 36. 6±3. 4mm , 46. 7±12. 1m l vs. 57. 5±10. 1m l, 6. 2±0. 6mm vs. 6. 9±0. 9mm; P lt; 0. 05). LVEDD, LVEDV , LVPW in OM I-BMMNC group were significantly less than those in OM I-control group (32. 8±4. 2 mm vs. 36. 8±4. 4mm , 48. 2±12. 9m l vs. 60.6±16.5m l, 7. 0±0. 4mm vs. 7. 3±0. 5mm; P lt; 0. 05). The value of eject fraction (EF) in OM I-BMMNC group were significantly higher than that in OM I-control group (53. 3% ±10. 3% vs. 44. 7%±10. 1% ). Compared with their control group in morphological measurement, the increase of infarct region thickness (7. 0 ± 1. 9mm vs. 5. 0 ±2.0mm , 6.0±0. 6mm vs. 4. 0±0. 5mm; P lt; 0. 05) and the reduction of infarct region length (25. 5±5. 2mm vs. 32. 1±612mm , 33. 6±5. 5mm vs. 39. 0±3. 2mm , P lt; 0. 05) were observed after transplantation in AM I-BMMNC group and OM I-BMMNC group, no ventricular aneurysm was found in AM I-BMMNC group, and the ratio between long axis and minor axis circumference of left ventricle increased in OM I-BMMNC group (0. 581±0. 013 vs. 0. 566±0.015; P lt; 0. 05). Both in AM I-BMMNC group and OM I-BMMNC group, fluorescence expressed in transplantation region was observed, the morphology of most nuclei with fluorescencew as irregular, and the differentiated cardiocyte with fluorescence was not found in myocardium after transplantation. The histological examination showed more neovascularization after transp lantation both in AMI and in OM I, and significant lymphocyte infiltration in AM I-BMMNC group. Conclusion BMMNC implantation into infarct myocardium both in AMI and OMI have a beneficial effect, which can attenuate deleterious ventricular remodeling in morphology and st ructure, and improve neovascularization in histology, and improve the heart function.
Objective o study the feasibility of homologous vascularized nerve transplantation after ultra deep cryopreservation. Methods Vascularized sciatic nerve from 12 female dogs was transplanted after ultra deep cryopreservation. Fortyeight male dogs were divided into 4 groups: ultra deep cryopreservation homologous vascularized nerve (group A), ultra deep cryopreservation homologous nerve (group B), fresh homologous vascularized nerve (group C), and fresh autologous vascularized nerve (group D). The gross appearance, patency rate of arteryand morphological transplanted nerve were observed 1, 4 and 12 weeks after transplantation respectively. Immunological analysis was performed using IL 2 assay and T lymphocyte subpopulations assay after 4 weeks. Image pattern analysis andelectromyogram were observed after 12 weeks. Results In groups A and D, no toe ulcer occurred, the atrophy of later limb and the sense of pain from skin of calf were restore significantly in the postoperative 12th week. In groups B and C, toe ulcer occurred, the atrophy of later limb and the sense of pain from skin of calf were not restored significantly in the postoperative 12thweek. The vessel patency rate of groups A and D was 83.3%, which was significantly higher than that of group C (50%,Plt;0.05). The changes of IL2 and Th, Ts in group C were significantly higher than that in groups A,B,D(Plt;0.01). There were increased vessel and regenerated nerve in transplanted nerve under optical microscope and image pattern analysis in groups A and D. There were shorter latent period of motor evoked potential, greater amplitude of action potenlial and faster motor nerve conducting velocity in groups A and D after 12 weeks. Conclusion The antigenicity of the homologous never and vessel may be reduced significantly by being frozen, and cryopreserved vascularized nerve can transferred successfully without the use of immunosuppressive agents. Vascularized nerve may restore good significantly for the thick nerve.
Objective To investigate the antibacterial and osteogenic capabil ities in vivo of hydroxyapatite (HA)/silver (Ag) coating. Methods HA/Ag coating (Ag qual ity percentage was 3%) and HA coating were deposited to external fixator Schanz screws. The tibial fracture model was establ ished in right hindl imb of 18 adult male Beagle dogs (weighing 15-20 kg). Thetibia was stabil ized with an external fixator and 2 Schanz screws of HA coating at proximal tibia (control group, n=18) and HA/Ag coating at distal tibia (experimental group, n=18), and every screw incision was infected with Staphylococcus aureus. Infection in screw holes and the changes of bone-screw interface were observed by wound grading and X-ray films. Results In control group, wounds infection became worse with time (χ2=13.492, P=0.001), while in experimental group, no obvious change was observed (χ2=0.208, P=0.901). The wound grading of experimental group was significantly better than that of the control group at 1, 2, and 3 weeks (P lt; 0.05). Laser scanning confocal microscope showed that there was bacterial adhesion on the surface of screws in 2 groups, viable becteria mainly in control group and non-viable becteria mainly in experimental group. The scanning electron microscope (SEM) observation results of the fractured sclerous tissue section showed that an obvious transparent boundary between screw and bone in control group, but no obvious boundary in experimental group. The osseointegration ratios were 76.23% ± 15.54% in control group and 93.42% ± 5.53% in experimental group, showing significant difference (t=8.843, P=0.000). The SEM observation showed that HA/Ag coating integrated with new bone and the surface of implant was filled with new bone in experimental group; obvious interspace was seen between the HA coating and new bone in control group. Conclusion HA/Ag coating has good antibacterial and osteogenic capabil ities, so it can take effects in preventing infection in screw holes and loosening of implants.
目的 建立犬开放性气胸海水浸泡的实验模型 ,探讨实验动物早期死亡原因。 方法 2 0条健康成年杂种犬随机分为两组。对照组 :实验动物受伤后直接观察 ;实验组 :动物受伤后置入人工配制的海水中。监测血流动力学、呼吸、血液渗透压、血液电解质、动脉血气变化以及肺部病理改变。 结果 实验组死亡率明显高于对照组 ,平均生存时间为 45分钟。实验组经海水浸泡后有急性呼吸和循环功能衰竭、严重电解质平衡紊乱、高渗血症、重度肺损伤以及严重代谢性和呼吸性酸中毒。 结论 开放性气胸后海水浸泡可引起一系列严重的病理生理变化 ,其结果是导致实验动物早期死亡的重要原因。