To evaluate the effect of technique combination of implant-retented titanium lattice with decalcified dental matrix (DDM) implanting. Methods Six healthy male dogs (weighing of 10-20 kg) were randomly divided into 3 groups. All the premolars were extracted on both sides of the jaw in dogs. After 2 weeks, titanium lattice and implant were implanted in the maxillary premolar region with DDM on one side (experimental group), but without on the other side (control group) of each dog. After 4, 9 and 14 weeks, respectively, 2 animals were individually killed each time, and the samples wereevaluated by general observation, X-ray examination, histological observation and histomorphometric analyses. Results General observation: Among the 6 dogs, there was no postoperative infection or death. The X-ray examination showed that the bone density of the experimental group was greater than the control group at 4 and 9 weeks, and had no significant difference as to the vicinity bone at 14 weeks. On the other hand, the density of the control group was very low under the titanium lattice and around the implant. The experimental group revealed a ridge augment of (1.93 ± 0.24) mm, and control group (-1.02 ± 1.20) mm (P lt; 0.05). Developed bone sponge could be found after 14 weeks. Histological observation showed that in the experimental group, the DDM surface was nearly absorbed at 4 weeks. A few new bones were formed at 9 weeks. The whole DDM was absorbed; the trabecular bone was thick and arranged regularly; and the intergradations of implant were observed at 14 weeks. In the control group, there were some inflammatory fibers around the neck of implant at 4 weeks. The inflammatory condition extended to the root of implant and the titanium lattice at 9 weeks. There was no newly-formed bone under the titanium lattice at 14 weeks. Histomorphometric analyses showed that the implant contact bone ratio approached 1 ∶ 1, and showed no significant difference between the new bone fragment and former bone fragment in the experimental group. Conclusion This augmentation of alveolar ridge evaluated by the study is appl icable, but further study is necessary.
Objective To explore the construction of a canine model of vascularized allogeneic spinal cord transplantation (vASCT) and preliminarily evaluate its therapeutic efficacy for spinal cord injury (SCI). Methods Sixteen female Beagle dogs aged 8-12 months were randomly selected, with 8 dogs serving as donors for the harvesting of spinal cord tissue with a vascular pedicle [dorsal intercostal artery (DIA) at the T10 level and accompanying vein]. The remaining 8 dogs underwent a 1.5-cm-length spinal cord defect at the T10 level, followed by transplantation of the donor spinal cord tissue for repair. Polyethylene glycol (PEG) was applied to both ends to spinal cord graft; then, using a random number table method, the dogs were divided into an experimental group (n=4) and a control group (n=4). The experimental group received immunosuppressive intervention with oral tacrolimus [0.1 mg/(kg?d)] postoperatively, while the control group received no treatment. The operation time and ischemia-reperfusion time of two groups were recorded. The recovery of hind limb function was estimated by Olby score within 2 months after operation; the motor evoked potentials (MEP) was measured through neuroelectrophysiological examination, and the spinal cord integrity was observed through MRI. ResultsThere was no significant difference in the operation time and ischemia-reperfusion time between the two groups (P>0.05). All dogs survived until the completion of the experiment. Within 2 months after operation, all dogs in the control group failed to regain the movement function of hind limbs, and Olby scores were all 0. In the experimental group, the movement and weight-bearing, as well as walking abilities of the hind limbs gradually recovered, and the Olby scores also showed a gradually increasing trend. There was a significant difference between the two groups from 3 to 8 weeks after operation (P<0.05). Neuroelectrophysiological examination indicated that the electrical signals of the experimental group passed through the transplanted area, and the latency was shortened compared to that at 1 month after operation (P<0.05), showing continuous improvement, but the amplitude did not show significant improvement (P>0.05). The control group was unable to detect any MEP changes after operation. MRI examination showed that the transplanted spinal cord in the experimental group survived and had good continuity with normal spinal cord tissue, while no relevant change was observed in the control group. ConclusionThe vASCT model of dogs was successfully constructed. This surgical procedure can restore the continuity of the spinal cord. The combination of tacrolimus anti-immunity is a key factor for the success of transplantation.
The purpose of the study was to observe effect of chinese medicine “Qing Yi Tang” on the repair of injury of intestinal mucosa in acute necrotizing pancreatitis (ANP). Dogs ANP model were induced by injection of 5% sodium taurocholate (0.5 ml/kg) with 3 000 u/kg trypsin into the pancreatic duct. Diamine oxidase and anylase activity in blood, protein and MDA levels of ileal mucosa were to be determined in ANP and after treatment of “Qing Yi Tang”. Intestinal permeability was also to be studied, LPS and bacteria translocation (BT) were obseved. All animals were sacrificed on day 7, the tissue of ileal mocosa was collected for histological and ultrastructural studies. The results showed that after treatment with Chinese medicine “Qing Yi Tang”, the injury of intestinal mucosa in ANP reduced. The length, height, area and protein of ileal mucosa increased significantly, intestinal permenbility decreased, the levels of LPS reduced in 1-2 times, and organ BT rate also reduce in 50%. The results indicated that chinese medicine “Qing Yi Tang” had good effect on improving repair of intestinal mucosa injury, protecting gut barrier function, reducing the incidence of LPS and bacteria translocation.
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.
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.
【Abstract】 Objective To measure the changes of bone mineral density and bone micro-architecture of thefemoral head that harvested from the three-foot bearing ethanol destroyed canine model for osteonecrosis of femoral head, and discuss the influences of local injection of ethanol and biomechanical loading to the structural properties of the femoral head. Methods Twenty-four Beagles were divided randomly into four-foot bearing canines and three-foot bearing canines. One fore-l imb was fixed randomly in three-foot bearing canines. Osteonecrosis was induced in all experimental animals by local injection of 5 mL pure ethanol into one side of the femoral head. The hind l imbs injected with NS were acted as control group, that of three-foot canines injected with ethanol were acted as three-foot canine group, and that of four-foot canines injected with ethanol were acted as four-foot canine group. The contralateral femoral head was injected into equal amount of NS. Animals were sacrificed at the time intervals of 1, 3, 6, and 12 weeks after the injection of ethanol. Quantitative microcomputedtomography was used to characterize changes in bone micro-architecture and bone mineral density of femoralhead. Results The clear three-dimensional model of trabecular bone of necrotic femoral head were obtained. There were no significant differences among 3 groups according to the time l ine by 1 week after ethanol injection(P gt; 0.05). At 3 weeks after injection of ethanol, in three-foot canine group and four-foot canine group, the volume of BMC, BMD, BVF, and BS/BV increased gradually as the distance to the drill ing canal increased. There were significant differences between 3 regions (P lt; 0.05). At 6 weeks, in three-foot canine group and four-foot canine group, the volume of BMC, BMD,BVF, and Tb.N of region I and II decreased significantly compared with region III (P lt; 0.05). At 12 weeks, there are no differences among 3 groups (P gt; 0.05). There were significant decreases in BMD values, BVF, BS/BV, Tb.N, Tb.Sp and Tb.Th after the injection of pure ethanol. And, the changes were more and more obvious by the time l ine. These changes were differentiable at 3 weeks after injection of ethanol, and became obvious at 6 weeks. These changes were more obvious at the part that near the injection canal. The changes in threefoot canine group were more obvious than that in four-foot canine group. Conclusion Resorption of necrotic compact bone trabecular may weaken the structural properties of the femoral head. Moreover, remodel ing and repairing process of necrotic bone trabecular may be hampered by constant biomechanical loading that presented in three-foot bearing canines, and thereby further weaken the structural properties of the femoral head. Biomechanical loading may be one of the critical reasons that lead to the collapse of femoral head.
Objective Using chemically extracted acellular methods to treat extracranial section of the canine whole facial nerve, to evaluated its effects on nerve structure and the removal extent of Schwann cells and myel in. Methods Twenty whole facial nerves were exposed from 10 canines [weighing (18 ± 3) kg]. The extracranial trunk of canine facial nerve and its branches (temporal branch, zygomatic branch, buccal branch, marginal mandibular branch, and cervical branch) were dissected under l ight microscope. Twenty facial nerves were divided into the experimental group (n=12) and control group (n=8) randomly. In experimental group, the nerve was extracted with the 3%TritonX-100 and 4% sodium deoxycholate. In control group, the nerve was not extracted. HE staining and immunofluorescence histological stainings for Hoechst33258, P75, Zero, and Laminin were performed. Results After histological staining, it was found that myel in and Schwann cells were removed from the facial nerve while the basal lamina tube remained intact. The whole canine facial nerves (one nerve trunk and multiple nerve branches) had the similar result. Conclusion The canine whole facial nerve has natural structure (one nerve trunk and multiple nerve branches) by extracted with chemically extracted acellular methods, so it is an available graft for repairing the defect of the whole facial nerve.
Objective To verify adhesion and growth ability of canine esophageal epithelial cells (EECs) on the poly (lactic-co-glycolic acid) (PLGA), a three-dimensional biodegradable polymer scaffold, and to reconstruct the canine esophagus by the tissue engineering. Methods Free canine EECs isolated from adult dogs by esophagoscopy were seeded onto the PLGA scaffolds precoated with collagen type Ⅳ after the first passage by the in vitro culture. Then, the composites of the cell-scaffold were respectively cultured invitro and in the abdominal cavity of the dog in vivo. After different periods, the cell-seeded scaffolds were assessed by histological HE staining, scanning electron microscopy, and immunohistochemical analysis. Results The cells displayed a cobblestone-shaped morphology that was characteristic of the epithelial cells and were stained to be positive for cytokeratin, which indicated that the cells were EECs. The canine EECs were well distributed and adhered to the PLGA scaffolds, and maintained their characteristics throughout the culture period. After the culture in vivo for 4 weeks, the cell-seeded scaffolds looked like tissues. Conclusion PLGA scaffolds precoated with collagen type Ⅳ can be suitable for adhesion and proliferation of EECs, and can be used as a suitable tissue engineering carrier of an artificial esophagus.
Objective To provide an ideal seed cell for tissue engineered urinary bladder and urethra by serially culturing canine smooth muscle cells from urinary bladder in vitro and compare biological characteristics of different passagesof cells. Methods Bladder smooth muscle cells of 12-month-old male dogs weighing 10-12 kg were isolated from adult dogs’ urinary bladders by collagenase and trypsin digestion and serially cultured in DMEM medium supplemented with 10% serum of newborn bovines. Morphology and prol iferation of the cells were observed and the serially-cultured cells were identified with the transmission electron microscope and immunohistochemistry. Results The cells appeared spindle in parallel rows when they grew to the degree of subconfluence, and showed the “peak-valley” structure under the inverted phase contrast microscope. The cells could be prol iferated serially to the 12th passage in vitro. The growth curve showed the cells before the 7th passage had the similar prol iferation characteristics and the growth cycle was about 40 hours. The TEM showed myofilament and the dense body in cytoplasm of smooth muscle cells. Smooth muscle actin was positive by immunohistochemical staining. After the 7th passage, the cells’ growth became slow, and myofilament and the dense body in cytoplasm vanished. Conclusion The canine smooth muscle cells from urinary bladder can be serially cultured in vitro and highly purified and largely prol iferated by the appropriate method. The cells before the 7th passage can be used as optimal seed cells for tissue engineered urinary bladder and urethra.