Some Chinese traditional medicines were found to inhibit rejection of graft. The antirejection effects of chuanxiong, LCH and HXI in thyroid allografts of rabbits were studied for selecting an immune depressor from Chinese traditional medicine with efficient and less sideeffect. The rabbits were divided into 5 groups in the study, with 7 in each group. Group I: The control group, no drug was used. Group II: dexamethason 0.25mg/kg/day, intramuscularly. Group III: chuanxiong water solution, 5g/kg/day, orally. Group Ⅳ： LCH water solution, 10g/kg/day, orally. Group Ⅴ： HXI water solution, 6g/kg/day, orally. The medication was given for 28 days. The grafted thyroids were removed for histopathological examination on the 28th day postoperatively and were scored and classified. The rejection and the survival of grafts were scored and classfied according to the La Rosa and Warrens criterion. The histopathological findings were as following: in Group I, follicles were badly damaged with much lymphocytes infiltration and fibrosis; in Gracup Ⅱ， two rabbits died, the other three showed damaged of the thyroid tissue and much lymphocytes infiltration; in group Ⅲ and Ⅴ, three cases showed damage of thyroid tissue, however, better revascularization was evident in Group Ⅲ; in Group Ⅳ, there was one case with much lymphocytes infiltration. It seemed that the degree of damage of grafts in the experimental groups was better than that in the control group, and had less lymphocytes infiltration, especially in Group Ⅳ. It was suggested that chuanxiong, LCH, HXI and dexamethason could protect the grafted thyroid, but the sideeffect of dexamethason was more than the other three. The antirejection of LCH was the best of the three. It was worth doing more research. HXI.
The damage effects of the pure tumor necrosis factor (TNF) on the normal animals were observed. Eighteeen rabbits were divided into two groups, eight in tested group and ten in control group. 0.5mg per kg of the pure rabbit TNF was given to each animal of the tested group. Results:The symptoms similar to that induced by endotoxin appeared after the TNF injection. The functions of the main organs were markedly damaged. The arterial blood pressure of most animal was low. The weight ratio of the orgen to the body was raised. The pathologic changes were similar to those of the multiple organ failure (MOF) model. Most of the animal died before the end of the experiment. The results suggest that pure TNF could indece multiple organ damages similar to those of MOF.
ObjectiveTo study the effect of chemical extraction of allogeneic tendon and allogeneic chondrocytes for reconstruction of anterior labrum of shoulder joint in rabbits.MethodsThe body weight of 45 adult New Zealand white rabbits ranged from 2.5 to 3.0 kg. The Achilles tendons of 15 rabbits were taken and the allogeneic tendons were prepared by chemical extraction with antigen inactivation. The extracted tendons were compared with untreated tendons by HE and Masson stainings. Chondrocytes were isolated and cultured by trypsin method and identified by immunohistochemical staining of collagen type Ⅱ. The remaining 30 rabbits were used to prepare the model of anterior labrum defect of shoulder joint. After the allogeneic tendon was transplanted to the damaged labrum, the rabbits was randomly divided into two groups (15 in each group). In group A, the allogeneic chondrocytes were injected into the joint immediately after transplantation, while in group B, no treatment was made. At 4, 6, and 8 weeks after operation, 5 transplanted tendons of each group were taken. After general observation, HE staining was used to observe the number of nuclei, Masson staining was used to observe the expression of collagen fibers in muscle fiber tissues, and AB staining was used to detect the glycosaminoglycan level after transplantation, to evaluate the cell growth in the tissues of the two groups of allogeneic tendon.ResultsBy HE and Masson stainings, the allogeneic tendon antigen prepared by chemical extraction method was inactivated and the fibrous tissue structure was intact; collagen type Ⅱ immunohisto-chemistry staining showed that the cultured cells were chondrocytes. After tendon transplantation, the content of glycosaminoglycan in group A was significantly higher than that in group B (P<0.05). At 6 weeks after operation, HE staining showed that the nuclear in tendon tissue of group A was significantly more than that of group B (t=20.043, P=0.000). Masson staining showed that the number of nuclei in tendon tissue of group A was significantly increased, the muscle fibers and collagen fibers were interlaced, the tissue structure was more compact, and the tendon tissue was mainly blue stained; while the number of nuclei in group B was less, mainly collagen fibers of the original graft.ConclusionThe allogeneic tendon inactivated by chemical extraction can be used to reconstruct the defect of anterior labrum of shoulder joint in rabbits, and the combination of allogeneic chondrocytes can promote the healing of tendon transplantation.
Objective To investigate the preparation of decellularized Achilles tendons and the effect of co-culture of human fibroblasts on the scaffold so as to provide a scaffold for the tissue engineered ligament reconstruction. Methods Achilles tendons of both hind limbs were harvested from 10 male New Zealand white rabbits (5-month-old; weighing, 4-5 kg). The Achilles tendons were decellularized using trypsin, Triton X-100, and sodium dodecyl sulfate (SDS), and then gross observation, histological examination, and scanning electron microscope (SEM) observation were performed; the human fibroblasts were seeded on the decellularized Achilles tendon, and then cytocompatibility was tested using the cell counting kit 8 method at 1, 3, 5, 7, and 9 days after co-culture. At 4 weeks after co-culture, SEM, HE staining, and biomechanical test were performed for observing cell-scaffold composite, and a comparison was made with before and after decellularization. ResultsAfter decellularization, the tendons had integrated aponeurosis and enlarged volume with soft texture and good toughness; there was no loose connective tissue and tendon cells between tendon bundles, the collagen fibers arranged loosely with three-dimensional network structure and more pores between tendon bundles; and it had good cytocompatibility. At 4 weeks after co-culture, cells migrated into the pores, and three-dimensional network structure disappeared. By biomechanical test, the tensile strength and Young’s elastic modulus of the decellularized Achilles tendon group decreased significantly when compared with normal Achilles tendons group and cell-scaffold composite group (P lt; 0.05), but no significant difference was found between normal Achilles tendons group and cell-scaffold composite group (P gt; 0.05). There was no significant difference in elongation at break among 3 groups (P gt; 0.05). ConclusionThe decellularized Achilles tendon is biocompatible to fibroblasts. It is suit for the scaffold for tissue engineered ligament reconstruction.
Objective We modeled superior vena cava (SVC) occlusion in rabbits to observe the effect of different blocking time on brains. Method Forty rabbits were randomly divided into four groups. Group Ⅰ was set as a control group (n=10). Group Ⅱ was set as a 30 minutes SVC blocking time group (n=10). Group Ⅲ was set as a 60 minutes SVC blocking time group (n=10). And group Ⅳ was set as a 90 minutes SVC blocking time group (n=10). We detected the patho- logical and physiological changes in the course of the experiment. After the intervention, malondialdehyde (MDA) and superoxide dismutase (SOD) of brain tissue homogenate in each group were detected. Brain sections were stained with hematoxylin-eosin (HE). And we observed the edema and damage of brain tissue under the microscope. Results There was no obvious change on the content of MDA and SOD within 30 minutes interruption (P>0.05). When the blocking time was longer than 60 minutes, the content of MDA increased significantly (P<0.05) and the SOD decreased significantly (P<0.05). Compared to the group Ⅰ and the group Ⅱ, the brain water content in the group Ⅲ and the group Ⅳ with a interruption time above 60 minutes increased significantly. And under the microscope, the cell edema and damage induced by ischemia and hypoxia increased significantly. Conclusion The blocking time of SVC within 30 minutes is relatively safe. But there would be significant brain edema and neurocyte degeneration when the blocking time is more than 60 minutes.
Objective To study the effect of chitosan (CS) mediated insul in-l ike growth factor 1 gene (igf-1) transfection on the repair of articular cartilage defect. Methods Twelve 3-month-old healthy male rabbits weighting 2.0-2.5 kg were randomly divided into 2 primary groups, control and intervention groups (n=6 per group). Control group was further divided into normal control (left knee) and normal saline (NS) control (right knee) groups. While, intervention group was divided into CS (left knee) and CS/igf-1 intervention (right knee) groups. Cartilage defects were created in the knee joints except normalcontrol. Intra-articular injections of CS/igf-1 complex was administrated 2 times a week for 4 weeks in CS/igf-1 interventiongroup, 0.5 mL CS in CS intervention group, and 0.5 mL sal ine solution in normal control and sal ine control groups. At 28days after treatments, the cartilage samples were collected for histological observation and collagen type II and aggrecan mRNA evaluation. Results HE staining and toluidine blue staining revealed that CS/igf-1 and CS intervention could significantly stimulated cartilage regeneration accompanied with fibrosis and inflammatory cell infiltration, however, CS/igf-1 treatment resulted in the best repair of cartilage defect. In contrast, sal ine control group only showed fibrous tissue prol iferation and inflammatory cell infiltration without significant cartilage repairing. In terms of collagen type II and aggrecan gene expression, significant differences were observed in each pairwised comparison among 4 groups in the order of CS/igf-1 gt; CS gt; NS gt; normal control (P lt; 0.05). Conclusion In situ CS/ifg-1 complex transfection can enhance the formation of mesochondrium by upregulating collagen type II or aggrecan expression, which might enhance the repair of articular cartilage defect.
Histological studies and morphometry quantitative analysis have been performed on trial rabbit’s dilated common bile duct(CBD),which does not dilate simultaneously.The results shows:①Epithelia of rabbit’s CBD have a ber reparable function,which is fairly significant to the prevention of bile duct’s further injure under the pathogenic situation.②The smooth muscle cell(SMC)of the CBD is the histological basis of contraction,some SMC can be seen in contracting state under light microscope.This indicates that the SMC in rabbit’s CBD possess contracting function.③The collagenous and elastic fibers have the normal histological morphometric characteristics and quantity in it’s dilatation process,and no breekdown and degeneration of the fibers can be detected.Because of the morphological structure of these sections is quite similiar with normal ones,theoretically,we suspect that when pathological change of bile duct’s distal portion is relieved and the bile pressure is normal again.It is possible for this dilating bile duct to return to its formal shape and size.
Objective To compare the difference of preparing the acellular larynx scaffold between perfusion method and immersion method, and find better way to make acellular larynx scaffold for tissue engineering. Methods Twenty 6-month-old male New Zealand rabbits, weighing 2.0-2.5 kg, were divided into perfusion group (n=10) and immersion group (n=10) at random. All the larynxes were excised in a sterile fashion. The acellular larynx scaffold was obtained by perfusionmethod and immersion method respectively, and then comparative examinations were performed by the macroscopicview, histological view, scanning electron microscope (SEM), cartilage vital ity assay and toluidine blue staining. ResultsMacroscopic view showed that the larynxes perfused by sodium dodecyl sulphate (SDS) became transparent after 2 hoursof perfusion, but the larynxes immersed by SDS over 16 hours still appeared pink-white. Histology and SEM indicated thatcompared with immersion group, perfusion group showed better acellular effect, more ventages and collagen fibers wereretained, no intact cell or nuclei remained in acellular matrix and chondrocytes were still survival. The porosity was 85.39% ± 3.16% in perfusion group and 34.72% ± 4.51% in immersion group, showing significant difference (P lt; 0.01). The chondrocyte vital ity rate of perfusion group (86.93% ± 1.52%) was higher than that of immersion group (77.73% ± 1.66%), showing significant difference (P lt; 0.01). Toluidine blue staining showed that the chondrocyte heterochromaty was ber in perfusion group than that in immersion group. Conclusion Compared with immersion method, perfusion method is a better way to construct acellular larynx scaffold because it can achieve better acellular effect and retain chondrocyte vital ity at the greatest extent in the acellular larynx scaffold.
【Abstract】 Objective Prostaglandin E2 (PGE2) production increases in human tendon fibroblasts after the tendon injuries and repetitive mechanical loading in vitro. To analyze the relations between PGE2 and tendinopathy by observing the changes of collagen content and proportion after the Achilles tendon of rabbits is repeatedly exposed to PGE2. Methods Twenty-four Japanese rabbits (aged 3-4 months, weighing 2.0-2.5 kg, and male or female) were equally randomized into 2 groups according to injection dose of PGE2: low dose group (50 ng) and high dose group (500 ng). Corresponding PGE2 (0.2 mL) was injected into the middle segment of the Achilles tendon of hindlimb, the same dose saline into the same site of the other side as controls once a week for 4 weeks or 8 weeks. The Achilles tendons were harvested at 4 and 8 weeks after injection. HE staining was used to observe the cell structure and matrix, and picric acid-sirius red staining to observe the distribution and types of collagen fibers, and transmission electron microscopy was used to measure the density of the unit area and diameter of collagen fibers. Results HE staining showed that collagen structural damage was observed in low dose and high dose groups. Picric acid-sirius red staining showed that the content of type I collagen significantly decreased while the content of type III collagen significantly increased in experimental side of 2 groups at 4 and 8 weeks after injection when compared with control sides (P lt; 0.05). The content of type I collagen was significantly lower and the content of type III collagen and ratio of type III to type I were significantly higher in high dose group than in low dose group (P lt; 0.05). Transmission electron microscopy showed that the collagen fibers density of unit area was significantly lower and the diameter was significantly smaller in high dose and low dose groups than in the controls (P lt; 0.05), and in high dose group than in low dose group (P lt; 0.05). Conclusion Repeat exposure of the Achilles tendon of rabbit to PGE2 can cause the decrease of type I collagen, the increase of type III collagen, the reverse ratio of type I to type III, reduced unit density of collagen fibers, and thinner collagen fibers diameter, which is related with tendinopathy.
Objective To develop a kind of biological artificial knee joint prosthesis with stereo mesh surface for rabbit, to observe its function after being implanted into rabbit knee joint and to evaluate its biomechanical property.Methods Thirty adult New Zealand rabbits were randomized into experimental and control groups (n=15), total left knee arthroplasty was performed in both groups, no patella replacement was performed. Biological artificial knee joint prosthesis with stereo mesh surface was self-designed. The adjacent 4/5 surface of femur and tibia stem of the prosthesis was covered by stainlesssteel stereo mesh, the inner surface of femur condyles and tibia plateau was welded with two layers of stainless steel stereo mesh, then the prosthesis underwent biological fixation in the experimental group. Meanwhile, prosthesis having smooth marrow internal stem, femoral condyle and tibial plateau internal surface and sharing the same shape and size with the experimental group were prepared and fixed with bone cement in the control group. The postoperative general condition of animal was observed. At 1, 3 and 6 months after operation, the rabbits were killed for gross observation, X-ray examination was conducted to observe the fixation condition of prosthesis and heal ing condition, the range of motion (ROM) of knee joints was tested, biomechanics test was carried out and the maximum shear strength of prosthesis bone interface was calculated. Results In each group, there was 1 rabbit died and new one was added during the second experiment. The others survived till the end of the experiment and crawled normally 7 days after operation. For the excellent and good rate concerning the recovery of ROM of the knee joint at 1, 3 and 6 months after operation, the experimental group was 60%, 80% and 80%, respectively, and the control group was 60%, 80% and 60%, respectively, indicating there were no significant differences between two groups (P gt; 0.05). For the experimental group, the gross observation showed large quantities of bone reconstruction, X-ray films indicated the prosthesis fitted well, with sol id fixation and without dislocation and lossening；while for the control group, the gross observation showed no bone reconstruction, X-ray films displayed the location of prosthesis was good, with sol id fixation and without dislocation and loosening. Radiolucent zone around the femur prosthesis and stress shileding occured 6 months after operation. For the maximum shear strength, the experimental group was less than the control group at 1 month after operation; and it was higher than the control group at 3 and 6 months after operation, indicating there were significant differences betweentwo groups (P lt; 0.01). Conclusion The fixation strength of the biological artificial knee joint prosthesis with stereo meshsurface is better than that of the bone cement prosthesis in rabbits at 3 and 6 months after operation.