Objective To investigate the osteoblasts effect, compl ications and influencing factors in the appl ication of small freeze-drying allogeneic bone plots mixed autologous bone fragments in spinal surgery, and to compare with autogenous bone graft. Methods From January 2003 to January 2007, 515 cases of spinal injuries were treated. A total of 324 cases weretreated with small freeze-drying allogeneic bone plots mixed with autologous bone grafts (group A), including 211 males and 113 females with an average age of 36 years (18-83 years). There were 182 cases of thoracolumbar vertebra fracture, 68 cases of lumbar spondylol isthesis, 47 cases of lumbar vertebral canal stenosis, 17 cases of cervical disc herniation, 5 cases of cervical spine fracture-dislocation and 5 cases of thoracolumbar vertebra tumor. The weight of bone graft was 10-60 g (mean 30 g). A total of 191 cases were treated with autogenous bone grafting (group B), including 135 males and 56 females with an average age of 32 years (23-78 years). There were 109 cases of thoracolumbar vertebra fracture, 23 cases of lumbar spondylol isthesis, 17 cases of lumbar vertebral canal stenosis, 19 cases of cervical disc herniation, and 23 cases of cervical spine fracture-dislocation. The weight of bone graft was 10-50 g (mean 25 g). Results In group A, effusion of wound increased in 4 cases and the result of bacterial culture was negative; effusion was absorbed after 2 weeks of local irrigation, drainege and cortin management. In group B, no obvious effusion was observed. The follow-up time was 10-36 months (mean 17.4 months) in group A and 8-36 months (mean 16.8 months) in group B. The bone heal ing was achieved in 308 cases within 4-10 months (mean 8.1 months) and in 184 cases within 4-10 months (mean 5.8 months), and the bone fusion rates were 95.06% and 96.34% in groups Aand B, respectively. There was no significant difference in bone fusion rate between groups (P gt; 0.05). According to Mankin and Komender evaluation standard, the response rates were 95.06% and 96.34% in groups A and B, respectively, showing no significant difference (P gt; 0.05). Conclusion Mix-bone grafting has the same effective to autologous bone grafting in bone fusion rate. It could be used as the supplement of the autologous bone inadequacy.
Objective To evaluate the immunological reaction and the outcome of allogeneic chondrocyte transplantation in repairing articular cartilage defects in porcins. Methods Full articular cartilage from the knee of two Shanghai white porcins about one-month-old was removed and cut mechanically, digested by 0.25% trypsin and 0.2% type Ⅱ collagenase and cultured in 10％ DMEM medium. Defects of 0.5 cm×0.5 cm involving the subchodral bone were created in both the left and right femur condyloid in 8 two-month-old Yunnai bama porcins. Allogeneic chondrocyte transplantation were implanted in defects at a density of (1.0-2.0)×106,0.2 ml. The lymphocytes from the receivers’ blood were collected before transplantation and after 3, 5, 7 and 12 weeks of transplantation, then mixed with allogeneic chondrocytes to determin the lymphocyte stimulation index(SI) in vitro. The histological observation in vivo was made after 5, 7 and 24 weeks of transplantation. Results Lymphocyte SI at 3, 5, 7 and 12 weeks(1.457±0.062,1.739±0.142,1.548±0.047，1.216±0.028) after transplantation was higher than that before transplantation(1.102±0.034，Plt;0.05). SI began to increase in the 3rd week and reached the peak value in the 5th week, then gradually declined at the 7th and 12th weeks, showing significant differences when compared with in the 5th week (Plt;0.05). Inflammation and lymphocytes infiltration could be seen in subchondral bone and the intergration area between repair tissue and normal cartilage in the 5th week, and then decreased and limited in subchondral bone in the 7th week. Defects were filled with cartilage tissue, which had good intergration with subchondral bone at 24 weeks after transplantation. Conclusion Immunological reactions can be found at early stage of allogeneic chondrocyte transplantation and then decreased with the time, the fullthickness articular cartilage defects could be repaired mainlywith hyaline cartilage by the allogeneic chondrocyte transplantation. This may provide a new method to repair articular cartilage defects clinically.
OBJECTIVE: To observe the difference of the fracture reparation using autogeneic-iliac bone and allogenic bone. METHODS: Comminuted fracture of humerus in two sides were made in rabbits. Autogeneic-iliac bone was implanted in one side, while allogenic bone of equal capacity was implanted in the other side. General observation, X-ray, and HE histologic section were done when the rabbits were put to death in different stages. RESULTS: One week after implantation, the graft had been enclosed by connective tissue without infiltration of the inflammatory cells. At the 2nd week, the graft had been enclosed in osteoplastic granulation tissue, and the cartilage callus had formed. At the 3rd week, there had been broken sequestrum among the callus; the cartilage had actively formed the bone; and the medulla had been making. At the 4th week, the sequestrum had disappeared, and the mature callus had appeared; the osteoblasts had arranged in a line around the edge of the mature callus. At the 5th week, the callus was b, compact and approached mature bones. At the 6th week, there had been the compact lamellar structures and the complete haversian’s systems. There was no significant difference between callus of two sides by using image quantitative analysis in the 3rd, 4th week (P gt; 0.05). CONCLUSION: The allogenic bone has good histocompatibility and bone conduction effect, and can be used for bone transplantation substitute with autogenous-iliac bone.
OBJECTIVE To investigate the effect of allogeneic decalcified bone graft in the treatment of nonunion in children. METHODS From April 1990 to September 1997, 17 children with nonunion were adopted in this study. Among them, there were 10 boys and 7 girls, the age ranged from 2 to 13 years. The allogeneic decalcified bone graft taken from fresh corpse by aseptic manipulation were used to repair 3 cases of congenital nonunion and 14 cases of acquired nonunion. RESULTS All cases were followed up 2 to 9 years after operation, 9 cases were cured successfully and 7 cases were cured after twice operations. One case of congenital pseudoarthrosis of the tibia was operated twice and there were callus grown half years after the second operation, but reoccurred after one year. Although there were 1.5 cm to 3 cm shortening deformity of extremity including another 2 cases of congenital pseudoarthrosis of the tibia and 5 cases of nonunion caused by chronic osteomyelitis, the function of joint was normal. CONCLUSION Treatment of nonunion in children by allogeneic decalcified bone graft is a valuable technique in clinical practice. It is characterised by high capacity of osteoinduction, low antigenicity, rapid bony union, plentiful source of bone graft and convenient utilization.
Objective To give a prel iminary experimental evidence and to prove chitosan and allogeneic morsel ized bone as potential bone substitutions in repairing rabbit radius segmental defect. Methods Chitosan and allogeneic morsel ized bone were mixed with various ratios (1 ∶ 5, 1 ∶ 10, 1 ∶ 25, 1 ∶ 50, and 1 ∶ 100). After preparation, the physicaland chemical properties of the composites were prel iminary detected; the composites at the ratios of 1 ∶ 50 and 1 ∶ 25 had good physical and chemical properties and were used for the animal experiment. The radius segmental defects of 15 mm in length were made in 50 adult New Zealand white rabbits (weighing 2.5-3.0 kg), then the animals were divided into 2 groups. In groups A and B, chitosan/allogeneic morsel ized bone composites were implanted at the ratio of 1 ∶ 50 and 1 ∶ 25, respectively. After 1, 2, 4, 8, and 12 weeks of operation, the gross, histological, immunohistochemical observations were performed. Before the rabbits were sacrified, X-ray films were taken; the serum calcium and alkal ine phosphatase (ALP) concentration were measured; and the biomechanical measurement was carried out at 12 weeks. Results The results of gross observation were essentially consistent with those of the X-ray films. The histological observation showed that the bone formation was earl ier in group A than in group B; the amount of new bone formation in group A was more than that in group B; and the bone forming area in group A was bigger than that in group B (P lt; 0.05) at 4 and 8 weeks after operation. The immunohistochemical staining showed that vascular endothel ial growth factor and insul in-l ike growth factor receptor II proteins expressed in the cytoplasm of 2 groups after 4 and 8 weeks, and the expression in group A was higher than that in group B (P lt; 0.05). There was no significant difference in the serum calcium concentration between 2 groups at each time point (P gt; 0.05). After 4 and 8 weeks, the ALP concentration in group A was significantly higher than that in group B (P lt; 0.05). After 12 weeks, the radius maximum bending loads of groups A and B were (299.75 ± 27.69) N and (278.54 ± 17.09) N, respectively, showing significant difference (t=4.045,P=0.002). Conclusion The composite of chitosan and allogeneic morsel ized bone has good osteogeneic activity and can beused as a bone tissue engineering scaffold, and the optimum ratio of chitosan to allogeneic morsel ized bone was 1 ∶ 50.
Objective To study the effect of the allogeneic bone sheet that has been treated by the freezedrying and radiation sterilization in preventing the epidural adhesion after laminectomy in sheep. Methods Laminectomy was performed on L3,4and L4,5 of 12 adult male sheep. Afteroperation, one site of L3,4 or L4,5was covered by the allogeneicbone sheet in “H” shape after the freeze-drying and radiation sterilization treatment; and the other site was used as a control. The sheep were killed and the specimens were retrieved at 4，8，12，16，20 and 24 weeks after operation to observe the scar formation process. X-ray andCT scan were performed in the segments of L3,4and L4,5at 4 and 24weeks. Results Four weeks after operation, the position and the shape of the allogeneic bone sheet were good in condition, and no lumbar spinal stenosis or compression of the dura was found in the experimental sections. Twentyfour weeks after operation theexamination on the experimental sections revealed that the vertebral canal reconstruction was completed, the allograft was absorbed almost completely, and inosculation with the lamina of the vertebra and the shape of the lumbar spine were good, with no fibroid tissues found in the epidural area. Compared with the control segment, the dura adhesion degree in the laminoplasty segment was significantly decreased (Plt;0.01), and the dura sac remained in a good shape. There was no obvious compression on the dura. Conclusion The allogeneic bone sheet after the treatment of freeze-drying and radiation sterilization can effectively reduce the scar formation after laminectomy and prevent postoperative recurrence of the spinal stenosis.
OBJECTIVE: To construct tissue engineering bone with bio-derived materials and bone marrow stromal cells (MSCs), and to investigate the effect of allogeneic engineering bone implants on healing of segmental bone defects. METHODS: MSCs being aspirated aseptically from tibial tuberosities of young rhesus monkeys were induced into osteoblasts in vitro and then were cultured and marked with 5-bromo-2-deoxyuridine (BrdU). Tissue engineering bones were constructed with these labeled osteoblasts being seeded onto bio-derived materials made from fresh human bones which were treated physically and chemically, Then the constructs were implanted in 15 allogeneic monkeys to bridge 2.5 cm segmental bone defects of left radius as experimental groups, bio-derived materials only were implanted to bridge same size defects of right radius as control group. and, 2.5 cm segmental bone defects of both sides of radius were left empty in two rhesus monkeys as blank group. Every 3 monkeys were sacrificed in the 1st, 2nd, 3rd, 6th and 12th weeks postoperatively and both sides of the implants samples were examined macroscopically, histologicaly, and immunohistochemicaly. The two monkeys in blank group were sacrificed in the 12th week postoperatively. RESULTS: Apparent inflammatory reactions were seen around both sides of the implants samples in the 1st, 2nd, 3rd weeks, but it weakened in the 6th week and disappeared at the 12th week. The labeled osteoblasts existed at the 6th week but disappeared at the 12th week. The bone defects in experimental group were repaired and the new bone formed in multipoint way, and osteoid tissue, cartilage, woven bone and lamellar bone occurred earlier when compared with control group in which the bone defects were repaired in ’creep substitution’ way. The bone defects in blank group remained same size at the 12th week. CONCLUSIONS: Engineering bones constructed with bio-derived materials and MSCs were capable of repairing segmental bone defects in allogeneic monkeys beyond ’creep substitution’ way and making it healed earlier. Bio-derived materials being constituted with allogeneic MSCs may be a good option in construction of bone tissue engineering.
Objective Tri ptol ide can suppress immunological rejection reaction. To investigate the effect of tri ptol ide on allogenic tendon transplantation in repairing tendon defect in chicken. Methods The defect model of the third toes tendon was establ ished in 64 healthy-cleaning male Leghorn chickens (4-month-old, weighing 1.9-2.3 kg), which underwent allogenic tendon transplantation for repairing and were divided into 2 groups randomly (n=32). Tri ptol ide feeding[100 μg/(kg·d)] was given for 3 weeks in the experimental group and normal feeding in the control group. General condition of the chickens was observed after operation. The transplanted tendons were harvested from 4 chickens in each group for gross observation at 1, 2, 3, and 4 weeks after operation; the histological observation was performed at 1 and 3 weeks, and transmission electron microscope observation at 2 and 4 weeks. The blood and tendon were harvested from another 8 chickens in each group for flow cytometry and biomechanical tests respectively at 3 and 6 weeks. Results All chickens survived to the experiment end. Gross observation: with time extending, hyperemia and edema around transplanted tendon were rel ieved. Rarefaction adhering zone was seen in experimental group, and pyknotic adhering zone in control group. Histological observation: inflammatory reaction in experimental group was sl ighter than that in control group at 1 and 3 weeks. Transmission electron microscope observation: at 2 and 4 weeks, fibroblasts had big cell nucleus, more euchromatin, and l ittle heterochromatin in experimental group; however, there were small amount of rough endocytoplasmic reticulums with gentle expanded capsular space in control group, which contained sparse content. Flow cytometry test: at 3 and 6 weeks, peri pheral blood contained less CD4+ and CD8+ T lymphocytes in experimental group than in control group, and the ratio of CD4+ to CD8+ T lymphocyte significantly decreased in experimental group when compared with control group (P lt; 0.05). Biomechanical examination: at 3and 6 weeks, the maximum tensile strength in experimental group was bigger than that in control group, and tensile adhesion power in experimental group was smaller than that in control group. There were significant differences in the indexes between 2 groups (P lt; 0.05). Conclusion Tri ptol ide can suppress immunological rejection reaction, strengthen tendon healing strength, and reduce tendon adhesion in allogenic tendon transplantation.
Objective To evaluate the cl inical effect of local autogenous bone chips extended with allogeneic bone grafts in the posterolateral lumbar fusion. Methods From March 2005 to April 2007, 22 cases which underwent posterolaterallumbar fusion with allograft bone mixed with local autograft bone were analyzed retrospectively. The postoperative temperature, drainage flow and heal ing time of the incision were analyzed; postoperative lumbar pain was evaluated by visual analog scale (VAS) and JOA score; the postoperative efficacy was assessed by MacNab criteria and Oswestry disabil ity index (ODI); the fusion rate was defined by Jorgenson fusion criteria. Results All cases were followed up for 17-35 months with an average of 21 months, the wound all healed by first intention; no red swell ing, exudation and infection occurred. The excellent and good rate was 81.8% for JOA score (excellent in 4 cases, good in 14 cases, fair in 4 cases), 77.3% for MacNab criteria (excellent in 4 cases, good in 13 cases, fair in 5 cases) and 90.9% for ODI index (excellent in 3 cases, good in 17 cases, fair in 2 cases). The postoperative X-ray fusion rate within 1 year was 90.9%. Conclusion Allograft bone mixed with local autograft bone can achieve good efficacy and fusion rate in posterolateral lumbar fusion.
Objective To study the repair and function reconstruction of complex soft tissue defect of posterior of hand and forearm. Methods From May 2001 to November 2003, 8 cases of soft tissue defect of posterior of hand and forearm were repaired with thoracico abdominal flaps with hilum for primary stage. The tendon transplantation and allogeneic tendon function reconstruction of hand were performed for secondary stage. The range of the flap was 9 cm×15 cm to 12cm×38 cm. Allogeneic tendon amounted to 6.Results All the flaps survived. The flap countour was good. The results of allogeneic tendon transplantation were satisfactory and the function of hand was good. Conclusion Repairing complex soft tissue defect of posterior of hand and forearm and reconstructing hand function by use of thoracico abdominal flaps with hilum and transplantation of allogeneic tendon have the satisfactory clinical results.