Objective To explore the feasibility of allogeneic marrow stromal stem cells(MSCs) as seed cells to construct tissue engineered bone bydetecting the expressions of interleukin 2(IL-2) and IL-2 receptor in rhesus monkeys after implanting these tissue engineered bones.Methods Engineered bones were constructed with osteoblasts which derived from allogeneic MSCs and bio-derived materials in vitro, and then were implanted to bridge 2.5 cm segmental bone defects of left radius in 15 rhesus monkeys as experimental group, bioderived materials only were implanted to bridge same size defects of right radius as control group. Every 3 monkeys were sacrificed in the 1st, the 2nd, the 3rd, the 6th andthe 12th weeks postoperatively and the expressions of IL-2 and IL-2 receptor in blood and graft samples were detected quantitatively by enzymelinked immuneosorbent assay (ELISA).Results There was no significant difference in the contents of IL-2 and its receptor between 2 groups（P＞0.05）. The contents ofIL-2 and its receptor increased from the 2nd week and maintained high level from the 2nd to the 6th week, but decreased after 6 weeks.ConclusionTissue engineered bones constructed with allogeneic MSCs and bio-derived materials show low immunogenicity. Allogeneic MSCs may be used as seed cells to construct tissue engineered bone.
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 investigate the efficacy of the pedicle screw fixation and allograft bone in posterior spinal fusion for the treatment of thoracolumbar vertebral fractures. Methods From September 2006 to March 2008, 105 cases of thoracolumbar vertebral fractures were treated with allograft bone in posterior spinal fusion after the pedicle screw fixation,including 75 males and 30 males aged 15-65 years (mean 37 years). The mixture which consisted of spinous process and vertebral plate sclerotin and homogeneity variant bone was used as bone graft to implant into articular process and processus transversus space or vertebral plate space. The time from injury to surgery varied from 8 hours to 21 days, with an average of 3 days. There were 52 cases of fall ing injuries from height, 35 cases of traffic accident injuries, 11 cases of bruise injuries and 7 cases of tumbl ing injuries. Before operation, the primary cl inical symptoms of patients included local pain combined with l imitation of activity, 30 cases compl icated by various degrees of spinal cord and nerve root functional disturbance. According to Mcaffee classification, there were 7 cases of flexion depressed fractures, 86 cases of blow-out fractures, 9 cases of Chance fractures and 3 cases of dislocation-fracture. According to Frankel grade, there were 11 cases of grade A, 2 cases of grade B, 7 cases of grade C, 10 cases of grade D and 75 cases of grade E. The X-ray examination of all patients denoted that the bodies of injuryed vertebra were compressed and wedge-shaped, and the CT scan showed that 98 cases had spinal stenosis. After 2 weeeks and 3, 6, 12 months, the X-ray films were taken to evaluate bone graft fusion. The Cobb angle was measured. The recovery of nerve function was analyzed. Results The operation time was 55-180 minutes (mean 90 minutes) and the blood loss was 100-900 mL (mean 200 mL). All patients achieved heal ing by first intention with no compl ication. After operation, 93 cases were followed up for 6-15 months with an average of 11 months. Except for 11 patients who were at grade A before operation, one to two grade recover was observed in other patients. The average Cobb angle of injury segment was improved from preoperative 32.1° to postoperative 5.2°. The height of anterior border of injuried vertebral body was recovered from the preoperative average compressed remaining height 61.5% to postoperative 96.8%. The vestigial degree of canal is spinal is anteroposterior diameter was recovered from preoperative 65.7% to postoperative 89.9%. Imageology examination showed that all the patient achieved bone union within 6 months. The fusion rate of bone graft in spinal fusion was 100%. No loosening and breaking of nails occurred. Conclusion Pedicle fixators can restore and fix the thoracolumbar fractures, and the combination of autograft and allograft bone transplantation is a safe, rel iable and effective method.
Objective To analyze the causes and managing methods of long bone fracture nonunion after allogeneic bone transplantation. Methods From December 1995 to December 2000, 43 cases of postoperative nonunion of long bone fracture were treated. These cases included 31 males and 12 females at the age of 19-57 years (40 years on average). The locations were femur in 11 cases, tibia in 21 cases, humerus in 8 cases and forearm in 3 cases. Bone nonunionafter allogeneic bone transplantation was caused by extensive soft tissue contusion and poor cover around bone fractue site in 6 cases, by incomplete debridement in 8 cases of osteomyel itis (including 6 without continual lavage and 4 only skin flap transfer), by secondary infection of wounds in 3 cases, by severe primary injury or secondary scar covering bone in 12 cases, instable internal fixation in 7 cases, and by earl ier weight bearing in 7 cases. Accroding to Weber classification, there were 4 cases of hypertrophic type and 39 cases of atrophic type. Basing on the type of nonunion, the methods such as refixation, bone retransplantation and transferring tissue flap were chosen to manage the cases. Results Incision healed by first intention in 37 cases, superficial infection occurred and cured after 2 weeks of dressing change in 5 cases, and sinus formed and cured after 2 months of dressing change and drainage in 1 case. All transferring tissue flap survived, and partial necrosis occurred at flap edge in 4 cases and cured after dressing change. All patients were followed up for 38-91 months with an average of 54.6 months. All the nonunion cases achieved bone union 5-11 months with an average of 7.6 months. But low extreme shorten monstrosity occured in 4 cases, malunion in 2 cases, functional l imitation near joints in 7 cases, and rotational l imitation of forearm in 1 case. Conclusion Fracture nonunion after allogeneic bone transplantation mainly shows atrophic type with overall or partial absorption of the allograft bone. The preconditions to guarantee fracture nonunion heal ing include stabil ity of fracture fixation, using transferring tissue flaps, controll ing infection, and adequate bone transplantion.
Objectives To investigate the curative effect of skin flap and allogenic tendon in reconstructing l imbs function of complex soft-tissue defect. Methods From May 2001 to December 2007, 43 cases of complex soft-tissue defect of l imbs were repaired by pedicled skin flaps, free skin flaps, cutaneous nerve nutrient vessel skin flaps and arterial island skin flaps for primary stage, then the function of the l imbs were reconstructed with allogeneic tendon after 2-3 months of skin flapoperation. There were 31 males and 12 females, aged 5-53 years(mean 25 years). Injury was caused by machine in 28 cases, by traffic accident in 14 cases and others in 1 case. There were 27 cases in upper l imb, 16 cases in lower l imb. Twenty-six cases compl icated by bone fracture, dislocation and bone defect, the most of bone defect were cortical bone defect. The sizes of skin and parenchyma defect were 9 cm × 4 cm to 37 cm × 11 cm, the length of tendon defect was 6 to 26 cm. The sizes of skin flaps were 10 cm × 5 cm to 39 cm × 12 cm. Allogeneic tendons were used 2-6 strips(mean 4 strips). Results Forty-three cases were followed up for 5-56 months (16 months on average), all flaps survived. The donor area healed by first intention, the incision healed by first intention in second operation, and no tendon rejection occurred. The cl inical heal ing time of fracture was 3-8 months, and the cl inical heal ing time of allograft was 6-8 months. Six cases were given tenolysis for adhesion of tendon after 3-5 months of tendon transplantation. The postoperative flexion of wrist joint was 20-50°, the extension was 20-45°. Articulatio metacarpophalangea and articulatio interphalangeae could extend completely. The flexion of articulatio metacarpophalangea of thumb was 20-45°, the flexion of articulatio interphalangeae was 30-70°. The flexion of articulatio metacarpophalangea and articulatio interphalangeae of the other fingers was 60-90°. The postoperative ankle can extend to neutral position, the neutral position of ankle was 30-50°. The flexion of articulatio metatarsophalangeae and articulatio interdigital was 20-40°. Theextension of articulatio metatarsophalangeae was 30-60°. Conclusion Through designing systematically treatment plan,practicing operation by stages, preventing adhesion of tendon actively and exercising function reasonably, the functions of l imbs reconstructed by allogenic tendon and skin flap can leads to satisfactory effect.
Objective To investigate the value of computer-aided design (CAD) in defining the resection boundary, reconstructing the pelvis and hip in patients with pelvis tumors. Methods Between November 2006 and April 2009, 5 cases of pelvis tumors were treated surgically using CAD technology. There were 3 males and 2 females with an average age of 36.4 years (range, 24-62 years). The cause was osteosarcoma, giant cell tumor of bone, and angiosarcoma in 1 case, respectively,and chondrosarcoma in 2 cases. According to the Enneking system for staging benign and mal ignant musculoskeletal tumors, regions I, I + II, III, IV, and I + IV is in 1 case, respectively. According to the principle of reverse engineering, 5 patients with pelvis tumors were checked with lamellar CT/MRI scanning, whose two-dimensional data were obtained in disease area. The three-dimensional reconstruction of pelvic anatomical model, precise resection boundary of tumor, individual surgical template, individual prosthesis, and surgical simulation were precisely made by computer with CAD software. Based on the proposal of CAD, the bone tumor was resected accurately, and allograft il ium with internal fixation instrument or allogeneic il ium with personal ized prosthetic replacement were used to reconstruct the bone defect after tumor was resected. Results The operation was successfully performed in 5 cases. The average operation time was 7.9 hours, and the average blood loss was 3 125 mL. Hemorrhage and cerebrospinal fluid leakage occurred in 1 case, respectively, and were cured after debridement. Five patients were followed up from 24 to 50 months (mean, 34.5 months). All patients began non-weight bearing walk with double crutches at 4-6 weeks after operation, and began walk at 3-6 months after operation. Local recurrence developed in 2 patients at 18 months after operation, and resection and radiotherapy were performed. According to International Society of Limb Salvage criteria for curative effectiveness of bone tumor l imb salvage, the results were excellent in 2 and good in 3. Conclusion The individual surgical template, individual prosthesis, and surgical simulation by CAD ensure the precision and rel iabil ity of pelvis tumors resection. The CAD technology promotes pelvis tumor resection and the reconstruction of pelvis to individual treatment stage, and good curative effectiveness can be obtained.
Objective It is difficult to treat chronic osteomyel itis due to the formation of the Staphylococcus aureus biofilms. Liposomal gentamicin-impregnated allogeneic cortical bone can inhibit the formation of the Staphylococcus aureusbiofilms. To explore the treatment of chronic osteomyel itis of rabbit by l iposomal gentamicin-impregnated allogeneic cortical bone. Methods The l iposomal gentamicin, l iposomal gentamicin-impregnated allogeneic cortical bone and gentamicinimpregnated allogeneic cortical bone were produced. Then the chronic Staphylococcus aureus osteomyel itis models of rabbit were made in left lower l imbs of 40 6-month-old rabbits and the right lower l imbs were used as controls. After 2 weeks, the observations of gross and X-ray were done. Four rabbits died within 10 days after the models were made and other 36 rabbits were devided into 6 groups: group A (no antibiotics), group B (intravenous injection of gentamicin), group C (intravenous injection of l i posomal gentamicin), group D (implantation of gentamicin-impregnated allogeneic cortical bone), group E (implantation of l i posomal gentamicin-impregnated allogeneic cortical bone), and group F (implantation of allogeneic cortical bone). After 2 weeks of treatment, the bacterial culture, X-ray and HE staining were done. Results The chronic Staphylococcus aureus osteomyel itis model of rabbit was made successfully. The X-ray showed dissolution of bone and periosteal reaction in groups A, B, C, and F, and no obvious dissolution of bone and periosteal reaction in groups D and E. The Norden scores were (2.5 ± 0.3), (2.1 ± 0.2), (1.5 ± 0.3), (1.5 ± 0.2), (0.9 ± 0.3), and (2.7 ± 0.3) points in groups A-F, respectively; showing significant differences between group A and groups B-E (P lt; 0.05), between groups B, E, F and other groups (P lt; 0.05). The results of blood and marrow cultures for Staphylococcus aureus were positive in groups A and F, and negative in other 4 groups; the results of bone marrow culture for Staphylococcus aureus were positive in 6 rabbits of group B, 4 rabbits of group C and 3 rabitts of group D; and the results were negative in group E. HE staining showed: in groups A and F, abscess and dead bone formed, and no new bone formation were observed; in groups B and C, different degrees of neutrophil accumulation was seen; in group D, some neutrophil accumulation occurred, and osteoprogenitor cells and osteoclasts were seen around implanted bone; and in group E, no neutrophil accumulation was observed, a lot of granulation tissues formed, and osteoprogenitor cells and osteoclasts were seen around implanted bone. Conclusion Implantation of l iposomal gentamicin-impregnated allogeneic cortical bone has remarkly better effect in treating chronic osteomyel itis than intravenous injection of l iposomal gentamicin and implantation of gentamicin-impregnated allogeneic cortical bone.
ObjectiveTo investigate the short-term effectiveness of allogeneic bone transplantation for pelvic reconstruction of large skeletal defects after tumor resection. MethodsBetween January 2008 and November 2009, 9 patients with pelvic tumor were treated, and the clinical data were retrospectively analyzed. There were 1 male and 8 females with an average age of 36.1 years (range, 18-44 years). There were 5 cases of giant cell tumor of bone, 2 cases of chondrosarcoma, 1 case of malignant giant cell tumor of bone, and 1 case of Ewing sarcoma. According to the Enneking system for staging musculoskeletal tumors, the lesion involved ilium (region I) in 4 cases, acetabulum (region II) in 6 cases, pubis and ischium (region III) in 3 cases, and sacrum (region IV) in 1 case. The average disease duration was 6.8 months (range, 1-36 months). After accurate resection of the tumor, allograft ilium with or without acetabulum was used in 6 cases and 2 cases respectively, and allogenous bone plate in 1 case for pelvic reconstruction; total hip arthroplasty was performed in 6 cases; sacrum screw was used in 1 case. ResultsInfection of incision and abdominal distention occurred in 1 case, and was cured after debridement; primary healing of incision was obtained in the other cases, without infection, lower limb deep venous thrombosis, or dislocation. The average follow-up was 11.8 months (range, 8-15 months). The X-ray films showed breakage of plate in 1 case at 8 months after operation and partial bone resorption in 2 cases; good position of the prosthesis, plate, and transplanted bone was observed in the other cases. No local recurrence was found. At last follow-up, the average musculoskeletal tumor score (MSTS) was 23 (range, 18-29). ConclusionAllogeneic bone transplantation for pelvic reconstruction of large skeletal defects after tumor resection can achieve satisfactory short-term surgical and functional effectiveness.
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 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.