ObjectiveTo investigate the effectiveness and technical key points of limb salvage surgery by allotransplantation of cryopreservated vascularized bone in children and adolescents with osteosarcoma.
MethodsA retrospective analysis was made on the clinical data of 21 children and adolescents with osteosarcoma receiving limb salvage surgery by allotransplantation of cryopreservated vascularized bone from their relatives between February 2004 and April 2012. There were 13 males and 8 females, aged from 7 to 16 years (mean, 12.6 years). According to Enneking stage system, 15 cases were rated as stage ⅡA and 6 cases as stage ⅡB. The tumors located at the distal femur in 10 cases, at the proximal femur in 1 case, at the proximal tibia in 8 cases, at the proximal humerus in 1 case, and at the distal radius in 1 case. Imaging examination showed that epiphyseal extension of malignant bone tumors in 7 cases. The iliac bone allograft with deep iliac vessels was obtained from their lineal consanguinity. After preservation by a twostep freezing schedule, the iliac bone allograft with deep iliac vessels was implanted into the bone defect area after tumor resection. The size of iliac bone flap was 8.0 cm×3.0 cm×2.0 cm-14.0 cm×5.0 cm×2.5 cm. Reserved joint surgery was performed on 16 cases and joint fusion surgery on 5 cases, and external fixation was used in all cases. The chemotherapy was given according to sequential high-dose methotraxate, adriamycin, and cisplatine before and after operation.
ResultsAll 21 cases were followed up from 5 months to 11 years (mean, 6.4 years). At 2 weeks after operation, the erythrocyte rosette forming cells accounted for 56.7%±3.9%, showing no significant difference when compared with that of normal control (58.3%±4.3%) (t=1.56, P=0.13), which suggested no acute rejection. At 4 weeks after operation, single photon emission computerized tomography bone scan indicated that the blood supply of bone graft was rich, and the metabolism was active. At 12 weeks after operation, the digital subtraction angiography showed the artery of iliac bone flap kept patency. X-ray films showed that malunion and non-union occurred at 5 and 6 months after operation in 1 case, respectively. The bone graft healed in the other patients, and the healing time was 3.2-6.0 months (mean, 4.4 months). At last follow-up, American Musculoskeletal Tumor Society (MSTS) score was significantly improved to 26.80±2.14 from preoperative value (17.15±1.86) (t=-4.15, P=0.00). The survival rate was 85.7% (18/21) and the recurrence rate was 9.5% (2/21).
ConclusionAllotransplantation of cryopreservated vascularized bone from the relatives provides a new method for the treatment of osteosarcoma in children and adolescents. A combination of allotransplantation and chemotherapy can achieve the ideal treatment effect. The correct cutting, preservation, and transplantation of the donor bone, and indication are the key to improve the effectiveness.
In order to observe stereological correlation between new vessels and surrounding tissue, 6 rabbits were used for the following experiments. The mandible defect model was made by cutting 1 cm x 0.5 cm bone from both right and left side of mandible. Then, the left defect was repaired by the bone segment from the right side, and the right defect was repaired by frozen allogenic bone segment. One month later, the metabolism of the bone segments was observed by nuclein scintiphotography. The revascularization of the bone segments was observed by vascular corrosion cast method. It was shown that new vessels from host soft tissue could penetrate the periosteum of allogenic transplanted bone, along the absorbing path, ingrowth into the bone. The metabolism of the bone was active. It was suggested that the vessel growth from host to the graft is one of the main patterns of revascularization.
Objective To evaluate the tissue response induced by three kinds of bone transplantation materials implanted in rat so as to provide proper evidence for their cl inical appl ication. Methods Thirty-six healthy mature Sprague- Dawly mice, weighing from 229 g to 358 g, were randomly assigned to groups A and B (n=18). Three kinds of materials wereimplanted into muscles of rats. Calcium sulfate (CS) granular preparations and allogeneic demineral ized bone matrix (DBM) were transplanted into the left (group A1) and right (group A2) thigh muscle pouches of group A. Respectively, whereas xenogenic DBM were transplanted into the left (group B1) thigh muscle pouches of group B and the right (group B2) sites were taken as control without implant. The samples (n=6) were collected to make the observation of gross and histology and to analyze histological score after 2, 4, and 6 weeks. Results The gross observation: implanted materials were gradually absorbed at late stage in group A1. No obvious degradation and absorption, but fibrosis of tissues were observed in group A2 and B1. The inflammatory reactions were more severe in groups A2 and B1. In group B2, only the changes of scar were seen at operative site. The histological observation: no obvious inflammatory reactions were seen in group A1, CS were gradually absorbed and completely absorbed at 6 weeks, while fibrosis of tissues increased at late stage. Inflammatory reactions in group A2 and group B1 were alleviated gradually, no obvious absorption and degradation were observed. The different two DBM could induce granulation tissues and bone formation at different sites and secondary fibrosis with no obvious immune response was observed. In group B2, there was an increase in collagen fiber density and angiogenesis at late stage. The scores of inflammatory infiltration were significantly higher in groups A2, B1 than in groups A1, B2 (P lt; 0.05), and the scores of fibrosis was larger in groups A1, A2 and B1 than in group B2 (P lt; 0.05). Conclusion CS has rapid dissolution and good biocompatibil ity. It is a good replaceable packing materials of bone defects in some upper l imb’s or acute bone fracture. Both of two DBM have biocompatibil ity and osteoinductive potential, which dissolution are very slow. Due to these capacity, they can be served as an ideal materials in treatment of lower l imb’s bone defect and nonunion.
Objective To investigate the application of rigid internal fixation in mandibular reconstruction with autogenous bone and to evaluate its efficacy. Methods From January 1994 to May 2004, 98 patients with mandibular defect received mandibular reconstruction with autogenous bone by using rigid internal fixation. Seventy-two cases of benign tumor and 26 cases of malignant tumor were included. Four hundred and ono rigid fixation plates were inserted.The clinical results and X-ray films were analyzed and the healing processes were evaluated. The functional and aesthetic results of the mandibular reconstruction were also evaluated according to Lopez assessment system. Results After a follow-up of 1 to 3 years, 95 patients（96.9%）achieved successful effect. The forms and function of the mandibleswere resumed. Eightyone （82.7%）patients were satisfied with the results ofoperations. Thirteen patients（13.3%） achieved acceptable results. Four patients (4.1%) were dissatisfied with the results of operations. Conclusion The rigid internal fixation is conductive to healing and remodeling ofthe transplant bone in mandibular reconstruction.
Objective To explore an effective way fortreating severe complicated distal femoral fractures. Methods Twenty-six patients with complicated distal femoral fracture who all belonged to 33C3.3type according to AO/ASIF lassification, were treated with a lateral condylar buttress plate or self-desinged aliform anatomical plate, and operated on with allogeneic bone grafting. Results All cases were followed up for an average of 14 months (ranging 5-25 months). Twenty-four wounds were primary healing postoperatively, 2 wounds were infected and healed after dressing change. Twenty-four had bone healing after 411 months, 2 needed to operate again because of earlier weight-bearing resulting in fixation failure. According to shelbourne and Brueckmann score, the excellent and good rate was 88.46%. Conclusion The internal fixation forcomplicated distal femoral fracture by self-designed aliform anatomical plate and lateral condylar buttress plate with a great deal of allograft bone is an effective surgical method. As it has long oval holes and the holes are consecutive ,the aliform anatomical plate is more suitable for severe complicated fractures. At the same time, autogenous-ilium transplantation can be substituted by the allograft bone.
Objective To study the immunological tolerance induced by blocking the second signal of T cell with extrinsic cytotoxic T lymphocyteassociated antigen 4 immuno globlin(CTLA4-Ig). Methods Fifty-four BALB/C mice, inbred strains, were employed as recipients of bone allografts, using a model of heterotopic muscle pouch. The 54 mice were divided into 3 groups and18 for each group. The first group, in which the donor was C57BL/6 with intraperitoneal injection ofL6(as a control), was named AL group. The second group,also C57BL/6 with injection CTLA4-Ig, was named AC group. The third group,homologous BALB/C with injection of PBS buffer solution, was named AB group.The serum antibody, lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor, the analysis of lymphocyte subsets, a regraft experiment and histology were determined 2, 4 and 6 weeks after transplantation. The second transplantation was to regraft C57BL/6(BC group) and C3H(BHgroup) mice respectively after first 12 mice being transplantated with C57BL/6 and injected with CTLA4-Ig as to detect donor-specificity of immunological tolerance. Results Compared with AB group, AL group created more intensive immune rejection: CD4 T cell subsets(Plt;0.05), the serum antibody(Plt;0.05) and lymphocyteproliferation of the second stimulation by splenic cell and bone supernatant ofdonor (Plt;0.01 and 0.05) were significantly increased. However, the results of AC group showed that CTLA4-Ig significantly inhibited the immune rejection: CD4T cell subsets(Pgt;0.05), the serum antibody (Pgt;0.05), and lymphocyte proliferation of the second stimulation(Pgt;0.05) were similar to those of AB group. Histological observation of AC group showed that lymphocyte infiltration disappeared,cartilage and new bone formed, and bone marrow cavities emerged. A regraft experiment showed that CD4 T cell subsets (Plt;0.05) and lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor(Plt;0.05), BC group was significantly lower than those of BH group. So theimmunological tolerance induced by CTLA4-Ig was of donor-specificity. Conclusion The immunological tolerance induced by CTLA4-Ig was prolonged for 6 weeks. This study provides a brand-new path for bone transplantation, which can be helpful to other organ transplantation.
OBJECTIVE To study the function of the composite of bone matrix gelatin(BMG) and plaster in the repairing process of bone defects. METHODS Sixteen New Zealand rabbits which were defected in corpus radii were made as implant zone of bone. Sixteen sides of radii were implanted with the composite of BMG and plaster as experimental group. Others were implanted with BMG(8 sides) and bone stored in alcohol(8 sides) as control groups. The repairing process in bone defects were observed by X-ray and histological examination. RESULTS There was an obvious osteogenesis in experimental group. The defects of radii were almost healed at 12th week after operation. There were osteogenesis in both control groups, but the repairing process was slower than that of the experimental group. CONCLUSION The composite of BMG and plaster is a good material for bone transplantation.
OBJECTIVE To compare the osteogenesis of recombination artificial bones, which are bovine deproteined bone (bDPB) and bovine bone morphogenetic protein (bBMP), combined with tumor necrosis factor alpha (TNF alpha), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) respectively. METHODS One hundred trephined skull bone defects in fifty rabbits were divided into four groups, which implanted with bDPB/bBMP/TNF alpha, bDPB/bBMP/bFGF, bDPB/bBMP/EGF, and bDPB/bBMP respectively. X-ray and histological changes were observed in the 1st, 2nd, 4th, 6th, 8th weeks after implantation. The content of 35S and 45Ca and ash weight were measured at 10 and 42 days after operation. RESULTS The osteogenesis of bDPB/bBMP/TNF alpha group was ber than that of bDPB/bBMP/bFGF group(P lt; 0.01), while bDPB/bBMP/bFGF group was ber than that of bDPB/bBMP/EGF(P lt; 0.01). No significant statistical difference were found between bDPB/bBMP/EGF and bDPB/bBMP(P gt; 0.05). CONCLUSION TNF alpha combined with bBMP and carrier can stimulate bone formation and increase the volume of new bone in vivo. It suggests that bDPB/bBMP/TNF alpha is a valuable biomaterial of bone graft.
ObjectiveTo investigate the effectiveness of tibial periosteal flap pedicled with intermuscular branch of posterior tibial vessels combined with autologous bone graft in the treatment of tibial bone defects.
MethodsBetween January 2007 and December 2013, 19 cases of traumatic tibia bone and soft tissue defects were treated. There were 14 males and 5 females, aged from 18 to 49 years (mean, 28 years). The tibial fracture site located at the middle tibia in 6 cases and at the distal tibia in 13 cases. According to Gustilo type, 4 cases were rated as type Ⅲ A, 14 cases as type Ⅲ B, and 1 case as type Ⅲ C (injury of anterior tibial artery). The length of bone defect ranged from 4.3 to 8.5 cm (mean, 6.3 cm). The soft tissue defects ranged from 8 cm×5 cm to 17 cm×9 cm. The time from injury to operation was 3 to 8 hours (mean, 4 hours). One-stage operation included debridement, external fixation, and vacuum sealing drainage. After formation of granulation tissue, the fresh wound was repaired with sural neurovascular flap or posterior tibial artery perforator flap. The flap size ranged from 10 cm×6 cm to 19 cm×11 cm. In two-stage operation, tibial periosteal flap pedicled with intermuscular branch of posterior tibial vessels combined with autologous bone graft was used to repair tibial defect. The periosteal flap ranged from 6.5 cm×4.0 cm to 9.0 cm×5.0 cm; bone graft ranged from 4.5 to 9.0 cm in length. External fixation was changed to internal fixation.
ResultsAll flaps survived with soft texture, and no ulcer and infection occurred. All incisions healed by the first intention. All patients were followed up 18-40 months (mean, 22.5 months). All graft bone healed, with the healing time from 3 to 9 months (mean, 6.5 months). No complication of implant loosening or fracture was observed. No pain and abnormal activity in the affected leg occurred. All patients resumed weight-bearing and walking function. The length of the limb was recovered and difference value was 0.5-1.5 cm between normal and affected sides. The function of the knee and ankle joint was good without infection, malunion, and equinus. According to the Johner standard at last follow-up, the results were excellent in 15 cases, good in 3 cases, and fair in 1 case, with an excellent and good rate of 94.7%.
ConclusionTibial periosteal flap pedicled with intermuscular branch of posterior tibial vessels combined with autologous bone graft is an effective method to treat bone defect of the tibia.
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.
