It is reported in this paper that defect of chest wall of dogs were repaired by hetcrotransplantation of pig bone. The shape of original chest wail were achieved following operation. Histologic study showed that the grown between trabeculae with immersion that method is not only to have same function of other reparing materials,but there is no rejection.
Objective To evaluate the feasibility of poly-L-lactide(PLLA)/porcinederived xenogeneic bone(PDXB) composite as a scaffold for the bone tissue engineering. Methods The film and the scaffold of the PLLA-PDXB composite were respectively prepared by a solution casting method and a solution casting-particle leaching method. The composite film and scaffold were further treated by the surface alkaline hydrolysis. The surface morphology of the composite was observed by the scanning electron microscopy, and hydrophilicity degree of the composite was measured. The OCT-1 osteoblastlike cells were cultured and amplified in vitro as the seeding cells, which werethen implanted on the film and scaffold. The adherence rate, adherence shape,proliferating activity, and growing morphology of the OCT-1 osteoblastlikecells were observed on the film. Results The PDXB particle 50 μm in diameter on average had a similar phase structure to that of hydroxyapatite. But its Ca/P ratio was lower than that of hydroxyapatite. After the surface alkaline hydrolysis, the PDXB particle could be exposed on the surface of the PLLA-PDXB composite. The surface roughness and hydrophilicity of the PLLAPDXB composite were obviously enhanced. The cell adherence rate and the cell proliferation activity of the PLLAPDXB composite were higher than those of the pure PLLA material. The cells tended to grow on the exposed surface of the PDXB particles. The cells seeded on the composite scaffold could migrate to the inside of the composite scaffold and grew well. Conclusion The PLLA-PDXB composite has a good cell affinity, and this kind of composite can hopefullybecome a new scaffold material to be used in the bone tissue engineering.
Objective To investigate the effect of anti-infective reconstitutedbone xenograft (ARBX) as primary grafting on repair of a segmental contaminateddefect in canine radius. Methods The contaminated segmentaldefects of 1.5 cm were made in both radius of 8 canine and 1 ml of staphylococal suspension was injected into the defect region at a concentration of 5×106 CFU/ml. ARBX(experimental side) or RBX(control side) was implanted into the two sides of the defects respectively as primary grafting followed by internal fixation. The results were compared between the two grafting materials in repairing the contaminated segmental defect. Results In ARBX side, the defects were repaired completely in 5 cases and partially in 1 case, and there existed no osteomyelitis in all cases; while in RBX side, the defects were repaired partially in 1 case and were not repaired in 5 cases after 6 months of operation, and there existed osteomyelitis in all cases. Conclusion Besides its b osteoinductive and osteoconductive activity, ARBX is highly antibacterial and can be used as primary grafting in repairing contaminated segmental defects.
Objective To study efficiency of vascularized bone graft combining with reconstituted bone xenograft (RBX) in repairing bone defect and the expression of the vascular endothelial growth factor (VEGF) in serum. Methods From January 1998 to December 2002, 27 cases of bones defects were treated and randomly divided into 3 groups according to different repair materials: group A (the vascularized bone graft-RBX group, n=9), group B (the vascularized bone graft group, n=10)and group C(the RBX group, n=8). The bone defect repair, the bone healing time and the bone graft resorption were observed by radiograph after 3 months, 6 months and 12 months of operation, and the expression of VEGF in serum was assayed with lumino-enzyme immunoassay before operation and after operative 2 weeks, 4 weeks, 6 weeks and 8 weeks respectively. Results The X-ray films showed that the bonehealing was achieved in 8 cases of group A, in 6 cases of group B and in 3 cases of group C after 3 months; in 1 case of group A, respectively in 3 cases of both group B and group C after 6 months. The bone graft resorption was observed in1 case of group B and in 2 cases of group C after 12 months. The serum VEGF values after operative 2 weeks and 4 weeks were higher than those before operation in all of 3 groups(Plt;0.05), and the VEGF values of groups A and B were higher than that group C(Plt;0.05) after 4 weeks. There were no significant differences (Pgt;0.05) in serum VEGF level between postoperative 6, 8 weeks and preoperation in 3 groups. Conclusion The expression of serum VEGF obviously increase in the early period of bone transplanting, it is value of clinical evaluation of reparative efficiency of bone defect.
From January 1984 to November 1997, the boiling xenogenic bone (porcine and bovine bone) was applied in 65 patients. The xenogenic bone was used to promote bone healing in 24 cases, intramedullary bone graft in 37 and osteomyelitis in 4. All of the patients were followed up for 2 to 35 months. The results showed that in five cases there was infection after operation, and all other the wounds had primary healing. The xenogenic bone seemed to induce rejection in vivo. In the sevious cases immunosuppressive treatment was often needed. The volume of the bone grafted and the extent of the periosteum being stripped seemed to be important whether xenogenic bone graft would be successful or. There were lots of problem needed investigation.
Objective To find out an effective technique torepair large segmental infected bony defect.Methods Calcium phosphate cement(CPC) incorporated with bone morphogenetic protein and gentamycin was embedded in the massive reconstituted bovine xenograft(MRBX), then CPC-MRBX was obtained after CPC’s solidification. In vivo test was applied to test the drug delivery capability of CPC-MRBX, in which it was implanted in the dorsal muscle pouch of 18 rabbits. The drug concentration of animal blood and surrounding soft tissue of the CPC-MRBX in the muscle pouch was measured 1, 2, 5, 10, 15, 20, 25, 30 and 35 d after operation, 2 rabbits each time. Large segmental infected femur defect in the rabbit model was created to test the repairing capability of CPC-MRBX. External fixation was done 1.5~2.0 cm above the knee, the most adjacent nail to fracture site was 0.5～0.8 cm away, and proper pressure was applied to the graft. In experimental group(n=25), the bony defect was replaced by CPC-MRBX, while in the control group(n=15) dissected bone block was re-implanted in original position. The animal was subjected to radiographic, histological examination at 4, 8, 16 and 24 weeks. The general condition was observed after the operation.Results CPC-MRBX was easily made under normal temperature and pressure. In viro drug delivery test showed that the drug concentration of the tissue remainedabove the minimal inhibitory concentration of staphylococcus 30 d after operation and no significant increase of blood drug concentration was observed. In experimental group, no adverse influence was observed. Four weeks after operation, the animal could bear load, bony callus around the graft was observed by X-ray, and abundant chondral tissues that grew into CPC-MRBX were observed by histological method. Eight weeks after operation, progressively increasing bony callus around the graft was observed, external fixation could be removed, normal function was restored, and CPC was degenerated dramatically while new bone tissues were growing. Sixteen weeks after the operation, more new bone tissues grew and CPC was degenerated furtherly while marrow tissues were taking shape. Twenty-four weeks after the operation, femur healed completely and CPC was degenerated completely. In the control group, the autograft remained unhealedon X-ray at 4 weeks, and osteomyelitis manifestation such as inflammatory cells infiltration and osteolysis was detected at 4 weeks. All the animals in the control group died before the 8th week, 4 of which showed positive hemoculture. Conclusion CPC-MRBX is readily available and can be applied to repairing large segmental infected bony defect.30 d after operation and no significant increase of blood drug concentration was observed. In experimental group, no adverse influence was observed. Four weeks after operation, the animal could bear load, bony callus around the graft was observed by X-ray, and abundant chondral tissues that grew into CPCMRBX were observed by histological method. Eight weeks after operation, progressively increasing bony callus around the graft was observed, external fixation could be removed, normal function was restored, and CPC was degenerated dramatically while new bone tissues were growing. Sixteen weeks after the operation, more new bone tissues grew and CPC was degenerated furtherly while marrow tissues were taking shape. Twenty-four weeks after the operation, femur healed completely and CPC was degenerated completely. In the control group, the autograft remained unhealedon X-ray at 4 weeks, and osteomyelitis manifestation such as inflammatory cells infiltration and osteolysis was detected at 4 weeks. All the animals in the control group died before the 8th week, 4 of which showed positive hemoculture.Conclusion CPC-MRBX is readily available and can be applied to repairing large segmental infected bony defect.
ObjectiveTo evaluate the physical and chemical properties, immunogenicity, and osteogenesis of two antigen-extracted xenogeneic bone scaffolds—decalcified bone matrix (DBM) and calcined bone.MethodsBy removing the inorganic and organic components of adult pig femus, xenogeneic DBM and calcined bone were prepared respectively. The density and pH value of the two materials were measured and calculated, the material morphology and pore diameter were observed by scanning electron microscope, and the surface contact angle was measured by automatic contact angle measuring instrument. The safety, osteogenic activity, and immunogenicity of the two materials were evaluated by cytotoxicity test, osteoblast proliferation test, DNA residue test, and human peripheral blood lymphocyte proliferation test. The two materials were implanted into the 5 mm full-thickness skull defect of 6-week-old male Sprague Dawley rats (the blank control group was not implanted with materials). The materials were taken at 4 and 8 weeks after operation, the repair effect of the materials on the rat skull was observed and evaluated by gross observation, Micro-CT scanning, and HE staining observation.ResultsCompared with calcined bone, DBM has lower density and poor hydrophilicity; the pH value of the two materials was 5.5-6.1, and the pore diameter was 160-800 μm. The two materials were non-cytotoxic and could promote the proliferation of osteoblasts. The absorbance (A) values of osteoblast proliferation at 1, 4, and 7 days in the DBM group were significantly higher than those in the calcined bone group (P<0.05). The DNA residues of the two materials were much lower than 50 ng/mg dry weight, and neither of them could stimulate the proliferation and differentiation of human peripheral blood lymphocytes. The results of animal experiments in vivo showed that the bone volume/total volume (BV/TV) in DBM group and calcined bone group were significantly higher than that in blank control group at 4 weeks after operation (P<0.05), and that in calcined bone group was significantly higher than that in DBM group (P<0.05); at 8 weeks after operation, there was no significant difference in BV/TV between groups (P>0.05). HE staining showed that at 4 and 8 weeks after operation, the defect in the blank control group was filled with fibrous connective tissue, the defect was obvious, and no bone growth was found; the defect in DBM group and calcined bone group had been repaired to varying degrees, and a large number of new bone formation could be seen. The material degradability of DBM group was better than that of calcined bone group.ConclusionThe physical and chemical properties and degradability of the two kinds of xenogeneic bone scaffolds were slightly different, both of them have no immunogenicity and can promote the repair and reconstruction of skull defects in rats.
Ceramiclike xenogeneic bone (CXB) was obtained from the fresh bone of pig ribs being treated by physical and chemical methods to deprive of its organic substance. The CXB possessed the same natural porous network system as that of the human. The CXB was cultured with the bone marrow stromal cells of rabit. When the marrow cells had integrated with the CXB, thus a new material was obtained. (CXB-BM), and was implanted sacro-spinal muscle of rabbit. The specimens were observed under phase microscope, light microscope and electronic scanning microscope. The results showed that: at the 2nd week after the implantation of CBX-BM composite material there began the new bone formation, and the rate of bone formation was increased with time. There was evident new bone formation after 24 weeks. The process of the new bone formation were quite similar to the composite graft of HAP red autogenous and marrow, but the former degraded faster and formed typical cancellous structure earlier. There was no new bone formation when CXB was implanted alone in the control. Both the mechanism of osteogenetic potential and its clinical application were discussed.
Objective To observe the changes of immune status in recipient after implanting with xenogeneic acellular bone matrix (ACBM). Methods Twenty rabbits were randomly divided into 4 groups. Autograft，ACMB and bone soaked in alcohol were implant into the 3 experimental groups separately, and No-treatment was done as control group. The CD4+,CD8+,CD25+T lymphocytes in blood were detected by flow cytometer at 1, 2, 4 and 6 weeks after operation. After 2 and 6 weeks of implantation, the changes of bone and tissue were observed by histology. Results After 2-6 weeks, CD4+ and CD8+ T cells were significantly higher in the implantedgroup of bone soaked in alcohol than that in the other 3 groups(Plt;0.05) and there wasno statistically significant difference in the other 3 groups(Pgt;0.05). After 2 weeks, CD25+ T cells were significantly higher in the implanted group of bone soakedin alcohol than that in the other groups. In the 2nd week, there were inflammatory infiltration with a predominance of granulocytes. In the 6th week, there were many fibroblasts instead of granulocytes with a few lymphocytes and cartilage island formed in the implanted groups of autograft and ACMB. Conclusion ACBM implanting has low influence on cellular immunity in recipient.
Objective To study the clinical application ofheterogenous bone graft combined with auto-marrow. Methods Deproteinated and degreased heterogenouscancellous bone combined with auto-marrow was used in 21 cases of anterior cervical interbody fusion. Among them, 2 cases were treated by bone graft only, and the other 19 cases were further treated by anterior plate fixation. Results The follow-up time was 12-36 months with an average of 21 months. After operation, posteroanterior and lateral radiograph of all the cases revealed that the reduction and the position of the grafting bone were good without inflammation or other complications. After 6 months of operation, the radiograph of 2 cases of cervical disk herniation, which were treated by bone graft only, showed the bones lost their height and the curve of the cervical spine returned to the state of preoperation. For the other cases, the grafting bonesremained their original figure without dislocation of the bone or fracture of the plate or the screw. The nervous function recovered variously. Before operation, 10 cases were ranked as grade A,7 cases grade C, 4 cases grade D according to Frankel classification. After operation, in grade A cases, 5 cases did not recover, 3 cases recovered to grade B, 2 cases to grade C; in grade C cases, 5 to grade D, 2 to grade E; all ofgrade D 4 cases to grade E. Conclusion Heterogenous bone combined with auto-marrow can be used as grafting material in the anterior cervical interbodyfusion, but its mechanical rigidity need to be improved or the fusion shouldbe aided with rigid internal fixation.