Objective To study the differentiation of the human osteoblasts during the construction of the tissue engineered periosteum with the human acellular amniotic membrane(HAAM).Methods To construct the tissue engineered periosteum (n=60) with HAAM, the human fetal osteoblasts were used. The fetal osteoblasts were cultured for 2, 4, 6, 8, and10 days, and then their total RNA was extracted, which were reversely transcripted to cDNA. The realtime PCR analysis was used to reveal Cbfal and Osterix, and the cycle threshold (Ct) was also measured. The simplycultured osteoblasts were used as the control group (n=20).Results The expression of Cbfa1 was higher in the experimental group on the 2nd day when compared with that on the 4th, 6th, and 8th day(P＜0.05). The same result existed on the 10th day when compared with that on the 4th and 8th day. The expression of Osterix increased and was highest on the 8th day when compared with the other results(P＜0.05). Both of the 2 gene expressions were decreased in the control group when compared with those in the experimental group, but with no significant difference(P＞0.05). Conclusion Cbfa1 and Osterix can be normally expressed by the osteoblasts after their integration with HAAM. As a scaffold, HAAM can be used to keep the osteoblast phenotype and differentiation with an osteoconductive ability. Such a cell-scaffold complex may provide a basis for the osteogenesis.
OBJECTIVE: To investigate the repairing effect of transplantation of allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft to bone defect. METHODS: Twenty Long-eared white male rabbits were chosen as experimental model of bilateral 12 mm combined bony and periosteal radial defect. Cryopreserved allograft periosteum with allogeneic fetal bone were implanted in the left defect as experimental side and fetal bone was simply transplanted in the right defect as control side. Bone repair process in the two groups were compared by macroscopy, microscopy, roentgenograms and the contents of calcium and phosphate in the defect area at 2, 4, 8 and 12 weeks after transplantation. RESULTS: There was significant statistic difference in the contents of calcium and phosphate between the experimental and control sides at 4, 8 and 12 weeks after transplantation (P lt; 0.05). With time passing by, the contents of calcium and phosphate have the increasing trends. In the experimental group, lamella bone was seen and medullary canal recanalized at 8 weeks postoperatively. The histological section showed the bone lacuna and lamella bone were formed. CONCLUSION: It suggests that allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft can promote bone repair, and allogeneic fetal bone is excellent bone substitute.
Objective To compare the effectiveness between the myo-periosteal fibular bone bridging and traditional transtibial amputation in the treatment of amputation below knee so as to provide theoretical basis for choosing transtibial amputation in clinical application. Methods Between November 2001 and November 2011, 38 patients with mangled lower extremity were treated by transtibial amputation. Among 38 patients, 17 (group A) underwent myo-periosteal fibular bone bridging (the operation techniques of an attached peroneal muscle myo-periosteal fibular strut bridge between the end of the tibia and fibula below knee amputation), and other 21 (group B) underwent traditional transtibial amputation. There was no significant difference in age, gender, injury cause, amputation cause, side, and disease duration between 2 groups (P gt; 0.05). The quality of life (QOL) was analyzed using 36-item short form health survey (SF-36), and prosthesis satisfaction by Trinity amputation and prosthesis experience scale (TAPES). Results Healing of incision by first intention was obtained in all patients of 2 groups; no necrosis, infection, or poor stumps was observed. The mean follow-up time was 22 months (range, 14-30 months) in group A, and 26 months (range, 15-30 months) in group B. The patients achieved good healing of bone bridging, no bone nonunion occurred. The healing time was (5.1 ± 1.1) months in group A and (3.3 ± 0.6) months in group B, showing significant difference between 2 groups (t=9.82, P=0.00). Spur occurred at the distal fibula in an 11-year-old boy of group B after 2 years of operation, which blocked use of prosthesis; prosthesis was well used in the other patients. After 12 months of operation, SF-36 score was 55.84 ± 14.01 in group A and 49.93 ± 12.78 in group B, showing significant difference (P lt; 0. 05); the physical functioning, social functioning, role-physical, vitality, body pain, general health scores in group A were significantly higher than those in group B (P lt; 0.05), but no significant difference was found in role-emotional and mental health scores between 2 groups (P gt; 0.05). TAPES score was 12.12 ± 2.23 in group A and 10.10 ± 2.00 in group B, showing significant difference (t=2.891, P=0.006). Conclusion It is a very effective method to treat traumatic amputation using an attached myo-periosteal fibular bone bridging between the end of the tibia and fibula below knee, which can afford better quality of life and prosthesis satisfaction.
Objective To estimate clinical effect ofspin iliac deep vascular pedicled periosteum flap in repairing traumatic femoral neck of theca inside fracture in young and middleaged. Methods From April 1993 to September 2001, 12 cases of traumatic femoral neck fracture were given diaplastic operation with fixation of 3 centre hollow pressed bolt and were conducted under os traction bed and "C" arm X-ray machine. Spin iliac deep vascular pedicled periosteum flap wasstripped off, and transferred to the front of femoral neck fundus，then transplanted to the narrow inside of fracture through outer open door of articular capsule.Results All patients were followed up for 17 years. All fracture healedwithout femoral head necrosis, but mild arthritis appeared in 7 cases.Conclusion Vascular pedicled periosteum flap transfer of young and middle-aged femoral neck fracture, by decompression of femoral neck and reconstruction of blood circulation, can promote the fracture healing and decrease the wound and blood circulation destroy.
ObjectiveTo evaluate the effect of tissue engineered periosteum on the repair of large diaphysis defect in rabbit radius, and the effect of deproteinized bone (DPB) as supporting scaffolds of tissue engineering periosteum. MethodsBone marrow mesenchymal stem cells (BMSCs) were cultured from 1-month-old New Zealand Rabbit and osteogenetically induced into osteoblasts. Porcine small intestinal submucosa (SIS) scaffold was produced by decellular and a series mechanical and physiochemical procedures. Then tissue engineered periosteum was constructed by combining osteogenic BMSCs and SIS, and then the adhesion of cells to scaffolds was observed by scanning electron microscope (SEM). Fresh allogeneic bone was drilled and deproteinized as DPB scaffold. Tissue engineered periosteum/DPB complex was constructed by tissue engineered periosteum and DPB. Tissue engineered periosteum was "coat-like" package the DPB, and bundled with absorbable sutures. Forty-eight New Zealand white rabbits (4-month-old) were randomly divided into 4 groups (groups A, B, C, and D, n=12). The bone defect model of 3.5 cm in length in the left radius was created. Defect was repaired with tissue engineered periosteum in group A, with DPB in group B, with tissue engineered periosteum/DPB in group C; defect was untreated in group D. At 4, 8, and 12 weeks after operation, 4 rabbits in each group were observed by X-ray. At 8 weeks after operation, 4 rabbits of each group were randomly sacrificed for histological examination. ResultsSEM observation showed that abundant seeding cells adhered to tissue engineered periosteum. At 4, 8, and 12 weeks after operation, X-ray films showed the newly formed bone was much more in groups A and C than groups B and D. The X-ray film score were significantly higher in groups A and C than in groups B and D, in group A than in group C, and in group B than in group D (P<0.05). Histological staining indicated that there was a lot of newly formed bone in the defect space in group A, with abundant newly formed vessels and medullary cavity. While in group B, the defect space filled with the DPB, the degradation of DPB was not obvious. In group C, there was a lot of newly formed bone in the defect space, island-like DPB and obvious DPB degradation were seen in newly formed bone. In group D, the defect space only replaced by some connective tissue. ConclusionTissue engineered periosteum constructed by SIS and BMSCs has the feasibility to repair the large diaphysis defect in rabbit. DPB isn't an ideal support scaffold of tissue engineering periosteum, the supporting scaffolds of tissue engineered periosteum need further exploration.
The repair of the long bone defects by combined grafting of homogenous deealcified bene matrix(DBM ) with centrally enveloped vascularized periosteum Was reported as a new techniqe. Theroentgenograms,bone mineral count and histologic examination were done. The results showed thatthis method was beneficial and had better effect on prornoting healing of the long bene defeets fromone stage operation The oporative proeedure was described on deatil It was considered that the homogenous DBM ...
In order to repair cartilage defect in joint with transplantation of cryopreserved homologous embryonic periosteum, 30 rabbits were used and divided into two groups. A 4 mm x 7 mm whole thickness cartilage defect was made in the patellar groove of femur of each rabbit. The homologous embryonic rabbit skull periosteum (ERSP), preserved in two-step freezing schedule, was transplanted onto the cartilage defect of joints of one group and autogenous periosteal graft was done in the joint defect of the other group. The knees were not immobilized, following operation and 16 weeks later, the newly formed tissue in the defects were assessed by gross observation, histochemical examination and biochemical analysis. The results showed that new hyaline-like cartilage was formed in the cryopreserved ERSP grafted knee, and had no significant difference from that of the knee receiving autogenous periosteal graft, but had significant difference from that of the fresh ERSP grafted knee and the non-grafted knee. Furthermore, the new hyaline-like cartilage had the biochemical characteristics of a fibrous cartilage. The conclusion was that this method might be feasible to repair articular cartilage defects.
It is very difficult to repair large articular cartilage defect of the hip. From May 1990 to April 1994, 47 hips in 42 patients of large articuler cartilage defects were repaired by allograft of skull periosteum. Among them, 14 cases, whose femoral heads were grade. IV necrosis, were given deep iliac circumflex artery pedicled iliac bone graft simultaneously. The skull periosteum had been treated by low tempreturel (-40 degrees C) before and kept in Nitrogen (-196 degrees C) till use. During the operation, the skull periosteum was sutured tightly to the femoral head and sticked to the accetabulum by medical ZT glue. Thirty eight hips in 34 patients were followed up for 2-6 years with an average of 3.4 years. According to the hip postoperative criteria of Wu Zhi-kang, 25 cases were excellent, 5 cases very good, 3 cases good and 1 case fair. The mean score increased from 6.4 before operation to 15.8 after operation. The results showed, in compare with autograft of periosteum for biological resurface of large articular defect, this method is free of donor-site morbidity. Skull periosteum allograft was effective for the treatment of large articular cartilage defects in hip.
ObjectiveTo investigate the feasibility of tissue engineered periosteum (TEP) constructed by porcine small intestinal submucosa (SIS) and bone marrow mesenchymal stem cells (BMSCs) of rabbit to repair the large irregular bone defects in allogenic rabbits. MethodsThe BMSCs were cultivated from the bone marrow of New Zealand white rabbits (aged, 2 weeks-1 month). SIS was fabricated by porcine proximal jejunum. The TEP constructed by SIS scaffold and BMSCs was prepared in vitro. Eighteen 6-month-old New Zealand white rabbits whose scapula was incompletely resected to establish one side large irregular bone defects (3 cm×3 cm) model. The bone defects were repaired with TEP (experimental group,n=9) and SIS (control group,n=9), respectively. At 8 weeks after operation, the rabbits were sacrificed, and the implants were harvested. The general condition of the rabbits was observed; X-ray radiography and score according to Lane-Sandhu criteria, and histological examination (HE staining and Masson staining) were performed. ResultsAfter operation, all animals had normal behavior and diet; the incision healed normally. The X-ray results showed new bone formation with normal bone density in the defect area of experimental group; but no bone formation was observed in control group. The X-ray score was 6.67±0.32 in experimental group and was 0.32±0.04 in control group, showing significant difference (t=19.871,P=0.001). The general observation of the specimens showed bone healing at both ends of the defect, and the defect was filled by new bone in experimental group; no new bone formed in the control group. The histological staining showed new bone tissue where there were a lot of new vessels and medullary cavity, and no macrophages or lymphocytes infiltration was observed in the defect area of experimental group; only some connective tissue was found in the control group. ConclusionTEP constructed by porcine SIS and BMSCs of rabbit can form new bone in allogenic rabbit and has the feasibility to repair the large irregular bone defects.
Abstract In order to find a new method to repair large bone defect, the free periosteum autograft was investigated in experiment, and then the method was used clinically. In the experiment, a 6mm×18mm×5mm bone defect was made at upper end of both tibiae of 42 rabbites. The periosteum of each rabbit was cut into 1mm cubes, and implanted randomly into the tbial bone defect on one side and the other side was used as control. After 2, 4, 8 weeks, the bone defects of each group were examined for bone formation by roentgenography, radionuclide and histology. The results showed that the defects treated by free periosteum autografts healed twice as fast as the controls (its natural healing). The reason probably was that the periosteum provided with many osteogenic cells. On thebasis of these results, 21 cases of bone defects (the largest was 10.5cm×4cm×4cm, the smallest was 2cm×2cm×2cm) including 17 cases of benign bone tumor and4 cases of chronic osteomyelitis, were treated by free periosteum autografts. The defects were all healed, and the function of the joints was restored.