OBJECTIVE: To study the treatment efficacy of vascularized periosteum graft and bone filling material for long bone defect. METHODS: Forty young and forty adult rabbits were divided into four groups respectively according to the bone filling materials. A 3 cm long segment was removed from the middle part of the rabbit radius to make a bone defect model. The periosteum was reserved and restored to set up a vascularized tubulate periosteum graft. On the left side, autogenous bone graft, decalcified allograft, tricalcium phosphate, and hydroxyapatite were used to fill in the bone defect respectively; on the right side, no bone filling material was used as controls. The repairing effect of bone defect was evaluated by roentogenography, biomechanical, and histological methods. RESULTS: In young rabbits, bone defects on both sides healed in the 6th week after operation. The bending strength of radius in the tricalcium phosphate group and in the hydroxyapatite group were lower in the 12th week and there was significant difference when compared with autogenous bone graft group, decalcified allograft group and control group (P lt; 0.05). The repairing mechanism included intramembranous and endochondral ossification, and intramembranous ossification was prevalent. In the adult rabbits, the repairing rates of bone defect were 50% in the autogenous bone graft group, 40% in the decalcified allograft group, 30% in the tricalcium phosphate group and in the hydroxyapatite group and 42.5% in the control group, respectively. CONCLUSION: In young rabbits, large bone defect can be repaired with vascularized tubulate periosteum graft with or without the combining use of bone filling materials. The bone filling material which will be substituted slowly is disadvantageous to the recovery of bone strength. In adult rabbits, vascularized tubulate periosteum graft combined with bone filling materials can not repair the large bone defect effectively.
Objective To investigate the advance in the management of skeletal trauma of the extremities. Methods The literature at home and abroad was reviewed, and the research findings withclinical experience in the therapeutic methods for fracture of the extremities were summarized.Results The concept on fracture management was renewed, the minimally invasive surgery (MIS) was developed and popularized, the implantation was improved, the navigation technique with computerassisted surgery was applied, and the tissue engineering was developed. The fracture mana gement was changed from the anatomical reduction with absolutely rigid fixation to the biological osteosynthesis with protection of the fracture environment. The minimally invasive surgical techniques included the minimally invasive plate osteosynthesis, intramedullary nailing, external fixation, arthroscopic surgery,and computer-assisted surgery. In concordance with the MIS principles, the newimplants, such as the locking compression plate, and the less invasive stabilization system were well designed and put into clinical practice so as to provide effective therapeutic results in treating osteoporotic fractures and complicated articular and/or metaphyseal fractures. In treatment of the delayed union or nonunion offractures, more effective techniques were employed, including the application of bone substitutes, which are degradable and have properties of bone conduction and induction. In the repair of segmental defects of the long tubular bone, the bonetransport and the vascularized bone grafts could work well. The investigation of the bone engineering revealed its great potentiality.Conclusion Fracture of the extremities is a common problem and its management should emphasize the recovery of the extremity function of the patient in addition to emphasis on the replacement and fixation of the biological structures. The combination of bone engineering and microsurgery represents the development tendency inthis field.
Objective To compare the cl inical results of two plating osteosynthesis techniques, open reduction and plating ostosynthesis (ORPO) and minimal invasive plating osteosynthesis (MIPO), in surgical treatment of mid-distal humeralshaft fractures. Methods From March 2004 to October 2006, 40 cases of closed unilateral mid-distal humeral shaft fractures were surgically treated with MIPO or ORPO. In the MIPO treated group (n=19), there were 14 males and 5 females, with a mean age of 39.05 years. Fractures involved in middle humeral shaft in 10 cases and distal fragment in 9 cases. According to OTA classification, there were 3 cases of type A, 13 cases of type B and 3 cases of type C. Four cases compl icated by radial nerve palsies. In the ORPO treated group (n=21), there were 13 males and 8 females with a mean age of 39.05 years, including 14 cases of type A and 7 cases of type B fractures according to OTA classification. The fractures involved in middle humeral shaft in 13 cases and distal fragment in 8 cases. Five cases compl icated by radial nerve palsies. The time from injury to operation in both groups were 2 to 14 days. For patients in the MIPO group, fractures were closely reduced and fixated with an anterior placed plate inserted through two small incisions made at the anterior side of arm, away from fracture sites. The radial nerves were not exposed. For patients in the ORPO group, fractures were exposed, reduced, and fixated with an anterolateral or a posterior positioned plate after careful dissection and protection of radial nerve through an anterolateral or a posterior approach. The operation time, the occurrence of iatrogenic radial nerve palsy and the bone heal ing time were recorded. The functions of the affected shouldersand elbows were evaluated with UCLA end-result score and Mayo elbow perform index (MEPI), respectively. Results All the wounds in both groups healed primarily. There was no iatrogenic radial nerve palsies in the MIPO group after surgery; however, 5 cases of transient iatrogenic radial nerve palsies were identified in the ORPO group after surgery, and the function of radial nerve recovered in these cases at the last follow-up. Eighteen cases were followed up 14-44 months (mean 25.44 months) in MIPO group, and 19 cases were followed up 13-48 months (mean 32.11 months) in ORPO group. The mean bone heal ing time was 17.06 (12-32) weeks in MIPO group and 16.11 (8-58) weeks in ORPO group, showing no significant difference between two groups (P gt; 0.05). There was no nonunion and hardware failure in both groups. The mean forward flexion of the shoulder was 166.94° (150-170°) in MIPO group and 164.74° (130-170°) in ORPO group. The mean UCLA shoulder score was 34.78 (33-35) points in MIPO group and 34.42 (30-35) points in ORPO group. The mean range of motion of the elbow in MIPO and ORPO groups was 133.33° (120-140°) and 136.7° (120-140°), respectively. The MEPI in these two groups was 99.44 (90-100) and 99.74 (95-100) points, respectively. There was no statistically significant difference between two groups in all indexes mentioned above. Conclusion The good results could be obtained when ORPO and MIPO technique are appl ied to treat mid-distal humeral shaft fractures. MIPO technique has advantages to not expose the radial nerve and to decrease the occurrence of iatrogenic radial nerve palsies.
Objective To investigate the effect of various concentration of platelet-rich plasma (PRP) on osteogenic differentiation of rabbit skeletal muscle-derived stem cells (SMSCs) cultured in vitro. Methods Blood drawn from the central ear arteries of 9 one-year-old New Zealand white rabbits weighing 2.5-3.0 kg (male and female) was used to prepare PRP (Landesberg method). Full blood count and platelet count in PRP were tested. Soleus muscle of right hindl imb in rabbit was obtained and used to culture SMSCs in vitro. The cells at passage 3 were randomly divided into different groups: the experimental groups in which the cells were treated by conditioned culture media with various concentrations of autologousPRP (6.25%, 12.50%, 25.00%, 50.00%), and the control group in which the cells were treated with the media without PRP. At different time points after intervention, osteogenetic activity of the cells was detected by ALP staining observation, ALP activity detection was conducted, al izarin red staining for calcium nodules and immunofluorescence staining for osteocalcin were performed, and core binding factor α1 (Cbfα1) of osteogenic gene expression was tested by RT-PCR. Results The full blood PRP count and the platelet count in PRP was (3.06 ± 0.46) × 105/μL and (18.08 ± 2.10) × 105/μL, respectively. ALP staining: the cells in all the experimental groups were positive for the staining with many black sediment particles in cytoplasm; the cells in the control group were negative staining. ALP activity: all the experimental groups were higher than the control group (P lt; 0.05), the experimental group at 12.50% was superior to other experimental groups at each time point (P lt; 0.05). Al izarin red staining: at 14 days after culture, orange-red calcium nodules were evident in all the experimental groups; no orange-red calcium nodules were observed in the control group with a mineral ization rate of zero; there were significant difference between the experimental groups and the control group in terms of mineral ization rate (P lt; 0.05), the experimental group at 12.50% had a higher mineral ization rate than other experimental groups (P lt; 0.05). Immunofluorescence staining for osteocalcin: at 7 days after culture, the experimental groups were positive for the staining with yellow fluorescence in cytoplasm, and the result of the control group was negative. RT-PCR detection: no obvious changes of the gene expression were noted at 4, 12, and 24 hoursafter culture in the control group; the gene expression in all the experimental groups was significant superior to that of control group, especially at 12 hours, and the expression in the experimental group at 12.50% was the highest. Conclusion PRP can obviously promote the osteogenic differentiation of SMSCs cultured in vitro in a concentration-dependent manner, and the 12.50% is proved to be the ideal concentration.
【Abstract】 Objective To measure the changes of bone mineral density and bone micro-architecture of thefemoral head that harvested from the three-foot bearing ethanol destroyed canine model for osteonecrosis of femoral head, and discuss the influences of local injection of ethanol and biomechanical loading to the structural properties of the femoral head. Methods Twenty-four Beagles were divided randomly into four-foot bearing canines and three-foot bearing canines. One fore-l imb was fixed randomly in three-foot bearing canines. Osteonecrosis was induced in all experimental animals by local injection of 5 mL pure ethanol into one side of the femoral head. The hind l imbs injected with NS were acted as control group, that of three-foot canines injected with ethanol were acted as three-foot canine group, and that of four-foot canines injected with ethanol were acted as four-foot canine group. The contralateral femoral head was injected into equal amount of NS. Animals were sacrificed at the time intervals of 1, 3, 6, and 12 weeks after the injection of ethanol. Quantitative microcomputedtomography was used to characterize changes in bone micro-architecture and bone mineral density of femoralhead. Results The clear three-dimensional model of trabecular bone of necrotic femoral head were obtained. There were no significant differences among 3 groups according to the time l ine by 1 week after ethanol injection(P gt; 0.05). At 3 weeks after injection of ethanol, in three-foot canine group and four-foot canine group, the volume of BMC, BMD, BVF, and BS/BV increased gradually as the distance to the drill ing canal increased. There were significant differences between 3 regions (P lt; 0.05). At 6 weeks, in three-foot canine group and four-foot canine group, the volume of BMC, BMD,BVF, and Tb.N of region I and II decreased significantly compared with region III (P lt; 0.05). At 12 weeks, there are no differences among 3 groups (P gt; 0.05). There were significant decreases in BMD values, BVF, BS/BV, Tb.N, Tb.Sp and Tb.Th after the injection of pure ethanol. And, the changes were more and more obvious by the time l ine. These changes were differentiable at 3 weeks after injection of ethanol, and became obvious at 6 weeks. These changes were more obvious at the part that near the injection canal. The changes in threefoot canine group were more obvious than that in four-foot canine group. Conclusion Resorption of necrotic compact bone trabecular may weaken the structural properties of the femoral head. Moreover, remodel ing and repairing process of necrotic bone trabecular may be hampered by constant biomechanical loading that presented in three-foot bearing canines, and thereby further weaken the structural properties of the femoral head. Biomechanical loading may be one of the critical reasons that lead to the collapse of femoral head.
Objective To explore the effects of the basic fibroblast growth factor（bFGF） gene transfection on the meniscal fibrochondrocytes with the reconstructed lentivirus and to observe the response of the meniscal fibrochondrocytes to the bFGF gene transfection. Methods The cultured meniscal fibrochondrocytes were isolated from the same 3-monthold New Zealand rabbit. The cultured first-generation meniscal fibrochondrocytes were divided into 3 groups:Group A (experimental group), Group B (control group), and Group C (blank group). Each group comprised the cells in a 24hole flask in which each hole contained 2×104 cells. At the confluence of 60%, the fibrochondrocytes in Group A were cultured with the reconstructed lentivirus carrying the bFGF gene. The fibrochondrocytes in Group B were cultured with the lentivirus carrying no bFGF gene. The fibrochondrocytes in Group C were cultured without any intervention. After 48 h, the cell cycle, the collagen synthesis ability, the expression of bFGF, and the cell proliferation ability in each group were investigated. Results In Group A, the bFGF expression of 870±60 pg/ml was detected in the cells 48 h afterthe co-culture; however, in Group B and Group C, no expression of bFGF was found. After the co-culture for 6 days, the results of the MTT colorimetry revealed that the cells in Group A had an absorbtance of 0.427±0.037, which had a significant difference when compared with that in Group B and Group C (0.320±0.042,0.308±0.034,Plt;0.01). The cell cycle was significantly shorter in GroupA than in Group B and Group C (Plt;0.05); The durations of G1, S and G2M of the cells in Group A were 16.28, 12.60 and 11.04 h, but those in Group B and Group C were 23.61, 16.90, 21.33 h and 21.56, 19.80, 21.41 h, respectively. The disintegration per minute of the cells was significantly greater in Group A than in Group B and Group C (7281.69±805.50 vs 5916.40±698.11 and 5883.57±922.63，Plt;0.05). Conclusion The lentivirus vector can transfer the bFGF gene into the meniscal fibrochondrocytes, resulting in an increase of the cell proliferation and the collagen synthesis.
Objective To investigate the possibility of constructing eukaryoticexpression vector for human angiopoietin 1（hAng-1），transfecting it to bonemarrow mesenchymal stem cells (MSCs) so as to repair bone defect. Methods The eukaryotic expression vector pcDNA3-hAng-1 was constructed by recombinant DNA technique, transfected into MSCs by liposome DOTAP, and selected with G418. The hAng-1 expression of mRNA and protein was detected by reverse transcript-PCR and Western Blot. Results After the recombinant eukaryotic expressionvector for hAng-1 was digested with Xho-I and BamH-I, electrophoresis revealed 1.4 kb fragment for hAng-1 gene and 5.4 kb fragment for pcDNA3 vector. In the transfected MSCs, the mRNA and protein expression of hAng-1 gene were detected with reverse transcriptPCR and Western Blot. Conclusion The constructed eukaryotic expression vector hAng-1 could be expressed in the transfected MSCs, thus to provide the basis for bone repair with tissue engineering.
Objective To assess the effect of basic fibroblast growth factor (bFGF) and 5-fluorouracil (5-FU) appl ied topically on the tendon adhesion and the heal ing process after the flexor tendon repair in Leghorn chickens. Methods Ninety male Leghorn chickens (weighing 3.0-3.5 kg) were randomly divided into 3 groups, with 30 chickens in each group. The flexordigitorum profundus tendons of the third right toes were transected and sutured directly. The repair site in group A was given 0.6 μL fibrin sealant (FS). In group B, the repair site was given 0.6 μL FS containing 500 ng bFGF. In group C, before the tendons were transected, they had been soaked in 5-FU solution, and then the same treatment as group B was given. Six specimens of the third toe were harvested to perform the macroscopical and histological examinations at 1, 2, 4, and 8 weeks, respectively, and to perform the biomechanical test at 8 weeks. Results All animals survived until the experiment was completed. All incisions healed smoothly. No rupture occurred in the reparied tendon. At 8 weeks, the adhesion degree was l ighter in group C than in group B (P lt; 0.05), but there was no significant difference in the adhesion degree between group A and groups B, C (P gt; 0.05). At 1, 2, and 4 weeks after operation, the number of fibroblast cells of group A was significantly less than that of group B (P lt; 0.05), and the number of fibroblast cells of group C was significantly less than that of group A and group B in the tendon sheath and epitenon (P lt; 0.05); however, it was significantly more than that of group A in the tendon parenchyma (P lt; 0.05), and no significant difference was observed when compared with that of group B (P gt; 0.05). At 8 weeks, no difference was found among 3 groups (P gt; 0.05). The collagen fiber content of group A was significantly less than that of group B at 4 and 8 weeks (P lt; 0.05). In the sheath and epitenon, the collagen fiber content of group A was significantly more than that of group C at 4 weeks (P lt; 0.05); however, no significant difference was found between 2 groups at 8 weeks (P gt; 0.05). The collagen fiber content of group A wassignificantly less than that of group C in the parenchyma at 4 and 8 weeks (P lt; 0.05). At all time points, the collagen fiber content of group B was significantly more than that of group C in the sheath and epitenon (P lt; 0.05), but no significant difference in the parenchyma was observed between 2 groups (P gt; 0.05). The biomechanical tests showed that the gl iding excursion of the tendon in groups A, B, and C was (3.51 ± 0.56), (2.84 ± 0.42), and (4.56 ± 0.59) mm, respectively; the work of flexion was (14.08 ± 1.85), (20.62 ± 3.52), and (10.91 ± 1.53) N.mm, respectively; and the ultimate tensile strength of the tendon was (11.26 ± 1.83), (15.02 ± 2.20), and (14.40 ± 1.57) N, respectively. There were significant differences in the gl iding excursion of the tendon and the work of flexion among 3 groups (P lt; 0.05) and in the ultimate tensile strength of the tendon between group A and groups B, C (P lt; 0.05), but there was no significant difference in the ultimate tensile strength of the tendon between group B and group C (P gt; 0.05). Conclusion Local single-use bFGF and 5-FU can not only effectively promote the heal ing of flexor tendon, but also significantly reduce tendon adhesion.