1.Experiment study on compound ink Dabble-Van Gieson dyeing applied to osseous tissue capillary vessel imaging
Yuekui JIAN ; Xuehui WU ; Xiaobin TIAN
Orthopedic Journal of China 2006;0(07):-
[Objective]To find a simple and direct feasible method to detect bone tissue vascularize from comparing advantage or disadvantage of ink intravascular Dabble-VG dyeing and other tissue dyeing in the tissue section capillary vessel display.[Methods]One side femoral artery of 6 normal goat was mixed with liquor composed of 10 percentink,10 percent formaldehyde and 20 percent mannitol,volume ratio in 7∶2∶1.Stopped femoral vein effluent was black.To get off goat tibia,10 percent formaldehyde was fixed with.EDTA decalcification,cedar oil douse,fiat 50 to 100 micrometer pachy-section and compound ink-VG dyeing to observe capillary vessel.[Results]Compound ink-VG dyeing ould display relations of capillary vessel and surrounding tissue.But in control group,single HE,Masson and Weigert Resorcinol-aniline red dyeing after-stain VG dyeing ould display few capillary vessel and surrounding tissue,and recognize difficultly.[Conclusion]Compound ink-VG dyeing can display capillary vessel of bone clearly,and can disclose relationships of surrounding tissue as well.This method can try on in tissue vascularization detect.
2.Evaluation of therapeutics for lumbar tuberculosis(analysis of 65 clinical cases)
Zhuojia ZHOU ; Yuekui JIAN ; Bo LI
Orthopedic Journal of China 2006;0(15):-
[Objective]To evaluate therapeutics for lumbar tuberculosis. [Method]Data from 65 patients treated with five different therapeutics were retrospectively reviewed.Group A(n=14) was treated with focal debridement.Group B(n=15) with focal debridement and bone grafting fusion.Group C(n=9) with posterior focal debridement plus bone grafting fusion and internal fixation.Group D(n=12) with posterior fixation and anterior focal debridement plus,bone grafting fhsion.Group E(n=15) with anterior focal debridement plus bone grafting fusion and anterior internal fixation.Patients all received routine support and anti-tuberculosis therapy before and after operation.Follow-up, erythrocyte sedimentation rate was studied,and bone fusion and kyphosis correction with X-ray plain film and 3D-CT reconstruction were evaluated.[Result]There were not damages occurring in the great vessels,nerves or ureters.No recurrence was found during the follow-up of 12~16 months(average 13 months).Grafting bone were all fused after 4~6 months with an average time of 3.8 months.There was no missing of angel of kyphosis correction,complications of looseness or break of internal fixation.[Conclusion]Efficient anti-tuberculosis therapy and the debridement is the key for treatment of tuberculosis.The selection of fusion or internal fixation is depended on the severity of tuberculosis.Reasonable selection may improve the therapeutic effect of lumbar tuberculosis.
3.Observation on vascularization of big segmental bone defect repaired by heterogeneous deproteinized bone
Yuekui JIAN ; Qihong LI ; Lei LIU
Orthopedic Journal of China 2006;0(10):-
0.05).[Conclusion]Composite of heterogeneous deproteinized bone+autogeneous MSCs+rhBMP2 has the same ability of autogeneous bone graft to repair big experimental segmental tibia defect,thus it can be used as a scaffold in tissue engineering and for further study of immunogenicity and ossification ability.
4.Modified heterogeneous deproteinizated bone composited with recombinant human bone morphogenetic protein for repair of large segmental long bone defects in goat tibia
Yi DING ; Yuekui JIAN ; Bo LI ; Zhuchuan ZHAO
Chinese Journal of Tissue Engineering Research 2010;14(16):3023-3027
BACKGROUND: For its extensive sources and specific biological characteristics, heterogeneous deproteinized bone (DPB) as the tissue-engineered scaffold material gets a good application prospect in repair of large segmental bone defects.OBJECTIVE: To prepare scaffold material by compositing modified heterogeneous DPB with recombinant human bone morphogenetic protein 2 (rhBMP2) and evaluate the osteogenic ability in repairing large segmental long bone defects in large-sized animals.METHODS: Twenty-four goats were randomly and evenly divided into three groups: DPB, autologous bone, and DPB+rhBMP2. Bone defects (20% in length) were made in the middle and lower segments of the right tibia of each goat, and according to group design,different bone grafts were implanted and fixed using semi-ring slot external fixer. At 4-24 weeks after surgery, X-ray examination was performed once every other 4 weeks. At 24 weeks after surgery, newly formed bones were taken out for determining the repair effects of bone defects through duat-energy X-ray analysis, histological observation, and biomechanical analysis.RESULTS AND CONCLUSION: The prepared DPB exhibited a three-dimensional porous structure, with a porosity of (78.5±6.45)% and pore size of (472.5±7.02) μm. Time-dependent bone repair was found in each group. Results regarding anti-compression test, three-point anti-bend test, and anti-torsion test were greatest in the autologous bone group, followed by the DPB+rhBMP2 group, and lastly the DPB group. There were no significant differences in tibial bone density, bone mineral content,anti-compression pressure and ultimate pressure, anti-bending load and ultimate toad, anti-torsion torque and ultimate torque between autologous bone group, DPB+rhBMP2 group and normal bone control group. These findings indicate that modified heterogeneous DPB composited with rhBMP2 has osteogenic ability equivalent to autologous bone in repairing large segmental long bone defects in goat tibias. Therefore, it can be applied as tissue-engineered scaffold material.
5.Risk Factors and Preventive Measures of External Fixator-related Infections in Orthopedics
Yang LUO ; Yuekui JIAN ; Xue ZHANG ; Yongzhi QI ; Weiling FU
Chinese Journal of Nosocomiology 2006;0(03):-
OBJECTIVE To analyze the major reasons of wound infection after external fixator application and then introduce management measures to prevent following wound infections. METHODS Totally 542 patients adopting external fixators between May 2005 and May 2007 were retrospectively reviewed. All the external fixator-related infections were inspected and the excretions from these infected wounds were collected to perform bacterial culturing. RESULTS The total infection rate of these 542 patients after external fixator application was 2.77%. Among them, six were infected with the bacteria in distraction osteogenesis group and the infection rate was 8.82%; three were infected in bone un-union and bone defect group and the infection rate was 5.36%; whilest the common fracture-fixing group got the lowest infection rate of 0.39%. CONCLUSIONS Wire-crossing positions are the most frequently infected sites after external fixation and the drug-resisted bacteria are the most commonly detected pathogens. Thus, increasing the stability of fixators, enhancing the infection supervision of operation environment, draining the wound thoroughly and using antibiotics rationally are the most effective managing measures to prevent external fixator-related infections in orthopedics.
6.Constructing a goat tibial defect model by half-ring sulcated external fixator
Yuekui JIAN ; Xiaobin TIAN ; Zhuchuan ZHAO ; Rongze AN ; Bo LI ; Bing QIU ; Zheng YANG
Chinese Journal of Tissue Engineering Research 2008;12(17):3393-3397
BACKGROUND:Half-ring sulcated external fixator is a bone external fixation device.Its structure is relatively simple,with multi-plane fixed,and small occupying space.So far,there are few experimental reports about construction of tibial defect model by it in large animals.OBJECTIVE:To validate the practicality and repeatability of half-ring sulcated external fixator in establishing tibial defect models in goats.DESIGN,TIME AND SETTING:The randomized controlled observation was performed at the Animal Laboratory of the Third Military Medical University from March 2005 to February 2007.MATERIALS:Nine healthy adult(Animal Center of Third Military Medical University);Half-ring sulcated external fixator,made by Professor Li of the Third Military Medical University;Osseous pin,ψ2.5 mm,Shanghai Medicai Instruments Co.,Ltd.METHODS:After animals were anesthetized,two needles were threaded in the same plane of cancellous bone about 1.5-2.0 cm above the superior articular surface of the tibia,named group 1.The angle between the two needles was 40°.One needle was threaded at the place of 3.0 to 5.0 cm under the group 1 and parallel with optional needle of the group 1,named group 2.Needles of group 4 were threaded at the cancellous bone that was 2.0 cm from the joint surface of inferior extremity of the tibia.One needle was threaded at the place of 3.0 to 5.0 cm from group 4 and parallel with optional needle of the group 4.The included angle between these two needles of group 2 and group 3 was 60°.There were totally 6 needles.Tibia was fixed by half-ring sulcated external fixator.According to the length of the tibia from X-ray,tibia and periosteum were amputated by wire saw between the two needles in the middle and inferior segment of the tibia to make segmental bone and periosteal defects of 20 percent of tibia.MAIN OUTCOME MEASURES:①Postoperative general state of the animals and effect of half-ring sulcated external fixator.②Self-repair of bone defects by X-ray and Lane-Sandhu scoring standards.RESULTS:Of the 9 selected goats,1 died of postoperative infection,and 1 developed pinhole infection and supplemented.All the others survived and were included in final analysis.The goats were awake within 2-6 hours after surgery and able to stand up to eat.The loosening nuts were screwed up.No external fixation failure or loosening was found.No skin was necrotic.Injured limb could touch the ground in 2 days,but could not bear load until 2 weeks.The goats could walk in three weeks with a slight limp,then moved freely without claudication in four weeks after the operation.At the 5th week after operation,the radiographs showed little periosteal reaction.At the 10th week after operation,sclerosis was found in the fracture ends,and medullary cavity started to close.Fifteen weeks after operation,the medullary cavity closed and periosteal reaction did not aggravate.Inaddition.Lane score was 0 at any time point.No bone formation was found in the histological examination at 5,10 and 15 weeks after operation,and Lane score was 0 at any time point.CONCLUSION:The goat model established in goat tibia with 20%defects by the half-ring sulcated external fixator shows no bone healing after 15 weeks by X-ray or histological exanimation,indicating this is a practical and repetitive method to establish animal models of large segmental defect for bone tissue engineering.
7.In vivo osteogenesis by implanting dual gene activated nanobone putty
Yi ZHANG ; Li SUN ; Yuekui JIAN ; Ruyin HU ; Xiaobin TIAN ; Bo LI ; Wei HAN
Chinese Journal of Tissue Engineering Research 2014;(3):329-334
BACKGROUND:The bone morphogenetic protein 2 (BMP2)/vascular endothelial growth factor (VEGF) dual gene activated nanobone putty has been constructed in the previous experiments.
OBJECTIVE:To investigate the effects of osteogenesis and osteogenic gene expression in mice by implanting BMP2/VEGF dual gene activated nanobone putty.
METHODS:Twenty-four Kunming mice (48 sides) were randomly divided into four groups. Animals in each group (12 samples) were injected different materials into the right thigh muscle pouches:nanobone putty+hBMP2/VEGF plasmid;nanobone putty+hBMP2 plasmid;blank plasmid+nanobone putty;nanobone putty only. The effects of osteogenesis were evaluated by radiography, histology and molecular biology analysis in 2, 4 weeks after operation.
RESULTS AND CONCLUSION:Bone-like tissues were observed in groups of nanobone putty+hBMP2/VEGF plasmid and nanobone putty+hBMP2 plasmid after operation. There was apparent BMP2 and VEGF mRNA expression in group of nanobone putty+hBMP2/VEGF plasmid. Group of nanobone putty+hBMP2/VEGF plasmid was significantly better than group of nanobone putty+hBMP2 plasmid in the alkaline phosphatase levels, the speed of osteogenesisas and amount of new bone (P<0.05). Groups of blank plasmid+nanobone putty and nanobone putty had no obvious osteogenesis performance. Either BMP2/VEGF dual gene activated nanobone putty or BMP2 gene activated nanobone putty had the osteogenic ability in vivo. And the former was significantly enhanced in the speed and quality of osteogenesis.
8.The investigation of BMP-2 and VEGF dual gene activated nanobone enhanced the vertebral bone of ovariectomized goat
Zhen YANG ; Xinglin WU ; Jianyang LI ; Wei PAN ; Yuekui JIAN ; Bo LI
Chinese Journal of Microsurgery 2016;39(5):469-473
Objective To verify BMP-2 and VEGF gene activated nanobone paste can effectively enhance the vertebral bone of ovariectomized goat.Methods From January,2011 to May,2016,the goats had been neutered by ovariectomy 6 months earlier to induceosteoporosis.Then surgery to established the model of vertebral bone defected with nanobone implanting,and the operation vertebrae divided randomly into 3 groups:control group,nanobone group and double gene activated nanobone group.Three months after operation the goats were sacrificed and removed the vertebrae.Micro CT analysis of micro three-dimensional structure of trabecular bone,scanning electron microscope (SEM) analysis of the two-dimensional structure of the vertebrae,the structure of trabecular bone was evaluated by movat pentachrome staining.Results The dual gene activated nanobone group compared with the nanobone group,the bone volume fraction (BV/TV) significantly increased (85% at 1.2 mm vs 43% at 1.2 mm,P < 0.05);the dual gene activated nanobone group compared with nanobone group,in the largest ROI (1.2 mm),TbTh increased 10.9% (374 ± 26.2 μm vs 337 ± 22.3 μm,P < 0.05);Trabecular distribution coefficient (TbPf) was significantly decreased (7.519 ± 0.184 mm-1 vs 7.529 ± 0.261 mm-1,P < 0.05);In the largest ROI (0.8 mm),trabecular distribution coefficient (TbPf) was significantly decreased (283 ± 36.4 μm vs 327 ± 31.2 μm,P < 0.05),In the largest ROI (0.8 mm),the trabecular bone volume (Tbn) was increased 20%(1.404 ± 0.283 mm-1 vs 1.173 ± 0.224 mm-1,P < 0.05);Cortical thickness over the implantation area showed asignificant increase of 43% in vertebrae(P < 0.05);The histological analysis revealed a more extensive osseointegration of the dual gene activated nanobone group,with the presence of anabundant osteoid tissue and an osteoblastic celllining.Conclusion BMP-2 and VEGF gene activated nano bone paste can effectively enhance the vertebral bone of ovariectomized goat.
9.One stage posteroanterior decompression and bone implant for treatment of severe lower cervical spinal bony canal stenosis
Chunshan LUO ; Bing QIU ; Bo LI ; Xiaobin TIAN ; Zhaojia ZHOU ; Zhi PENG ; Yuekui JIAN ; Weifeng ZHAO
Chinese Journal of Trauma 2011;27(8):684-687
ObjectiveTo explore the clinical outcome of one stage posteroanterior decompression and bone implant in the treatment of severe lower cervical spinal bony canal stenosis. Methods The study involved 29 patients with severe lower cervical spinal bony canal stenosis treated with one stage posteroanterior decompression and bone implant from April 2006 to March 2009. There were 11 patients with old fractures, seven with posterior longitudinal ligament ossification and 11 with cervical disc calcification. The course of disease ranged from 2 months to 3.2 years, average 1.4 years. The nerve function was rated as grade B in two patients, grade C in 19 and grade D in eight according to Frankel scale. The average Japanese Orthopaedic Association (JOA) score was 9.8. ResultsAll patients were followed up for 7-28 months (average 15.2 months), which showed bony fusion five months after operation, with fusion rate of 100%. The Frankel grade was increased for average 1.2 grades and the nervous symptoms alleviated remarkably. Mean postoperative JOA score was 13.8 and increased for mean 4.0, with mean amehoration rate of 55.6%. ConclusionsOne stage posteroanterior decompression and bone implant is a safe and effective method for treatment of lower cervical spinal bony canal stenosis, when the intraoperative electrophysiological monitoring can assure the operative safety.
10.Observation on vascularization of large segmental bone defect repaired by heterogeneous deproteinized bone
Yuekui JIAN ; Xiaobin TIAN ; Bo LI ; Zhuojia ZHOU ; Weifeng ZHAO ; Yi ZHANG ; Zheng YANG ; Yuanzheng WANG
Chinese Journal of Tissue Engineering Research 2008;12(23):4549-4553
BACKGROUND: There are few reports about vascularization in the repairing of bone defect by heterogeneous deproteinized bone.OBJECTIVE: To verify the vascularization characteristics of heterogeneous deproteinized bone, tissue engineering scaffold material, in the repairing of large segmental bone defect.DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed between March 2005 and February 2007 at the Third Military Medical University, Chongqing, China.MATERIALS: Twenty-four 10 to 12 months old goats, weighing (22.5±2.5)kg, were obtained from the Animal Center of the Third Military Medical University, Chongqing, China. Segmental bone defects of 20 percent right tibia middle and inferior diaphysis of the 24 goats were made.METHODS: The 24 goats were divided into test group (n=16) and control group (n=8) randomly. Goats in test group were implanted with deproteinized bone+autologous MSCs+recombinant human bone morphogenetic protein-2 (rhBMP-2), goats in control group were implanted with autograft bone, and all fixed with half-ring sulcated external fixator. Every 4 weeks, 3 goats were killed after ink perfusing through femoral artery. A thick slice of new bone tissue was made to observe the vascularization.MAIN OUTCOME MEASURES: Vascularization of new bone observed through gross anatomy and imaging; vascular network of new bone observed through thick section, blood vessel amount and area measured by Image-proplus really image analysis software.RESULTS: No goat was infected or dead. Animal soft tissue was dyed black, blood vessels'size, ditribution and network structure were observed in subcutaneous tissue, fascia and periosteum. At 4 weeks postoperation implant margin became crude in the defect area; at 8 weeks postoperation transparent bone absorbing area of different size and irregular shape appeared; after 12 weeks postoperation high-dense calcification shadow appeared at the ends of defect bone and new bone connected with the ends completely. On 4 to 24 weeks postoperation, the blood vessel amount became large, alignment became regular, and their size and distribution became uniform. It showed no significant difference in blood vessel amount and area between 2 groups (P>0.05).CONCLUSION: Composite of heterogeneous deproteinized bone+autologous MSCs+rhBMP-2 has the same vascularization degree. of autogeneous bone graft in repairing of large segmental tibia defect.