摘要
一实验目的
临床高能量损伤造成的开放性骨折常发生骨髓炎并骨质外露坏死,手术清创后常造成节段性骨缺损。感染性骨缺损的修复重建非常困难。我们试图通过模拟临床创伤后骨髓炎的发病机制建立动物模型,在此模型基础上以骨髓炎坏死皮质骨为研究对象,制备出部分脱钙骨基质并设法恢复其血运,同时结合显微外科技术来修复胫骨感染性骨缺损。
二实验方法:
(一)第一部分
伴软组织缺损的创伤后骨髓炎皮质骨坏死动物模型的创建。以雄性Wistar大鼠为研究对象,随机分为两组,实验组和对照组各10只,共20只。在大鼠胫骨干制造4mm×8mm皮质骨缺损,将3×10~6cfu金黄色葡萄球菌和异物(纱布)接种到胫骨骨缺损处髓腔内,并将游离骨块回植到缺损区,外翻缝合软组织使骨质外露,对照组不接种金葡菌。术后2W、4W观察记录一般情况,行放射学、病理组织学检查。定量数据采用SPSS 12.0统计软件处理。术后通过病理组织学和放射学方法评价骨髓炎及骨坏死的情况。
(二)第二部分
大鼠骨髓炎坏死皮质骨脱钙骨基质制备及理化性质和生物力学测定。以大鼠骨髓炎坏死皮质骨和正常新鲜皮质骨作为研究对象,按改良Urist法将骨髓炎坏死皮质骨经脱钙脱脂处理得到部分脱钙骨基质,扫描电镜观察测量其孔隙度,全自动生化分析仪测量其钙含量并计算脱钙率,生物力学仪测量其形变—负荷曲线。
(三)第三部分
自体骨髓炎坏死皮质骨脱钙骨基质再血管化并修复骨缺损的实验研究。以骨髓炎骨坏死模型造模后的雄性Wistar大鼠为研究对象,随机分成两大组,腹部包埋组:其中实验组和对照组各9只,共18只;骨缺损修复组22只,共40只。所有大鼠取腹部切口显露腹壁浅动脉,将骨髓炎坏死皮质骨部分脱钙骨基质灭菌后,无菌条件下包埋于双侧腹壁浅动脉肌瓣中,对照组将未脱钙坏死皮质骨包埋于相同肌瓣中。腹部包埋组术后1W、2W、4W实验组和对照组各处死3只动物,取材行病理组织学检查观察血管化情况。骨缺损修复组22只大鼠于腹壁浅动脉肌瓣包埋后2W,将部分血管化的坏死皮质骨脱钙骨基质连同腹壁浅动脉带蒂肌瓣转移填充于胫骨骨髓炎骨缺损处,术后4W、8W、12W行经放射学和病理组织学检查骨髓炎及缺损修复情况。
三结果
(一)第一部分
实验组大鼠术后体重下降,体温和WBC升高,放射学检查见髓腔内骨质破坏,骨皮质变薄,骨膜反应和软组织肿胀阴影,部分出现病理性骨折。病理组织学见髓腔内大量炎细胞浸润,骨小梁部分溶解、吸收、稀疏、坏死,骨细胞坏死,骨陷窝空虚。对照侧仅表现为轻微的炎症反应,无明显骨髓炎征象。
(二)第二部分
骨髓炎坏死皮质骨经脱钙脱脂后形成部分脱钙骨基质,平均钙含量为92.17±0.54 mg/g,脱钙率为38.1%。扫描电镜显示:坏死皮质骨脱钙骨基质表面凹凸不平,包含两种微孔结构,直径在7～15μm和0.1～0.4μm之间,微孔之间相互沟通共同形成筛孔样结构,与正常新鲜皮质骨DBM比较,无明显差异。生物力学测定:形变与负荷呈函数关系,随着负荷增加,形变逐渐增加,形变一负荷表现为一条平缓的上升曲线,与新鲜皮质骨形变-负荷曲线比较接近。
(三)第三部分
腹部包埋实验组1W时,部分脱钙骨基质内大量骨细胞坏死,骨陷窝空虚,周围肌肉组织中炎细胞浸润。2W时,哈弗氏管内偶见血管长入,主要集中在表层组织内,骨陷窝内仍空虚,无细胞或血管。4W时少许血管长入,仍以集中在表层为主,血管直径不等,分布不均,骨陷窝内可见成骨细胞,分布密度不均,无特征性骨结构。对照组2W,4W坏死皮质骨组织内均未见血管长入。骨缺损修复组术后4W,脱钙骨基质内出现了软骨化生,周围被纤维细胞包绕,靠近骨髓腔一侧可见炎细胞浸润。术后8W:部分脱钙骨基质大量软骨化,外层包被纤维组织和肌肉。术后12W:部分脱钙骨基质出现不同程度的软骨化和骨化,靠近干骺端一侧骨化明显,出现部分类似骨小梁的结构。放射学显示胫骨缺损修复区4W时,胫骨缺损清晰可见,移植组织密度低,无钙化征象。8W时,胫骨缺损仍清晰可见,移植组织密度低,无明显钙化征象,周围组织硬化,骨膜增生。12W时,胫骨缺损稍模糊,移植组织密度稍增高,与周围骨质有部分连接。
四结论
(一)第一部分
大鼠胫骨造成皮质骨缺损并接种金葡菌和异物,骨块回植并骨外露,可以成功建立伴软组织缺损的创伤后骨髓炎皮质骨坏死动物模型。细菌、异物、创伤是建立此模型的必要条件,软组织外翻缝合骨外露更接近临床伴有软组织缺损的创伤后骨髓炎的实际情况。
(二)第二部分
创伤后骨髓炎坏死皮质骨经脱钙脱脂处理后可得到部分脱钙骨基质,既有三维的网状孔隙结构,又保持了一定的生物力学强度,既脱去部分钙质,暴露出骨基质中的细胞因子,又因源于自体组织不存在免疫排斥反应,具备了DBM的基本特性,并有独特的优点,具备再血管化可能和修复骨缺损的潜能。
(三)第三部分
自体骨髓炎坏死皮质骨部分脱钙骨基质包埋于自体腹壁浅动脉肌瓣中可以再血管化。部分血管化的坏死皮质骨脱钙骨基质连同腹壁浅动脉带蒂肌瓣转移填充于胫骨骨髓炎骨缺损,可以改善局部血运并修复骨髓炎骨缺损。
一Objective
The segmental bone defect often comes from debridement of necrosis cortical bone in post-trauma osteomyelitis.It is very difficult to repair the infect bone defect.We tried to create an animal model of post-trauma osteomyelitis according to clinical pathogenesy.Based on the animal model,the dead bone was prepared to be an partial decalcified bone matrix(DBM).Then we tried to revascularize the DBM to repair the infected tibia defect.
二Methods
The first part:Founding of animal model of necrosis cortical bone in post-trauma osteomyelitis with soft tissue defect.Staphylococcus aureus and foreign body were inoculated into the rat tibia medullary space through the bone defect.The bone block from the tibia defect was replanted onto the defect and was revealed by outpouching the soft tissue.Pathohistology and radiology were used to evaluated the osteomyelitis and bone necrosis.The second part:Preparation of decalcified bone matrix of necrosis cortical bone from the rat infect tibia and determination of the physico-chemical and vitodynamics.property.According to reforming Urist method,the partial decalcified bone matrix (DBM) was obtained from necrosis cortical bone of osteomyelitis by decalcification and defat.The porosity and calcium contents and deformation-lord curve were evaluated by scanning electron microscope and automatic biochemistry analyzer and vitodynamics machine created with of S.aureus and foreign body,while the bone was exposed in the air.It was evaluated with X-ray and pathology.After decalcified and defatted,the necrosis cortical bone was scanned by scanning electric-microscopy,measured the calcification by automatic biochemical machine,measured the biomechanics characteristic by biomechanics machine.The third part: The experimental study of autologous revascularized partial decalcified bone matrix to repair the osteomyelitis bone defect.The sterilized partial decalcified bone matrix was embedded into the superficial epigastric artery muscle flap to revascularize.Combined with the muscle flap vessel pedicle,it was transferred into the infected tibia defect.the revascularization and bone defect reparation,were observed by radiology and pathohistology
三Result
The test tibia revealed feature of osteomyelitis.And the replanted bone was necrosis. The animal model of necrosis cortical bone in post-trauma osteomyelitis with soft tissue defect was founded.After decalcification and defat,the partial decalcification bone matrix was obtained from the necrosis cortical bone.Its calcium contents is 92.17±0.54 mg/g and decalcification rate 38.1%.Two kinds of cellular structure communicating each other were observed through scanning electron microscope with 7～15μm and0.1～0.4μm in diameters. Deformation and lord is functional relation.the deformation raised with the increased lord.Deformation-load curve was similar to the fresh cortical bone curve.Four weeks later, the necrosis cortical bone matrix embedded in the superficial epigastric artery muscle flap were revascularized and decalcified in the infected tibia defect during the later time.
四Conclusion
The animal model of necrosis cortical bone in post-trauma osteomyelitis with soft tissue defect was successfully created in our method.After decalcification and defat,the necrosis cortical bone revealed porous structure and vitodynamics intensity.The necrosis cortical bone matrix embedded in the muscle flap was able to revascularize and repair the infected bone defect.
引文
[1]Mader JT,Cripps MW,Calhaun JH.Adult posttraumatic osteomyelitis of the tibia.Clin Orthop,1999,36 0:14
[2]Gustilo RB,Anderson JT:Prevention of infection in the treatment of one thousand and twenty five open fractures of long bones:Retrospective and prospective analyses.J Bone Joint Surg Am 1976,58:453-458
[3]Kelly PJ:Infected nonunion of the femur and tibia.Orthop Clin North Am 1984,15:481-490
[4]Cierny G,Mader JT,Pennick H:A clinical staging system of adultosteomyelitis.Contemp Orthop 1985,10:17-37
[5]Norden CW.Experimental osteomyelitis Ⅰ.A description of the model.J Infect Dis 1970;122:410-8
[6]Norden CW,Myerowitz RL,Keleti E.Experimental osteomyelitis due to Staphylococcus aureus or Pseudomonas aeruginosa:A radiographic-pathological correlative analysis.Br J Exp Pathol 1980;61:451-60
[7]May JW,Jupiter JB,Weiland AJ,et al:Current concepts review:Clinical classification of post traumatic tibial osteomyelitis.J BoneJoint Surg Am 1989,71:1422-1428
[8]Gazdag AR,Lane JM,Glaser D,et al.Alternatives to autogenous bone graft:efficacy and indications[J].JAAOS,1995,3:1
[9]Goulet JA,Sedudae LE,Deeilra GT,et al.Autogenous iliac crest bone graft-complication and functional assessment[J].Clin Othop,1997,339,76.440.
[10]Tsuchida H,Hashimoto J,Crawford E,et al.Engineered allogeneic mesenchymal stem cells repair femoral segmental defect in rats[J].J Orthop Res,2003,21(1):44-53
[11]Quarto R,Mastrogiacomo M,Cancedda R,et al.Repair of large bone defects by autologous human bone marrow stromal cells[J].N Engl J Med,2001,344(5):385-386
[12]Stock W,Hierner R.Applications and techniques of vascularized bone transfer[J].Injury,1994,25(suppl1):35
[13]刘兴炎,葛宝丰,文益民,等。游离腓骨复合移植重建上肢关节功能及骨缺损[J].中华显微外科杂志,1997,20(2):97-98
[14]徐中和,蔡维山,郭奇峰。带监测皮岛的腓骨移植[J]。中华显微外科杂志。2000,23(1):29-30
[15]杨志明,温玉明。带血管蒂组织瓣移位手术学。四川科学技术出版社,2005.1
[16]朱盛修。有血管循环的骨膜移植回顾与进展[J]。中华显微外科杂志,1992,15(2):139-140
[17]蒋祖言,王爱民,杜全印,等。带血管的骨膜中央包裹与同种脱钙骨基质联合移植修复长段骨缺损的实验研究[J]。中国修复重建外科杂志,1994,8(1):68
[18]One I,Gunji H.Bone induction of hydroxyapatite combined with bone morphogenetic protein and covered with periosteum[J].Plast Reconstr Surg,1995,95:1265.
[19]袁志,马平,胡蕴玉,等。复合rhBMP2的异种骨(rhBMP2/BCB)与骨膜联合移植修复兔桡骨节段性骨缺损[J]。中华骨科杂志,1999,19(9):557-561
[20]HassHJ,KrauseH,KrokerS,eta.l Implantation of human demineralized bone matrix(DBM) for the treatment of juvenile bonecysts[J].OperOrthop Traumato,l 2006,18(1):19-33
[21]Mauney JR,JaquieryC,VollochV,eta.l In vitro and in vivo evaluation of differentially demineralized cancellous bones scaffolds combined with human bone marrow stromal cells for tissue engineering[J].Biomaterials,2005,26(16):3173-3185
[22]Wolfe MW,Cook SD.Use of osteoinductive implants in treatment of bone defects[J].Med Teckol,1994,20:155
[23]Urist MR,Delange RJ,Finerman GA,Bone cell differentiation and growth factors[J].Science,1983,220:680
[24]LG,Naughton G.Tissue engineering—current challenges and expanding opportunities[J].Science,2002,295(5557):1009-1014
[25]Urist MR,Bone Formation by Autoinduction,Science.1965;150:893-898
[26]HassHJ,Krause H,KrokerS,eta.l Implantation of human demineralized bone matrix(DBM) for the treatment of juvenile bonecysts[J].OperOrthop Traumato,1 2006,18(1):19-33
[27]Mauney JR,JaquieryC,VollochV,eta.l In vitro and in vivo evaluation of differentially demineralized cancellous bones scaffolds combined with human bone marrow stromal cells for tissue engineering[J].Biomaterials,2005,26(16):3173-3185
[28]刘振东,范清宇,马宝安,等。成年狗股骨微波高温灭活后的再血管化研究.中华骨科杂志。1998.11:682-685
[29]Stringa G.Studies of the vascularization of bone grafts.J Bone Joint Surg(Br),1957,39:395-420
[1]Mader JT,Cripps MW,Calhaun JH.Adult posttraumatic osteomyelitis of the tibia.Clin Orthop,1999,360:14
[2]Gustilo RB,Anderson JT:Prevention of infection in the treatment of one thousand and twenty five open fractures of long bones:Retrospective and prospective analyses.J Bone Joint Surg Am 58:453-458,1976
[3]Urist MR(1965):Bone:Formation by Autoinduction,Science 150:893-898
[4]HassHJ,KrauseH,KrokerS,et al.Implantation ofhuman demineralized bone matrix(DBM) for the treatment of juvenile bonecysts[J].OperOrthop Traumato,l 2006,18(1):19-33
[5]Mauney JR,JaquieryC,VollochV,et al.In vitro and in vivo evaluation of differentially demineralized cancellous bones scaffolds combined with human bone marrow stromal cells for tissue engineering[J].Biomaterials,2005,26(16):3173-3185
[6]刘振东,范清宇,马宝安,蒋维中,裘秀春.成年狗股骨微波高温灭活后的再血管化研究.中华骨科杂志,1998,11:682-685
[7]Stringa G.Studies of the vascularization of bone grafts.J Bone Joint Surg(Br),1957,39:395-420.
[8]Smeltzer MS,Thomas JR,Hickman SG,et al.Characterization of a rabbit model of staphylococcal osteomyelitis.J Orthop Res,1997,15:414
[9]Rissing JP,Buxton TB,Weinstein RS,Shockley RK.Model of experimental chronic osteomyelitis in rats.[J]Infect Immun,1985,47:581-586
[10]Y.H.An,Q.K.Kang,C.R.Arcilla.Animal models of osteomyelitis.The international journal of artifical organs,2006,29:407-420
[11]沈霖,杜靖远,杨家玉等.慢性化脓性骨髓炎的造模方法及相关指标测定。中国中医骨伤科杂志,1989;5:3
[12]Scheman L,Janota M,Lewin P.The prodution of experimental osteomyelitis:A preliminary report.JAMA 1941;117:1525-9
[13]Norden CW,Experimental osteomyelitis.I.A discription of the model.J Infect Dis 1970;122:410-8.
[14]Einhom TA,Lane JM,Burstein AH,et al.The healing of segmental bone defects induced by demineralized bone matrix A radiographic and biomechanical stud-y.Bone Joint Surg(AM),.1984;66:274-279
[15]孙靖。实验动物学基础。北京:北京科学技术出版社。2005,4:55
[16]Cirny IG,Mader JT,Adult chroic osteomyelitis:An over-view.InD.Am-brosia RD.Marier RL(eds)Orthopedic infections thorofare NJ Slack,1989,31
[17]Worlock P,Slack R,Harvey L,et al.The prevention of infection in open fractures:An experimental study of the effect of fracture stability.Injury,1994 25:31-38
[18]Mackowiak PA,Jones SR,et al.Diagnostic value of sinus-tract cultures in chronic osteomyelitis,JAMA,1978;239:2772-2775
[19]Jauregui LE,Senour CL.Diagnosis and management of bone infections.New York:Marcel Dekker,1995:37-108
[20]刘立峰,张强,王平山,等。胫骨骨感染的分期与治疗。中华创伤骨科杂志,2007,1:27-31
[1]Urist MR,Bone Formation by Autoinduction,Science,1965,150:893-898
[2]Osbon DB,Lilly GE,Thompson CW,et al.Bone grafts with surface decalcified allogeneic and particulate autologous bone:report of cases.J Oral Surg.1977 Apr;35(4):276-84
[3]Pike RL,Boyne PJ.Composite autogenous marrow and surface-decalcified implants in mandibular defects.J Oral Surg.1973 Dec;31(12):905-12.No abstract available.
[4]赵廷宝,范清宇,张殿忠,等。人脱钙骨基质颗粒的制备及应注意的问题。滨州医学院学报。2001,24:6-7
[5]Niyibzi C,Baltzer A,Lattermann C,et al.Potential role for ene therapy in the enhancement of fracture healing[J].Clin Orthop,1998,355:148
[6]Johnson E E,Urist MR.Homan bone morphogenetic protein allografting for reconstruction of femoral nonunion[J].Clin Orthop,2000,371:61
[7]ZhangM,Powers RM,Wolfinbarger L,et a.l Effect(s) of the demineralization process on the osteoinductivity of demineralized bonematrix[J].J Periodonto,l 1997,68(11):1085-92
[8]Ma ZG,Zhou SX,Liu BL,et a.l The mechanical and biological properties ofdemineralized corticalbone allografts in animals[J].J Bone JointSurg(Br),1991,73:791-801
[9]蔡贤华,罗芝荣,徐永年,等。表面脱钙骨基质明胶修复节段性骨缺损的实验研究[J]。中华研究外科杂志。 1995,12(4):219
[10]李东,崔磊,等。脱钙骨基质骨诱导活性评价方法的研究进展[J]。山东医药。2004,44(8):65-66
[11]Osbon DB,et al.Bone grafts with surface decalcified allogeneic and particulate autologous bone.J Oral Surg,1973,31:905
[12]马振国,周树夏,刘宝林。不同脱矿时间皮质骨强度矿物质含量及骨诱导能力的关系。中华骨科杂志,1993,13:117-120
[13]Lin FH,LiaoCJ,ChenKS,et al.Preparation of a biphasic porous bioceramic by heating bovine cancellous bone with Na4P20710H20 addition[J].Biomaterials,1999,20(5):475-484
[14]钱鏊,沈尊理,张兆峰,等。运用组织工程原理结合纳米技术构建骨组织的实验研究[J]。中国修复重建外科杂志。2006,20(5):560-564
[15]李明善,王葆芳,皇甫超申。微孔对同种异体表面脱钙骨基质明胶诱导成骨能力的影响[J]。解剖学研究。2001,23(1):46-47,34
[16]Joseph A,Buckwalter,Orthopaedic.Basic Science:Biology and Biomechanics of the Musculoskeletal System.American Academy of Orthopaedic Surgeons.Second Edition:56-59
[17]Thoren K,Aspenbery P,Thorngren KG.Lipid extraction decreases the specific immunologic response to bone allografts in tabbits[J].Acta Orthop Scand,1993,64(1):44-46
[18]Thoren K,Aspenberg P,Thorngren KG.Lipid extracted bank bone.Bone biomechanical properties [J].Clin Orthop,1995,311:232-246
[19]孙磊,胡蕴玉,宁志杰,等.不同方法处理的同种异体骨移植免疫学比较[J].中华外科杂志,1996,34(8):460
[1]蔡锦方,刘立峰,邹林。复合组织移植治疗创伤性慢性跟骨骨髓炎。组织工程与重建外科。2006,03:12-14
[2]刘立峰,张强,王平山,等。胫骨骨感染的分期与治疗。中华创伤骨科杂志。2007,1:27-31
[3]蔡锦芳,丁自海,陈中伟,主编。显微足外科学。济南:山东科技出版社,2002:262-270
[4]蒋祖言,王爱民,杜全印,等。带血管的骨膜中央包裹与同种脱钙骨基质联合移植修复长段骨缺损的实验研究[J]。中国修复重建外科杂志。1994,8(1):68
[5]One I,Gunji H.Bone induction of hydroxyapatite combined with bone morphogenetic protein and covered with periosteum[J].Plast Reconstr Surg,1995,95:1265
[6]袁志,马平,胡蕴玉,等。复合rhBMP2的异种骨(rhBMP2/BCB)与骨膜联合移植修复兔桡骨节段性骨缺损[J]。中华骨科杂志。1999,19:557-561
[7]JIANG Bao-guo.Treatment Development of Bone Defects.Continuing medical education.2005;(2):4-12
[8]杨明,姜保国。长骨骨不连的治疗进展。中国矫形外科杂志。2003;11(92):119-121
[9]徐达传,钟世镇。实验临床骨缺损修复应用解剖学。中国医药科技出版社。2000;(1):38-72
[10]Lee SJ.Cytokine delivery and tissue engineering.Yonsei Med J.2000;(41)6:704-719
[11]Stock W,Hiemer R.Application and techniques of vascularized bone transfer.Injury,1994,25,(suppl1) G:35
[12]吴宗键,王继芳,卢世璧。诱导成骨的局部基因治疗。中华骨科杂志。2001;12(21)12:760-762
[13]Griffith LG,Naughton G.Tissue engineering-current challenges and expanding opportunities [J].Science,2002,295:1009-1014
[14]Bianco P.Gehron Robey P.Marrow stromal stern cells.J Clin Invest.2000,105..1663-1668.
[15]Iwata H,S akano S,hoh T,et al.Demineralized bone matrix and native bone morphogenetic protein in oethopaedic surgery.Clin Orthop.2002,395:99-109
[16]黄粹业,庞发水。骨缺损的显微外科修复治疗进展。中国解剖与临床。2001;6:183-185
[17]蔡锦方。小腿外侧岛状皮瓣转移治疗胫骨慢性骨髓炎。修复重建外科杂志。1988(2)2:142-143
[18]蔡锦方,刘晓平。应用小腿外侧复合组织瓣治疗下肢慢性骨髓炎。中华显微外科杂志。1990;13:141-142.
[19]Viateau V,Guillemin G,Bousson V.Long-bone critical-size defects treated with tissue-engineered grafts:a study on sheep.J Orthop Res.2007,25:741-9
[20]BrennanPA,Umar T,Wilson AW,et al.Expression of type 2 nitric oxide synthase and vascular endothelial growth factor in oral dysplasia[J].J Oral Maxillofac Surg,2002,60:1455-1460
[21]黄粹业,庞发水。骨缺损的显微外科修复治疗进展。中国解剖与临床。2001;6:183-185
[22]刘振东,范靖宇,马保安,等。成年狗股骨微波高温灭活后的再血管化研究。中华骨科杂志。1998:18:682-685
[23]Kang R,Ghivizzani SC,Muzzonigro TS,et al.Orthopaedic applications of gene therapy.Clin Orthop.2000;375:324-327
[24]侯春林。自体组织移植进展。中国修复重建外科杂志。2006;20(4):309-312
[25]王成琪,王剑利,张敬良,等。皮瓣移植术的回顾与展望。中华显微外科杂志。2000;23:12-13.
[26]Stringa G.Studies of the vascularization of bone grafts.J Bone Joint Surg(Br.),1957,39:395-420
[27]Nicholash H.Mast,MD,Daniel Horwitz,MD.Ostomyelitis:a review of current literature and concepts.Operative Techniques in Orthopaedics.2002,12:232-241
[28]Carl L.Nelsona,Sandra G.McLarena,Robert.The treatment of experimental osteomyelitis by surgical debridement and the implantation of calcium sulfate tobramycin pellets.Journal of Orthopaedic Research.2002,20:643-647
[29]许宋锋,王臻,李涤尘。组织工程化大段人工骨的成骨性能及修复机制。中华实验外科杂志。2005.22:6
[30]Mark E.Jones,James A.Karlowsky,Deborah C.Draghi,et al.Antibiotic susceptibility of bacteria most commonly isolated from bone related infections:the role of cephalosporins in antimicrobial therapyInternational Journal of Antimicrobial Agents,2004,23:240-246
[31]Luca Lazzarini,Benjamin A.Lipsky,Jon T.Mader,et al.Antibiotic treatment of osteomyelitis:what have we learned from 30 years of clinical trials? International Journal of Infectious Diseases.2005,9:127-138
[32]Mader JT,Shirtiliff ME,Bergquist SC,et al:Antimicrobial treatment of chronic osteomylitis.Clin Orthop,1999,36 0:47
[33]Fitzgerald,R.H.Experimental osteomyelitis:description of a canine model and the role depot administration of antibiotics in the prevention and treatment of sepsis.Bone and Joint Surg,1983;65-A(3):371
[1] Ykan Q.K, Kang CR, ARCIOLA. Animal models of osteomyelitis. The International Journal of Artificial Organs; 2006, 29 :407-420
[2] Norden CW. Experimental osteomyelitis I. A description of the model. J Infect Dis 1970; 122:410-8
[3] Norden CW, Myerowitz RL, Keleti E. Experimental osteomyelitis due to Staphylococcus aureus or Pseudomonas aeruginosa: A radiographic-pathological correlative analysis. Br J Exp Pathol 1980; 61: 451 -60
[4] Crane LR, Kapdi CC, Wolfe JN, et al. Xeroradiographic, bacteriologic, and pathologic studies in experimental Staphylococcus osteomyelitis. Proc Soc Exp Biol Med 1977; 156:303-14
[5] Andriole VT, Nagel DA, Southwick WO. A paradigm forhuman chronic osteomyelitis. J Bone Joint Surg Am 1973;55: 1511-5
[6] Borzsei L, Mintal T, Koos Z, et al. Examination of a novel, specified local antibiotic therapy through polymethylmethacrylate capsules in a rabbit osteomyelitis model. Chemotherapy,2006; 52:73-9
[7] Rodeheaver GT, Rukstalis D, Bono M, et al. A new model of bone infection used to evaluate the efficacy of antibiotic-impregnated polymethylmethacrylate cement Clin Orthop Relat Res ,1983; 178: 303-11
[8] Jacob E, Arendt DM, Brook I, et al. Enzyme-linked immunosorbent assay for detection of antibodies to Staphylococcus aureus cell walls in experimental osteomyelitis. J Clin Microbiol 1985; 22: 547-52
[9] Schulz S, Steinhart H, Mutters R. Chronic osteomyelitis in a new rabbit model. J Invest Surg 2001; 14:121-31
[10] Rutledge B, Huyette D, Day D, et al. Treatment of osteomyelitis with local antibiotics delivered via bioabsorbable polymer. Clin Orthop Relat Res 2003; 411:280-7
[11] Evans RP, Nelson CL, Harrison BH. The effect of wound environment on the incidence of acute osteomyelitis. Clin Orthop Relat Res, 1993; 286: 289-97
[12] Poelstra KA, Barekzi NA, Grainger DW, et al. A novel spinal implant infection model in rabbits. Spine 2000;25:406-10
[13] Kondell PA, Granstrom M, Heimdahl A, et al. Experimental mandibular Staphylococcus aureus osteomyelitis; antibody response and treatment with dicloxacillin. Int J Oral Maxillofac Surg,1986; 15: 620-8
[14] Poelstra KA, Barekzi NA, Grainger DW, et al. A novel spinal implant infection model in rabbits. Spine,2000;25:406-10
[15] Buxton TB, Walsh DS, Harvey SB, et al. Bisphosphonate-ciprofloxacin bound to Skelite is a prototype for enhancing experimental local antibiotic delivery to injured bone Br J Surg 2004; 91: 1192-6
[16] Nelson DR, Buxton TB, Luu QN, et al. The promotional effect of bone wax on experimental Staphylococcus aureus osteomyelitis. J Thorac Cardiovasc Surg 1990; 99: 977-80
[17] Rissing JP, Buxton TB, Fisher J, et al. Arachidonic acid facilitates experimental chronic osteomyelitis in rats. Infect Immun,1985; 49:141-4
[18] Lucke M, Schmidmaier G, Sadoni S, et al. Gentamicin coating of metallic implants reduces implant-related osteomyelitis in rats. Bone 2003;32:521-31
[19] Chen X, Tsukayama DT, Kidder LS, et al. Characterization of a chronic infection in an internally-stabilized segmental defect in the rat femur J Orthop Res 2005; 23: 816-23.
[20] Hienz SA, Sakamoto H, Flock JI, et al. Development and characterization of a new model of hematogenous osteomyelitis in the rat. J Infect Dis 1995; 171: 1230-6
[21] Lucke M, Wildemann B, Sadoni S, et al. Systemic versus local application of gentamicin in prophylaxis of implanWelated osteomyelitis in a rat model. Bone-2005; 36: 770-8
[22] Philipov JP, Pascalev MD, Aminkov BY, et al. Changes in serum carboxyterminal telopeptide of type I collagen in an experimental model of canine osteomyelitis. Calcif Tissue Int 1995;57:152-4
[23] Lambe DW Jr, Ferguson KP, Mayberry-Carson KJ, et al. Foreign-body-associated experimental osteomyelitis induced with Bacteroides fragilis and Staphylococcus epidermidis in rabbits. Clin Orthop Relat Res, 1991; 266: 285-94
[24] Horn J, Schlegel U, Krettek C, et al. Infection resistance of unreamed solid, hollow slotted and cannulated intramedullary nails: An in vivo experimental comparison. J Orthop Res, 2005;23:810-5
[25] Arens S, Schlegel U, Printzen G, et al. Influence of materials for fixation implants on local infection. An experimental study of steel versus titanium DCP in rabbits. J Bone Joint Surg Br ,1996; 78:647-51
[26] Costerton JW. Biofilm theory can guide the treatment of device-related orthopaedic infections. Clin Orthop Relat Res, 2005; 437: 7-11
[1]Han SN,Cammisa FP,Sandhu HS,et al.The biology of bone grafting.J Am Acad Orthop Surg,2005,13:77-86
[2]Urist MR.Bone Formation by Autoinduction.Science,1965;150:893-898.
[3]Senn N.On the healing of aseptic bone cavities by implantation of antiseptic decalcified bone.Am J Med Sci,1889;98:219
[4]Sharrard.WJW and Collins DH:The fate of human decalcified bone grafts.Proc Roy Soc Med,1961;54:1101
[5]Wolfinbarger,L.Process for production osteoinductive bone and osteoinductive bone produced Thereby.U.S.patent,2001,6,189,537
[6]Reddi AH and Huggins.Biochemical sequences in transformation of normal fibroblasts in adolescent rats.Proc Nat Acad Sci,1972,69:1601-1605
[7]Leonard B,Kaban,Julie Glowacki.Induced osteogenesis in the repair of experimental mandibular defects in rats.J Dent Res,1981 60(7):1356-1364
[8]孙磊,胡蕴玉,宁志杰,等。不同方法处理的同种异体骨移植免疫学比较[J]。中华外科杂志。1996,34(8):460.
[9]ZhangM,Powers RM,Wolfinbarger L,et al.Effect(s) of the demineralization process on the osteoinductivity of demineralized bonematrix[J].J Periodonto,1997,68(11):1085-92
[10]HassHJ,KrauseH,KrokerS,et al.Implantation ofhuman demineralized bone matrix(DBM) for the treatment of juvenile bone cysts[J].OperOrthop Traumato,2006,18(1):19-33
[11]Ma ZG,Zhou SX,Liu BL,et al.The mechanical and biological properties ofdemineralized corticalbone allografts in animals.J Bone JointSurg(Br),1991,73:791-801
[12]蔡贤华,罗芝荣,陈庄洪,等。表面脱钙骨基质明胶修复大块骨缺损的实验与临床研究。中国骨伤。2003,16:14-16
[13]钱鋆,沈尊理,张兆锋,等。运用组织工程原理结合纳米技术构建骨组织的实验研究.中国修复重建外科杂志,2006。20:560-564
[14]刘俊,余国荣,谭金海,等。带血供骨膜瓣与同种异体脱钙骨联合修复大段骨缺损的实验研究.医学新知杂志.2003,3:145-147
[15]Reddi A H,Wientroub S,Muthukumaran N.Biologic priniciples of bone induction.Orthop Clin North America,1987,18(4):207
[16]RabieAB,DengYM,Samman N,et al.The effect of demineralized bonematrix on the healing of intramembranous bone grafts in rabbit skull defects.J Dent Res,1996,75:1045
[17]Connolly JF.Injectable bone marrow preparations to stimulate osteogenic repair.Clin Orthop,1995, 313(4):8-18
[18]Tiedeman JJ,GarvinKL,KileTA,et al.The role of a composite demineralized bonematrix and bonemarrow in the treatmentofosseous defects[J].Orthopedics,1995,18:1153-1158
[19]KanellopoulosAD,YiannakopoulosCK,Soucacos,et al.Percutaneous reaming of simple bone cysts in children followed by injec-tion ofdemineralized bonematrix and autologous bonemarrow.J PediatrOrthop,2005;25(5):671-675
[20]谭祖健,林贵德。局部注射DBM、BMG修复骨缺损的实验研究。中华创伤杂志。1994:10:171-172
[21]Liu ZT,WenH,FengYB,et al.A experimental study of the efficacy of a demineralized bone matrix allograft/bBMP composite graft in healing of rabbit segmental radial defect in rabbits.Journal of Practical Orthopaedics,2005,11(6):518-521
[22]Moxham JP,Kibblewhite DJ,Bruce AG,et al.Transforming growth factor-beta 1 in a guanidine-extracted demineralized bone matrix carrier rapidly closes a rabbit critical calvarial defect.J Otolaryngo,1996,25(2):82-87
[23]李峰,赵信义,吴军正,等。异体脱矿骨复合碱性成纤维细胞生长因子修复骨缺损的实验研究[J]。中国修复重建外科杂志。2005;19(6):431-434
[24]HollisterSJ,MsddoxRD,Taboas JM,et al.Optimal design and fabrication ofscaffolds tomimic tissue properties and satisfy biological constraints.Biomaterials,2002,23(20):4095-4103
[25]Mauney JR,JaquieryC,VollochV,et al.In vitro and in vivo evaluation of differentially demineralized cancellous bones scaffolds combined with human bone marrow stromal cells for tissue engineering.Biomaterials,2005,26(16):3173-3185
[26]Lin FH,LiaoCJ,ChenKS,et al.Preparation ofa biphasic porous bioceramic by heating bovine cancellous bone with Na4P20710H20 addition[J].Biomaterials,1999,20(5):475-484
[27]Kasten P,LuginbuhlR,Van GriensvenM,et al.Comparison of human bone marrows tromal cells seeded on calcium deficienthydroxyapatite,B-tricalcium phosphate and demineralized bone matrix.Biomaterials,2003,24(15):2593-2603
[28]刘杰,许建中,王序全,等。脱钙骨基质支架构建组织工程骨的实验研究。第三军医大学学报。2005,27(9):888-891
[29]闫冰,闫景龙,张涛。骨组织工程学研究进展。中国骨肿瘤骨病。2004,3(2):76-79
[1] Mader JT,Cripps MW, Calhaun JH. Adult posttraumatic osteomyelitis of the tibia.Clin Orthop,1999,36 0:14
[2] Gustilo RB, Anderson JT: Prevention of infection in the treatment of one thousand and twenty five open fractures of long bones: Retrospective and prospective analyses. J Bone Joint Surg Am 58:453-458, 1976
[3]. Seligson D, Klemm K. Adult posttraumatic osteomylitis of the tibial diaphysis of the tibial shaft. Clin Orthop, 1999,36 0:30
[4] Cierny G, Mader JT, Pennick H: A clinical staging system of adult osteomyelitis. Contemp Orthop 10:17-37, 1985
[5] Daniel P Plew,Francis A Waldvogel, Osteomyelitis: Lancet 2004 (364) : 369-79
[6}Gotz F: Staphylococcus and biofilms. Mol Microbiol. 2002 (43) : 1367-1378
[7] Souli M, Giamarellou H: Effects of slime produced by clinical isolates of coagulase-negative staphylococci on activities of various antimicrobial agents. Antimicrob Agents Chemother 1998 (42) :939-941
[8] Konig DP, Perdreau-Remington F, Rutt J, et al: Slime production of Staphylococcus epidermidis: Increased bacterial adherence and accumulation onto pure titanium. Acta Orthop Scand 69:523-526, 1998
[9] Gristina AG, Hobgood CD, Webb LX, et al: Adhesive colonization of biomaterials and antibiotic resistance. Biomaterials 1987 (8) :423-426
[10] Cierny G, Mader JT: Adult chronic osteomyelitis. Orthopedics 1984 (7) : 1557-1564
[11] May JW, Jupiter JB, Weiland AJ, et al: Current concepts review: Clinical classification of post traumatic tibial osteomyelitis. J Bone Joint Surg Am 1989 (71): 1422-1428
[12].Mader JT, Shirtiliff ME, Bergquist SC,et al :Antimicrobial treatment of chronic osteomylitis . Clin Orthop, 1999,36 0:47
[13] Tetsworth K, Cirny G. Osteomyelitis debridement technique .Clin Orthop 1999 ,36 0:8 7
[14] Peter J, Lorim Dickerson, Jonathan L. Diagnosis and Management of Osteomyelitis. American family physician, 2001 (63) : 2413-2420
[15] Brad Parsons,Elton Strauss. The American Journal of Surgery Surgical. Management of chronic osteomyelitis. 2004,188 (Suppl to July) 57S-66S
[16] Mark E. Jones, James A. Karlowsky, Deborah C. Draghi et al Antibiotic susceptibility of bacteria most commonly isolated from bone related infections: the role of cephalosporins in antimicrobial therapy.International Journal of Antimicrobial Agents. 2004, (23): 240-246
[17]Luca Lazzarini,Benjamin A.Lipsky,Jon T.Mader,Antibiotic treatment of osteomyelitis:what have we learned from 30 years of clinical trials? International Journal of Infectious Diseases(2005) 9,127-138
[18]赵廷宝,范清宇。复合生物活性材料修复骨缺损的研究进展。中国矫形外科杂志。2001(8):270-274
[19]胥少汀,葛宝丰,徐印坎。实用骨科学。北京:人民军医出版社。2003:1119-1120
[20]朱守荣,王继芳,刘玉杰。抗生素局部灌注负压吸引治疗骨髓炎的探讨。军医进修学院学报。1994(15):266-267
[21]Kevin O'Toole.Hyperbaric oxygen therapy and chronic refractory osteomyelitis.Operative Techniques in Orthopaedics,2002(12):282-285