纳米生物陶瓷人工听骨的实验研究
|
摘要
目的:制备纳米多孔羟基磷灰石(HA)陶瓷支架;研究新型陶瓷支架的生物相容性。方法:用水热法法制备纳米级HA粉体,有机泡沫浸渍法制备多孔陶瓷支架,观察其形貌特征,组成成份及力学性能。采用鼠成骨细胞(periosteal-derived osteoblast cells,POB)与两种陶瓷支架的体外复合培养,观察细胞在材料上的生长及功能表达。本研究通过不同理化方法的加工工艺,制备纳米级的多孔羟基磷灰石(HA)支架,采用与鼠骨膜成骨细胞的体外复合培养,体内动物植入实验,评价多孔的HA和AO,这两种骨组织工程支架材料的生物相容性。论文研究纳米多孔HA陶瓷的制备及性能测试并对纳米多孔HA陶瓷的生物相容性进行评价。采用纳米HA颗粒悬液直接作用于成骨前体细胞。运用四吟盐比色法、碱性磷酸酶的方法分析悬液对成骨前体细胞增殖和分化影响。探讨了钙离子溶出的培养液和条件培养液吸附的对细胞增殖、分化的影响。在体内行豚鼠听泡内植入实验。选择健康、白色豚鼠30只(60耳),采用耳后进路,听泡后外侧骨壁钻孔,左侧听泡植入AO普通陶瓷块作为对照组,右侧听泡植入纳米羟基磷灰石(HA)复合多孔陶瓷人工听骨做为实验组,分别于术后1、2、4、8、12、16周处死,行植入物表面接触缘大体形态学及扫描电镜观察。结果:制备出具有良好孔隙结构和一定强度的纳米多孔陶瓷支架材料;豚鼠POB在HA和AO表面良好的生长,并发挥成骨功能。30只豚鼠术后均无感染,听泡光镜及扫描电镜检查,植入物界面形态良好,结构完整,边界清晰。4周时,材料周围被薄层纤维组织包裹,因轻微炎性反应,有少量巨噬细胞和多核巨细胞存在,有纤维组织长入生物陶瓷边缘区域。12周时,均无炎性反应,可见大量胶原纤维组织长入陶瓷中央。纳米HA复合多孔陶瓷可见少量的骨组织。术后16周,有新骨形成,而对照耳仅有粘膜覆盖,无新骨形成。结论:以HA的纳米级粉体为原料,采用有机泡沫浸渍法可制备出具有良好孔隙结构和较好力学强度的多孔陶瓷支架;新型多孔陶瓷支架有良好的生物相容性。
Objective:To make porous nano-bioceramics and to study their biocompatibility as bone tissue engineerings caffolds.Methods:The nanometer powders of hydroxyapatite(HA) and tricalcium phosphate(AO) were prepared by Sol-gel methods and chemical deposition methods and the porous nano-bioceramics were made by foam sockage technics. The biocompatibility of new porous nano- bioceramics were evaluated by co-culture with mice periosteal-derived osteoblast cells (POB) in vitro. The attachment and the growth of POB on the scaffolds were investigated by morphologic, proliferative abilities Osteogenic activity. In the animal experiment nano-HA prostheses were put in the right ateralbullae as ossicular prothesis of the ears of 30 healthy white guinea pigs (totally 60 ears) , AO ceramic was in the left as the control group .By retroauricular approach ,a hole was made in poster lateral bone wall of the acoustic vesicle . Each group animals executing on 1 . 2 . 4 . 8 . 12 . 16 weeks after operation ,investigated responses of tissue to materials by general and common histology. Results:HA and AO porous nano-bioceramics scaffolds had a good network pore system and mechanic properties. POB can fully attached to, extended on both HA and AO, and excreted extracellular matrix. The nano-HA prostheses taken out from 60 ears of the guinea pigs were normal and complete structure, and clear margin in gross anatomy and microscopically. In each case a layer of fibrous tissue surrounding the frame work ,a moderate inflammatory response and foreign body reaction were observed at 4 weeks after surgery. At 12 weeks ,the inflammatory response disappeared . Dense fibrous tissue was grown into the center of the bioceramic . . A little bone tissue growed into the nano-bioceramic. the new bone was found binding with the grafted material closely in group of nano-HA in 16 weeks. In the group of control, the new bone tissue were not found. Conclusion:With the nanometer powders of HA and AO, the porous nano-bioceramics can be produced by foam sockage technics. The new porous nano-bioceramics have good biocompatibility with POB.
Objective:To make porous nano-bioceramics and to study their biocompatibility as bone tissue engineerings caffolds.Methods:The nanometer powders of hydroxyapatite(HA) and tricalcium phosphate(AO) were prepared by Sol-gel methods and chemical deposition methods and the porous nano-bioceramics were made by foam sockage technics. The biocompatibility of new porous nano- bioceramics were evaluated by co-culture with mice periosteal-derived osteoblast cells (POB) in vitro. The attachment and the growth of POB on the scaffolds were investigated by morphologic, proliferative abilities Osteogenic activity. In the animal experiment nano-HA prostheses were put in the right ateralbullae as ossicular prothesis of the ears of 30 healthy white guinea pigs (totally 60 ears) , AO ceramic was in the left as the control group .By retroauricular approach ,a hole was made in poster lateral bone wall of the acoustic vesicle . Each group animals executing on 1 . 2 . 4 . 8 . 12 . 16 weeks after operation ,investigated responses of tissue to materials by general and common histology. Results:HA and AO porous nano-bioceramics scaffolds had a good network pore system and mechanic properties. POB can fully attached to, extended on both HA and AO, and excreted extracellular matrix. The nano-HA prostheses taken out from 60 ears of the guinea pigs were normal and complete structure, and clear margin in gross anatomy and microscopically. In each case a layer of fibrous tissue surrounding the frame work ,a moderate inflammatory response and foreign body reaction were observed at 4 weeks after surgery. At 12 weeks ,the inflammatory response disappeared . Dense fibrous tissue was grown into the center of the bioceramic . . A little bone tissue growed into the nano-bioceramic. the new bone was found binding with the grafted material closely in group of nano-HA in 16 weeks. In the group of control, the new bone tissue were not found. Conclusion:With the nanometer powders of HA and AO, the porous nano-bioceramics can be produced by foam sockage technics. The new porous nano-bioceramics have good biocompatibility with POB.
引文
[1]张少容,熊科亮,邓凌,等.自体软骨-软骨膜复合物在鼓室成形术中的应用[J].临床耳鼻咽喉科杂志,2006,20(10):469-469.
[2] Alam M I,Asahina I,O hmamiuda K, et al.Evaluation of ceramics composed of different hydroxyapatite totricalcium phosphateration ascarriers for rhBMP2. Biomaterial,2001,22(12):1643-1651.
[3]厚清,李佛保,王迎军等.三种钙磷陶瓷材料复合重组人骨形成蛋白2体内成骨的研究.中国修复重建外科杂志,2003,17(3):180-184.
[4]贺小兵,卢卫忠,唐康来等.骨形成蛋白和转化生长因子2B对兔尺骨骨折愈合后生物力学性能的影响.中国修复重建外科杂志,2003,17(3):185- 188.
[5] Kokubo T,Kim HM,Kawashita M,et al.Bioactive metals:preparation and Properties [J].J Mater Sci Mater Med,2004,15(2):99-107.
[6] Lee TM,Yang CY,Chang E,Tsai RS,et al.Comparision of plasma-sprayed Hydroxyapatitle coatings and zirconia-reinforced hydroxyapatitle composite coatings:in vivo study[J].J Biomed Mater Res A.2004 Dec 15; 71(4): 652 - 660.
[7] Larry L Hench.Bioceriamics[J].J.Am.Ceram.Soc,2004,81(7):1705-1728.
[8]杨仕明,宇雅苹,韩东一.人工听骨在鼓室成形术中的应用[J].中华耳科学杂志,2007,5(2):141-144.
[9]黎可华,林湘如,邓元平等.用羟基磷灰石人工听小骨实施改良听骨链重建术的疗效分析[J].中国耳鼻咽喉颅底外科杂志,2004,10(5):285-287.
[10] Abukawa H,Shin M,Williams WB, et al.Reconstruction of mandibular defects with autologous tissue-engineered bone.J Oral Maxillofac Surg,2004,62(5): 601-606.
[11] Frayssinet P,Gineste L,Conte P,et al.Short2term implantation effect of a DCPD based calciumphosphate cement[J].Biomaterials,1998,19(11212):971- 977.
[12] Li DJ,Ohsaki K,Cui PC,et al.Thickness of fibrous capsule after implantation of hydroxyapatite in subcutaneous tissue in rats[J].J Biomed Mater Res, 1999,45 (4):322-326.
[13]叶青,大崎胜一郎,伊井邦雄等.人工听骨周围组织细胞反应的实验研究[J].耳鼻咽喉头颈外科,2002,9(3):175-177.
[14]张龙城,宋为明,唐增福等.BMP-HA人工听骨在显微外科鼓室成形术中的应用[J].中华显微外科杂志,2005,28(1):30-31.
[15]刘阳,孙建军,林勇生等.纳米生物陶瓷在听小骨重建及乳突修复中的应用[J].中华医学杂志,2006,86(1):45-47.
[16]任卫,曹献英,纳米羟基磷灰石合成及表面改性的途径及方法[J].硅酸盐通报,2002 (1):38-43.
[17] Wang Xuejiang,Li Yubao,et al.Development of biomimetic nano-hydroxy apatite/poly ( hexamethylene adipamide) composites [J]. Biomaterials,2002, (23): 4787- 4791.
[18] Lu HH,Kofron MD,El2Amin SF,et al.In vitro bone formation using muscle derived cells:a new paradigm for bone tissue engineering using polymer bone morphogenetic protein matrices.Biochem Biophys Res Commun 2003,305: 882-889.
[19] Li J,Han D,Sun H.Vascularization in transplantation of gene modified tissue engineered bone for repairing bone defect.Zhongguo Xiufu Chongjian Waike Zazhi 2006,20(9):931-935.
[20] Jalota S,Bhaduri SB,Tas AC.In vitro testing of calcium phosphate (HA, TCP,and biphas ic HA-TCP) whiskers.J Biomed Mater Res A 2006,78 (3):481-490.
[21]肖建得,熊建义,欧阳侃等.纳米陶瓷人工骨修复骨缺损的实验研究[J].中华创伤骨科杂志,2006,(11),1048-1052.
[22]张官萍,巫爱霞.生物陶瓷及钛金属人工听骨在鼓室成形术中的短期临床疗效分析[J].中华耳科学杂志,2007,5(2):136-140.
[23] Yuan HY,De Brurjin JD,Li YB,et al.Bone formation induced by calcium phosphate ceramics in soft tissue of dogs:a comparative study between porous a TCP and bTCP. J Mater Sci:Master Med,2001,12:72-75.
[24]唐月军,吕春堂.医用复合生物陶瓷的研究进展[J].国际生物医学工程杂志,2006,29(12):114-116.
[25]曾立刚,李江平.人工听骨重建材料的生物相容性及其应用特点[J].中国组织工程研究与临床康复,2007,48(11):9789-9792.
[26] Habibovic P,Yuan H,van der Valk CM,et al.3D microenvironment as essential element for osteoinduction by biomaterials.Biomaterials 2005,26(17):3565-3575.
[27] Wen B,Huang Y,Xu YZ. Cytocompatibility of alumina ceramics. Journal of Jilin University (Medicine Edition),2003,29(2):158.
[28] Wu HY,Kong H.Nano-structured materials and the preliminary applications in the biomedical field.Basic Medical Sciences and Clinic,2002,22(2):97.
[29] Nanotechnology.The next small thing.The Economist,July 7th 2001.
[30] Lu H H,Kofron M D,El-Amin S F,et al.In vitro bone formation using muscle derived cells:a new paradigm for bone tissue engineering using polymer-bone morphogenetic protein matrices[J].Biochem Biophys Res Commun,2003,305(4):882-889.
[31] Elgendy,H M,Norman,M. E,Keaton,A. R and Laurencin, C. T. Osteoblast- like cell (MC3T3-E1) proliferation on bioerodible polymers:An approach towards the development of a bone bioerodible polymer composite material. Biomaterials 14:263,1993.
[32] Webb, K.,Hlady,V. and Tresco, P. A. Relationships among cell attachment, spreading,cytoskeletal organization and migration rate for anchorage–dependent cells on model surfaces. J. Biomed. Mater. Res. 49: 362, 2000.
[33] Shea, L. D., Wang, D., Franceschi, R. T., and Mooney, D. J. Engineered bone development from a pre-osteoblast cell line on three-dimensional scaffolds. Tissue Eng.6:605,2000.
[34] Oshima H,Nakamura M.A study on reference standard for cytotoxicity assay of biomaterials.Biomed Mater,1994,4(4):327.
[35] LiangWeidong,Shi Yingkang.The research of evaluation the compatibility of biotic material in cell-culturingmethod[J].J Biomed Eng,1999,16(1): 86- 90.
[36] Wergadel J E.Fluoride and bovine bone extract influence cell proliferation and phosphatase activities in human bone cell cultures [J].Clin Orthop Res,1988, 233: 274-282.
[37]江捍平,王大平,阮建明等.纳米羟基磷灰石人工骨的毒性与细胞相容性实验研究[J].中国医学工程,2005,13(5):458-461.
[38] Herbertson A,Aubin J.Cell sorting enriches osteogenic population in rat bone marrow stromal cell cultures [J]. Bone,1997,21(6):491-500.
[39] Lopes MA,Knowles J C,Kuru L,et al.Flow cytometry for assessing biocompatibility [J].J Biomed Mater Res,1998,41(4):649-656.
[40] Merket D E,Mcguire W I.Ploidy,proliferation activity and prognosis: DNA flow cytometry of solid tumors [J].Cancer,1990,65(5):1194-1205.
[41]许安廷.听骨链植入物的病理学研究[J].中华耳鼻咽喉科杂志, 2003 ,6(3):171-172.
[42]陈光.羟基磷灰石复合材料人工骨的研究进展[J].国外医学口腔医学分册,1992,19(5):288-291.
[43] Jarcho M.Calcium Phosphate Ceramics as Hard Tissue Prosthetics[J]. Clinorthop, 1981,157:259.
[44] Jarcho M.Biomaterial Aspects of Calcium Phosphate[J].Dental Clinical of North America,1986,30(1):25.
[45]黄桂林,李华林,谢文杨,等.羟基磷灰石人工骨微粒植入牙槽窝的临床应用研究[J].临床口腔医学杂志,2004,20(2):91-92.
[46] Piecuch J F,Ponichtera A,Nikoulcari H.Long-term Evaluation of Porous Hydroxy- apatite Blocks for Ridge Augmentation[J].Int J Oral Maxillofac Surg, 1990, 19(3):147-150.
[47]董青山,李祖兵,王虎中,等.纳米羟基磷灰石修复鼠下颌骨缺损[J].第四军医大学学报,2005,26(13):1180-1184.
[48]冯庆玲,崔福斋,张伟.纳米羟基磷灰石/胶原骨修复材料[J].中国医学科学院学报,2002,24(2):124-128.
[49] Bonfield W.Composites for Bone Replacement [J]. Biomedical Eng, 1988,10:522.
[50] Arinzeh TL,Tran T,Mcalary J,et al.A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials 2005,26(17):3631-3638.
[51] Schopper C,Ziya-Ghazvini F,Goriwoda W,et al.HA/TCP compounding of a porous CaP biomaterial improves bone formation and scaffold . degradation-- along-term his tological study.J Biomed Mater Res B Appl Biomater 2005,74 (1):458-467.
[52] Blitterswijk CA,Hesseling SC,Grote JJ,et al. The biocompatibility of hydroxyapatite ceramic:a study of retrieved human middle ear implants. J BiomedMaterRes, 1990,24:433-453.
[53] Shinohara T,Gyo K,Saiki T,et al. Ossiculoplasty using hydroxyapatite prostheses: long-term results. Clin Otolaryngo,l 2000,25: 287-292.
[54] Grote JJ.Results of cavity reconstructionwith hydroxyapatite implants after15 years. Am JOto,l 1998, 519:565-568.
[1]张龙城,宋为明等.BMP-HA人工听骨在显微外科鼓室成形术中的应用[J].中华显微外科杂志,2005,2(1):34-35.
[2]马建军,刘国惠,高秀云等.NPHA/BMP复合物在盖髓治疗中的动物实验及临床应用,口腔医学,2004,(03):154-156.
[3] Friedman MS, Long MW, Hankenson KD.Osteogenic differentiation mesenchymal stem cells is regulated by bone morphogenetic protein. Cell Biochem, 2006, 8(3): 538-540.
[4] Tsiridis E, Bhalla A, et al.Enhancing the osteoinductive properties of hydroxyapatite by the addition of human mesenchymal stem cells, and recombinant human osteo genic proteinin vitro.Ingure,2006, 37(3):25-32.
[5] Elias KL,Price RL,Webster T.J. Enhanced Functions of osteoblasts on nanometer diameter carbon fiber. Biomaterial,2002,23(15):3279-3287.
[6] Shimaoka H,Dohi Y,et al.Recombinant Growth/differentiation factor-5 stimulates osteogenic differentiation of marrow mesenchymal stem cells in porous hydroxyl- apatite ceramic.Biomed Mater.Res. A, 2004,68(1):168-176.
[7]张建,陈婷梅,安洪等.纳米羟基磷灰石/聚酰胺复合材料的人工肱骨头体内成骨中BMP-2及其基因表达研究.激光杂志,2006,(03):92-93.
[8]王晓云,丛笑倩.小鼠胚胎干细胞定向诱导分化为软骨样细胞的初步研究.上海第二医科大学学报,2004,24(4):250-253.
[9]朱明华,曾怡,孙涛等.新型生物活性陶瓷复合人工听骨成股效应的实验研究.中华修复重建外科杂志,2005,15(3):174-177.
[10] Na K, Kim SW, Sun BK, et al.Osteogenic differentiation of rabbit mesenchymal stem cells in thermo-reversible hydrogel constructs containing hydroxyapatite and BMP-2. Biomaterials,2007,28(16): 2631-2637.
[11]彭超,杨天府.纳米羟基磷灰石/壳聚糖复合人工骨的成骨及再血管化.中国组织工程研究与临床康复,2007, (40):8025-8029.
[12] Li C,Vepari C, Jin HJ, Kim HJ, et al. Electrospun silk-BMP-2scaffolds for bone tissue engineering.Biomaterials,2006,27(16): 3115-3124.
[13] Suuronen R,Kontion R, Ashammakhi N, et al.Bioabsorbable self-reinforced plates and screws in craniomaxillofacial surgery.Biomed Mater. Eng.,2004,14(4):517-524.
[14] Ton WS,Yang Z,Liu H,et al.Effects of culture conditions and bone morpho genetic protein 2 on extent of chondrogenesis from human embryonic stem cells.Stem cells,2007,25(4):950-960.
[15] Liao SS,Cui FZ,Zhang W,et al.Hierarchically biomimetic bone scaffold ma terials: nano - HA / collagen / PLA composite [J].Biomed Mater Res,2004,69:158– 165.
[16]杨志明.组织工程.第一版.北京:化学工业出版社, 2003, 82-94.
[17]陶源,宋为明.新型HA-BMP复合人工听小骨临床应用观察.临床耳鼻咽喉头颈外科杂志, 2007, 17(16): 738-740.
[18] Rocha LB,Adam RL,Leite NJ,et al.Bio-mineralization of polyanionic collagen- elastin matrices during cavarial bone repair.J Biomed Mater Res A.2006, 79(2):237-245. .
[19]修晓光,李彦林,李雅娜等.双相陶瓷生物骨的制备和细胞相容性研究[J].昆明医学院学报,2005,26(2):59.
[20] Jalota S, Bhaduri SB, Tas AC. In vitro testing of calcium phosphate (HA,TCP, and biphasicHA-TCP) whiskers.J Biomed Mater Res A,2006,78(3):481-490.
[21] Habibovic P,Yuan H,van der Valk CM,et al. 3D Micro environment as essential element for osteoinduction by biomaterials.Biomaterials,2005,26(17):3565 -3575.
[22] Liu L,Zhang L,Ren B,et al.Preparation and characterization of collagen-hydroxy apatitecomposite used for bone tissue engineering scaffold.Artif Cells Blood Substit Immobil.Biotechnol,2003,31(4):435-448.
[23]胡晓波,吴多能.猪生物骨载体成份分析和对比研究[J].生物医学工程学杂志,1996,13(2):187.
[24] Wooley PH,Nasser S,Fitzgerald RH Jr.The immune response to implant materials in humans.Clin Orthop Relat Res 1996,(326):63-70.
[25] Schopper C, Ziya-Ghazvini F, Goriwoda W,et al. HA/TCP compounding of a porous CaP biomaterial improves bone formation and scaffold degradation—a long-term histological study.J Biomed Mater Res B Appl Biomater.2005,74 (1):458-467.
[26] Brodie JC,Goldie E,Conne lG,et al.Osteoblast interactions with calcium phosphate ceramics modified by coating with type I collagen.J Biomed Mater Res A 2005,73(4):409-421.
[27] Williams JT,Southerland SS,Souza J,et al. Cells is olated from adult human skeletal muscle capable of differentiating into multiple mesodermal pheno types.Am Surg 1999,65(1):22-26.
[28] Nakagawa T,Tagawa T.Ultrastructural study of direct bone formation induced by BMPs-collagen complex implanted into an ectopic site.Oral Dis 2000,6(3):172-179.
[29] Arinzeh TL,Tran T,Mcalary J,et al.A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials 2005,26(17):3631-3638.
[30] Li J,Han D,Sun H.Vascularization intransplantation of gene modified tissue engineered bone for repairing bone defect.Zhongguo Xiufu Chongjian Waike Zazhi 2006,20(9):931-935.
[31] Zollner J,Van Koo ten TG,Eysel P,et al.Artificial disc biomechanical and biological evaluation in vit ro.Abst ract in Six World Biomaterials Congress Transactions. Hawaii,2000.
[32] Abdelmagid SM,Barbe MF, Arango-Hisijara I,et al.Osteoactivin acts as downstream mediator of BMP-2 effects on osteoblast function.J Cell Physiol 2007,210(1):26-37.
[33] Li JJ,Zhao Q,Sun HB,et al.Effects of BMP-2 genetherapy on vascularization in repairing bone defects.Zhonghua ZhengxingWaike Zazhi 2006,22(4):303-305.
[34] Pekkarinen T, Lindholm TS,Hietala O,et al.The effect of different mineral frames on ectopic bone formation in mouse hind leg muscles induced by native reindeer bone morphogenetic protein.Arch Orthop Trauma Surg 2005,125(1):10-15.
[2] Alam M I,Asahina I,O hmamiuda K, et al.Evaluation of ceramics composed of different hydroxyapatite totricalcium phosphateration ascarriers for rhBMP2. Biomaterial,2001,22(12):1643-1651.
[3]厚清,李佛保,王迎军等.三种钙磷陶瓷材料复合重组人骨形成蛋白2体内成骨的研究.中国修复重建外科杂志,2003,17(3):180-184.
[4]贺小兵,卢卫忠,唐康来等.骨形成蛋白和转化生长因子2B对兔尺骨骨折愈合后生物力学性能的影响.中国修复重建外科杂志,2003,17(3):185- 188.
[5] Kokubo T,Kim HM,Kawashita M,et al.Bioactive metals:preparation and Properties [J].J Mater Sci Mater Med,2004,15(2):99-107.
[6] Lee TM,Yang CY,Chang E,Tsai RS,et al.Comparision of plasma-sprayed Hydroxyapatitle coatings and zirconia-reinforced hydroxyapatitle composite coatings:in vivo study[J].J Biomed Mater Res A.2004 Dec 15; 71(4): 652 - 660.
[7] Larry L Hench.Bioceriamics[J].J.Am.Ceram.Soc,2004,81(7):1705-1728.
[8]杨仕明,宇雅苹,韩东一.人工听骨在鼓室成形术中的应用[J].中华耳科学杂志,2007,5(2):141-144.
[9]黎可华,林湘如,邓元平等.用羟基磷灰石人工听小骨实施改良听骨链重建术的疗效分析[J].中国耳鼻咽喉颅底外科杂志,2004,10(5):285-287.
[10] Abukawa H,Shin M,Williams WB, et al.Reconstruction of mandibular defects with autologous tissue-engineered bone.J Oral Maxillofac Surg,2004,62(5): 601-606.
[11] Frayssinet P,Gineste L,Conte P,et al.Short2term implantation effect of a DCPD based calciumphosphate cement[J].Biomaterials,1998,19(11212):971- 977.
[12] Li DJ,Ohsaki K,Cui PC,et al.Thickness of fibrous capsule after implantation of hydroxyapatite in subcutaneous tissue in rats[J].J Biomed Mater Res, 1999,45 (4):322-326.
[13]叶青,大崎胜一郎,伊井邦雄等.人工听骨周围组织细胞反应的实验研究[J].耳鼻咽喉头颈外科,2002,9(3):175-177.
[14]张龙城,宋为明,唐增福等.BMP-HA人工听骨在显微外科鼓室成形术中的应用[J].中华显微外科杂志,2005,28(1):30-31.
[15]刘阳,孙建军,林勇生等.纳米生物陶瓷在听小骨重建及乳突修复中的应用[J].中华医学杂志,2006,86(1):45-47.
[16]任卫,曹献英,纳米羟基磷灰石合成及表面改性的途径及方法[J].硅酸盐通报,2002 (1):38-43.
[17] Wang Xuejiang,Li Yubao,et al.Development of biomimetic nano-hydroxy apatite/poly ( hexamethylene adipamide) composites [J]. Biomaterials,2002, (23): 4787- 4791.
[18] Lu HH,Kofron MD,El2Amin SF,et al.In vitro bone formation using muscle derived cells:a new paradigm for bone tissue engineering using polymer bone morphogenetic protein matrices.Biochem Biophys Res Commun 2003,305: 882-889.
[19] Li J,Han D,Sun H.Vascularization in transplantation of gene modified tissue engineered bone for repairing bone defect.Zhongguo Xiufu Chongjian Waike Zazhi 2006,20(9):931-935.
[20] Jalota S,Bhaduri SB,Tas AC.In vitro testing of calcium phosphate (HA, TCP,and biphas ic HA-TCP) whiskers.J Biomed Mater Res A 2006,78 (3):481-490.
[21]肖建得,熊建义,欧阳侃等.纳米陶瓷人工骨修复骨缺损的实验研究[J].中华创伤骨科杂志,2006,(11),1048-1052.
[22]张官萍,巫爱霞.生物陶瓷及钛金属人工听骨在鼓室成形术中的短期临床疗效分析[J].中华耳科学杂志,2007,5(2):136-140.
[23] Yuan HY,De Brurjin JD,Li YB,et al.Bone formation induced by calcium phosphate ceramics in soft tissue of dogs:a comparative study between porous a TCP and bTCP. J Mater Sci:Master Med,2001,12:72-75.
[24]唐月军,吕春堂.医用复合生物陶瓷的研究进展[J].国际生物医学工程杂志,2006,29(12):114-116.
[25]曾立刚,李江平.人工听骨重建材料的生物相容性及其应用特点[J].中国组织工程研究与临床康复,2007,48(11):9789-9792.
[26] Habibovic P,Yuan H,van der Valk CM,et al.3D microenvironment as essential element for osteoinduction by biomaterials.Biomaterials 2005,26(17):3565-3575.
[27] Wen B,Huang Y,Xu YZ. Cytocompatibility of alumina ceramics. Journal of Jilin University (Medicine Edition),2003,29(2):158.
[28] Wu HY,Kong H.Nano-structured materials and the preliminary applications in the biomedical field.Basic Medical Sciences and Clinic,2002,22(2):97.
[29] Nanotechnology.The next small thing.The Economist,July 7th 2001.
[30] Lu H H,Kofron M D,El-Amin S F,et al.In vitro bone formation using muscle derived cells:a new paradigm for bone tissue engineering using polymer-bone morphogenetic protein matrices[J].Biochem Biophys Res Commun,2003,305(4):882-889.
[31] Elgendy,H M,Norman,M. E,Keaton,A. R and Laurencin, C. T. Osteoblast- like cell (MC3T3-E1) proliferation on bioerodible polymers:An approach towards the development of a bone bioerodible polymer composite material. Biomaterials 14:263,1993.
[32] Webb, K.,Hlady,V. and Tresco, P. A. Relationships among cell attachment, spreading,cytoskeletal organization and migration rate for anchorage–dependent cells on model surfaces. J. Biomed. Mater. Res. 49: 362, 2000.
[33] Shea, L. D., Wang, D., Franceschi, R. T., and Mooney, D. J. Engineered bone development from a pre-osteoblast cell line on three-dimensional scaffolds. Tissue Eng.6:605,2000.
[34] Oshima H,Nakamura M.A study on reference standard for cytotoxicity assay of biomaterials.Biomed Mater,1994,4(4):327.
[35] LiangWeidong,Shi Yingkang.The research of evaluation the compatibility of biotic material in cell-culturingmethod[J].J Biomed Eng,1999,16(1): 86- 90.
[36] Wergadel J E.Fluoride and bovine bone extract influence cell proliferation and phosphatase activities in human bone cell cultures [J].Clin Orthop Res,1988, 233: 274-282.
[37]江捍平,王大平,阮建明等.纳米羟基磷灰石人工骨的毒性与细胞相容性实验研究[J].中国医学工程,2005,13(5):458-461.
[38] Herbertson A,Aubin J.Cell sorting enriches osteogenic population in rat bone marrow stromal cell cultures [J]. Bone,1997,21(6):491-500.
[39] Lopes MA,Knowles J C,Kuru L,et al.Flow cytometry for assessing biocompatibility [J].J Biomed Mater Res,1998,41(4):649-656.
[40] Merket D E,Mcguire W I.Ploidy,proliferation activity and prognosis: DNA flow cytometry of solid tumors [J].Cancer,1990,65(5):1194-1205.
[41]许安廷.听骨链植入物的病理学研究[J].中华耳鼻咽喉科杂志, 2003 ,6(3):171-172.
[42]陈光.羟基磷灰石复合材料人工骨的研究进展[J].国外医学口腔医学分册,1992,19(5):288-291.
[43] Jarcho M.Calcium Phosphate Ceramics as Hard Tissue Prosthetics[J]. Clinorthop, 1981,157:259.
[44] Jarcho M.Biomaterial Aspects of Calcium Phosphate[J].Dental Clinical of North America,1986,30(1):25.
[45]黄桂林,李华林,谢文杨,等.羟基磷灰石人工骨微粒植入牙槽窝的临床应用研究[J].临床口腔医学杂志,2004,20(2):91-92.
[46] Piecuch J F,Ponichtera A,Nikoulcari H.Long-term Evaluation of Porous Hydroxy- apatite Blocks for Ridge Augmentation[J].Int J Oral Maxillofac Surg, 1990, 19(3):147-150.
[47]董青山,李祖兵,王虎中,等.纳米羟基磷灰石修复鼠下颌骨缺损[J].第四军医大学学报,2005,26(13):1180-1184.
[48]冯庆玲,崔福斋,张伟.纳米羟基磷灰石/胶原骨修复材料[J].中国医学科学院学报,2002,24(2):124-128.
[49] Bonfield W.Composites for Bone Replacement [J]. Biomedical Eng, 1988,10:522.
[50] Arinzeh TL,Tran T,Mcalary J,et al.A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials 2005,26(17):3631-3638.
[51] Schopper C,Ziya-Ghazvini F,Goriwoda W,et al.HA/TCP compounding of a porous CaP biomaterial improves bone formation and scaffold . degradation-- along-term his tological study.J Biomed Mater Res B Appl Biomater 2005,74 (1):458-467.
[52] Blitterswijk CA,Hesseling SC,Grote JJ,et al. The biocompatibility of hydroxyapatite ceramic:a study of retrieved human middle ear implants. J BiomedMaterRes, 1990,24:433-453.
[53] Shinohara T,Gyo K,Saiki T,et al. Ossiculoplasty using hydroxyapatite prostheses: long-term results. Clin Otolaryngo,l 2000,25: 287-292.
[54] Grote JJ.Results of cavity reconstructionwith hydroxyapatite implants after15 years. Am JOto,l 1998, 519:565-568.
[1]张龙城,宋为明等.BMP-HA人工听骨在显微外科鼓室成形术中的应用[J].中华显微外科杂志,2005,2(1):34-35.
[2]马建军,刘国惠,高秀云等.NPHA/BMP复合物在盖髓治疗中的动物实验及临床应用,口腔医学,2004,(03):154-156.
[3] Friedman MS, Long MW, Hankenson KD.Osteogenic differentiation mesenchymal stem cells is regulated by bone morphogenetic protein. Cell Biochem, 2006, 8(3): 538-540.
[4] Tsiridis E, Bhalla A, et al.Enhancing the osteoinductive properties of hydroxyapatite by the addition of human mesenchymal stem cells, and recombinant human osteo genic proteinin vitro.Ingure,2006, 37(3):25-32.
[5] Elias KL,Price RL,Webster T.J. Enhanced Functions of osteoblasts on nanometer diameter carbon fiber. Biomaterial,2002,23(15):3279-3287.
[6] Shimaoka H,Dohi Y,et al.Recombinant Growth/differentiation factor-5 stimulates osteogenic differentiation of marrow mesenchymal stem cells in porous hydroxyl- apatite ceramic.Biomed Mater.Res. A, 2004,68(1):168-176.
[7]张建,陈婷梅,安洪等.纳米羟基磷灰石/聚酰胺复合材料的人工肱骨头体内成骨中BMP-2及其基因表达研究.激光杂志,2006,(03):92-93.
[8]王晓云,丛笑倩.小鼠胚胎干细胞定向诱导分化为软骨样细胞的初步研究.上海第二医科大学学报,2004,24(4):250-253.
[9]朱明华,曾怡,孙涛等.新型生物活性陶瓷复合人工听骨成股效应的实验研究.中华修复重建外科杂志,2005,15(3):174-177.
[10] Na K, Kim SW, Sun BK, et al.Osteogenic differentiation of rabbit mesenchymal stem cells in thermo-reversible hydrogel constructs containing hydroxyapatite and BMP-2. Biomaterials,2007,28(16): 2631-2637.
[11]彭超,杨天府.纳米羟基磷灰石/壳聚糖复合人工骨的成骨及再血管化.中国组织工程研究与临床康复,2007, (40):8025-8029.
[12] Li C,Vepari C, Jin HJ, Kim HJ, et al. Electrospun silk-BMP-2scaffolds for bone tissue engineering.Biomaterials,2006,27(16): 3115-3124.
[13] Suuronen R,Kontion R, Ashammakhi N, et al.Bioabsorbable self-reinforced plates and screws in craniomaxillofacial surgery.Biomed Mater. Eng.,2004,14(4):517-524.
[14] Ton WS,Yang Z,Liu H,et al.Effects of culture conditions and bone morpho genetic protein 2 on extent of chondrogenesis from human embryonic stem cells.Stem cells,2007,25(4):950-960.
[15] Liao SS,Cui FZ,Zhang W,et al.Hierarchically biomimetic bone scaffold ma terials: nano - HA / collagen / PLA composite [J].Biomed Mater Res,2004,69:158– 165.
[16]杨志明.组织工程.第一版.北京:化学工业出版社, 2003, 82-94.
[17]陶源,宋为明.新型HA-BMP复合人工听小骨临床应用观察.临床耳鼻咽喉头颈外科杂志, 2007, 17(16): 738-740.
[18] Rocha LB,Adam RL,Leite NJ,et al.Bio-mineralization of polyanionic collagen- elastin matrices during cavarial bone repair.J Biomed Mater Res A.2006, 79(2):237-245. .
[19]修晓光,李彦林,李雅娜等.双相陶瓷生物骨的制备和细胞相容性研究[J].昆明医学院学报,2005,26(2):59.
[20] Jalota S, Bhaduri SB, Tas AC. In vitro testing of calcium phosphate (HA,TCP, and biphasicHA-TCP) whiskers.J Biomed Mater Res A,2006,78(3):481-490.
[21] Habibovic P,Yuan H,van der Valk CM,et al. 3D Micro environment as essential element for osteoinduction by biomaterials.Biomaterials,2005,26(17):3565 -3575.
[22] Liu L,Zhang L,Ren B,et al.Preparation and characterization of collagen-hydroxy apatitecomposite used for bone tissue engineering scaffold.Artif Cells Blood Substit Immobil.Biotechnol,2003,31(4):435-448.
[23]胡晓波,吴多能.猪生物骨载体成份分析和对比研究[J].生物医学工程学杂志,1996,13(2):187.
[24] Wooley PH,Nasser S,Fitzgerald RH Jr.The immune response to implant materials in humans.Clin Orthop Relat Res 1996,(326):63-70.
[25] Schopper C, Ziya-Ghazvini F, Goriwoda W,et al. HA/TCP compounding of a porous CaP biomaterial improves bone formation and scaffold degradation—a long-term histological study.J Biomed Mater Res B Appl Biomater.2005,74 (1):458-467.
[26] Brodie JC,Goldie E,Conne lG,et al.Osteoblast interactions with calcium phosphate ceramics modified by coating with type I collagen.J Biomed Mater Res A 2005,73(4):409-421.
[27] Williams JT,Southerland SS,Souza J,et al. Cells is olated from adult human skeletal muscle capable of differentiating into multiple mesodermal pheno types.Am Surg 1999,65(1):22-26.
[28] Nakagawa T,Tagawa T.Ultrastructural study of direct bone formation induced by BMPs-collagen complex implanted into an ectopic site.Oral Dis 2000,6(3):172-179.
[29] Arinzeh TL,Tran T,Mcalary J,et al.A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials 2005,26(17):3631-3638.
[30] Li J,Han D,Sun H.Vascularization intransplantation of gene modified tissue engineered bone for repairing bone defect.Zhongguo Xiufu Chongjian Waike Zazhi 2006,20(9):931-935.
[31] Zollner J,Van Koo ten TG,Eysel P,et al.Artificial disc biomechanical and biological evaluation in vit ro.Abst ract in Six World Biomaterials Congress Transactions. Hawaii,2000.
[32] Abdelmagid SM,Barbe MF, Arango-Hisijara I,et al.Osteoactivin acts as downstream mediator of BMP-2 effects on osteoblast function.J Cell Physiol 2007,210(1):26-37.
[33] Li JJ,Zhao Q,Sun HB,et al.Effects of BMP-2 genetherapy on vascularization in repairing bone defects.Zhonghua ZhengxingWaike Zazhi 2006,22(4):303-305.
[34] Pekkarinen T, Lindholm TS,Hietala O,et al.The effect of different mineral frames on ectopic bone formation in mouse hind leg muscles induced by native reindeer bone morphogenetic protein.Arch Orthop Trauma Surg 2005,125(1):10-15.