纯钛表面GRGDSP多肽对骨结合能力的影响
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摘要
目的:将钛合金种植体微弧氧化(MAO)后,以多巴胺(PDA)为偶联,加载GRGDSP多肽,形成复合活性涂层,评价其骨结合能力。方法:选取18只成年新西兰大白兔,建立单侧下颌骨模型,并将A组(Ti-MAO)、B组(Ti-MAOPDA)、C组(Ti-MAO-PDA-GRGDSP)分别植入到新西兰大白兔一侧下颌骨,选取2周、4周、8周下颌骨植入样本,应用锥形束CT(CBCT)观察种植体周围骨结合情况。对4周样本以扫描电镜(SEM)观察界面形貌,激光共聚焦显微镜观察样本成骨活跃程度,扭力实验观察骨结合力。采用SPSS19.0软件包对数据进行单因素方差分析。结果:CBCT观察,C组接触区骨结合情况好于B组和A组,且B组优于A组。扫描电镜观察,C组在种植体与骨接触区聚集了更多新骨。激光共聚焦显微镜观察交界处,C组与B组较A组荧光强度明显,其中C组最为明显。扭力观察,8周时B、C组扭力值为A组的2倍,C组较B组多8 N·cm;C组、B组与A组相比,差异有统计学意义(P<0.05)。结论 :TiMAO-PDA与Ti-MAO-PDA-GRGDSP涂层对骨结合均有促进作用,其中Ti-MAO-PDA-GRGDSP涂层对骨结合的作用最明显。
PURPOSE: Micro-arc oxidation(MAO) of titanium alloy was implemented, then GRGDSP polypeptide was loaded by coupling of dopamine(PDA) to form a composite active coating,in order to enhance esteointegration.METHODS: Eighteen adult New Zealand white rabbits were selected to establish a unilateral mandibular model. Ti-MAO(group A), Ti-MAO-PDA(group B) and Ti-MAO-PDA-GRGDSP(group C) were implanted into the mandible of rabbits. The mandibular bone on one side was selected at 2, 4, 8 weeks to harvest sample. Bone allografts were observed on cone-beam CT(CBCT). The interface of sample at 4 w was observed under scanning electron microscopy(SEM), laser scanning confocal microscope was used to observe the degree of osteoblast activity, torsion test was applied to observe osseointegration. The date were compared between groups with one-way ANOVA using SPSS19.0 software package.RESULTS: For osseointegration at contact region, group C was better than group A and group B, and group B was better than group A on CBCT. SEM showed that group C had more new bone aggregated in the contact area between implant and bone. Laser confocal microscopy showed that the fluorescence intensity of group C was significantly higher than that of group B at the junction. The torsional force of group B and C at 8 w was 2 times of that of group A, and group C was 8 N·cm greater than group B. The difference was statistically significant between group C and group B, comparing to group A(P<0.05). CONCLUSIONS: Ti-MAO-PDA and Ti-MAO-PDA-GRGDSP coatings can promote osseointegration, of which Ti-MAO-PDA-GRGDSP coating has the strongest osseointegration effect.
PURPOSE: Micro-arc oxidation(MAO) of titanium alloy was implemented, then GRGDSP polypeptide was loaded by coupling of dopamine(PDA) to form a composite active coating,in order to enhance esteointegration.METHODS: Eighteen adult New Zealand white rabbits were selected to establish a unilateral mandibular model. Ti-MAO(group A), Ti-MAO-PDA(group B) and Ti-MAO-PDA-GRGDSP(group C) were implanted into the mandible of rabbits. The mandibular bone on one side was selected at 2, 4, 8 weeks to harvest sample. Bone allografts were observed on cone-beam CT(CBCT). The interface of sample at 4 w was observed under scanning electron microscopy(SEM), laser scanning confocal microscope was used to observe the degree of osteoblast activity, torsion test was applied to observe osseointegration. The date were compared between groups with one-way ANOVA using SPSS19.0 software package.RESULTS: For osseointegration at contact region, group C was better than group A and group B, and group B was better than group A on CBCT. SEM showed that group C had more new bone aggregated in the contact area between implant and bone. Laser confocal microscopy showed that the fluorescence intensity of group C was significantly higher than that of group B at the junction. The torsional force of group B and C at 8 w was 2 times of that of group A, and group C was 8 N·cm greater than group B. The difference was statistically significant between group C and group B, comparing to group A(P<0.05). CONCLUSIONS: Ti-MAO-PDA and Ti-MAO-PDA-GRGDSP coatings can promote osseointegration, of which Ti-MAO-PDA-GRGDSP coating has the strongest osseointegration effect.
引文
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[2]Oshida Y,Tuna EB,Aktoren O,et al.Dental implant systems[J].Int Mol Sci,2010,11(4):1580-1678.
[3]唐仕光,陈泉志,李少波,等.金属复合材料微弧氧化研究进展[J].表面技术,2016,45(11):23-31.
[4]Spriano S,Ferraris S,Pan G,et al.Multifunctional titanium:surface modification process and biological response[J].J Mech Med Biol,2015,15(2):374-383.
[5]Madry H,Rico AR,Venkatesan JK,et al.Transforming growth factorβ-releasing scaffolds for cartilage tissue engineering[J].Tissue Eng Part B Rev,2014,20(2):106-125.
[6]Hwang DS,Sim SB,Cha HJ.Cell adhesion biomaterial based on mussel adhesive protein fused with RGD peptide[J].Biomaterials,2007,28(28):4039-4046.
[7]Boxus T,Touillaux R,Dive G,et al.Synthesis and evaluation of RGD peptidomimetics aimed at surface bioderivatization of polymer substrates[J].Bioorg Med Chem,1998,6(9):1577-1595.
[8]Chen X,Sevilla P,Aparicio C.Surface biofunctionalization by covalent co-immobilization of oligopeptides[J].Colloids Surf B Biointerfaces,2013,107:189-197.
[9]Anselme K.Osteoblast adhesion on biomaterials[J].Biomaterials,2000,21(7):667-681.
[10]Yang GL,He FM,Yang XF,et al.In vivo evaluation of bonebonding ability of RGD-coated porous implant using layer-bylayer electrostatic self-assembly[J].J Biomed Mater Res A,2009,90(1):175-185.
[11]Mavrogenis AF,Dimitriou R,Parvizi J,et al.Biology of implant osseointegration[J].J Musculoskeletal Neuronal Interact,2009,9(2):61-71.
[12]田垒,李德超,李慕勤,等.c(RGDf K)肽影响成骨细胞黏附、增殖的体外实验[J].中国体视学与图像分析,2016,21(4):430-435.
[13]林野,邱立新,胡秀莲.口腔种植学[M].北京:北京大学医学出版社,2015:8.
[14]宿玉成.口腔种植学[M].第2版.北京:人民卫生出版社,2014:29-30.
[15]吴大怡.微创种植新概念[A]//口腔种植骨量不足治疗技术与方案研讨会论文集[C].西安:第四军医大学,2014:25-26.
[16]孟玲娜,沈兰花,王鑫,等.兔下颌骨种植体即刻种植动物模型的建立[J].中国口腔种植学杂志,2015,10(1):4-10
[17]Shin YM,Jo SY,Park JS,et al.Synergistic effect of dual functionalized fibrous scaffold with BCP and RGD containing peptide for improved osteogenic differentiation[J].Macromol Biosci,2014,14(8):1190-1198.
[18]He J,Huang T,Gan L,et al.Collagen-infiltrated porous hydroxyapatite coating and its osteogenic properties:in vitro and in vivo study[J].J Biomed Mater Res A,2012,100(7):1706-1715.
[19]Jager M,Boge C,Janissen R,et al.Osteoblastic potency of bone marrow cells cultivated on functionalized bio metals with cyclic RGD-peptide[J].J Biomed Mater Res A,2013,101(10):2905-2914.
[20]Hersel U,Dahmen C,Kessler H.RGD modified polymers:biomaterials for stimulated cell adhesion and beyond[J].Biomaterials,2003,24(24):4385-4415.
[21]Motoyoshi M,Matsuoka M,Shimizu N.Application of orthodontic mini-implants in adolescents[J].Int J Oral Maxillofacial Surg,2007,36(8):695-699.
[22]Black J.Push-out test[J].J Biomed Mater Res,1989,23(11):1243-1245.