脱矿牙本质基质—富血小板血浆凝胶构建牙周骨组织工程支架的初步研究
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摘要
目的:评价自制脱矿牙本质基质(DDM)对富血小板血浆(PRP)作为生长因子缓释系统的影响。方法:二次离心法制备PRP凝胶,分为3组:A:PRP/DDM,B:PRP,C:全血。通过酶联免疫吸附法(ELISA)分别测定2周内单位体积各组凝胶释放的血小板衍化生长因子(PDGF)、转化生长因子-β(TGF-β)的水平。扫描电子显微镜下观察PRP/DDM凝胶的空间结构。将DDM植入SD大鼠股部肌肉陷窝内,1、3周后取材行组织学切片,观察有无毒性反应。结果:PRP/DDM凝胶较其两组持续释放PDGF和TGF-β的时间较长,组间差异有显著的统计学意义(P<0.05);经扫描电镜下观察及组织切片评估PRP与DDM结合紧密、有序;DDM具有很好的生物相容性。结论:DDM与PRP结合能够更好的促进生长因子的缓慢释放。
Objective:To evaluate the effects of demineralized dentin matrix(DDM)to the platelet-rich plasma(PRP)gel delivery system for growth factors.Methods:After double centrifugation for preparation of PRP,the concentrations of platelet-derived growth factor(PDGF)and transforming growth factor-β(TGF-β)in the whole blood gel(group C),PRP gel(group B),and PRP/DDM gel(group A)were measured in two weeks by enzyme-linked immunosorbent assay(ELISA).The structure of prepared PRP/DDM gel was observed by scanning electron microscopy(SEM).DDM was implanted into the pit of femoral muscles of SD rats and tissue sections were performed to study the toxicity reaction.Results:Compared with other two groups,PRP/DDM had a much longer time to continuously release the growth factors(P<0.05).Through SEM and tissue section,it was found that PRP and DDM combined with each other closely and orderly and DDM had a fine biocompatibility.Conclusion:The combination of PRP and DDM can better slower the release of growth factors.
Objective:To evaluate the effects of demineralized dentin matrix(DDM)to the platelet-rich plasma(PRP)gel delivery system for growth factors.Methods:After double centrifugation for preparation of PRP,the concentrations of platelet-derived growth factor(PDGF)and transforming growth factor-β(TGF-β)in the whole blood gel(group C),PRP gel(group B),and PRP/DDM gel(group A)were measured in two weeks by enzyme-linked immunosorbent assay(ELISA).The structure of prepared PRP/DDM gel was observed by scanning electron microscopy(SEM).DDM was implanted into the pit of femoral muscles of SD rats and tissue sections were performed to study the toxicity reaction.Results:Compared with other two groups,PRP/DDM had a much longer time to continuously release the growth factors(P<0.05).Through SEM and tissue section,it was found that PRP and DDM combined with each other closely and orderly and DDM had a fine biocompatibility.Conclusion:The combination of PRP and DDM can better slower the release of growth factors.
引文
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[11]罗涛,李放,张宁,等.不同抗凝剂和激活剂联合应用对富血小板血浆凝胶释放生长因子影响的比较[J].中国组织工程研究,2012,16(16):2893-2897
[12]康健,袁文.富血小板凝胶制备方法的比较与优选[J].中国组织工程研究,2014,18(3):476-481
[13]刘琳,包广洁.牙髓牙本质组织工程生物支架材料的研究进展[J].中国组织工程研究,2013,17(8):1455-1460
[14]Harrison S,Vavken P,Kevy S,et al.Platelet activation by collagen provides sustained release of anabolic cytokines[J].Am J Sports Med,2011,39(4):729-734
[15]李康婧,黃杨,陈文霞.牙本质支架体外诱导兔骨髓干细胞分化[J].口腔医学研究,2012,4(28):291-294
[16]李成林,岑远坤,李文,等,脱钙牙本质基质在肌肉中诱导成骨现象的观察[J].中国口腔种植学杂志,1998.6(3):p.59-61
[17]李思敏,周晋,颜承伟,等.富血小板血浆缓释载体在骨组织工程中的研究进展[J].口腔医学研究,2015,31(6):632-634
[18]吕敏,裴国献,刘勇,等.富血小板血浆的制备现状及研究进展[J].现代生物医学进展,2013,13(13):2574-2577
[2]Uris t M R,Silv erman BF,Bu ring K,et al.The bone inducti on principle[J].Clin Orthop,1967,53:23
[3]Katz RW,Hol linger JO,Reddi HA.The functional equivalence of demineralized dentin mat rices in ectopic bone induction[J].Biomed Mater Res,1993,27:239
[4]Peng L,Ye L,Zhou XD.Mesenchymal stem cells and tooth engineering[J].Int J Oral Sci,2009 1(1):6-12
[5]吕欣欣,寇育荣,潘亚萍.富血小板纤维蛋白在GTR联合植骨术中促进牙周组织再生作用的临床研究[J].口腔医学研究,2013,29(12):1174-1178
[6]张利,李玉宝,杨爱萍,等.骨组织工程用纳米羟基磷灰石/壳聚糖多孔支架材料的制备及性能表征[J].功能材料,2005,36(2):314-316
[7]孙洁,张剑明,李彦秋.富血小板纤维蛋白超微结构的观察与探讨[J].口腔医学研究,2010,26(1):98-101
[8]Ma J,Li F,Ren DJ,et al.Zhongguo Kangfu Lilun yu Shijian,2011,17(3):223-225
[9]Pinbolt EM,Bang G,Hanuaes HR.Alveolar ridge augmentation of demineralized dentin mat rices in ectopic bone induction.by osteoinduction in rats[J].Scand J Dent Res,1990,98(5):434
[10]谭剑斌,赵敏.体内外代替方法在急性毒性评价中的研究进展[J].毒理学杂志,2010,24(6):479-482
[11]罗涛,李放,张宁,等.不同抗凝剂和激活剂联合应用对富血小板血浆凝胶释放生长因子影响的比较[J].中国组织工程研究,2012,16(16):2893-2897
[12]康健,袁文.富血小板凝胶制备方法的比较与优选[J].中国组织工程研究,2014,18(3):476-481
[13]刘琳,包广洁.牙髓牙本质组织工程生物支架材料的研究进展[J].中国组织工程研究,2013,17(8):1455-1460
[14]Harrison S,Vavken P,Kevy S,et al.Platelet activation by collagen provides sustained release of anabolic cytokines[J].Am J Sports Med,2011,39(4):729-734
[15]李康婧,黃杨,陈文霞.牙本质支架体外诱导兔骨髓干细胞分化[J].口腔医学研究,2012,4(28):291-294
[16]李成林,岑远坤,李文,等,脱钙牙本质基质在肌肉中诱导成骨现象的观察[J].中国口腔种植学杂志,1998.6(3):p.59-61
[17]李思敏,周晋,颜承伟,等.富血小板血浆缓释载体在骨组织工程中的研究进展[J].口腔医学研究,2015,31(6):632-634
[18]吕敏,裴国献,刘勇,等.富血小板血浆的制备现状及研究进展[J].现代生物医学进展,2013,13(13):2574-2577