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1、磷酸三钙表面吸附P-15多肽对细胞相容性的影响天津医药2011年5月第39卷第5期doi:lO.3969~.issn.0253-9896.2011.05.018I3一磷酸三钙表面吸附P一15多肽对细胞相容性的影响赵军沈文文樊洪445摘要目的:在体外研究B一磷酸三钙(13-TCP)吸附15肽(P一15)后对细胞相容性的影响.方法:将冻存的鼠成肌细胞系C2C12进行复苏,待细胞正常生长后向培养基中加人骨形成蛋白2(BMP2)刺激C2C12细胞向成骨细胞转化,使用200g,LBMP2刺激的C2C12细胞;在13-TCP/P一15和13-TCPI-分别培养B
2、MP2诱导的C2C12细胞6d,分别于培养2,4和6d取材,CCK一8法检测细胞增殖情况;BCA法检测蛋白质含量;碱性磷酸酶试剂盒检测碱性磷酸酶活性.结果:13-TCP组与B—TCP/P一15组的细胞增殖情况与蛋白含量随时间的延长而呈增加趋势,p—TcP,P一15组较p—TCP组各相同时点细胞增殖及蛋白含量明显上升,差异有统计学意义(P<O.05);2组在细胞与材料复合培养2d时均未检测出碱性磷酸酶的活性,培养4,6dB—TCP/P一15组较p—TCP组各相同时点碱性磷酸酶的活性明显升高,差异均有统计学意义(P<0.05).结论:13-T
3、CP吸附P一15后生物相容性提高.关键词碱性磷酸酶磷酸钙类肽类材料试验生物相容性材料成肌细,骨骼肌骨形态发生蛋白质类EffectofP-15PeptideAttached~-TricalciumPhosphateSurfaceonCellularCompatibilityZHAOJun,SHENWenwen,FANHongTianjinMedicalUniversity,Tianjin300070,ChinaAbstractObjective:ToevaluatetheeffectofP一15peptideattachedp—tricalciumph
4、osphate(p—TCP)surfaceoncellularcompatibilityinvitro.Methods:B—TCPwasputintoP一15peptidesolution.Incubationwascarriedoutatroomtern.peraturefor24hourswithgentleshakingtoensurethepeptidefullabsorptioninto13-TCP.Theun-adsorbedpeptidewasre-movedfromthe13-TCPbyPBS.The13-TCP/P-15compos
5、itewasdriedandthenstoredatroomtemperatureinmois—ture—proofcontainers.ThemousemyoblastcelllineC2C12wasrecovered.Bonemorphogeneticprotein2(BMP2)wasaddedinmediumtostimulateC2C12cellsintoosteoblastswhenC2Cl2cellsgrewnormally.C2C12cellsstimulatedbyBMP2of200g,Lwereusedinthefollow-upe
6、xperiment.C2C12cellstreatedbyBMP2wereculturedonthesurfaceof13-TCP/P-15and13-TCPfor6days.Sampleswereobtained2,4and6daysafterculture.CellproliferationwasdetectedbyCCK一8meth-od.TotalcellproteinwasexaminedbyBCAmethod.Cellalkalinephosphatase(ALP1activitieswereassayedbyALPkit.Re-suit
7、s:BothcellproliferationandtotalamountofproteinofC2C12cellsincreasedwithtimein13-TCPgroupand13-TCPIP一15group.ThecellproliferationandtotalamountofproteinwereobviouslyhigherinC2C12adheredto3-TCP/P一15thanthatofC2Cl2adheredtoB—TCP(尸<0.05).After2dayscultureALPactivitieswerenotdete
8、ctedinC2C12oftwogroups.Af-ter4daysand6daysofcuhure,ALP
