人工生物心脏瓣膜材料及制备方法和用图
【技术领域】
[0001] 本发明涉及心脏瓣膜材料领域，尤其涉及一种原花青素（PC)与戊二醛（GA)共交 联的心脏瓣膜材料及其制备方法。
【背景技术】
[0002] 世界范围内心脏瓣膜疾病发病率和致死率都很高，美国毎年要进行6万例人 エ心脏瓣膜置换手术，同时大约有2万人直接死于此病[Fuchs,J.R.，B.A.Nasseri,and J.P.Vacanti,Tissueengineering:a2丄stcenturysolutiontosurgical reconstruction.TheAnnalsofThoracicSurgery, 2001? 72(2):p. 577-591.]〇 临床 上人工心脏瓣膜主要采用机械瓣膜和异种生物瓣膜移植替换治疗。低温贮藏的人同种 异体生物瓣膜也被用于临床，但由于来源不足应用极少。异种生物瓣膜结构如同同种生 物瓣膜，具有类似人体中心血流特点，因几乎不引起凝血，患者术后不需象机械瓣膜术后 那样服用价格高昂的抗凝血药物[Fuchs,J.R.，B.A.Nasseri,andJ.P.Vacanti,Tissue engineering:a21stcenturysolutiontosurgicalreconstruction.TheArmaisoi ThoracicSurgery, 2001? 72 (2) :p. 577-591.]。在尺寸大小和解剖结构等方面，猪的心脏瓣 膜因结构和尺寸与人的心脏瓣膜最为相似，所以猪心脏主动脉瓣膜是制备异种生物瓣膜的 理想材料之一 [Fuchs,J.R. ,B.A.Nasseri,andJ.P.Vacanti,Tissueengineering:a21st centurysolutiontosurgicalreconstruction.TheAnnalsofThoracic Surgery, 2001. 72 (2) :p. 577-591.]。为提高所植入异种生物瓣膜的稳定性和持久性，降低 免疫原性，提高抗菌性，临床上所用的人工生物心脏瓣膜均采用戊二醛交联处理。但戊二 醛本身具有细胞毒性，抑制生物瓣膜在宿主体内内皮化，容易引起瓣膜钙化、变脆、易撕裂， 致使瓣膜工作平均寿命仅机械瓣膜的1/3左右[Schmidt,C.E.andJ.M.Baier，Acellular vasculartissues:naturalDiomaterialsfortissuerepairandtissueengineering. Biomaterials，2000? 21 (22) :p. 2215-2231.]。另外，尽管戊二醛有杀菌和降低免疫原性的 能力，但效果有限。
[0003] 原花青素（Procyanidins,PC)是ー种黄酮类化合物，广泛存在于各种食 物中，如水果、蔬菜、坚果、种子、花朵、树皮、茶叶等[Dixon,R.A.，D. -Y.Xie,and S.B.Sharma,Proanthocyaniains-afinalfrontierinflavonoidresearchNew Phytologist, 2005. 165(1) :p. 9-28. ;Packer，L，G.Rimbach，andF.VirgiIi，Antioxidant activityandbiologicpropertiesofaprocyanidin-richextractfrompine(pinus maritima)bark，pycnogenol.FreeRadicalBiologyandMedicine, 1999. 27 (5-6):p.70 4-724.]，具有抗氧化、清除自由基[Packer，L，G.Rimbach，andF.Virgili，Antioxidant activityandbiologicpropertiesofaprocyanidin-richextractfrompine(pinus maritima)bark,pycnogenol.FreeRadicalBiologyandMedicine, 1999. 27 (5-6):p.7 04-/24. ；Andry,M.C.,etal. ,Proanthocyanidinmicrocapsules:preparation，proper tiesandfreeradicalscavengingactivity.InternationalJournalofPharmace utics，1998. 171 (2) :p. 217-226.]、抗癌[Zhang,X. -Y. ,etal. ,Proanthocyanidinfrom grapeseedspotentiatesanti-tumoractivityofdoxoruoicinviaimmunomodulatory mechanism.InternationalImmunopharmacology，2005? 5 (7-8) :p. 1247-1257.]、保护心血 ^ [Sato,M. ,etal. ,Grapeseedproanthocyanidinreducescardiomyocyteapoptosis byinhibitingischemia/reperfusion-inducedactivationoijNK-IandC-jUN.；Free RadicalBiologyandMedicine，2001? 31 (6) :p. 729-737.Shao,Z. -H.，etal.，Grape seedproanthocyanidinextractattenuatesoxidantinjuryincardiomyocytes. PharmacologicalResearch, 2003. 47 (6) :p. 463-469. ；Du,Y.andH.Lou,Catechinand proanthocyanidinB4fromgrapeseedspreventdoxorubicin-inducedtoxicityin cardiomyocytes.EuropeanJournalofPharmacology，2008? 591 (1-3):p.96-101.]、 抗 柄-母[Gescher，K.，etal. ,Proanthocyanidin-enrichedextractfromMyrothamnus flabelliioliaWelw.exertsantiviralactivityagainstherpessimplexvirus typelbyinhibitionofviraladsorptionandpenetration.JournalofEthnopharma cology,2011.134(2):p. 468-474. ;Gescher，K.，etal. ,Oligomericproanthocyaniains fromRumexacetosaLinhibittheattachmentofherpessimplexvirustype-1. AntiviralResearch，2011.89(1):p. 9-18.]、抗菌消炎[Baydar，N.G.，G.Ozkan，and 0.Oagdi^9Totalphenoliccontentsandantibacterialactivitiesofgrape(Vitis viniferaL. )extracts.FoodControl, 2004. 15 (5) :p. 335-339. ；Mvakumaran,S. ,et al. ,VariationinantimicrooialactionofproanthocyanidinsfromDorycnium rectumagainstrumenbacteria.Phytochemistry,2004.65 (17):p.2485-2497.； Furiga,A. ,A.Lonvaud-FuneI,andC.Badet,Invitrostudyofantioxidantcapacity andantibacterialactivityonoralanaerobesofagrapeseedextract.FoodC hemistry, 2009. 113 (4) :p. 1037-1040.；RodriguezVaquero,M.J. ,etal. ,Antioxidant capacityandantibacterialactivityofphenoliccompoundsfromargentmean herbsinfusions.FoodControl，2010.21(5) :p. 779-785.]等多种生物活性。尤其是近 年来发现原花青素具有抗胶原降解[Green，B.，etal.，Grapeseedproanthocyanidins increasecollagenDiodegradationresistanceinthedentin/adhesiveinterface whenincludedinanadhesive.JournaloiDentistry，2010? 38(11):p. 908-915. ]、抗血栓形成[Sano,T.，etal. ,Anti-thromboticeffectofproanthocyanidin,a purifiedingredientofgrapeseed.ThrombosisResearch,2005. 115 (1-2):p. 115-121. ] ^ 抑制?丐化[Zhai，W.，etal. ,Procyanidins-crosslinkedheartvalve matrix:Anticalcificationeffect.JournalofBiomedicalMaterialsResearch PartB:AppliedBiomaterials，2009? 90B(2) :p. 913-921.]、降低免疫原性[Liu，Y.Z.，et al. ,ImmunomodulatoryeffectsofproanthocyanidinA-Iderivedinvitrofrom Rhododendronspiciferum.Fitoterapia，2010.81(2):p. 108-114.]的特点，使得原花青素 可能成为ー种潜在的人工生物辧膜交联剂。Zhai等[Zhai，W.，etal.,Crosslinkingof decellularizedporcineheartvalvematrixbyprocyanidins.Biomaterials, 2006. 2 7 (19) :p. 3684-3690. ] 2006年发现，PC用作交联剂交联猪脱細胞心脏瓣膜，具有力学性能 好、稳定性强、抑制钙化生成等优点。与戊二醛交联相比，其突出优点即有效抑制瓣膜钙化、 无细胞毒性。但也存在着一