CN113546213A - 一种生物能量活性材料及其应用 - Google Patents
一种生物能量活性材料及其应用 Download PDFInfo
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- CN113546213A CN113546213A CN202110875054.2A CN202110875054A CN113546213A CN 113546213 A CN113546213 A CN 113546213A CN 202110875054 A CN202110875054 A CN 202110875054A CN 113546213 A CN113546213 A CN 113546213A
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- active material
- bioenergy
- acid
- hydroxybutyrate
- bone tissue
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Abstract
本发明公开了一种生物能量活性材料及其应用,具体公开一种生物能量活性材料,其为生物可降解聚合物;所述生物能量活性材料的降解产物为三羧酸循环和/或糖酵解途径中的代谢中间产物,或者为能够转换生成三羧酸循环和/或糖酵解途径中代谢中间产物的聚合物单体,或者为能够转换生成乙酰辅酶A的聚合物单体。该生物能量活性材料的降解产物经过三羧酸代谢循环或者糖酵解途径为组织细胞提供生物能量,解决传统生物可降解材料无法持续提高细胞中ATP的稳定性和相关生物质活性的问题,在骨组织再生领域尤其是在大段骨缺损修复方面表现出广阔的应用前景。
Description
技术领域
本发明属于生物医学组织工程,具体涉及一种生物能量活性材料及其应用。
背景技术
在临床上,因创伤、感染、骨肿瘤切除等原因造成的各种类型骨缺损十分常见。全世界范围内每年进行骨移植患者的数量达到220万【J.Van der Stok,E.M.Van Lieshout,Y.El-Massoudi,G.H.Van Kralingen,P.Patka,Bone substitutes in the Netherlands-asystematic literature review,Acta Biomater 7(2)(2011)739-50.】。据美国骨科医师协会(AAOS)统计,在美国每年有630万人发生骨折,其中需要接受骨移植物的患者就有50万之多,每年仅在骨折治疗方面的费用支出高达2000亿美元【D.C.Lobb,B.R.DeGeorge,Jr.,A.B.Chhabra,Bone Graft Substitutes:Current Concepts and Future Expectations,JHand Surg Am 44(6)(2019)497-505e2.】。我国每年骨损伤患者超过350万,每年新增病例达到数十万之多,创伤住院年增长率达7.2%,高居住院人数的第二位,骨缺损的高发生率使得骨移植物成为仅次于输血之后需求量最大的医疗耗材【G.H.Brundtland,A WHOScientific Group on the Burden of Musculoskeletal Conditions at the Start ofthe New Millennium met in Geneva from 13to 15January 2000.,Who Tech Rep Ser919(2003)1-218.】,给社会带来沉重的医疗负担。
自体骨的移植可以较好的进行骨修复,但是“供体有限”限制了其更好的应用。生物可降解高分子材料因具有良好的生物相容性和降解性而广泛用于骨组织工程研究。生物可降解高分子材料包括天然可生物降解聚合物(如胶原、壳聚糖)和合成生物可降解聚合物(如PLA、PLGA、PCL),因具有良好的生物相容性和降解性,是目前常用的骨组织工程支架材料【F.Asghari,M.Samiei,K.Adibkia,A.Akbarzadeh,S.Davaran,Biodegradable andbiocompatible polymers for tissue engineering application:a review,ArtifCellsNanomed Biotechnol 45(2)(2017)185-192.】。虽然可生物降解聚合物具有一定的骨缺损修复功能,但仍然存在降解速率不可控、力学性能较差、酸性降解产物引起炎症反应等系列自身难以克服的缺点【Y.X.Lai,Y.Li,H.J.Cao,J.Long,X.L.Wang,L.Li,C.R.Li,Q.Y.Jia,B.Teng,T.T.Tang,J.Peng,D.Eglin,M.Alini,D.W.Grijpma,G.Richards,L.Qin,Osteogenic magnesium incorporated into PLGA/TCP porous scaffoldby 3D printingfor repairing challenging bone defect,Biomaterials 197(2019)207-219.】,阻碍了其在临床上的广泛应用。
骨再生过程是一个能量消耗的过程,细胞能量代谢对组织修复和再生起着至关重要的作用。三磷酸腺苷(ATP)是细胞能量的主要来源,在许多生物过程中发挥作用,包括细胞的增殖、迁移及分化【I.Gadjanski,S.Yodmuang,K.Spiller,S.Bhumiratana,G.Vunjak-Novakovic,Supplementation of Exogenous Adenosine 5-Triphosphate EnhancesMechanical Properties of 3D Cell-Agarose Constructs for Cartilage TissueEngineering,Tissue Eng Pt A 19(19-20)(2013)2188-2200.】。研究表明,生物能量作为一种潜在的治疗方法,已在体外模型或相对薄的浅表组织(如皮肤)再生方面取得了成功。到目前为止,用于复杂骨组织缺损修复的具有长期生物能量释放效应的3D支架鲜有报道。这在很大程度上是由于使用现有的生物支架材料无法持续提高细胞中ATP的稳定性和相关生物质活性,阻碍了生物能量活性材料在骨组织工程中的应用和发展。
发明内容
本发明为了解决现有技术中的不足和缺点,提出一种生物能量活性材料及其应用。具体方案如下:
本发明第一方面提供一种生物能量活性材料,所述生物能量活性材料为生物可降解聚合物;所述生物能量活性材料的降解产物为三羧酸循环和/或糖酵解途径中的代谢中间产物;
或者所述生物能量活性材料的降解产物为能够转换生成三羧酸循环和/或糖酵解途径中代谢中间产物的聚合物单体;
或者所述生物能量活性材料的降解产物为能够转换生成乙酰辅酶A的聚合物单体。
进一步地,本发明的上述技术方案中,所述三羧酸循环的代谢中间产物包括柠檬酸、顺乌头酸、异柠檬酸、草酰琥珀酸、a-酮戊二酸、琥珀酰辅酶A、琥珀酸、延胡索酸、苹果酸和三磷酸腺苷中的一种以上;
所述糖酵解途径的代谢中间产物包括葡萄糖-6-磷酸、果糖-6-磷酸、果糖-1,6-二磷酸、3-磷酸甘油醛、磷酸二羟丙酮、1,3-二磷酸甘油酸、3-磷酸甘油酸、2-磷酸甘油酸、磷酸烯醇式丙酮酸PEP和丙酮酸中的一种以上;
所述能够转换生成乙酰辅酶A的聚合物单体为3-羟基丁酸。3-羟基丁酸会在3-羟基丁酸脱氢酶(3-hydroxybutyrate dehydrogenase)的作用下生成乙酰乙酸,再和琥珀酰辅酶A(succinyl-CoA)在3-琥珀酰辅酶A转移酶(3-oxoacid CoA-transferase)酶的作用下合成乙酰乙酰辅酶A(Acetoacetyl-CoA),乙酰乙酰辅酶A与一个辅酶A(CoA)反应,在乙酰辅酶A-酰基转移酶(acetyl-CoAC-acetyltransferases)生成两个乙酰辅酶A(acetyl-CoA),从而进入三羧酸循环,为组织细胞提供生物能量。
优选地,本发明的上述技术方案中,所述生物能量活性材料为降解产物为3-羟基丁酸的聚羟基脂肪酸酯。
进一步地,所述聚羟基脂肪酸酯包括3-羟基丁酸-4-羟基丁酸共聚酯(P34HB)、聚-3-羟基丁酸酯(PHB)、3-羟基丁酸-3-羟基戊酸共聚酯(PHBV)和3-羟基丁酸-3-羟基己酸共聚酯(PHBHHx)中的一种以上。
本发明第二方面提供所述的生物能量活性材料在骨组织再生及修复领域中的应用。
本发明第三方面提供所述的生物能量活性材料在制备骨组织修复多孔支架中的应用。
本发明第四方面提供一种骨组织修复多孔支架,由所述的生物能量活性材料制备得到。进一步地,骨组织修复多孔支架可以通过传统制备方法,也可以通过3D打印技术制备。
本发明第五方面提供一种骨组织修复3D多孔支架的制备方法,包括如下步骤:
(1)合成所述的生物能量活性材料;
(2)结合3D打印技术制备骨组织修复3D多孔支架。
进一步地,本发明的上述制备方法中,所述生物能量活性材料通过微生物法或化学合成法合成,也可以通过其他方式合成。例如,可以通过微生物嗜盐单胞菌发酵生产3-羟基丁酸-4-羟基丁酸共聚酯。
本发明第五方面提供上述制备方法制备的骨组织修复3D多孔支架。
本发明的有益效果:
1、本发明提供的生物能量活性材料的降解产物为三羧酸循环和/或糖酵解途径中的代谢中间产物,或者为能够转换生成三羧酸循环和/或糖酵解途径中代谢中间产物的聚合物单体,或者为能够转换生成乙酰辅酶A的聚合物单体,该生物能量活性材料的降解产物经过三羧酸代谢循环或者糖酵解途径为组织细胞提供生物能量,解决传统生物可降解材料无法持续提高细胞中ATP的稳定性和相关生物质活性的问题,在骨组织再生领域尤其是在大段骨缺损修复方面表现出广阔的应用前景。
2、本发明提供的生物能量活性材料3-羟基丁酸-4-羟基丁酸共聚酯(P34HB)是一类可降解高分子聚酯,具有独特的生物能量活性、机械力学特性、生物可降解性和生物相容性,其降解产生的主要产物3-羟基丁酸(3HB)是哺乳动物体内酮体的主要组成之一,不仅对机体无毒性作用,而且可作为一种能量物质促进细胞的黏附、增殖以及分化。此外,与聚乳酸相比,在相同条件下,3-羟基丁酸-4-羟基丁酸共聚酯材料可以维持较长的降解时间,根据组织修复的要求调整材料中3-羟基丁酸的比例可以实现6-12月较长时间的降解。3-羟基丁酸-4-羟基丁酸共聚酯材料能够解决传统生物可降解材料无法持续提高细胞中ATP的稳定性和相关生物质活性,以及酸性降解产物引起炎症反应的问题,是一种性能优异的具有生物能量活性兼具骨形成促进功能的骨修复支架活性材料。
3、本发明提供的骨组织修复多孔支架基于生物能量活性材料和3D打印技术,该多孔支架结构可控,且具有良好的生物相容性、生物可降解性和机械力学特性,支架植入体内后随着自身不断的降解,其降解产生的生物能量活性物质可以促进骨髓间充质干细胞(hBMSCs)细胞的增殖、分化及矿化,具有骨形成促进功能,是传统高分子支架材料所不具备的。
4、本发明进一步提供的骨组织修复3D多孔支架基于生物能量活性材料3-羟基丁酸-4-羟基丁酸共聚酯(P34HB)和3D打印技术制备,该多孔支架结构可控,且具有良好的生物相容性、生物可降解性和机械力学特性,植入体内后随着自身不断的降解,降解产生的生物能量活性物质3-羟基丁酸(3HB),经过三羧酸代谢循环不仅为组织细胞提供生物能量,促进骨髓间充质干细胞(hBMSCs)细胞的增殖、分化及矿化,而且产生的中间代谢物以柠檬酸盐的形式参与骨形成,有利骨修复过程中成骨和成血管功能,缩短了骨缺损修复时间,在骨组织再生及修复领域尤其是在大段骨缺损修复方面表现出巨大的应用前景。
附图说明
图1为3-羟基丁酸-4-羟基丁酸共聚酯的凝胶渗透色谱图;其中,A:标准曲线,B:3-羟基丁酸-4-羟基丁酸共聚酯;
图2为3D打印制备的多孔支架(A)及多孔支架横截面电镜扫描图(B);
图3为支架降解产物3-羟基丁酸对人源骨髓间充质干细胞(hBMSCs)增值的影响结果;
图4为支架降解产物3-羟基丁酸对hBMSCs碱性磷酸酶表达活性的影响结果;
图5为支架降解产物3-羟基丁酸对hBMSCs胞外钙结节形成的影响结果;
图6为激光共聚拉曼光谱分析hBMSCs胞外磷灰石的形成情况;
图7为支架降解产物3-羟基丁酸对hBMSCs成骨分化相关基因表达的影响结果;
图8为支架降解产物3-羟基丁酸对细胞线粒体膜势能的影响结果。
具体实施方式
为了更清楚地理解本发明,现参照下列实施例及附图进一步描述本发明。实施例仅用于解释而不以任何方式限制本发明。实施例中,各原始试剂材料均可商购获得,未注明具体条件的实验方法为所属领域熟知的常规方法和常规条件,或按照仪器制造商所建议的条件。
实施例1
(1)将能合成3-羟基丁酸-4-羟基丁酸共聚酯的微生物嗜盐单胞菌在60MMG培养基中,37℃,400-800rpm条件下发酵培养72小时,72小时后收集菌体,置于70℃对菌体进行通风干燥,得到含有3-羟基丁酸-4-羟基丁酸共聚酯的干菌体粉末。
60MMG培养基组成为:葡萄糖30g/L,酵母提取物1g/L,硫酸铵0.25g/L,硫酸镁0.2g/L,磷酸氢二钠9.65g/L,磷酸二氢钾1.5g/L,微量元素I10ml/L,微量元素II 1ml/L。
(2)用氯仿对干菌体粉末中的3-羟基丁酸-4-羟基丁酸共聚酯进行提取(1g干菌体粉末加入20ml氯仿),搅拌均匀后装入高压反应釜中,在100℃条件下反应4小时。
(3)待高压反应釜冷却后,采用过滤或者抽滤的方法去掉细胞碎片,得到澄清氯仿溶液。
(4)在60℃条件下将氯仿溶液浓缩(按照100ml氯仿溶液浓缩到60ml的比例进行),然后加入到15倍体积的预冷无水乙醇中,然后置于4℃冰箱中过夜,进行沉淀析出。
(5)过滤收集步骤(4)中的沉淀析出物,并将收集到的沉淀析出物置于40℃真空干燥箱中24h,待溶剂挥发完全后,即可得3-羟基丁酸-4-羟基丁酸共聚酯。
称取50mg 3-羟基丁酸-4-羟基丁酸共聚酯溶解于氯仿中,静置1小时形成均匀的溶液,然后取10ul进行凝胶渗透色谱,测试分子量。结果如图1所示。
分子量计算结果如下:
实施例2
将实施例1中合成得到的3-羟基丁酸-4-羟基丁酸共聚酯材料装入熔融3D打印机(180℃),进行骨组织修复多孔支架制备。
结果如图2所示,其中,图2-A为3D打印制备的多孔支架,图2-B为支架横截面电镜扫描图,多孔支架的孔径大约在350-400μm。
实验例
将实施例2中3D多孔支架浸泡于磷酸盐缓冲液8周,收集降解产物。在磷酸盐缓冲液中,3-羟基丁酸-4-羟基丁酸共聚酯的降解产物主要是3-羟基丁酸(3HB)。对收集到的降解产物进行浓度测定,然后进行体外实验。
磷酸盐缓冲液的组成为:氯化钠7.9g/L,氯化钾0.2g/L,磷酸二氢钾0.24g/L。
(1)取生长状态良好的人源骨髓间充质干细胞(hBMSCs),按照2×104细胞密度接种于48孔板,4小时后向上述细胞中加入不同浓度的3-羟基丁酸(0μM,10μM,40μM,80μM,160μM,320μM),在第1,5,7天用CCK-8分别测定细胞增殖情况。同时以乳酸(LA)作为对照组。
结果如图3所示。图3为支架降解产物3-羟基丁酸对人源骨髓间充质干细胞(hBMSCs)增值的影响结果,与对照组(LA)相比,在第5天,第7天3HB刺激后显著促进了骨髓间充质干细胞的增殖,表明支架降解产物能够促进hBMSCs增值。
(2)将人源骨髓间充质干细胞(hBMSCs),按照1×105细胞密度接种于6孔板,培养12小时后,向上述细胞中加入含有不同浓度的3-羟基丁酸(0μM,40μM,160μM,320μM)的成骨诱导分化液,在第7,14天分别检测碱性磷酸酶的表达情况。根据碱性磷酸酶活性检测试剂盒的使用说明进行碱性磷酸酶表达的检测。
成骨诱导分化液的组成:低糖DMEM培养基+10%胎牛血清+2mM L-谷氨酰胺+100U/mL青霉素+100μg/mL链霉素+100nM地塞米松+0.2mML-抗坏血酸+10mMβ-甘油磷酸钠。
结果如图4所示。图4为支架降解产物3-羟基丁酸对hBMSCs碱性磷酸酶表达活性的影响结果,结果表明支架降解产物能够促进hBMSCs碱性磷酸酶表达。且与对照组(LA)相比,3HB刺激骨髓间充质干细胞诱导分化14天后,显著增强了碱性磷酸酶的表达,而对照组则表现出抑制作用。
(3)将人源骨髓间充质干细胞(hBMSCs),按照1×105细胞密度接种于6孔板,培养12小时后,向上述细胞中加入含有不同浓度的3-羟基丁酸(0μM,40μM,160μM,320μM)的成骨诱导分化液,在第10,14天分别用茜素红然检测胞外钙结节的形成。
结果如图5所示。图5为支架降解产物3-羟基丁酸对hBMSCs胞外钙结节形成的影响结果,与对照组(LA)相比,3HB刺激骨髓间充质干细胞诱导分化10,14天后显著增强了胞外钙结节的沉积,表明3-羟基丁酸促进了骨髓间充质干细胞成骨分化标志物钙结节的形成。
(4)将人源骨髓间充质干细胞(hBMSCs),按照1×105细胞密度接种于6孔板,培养12小时后,向上述细胞中加入含有不同浓度的3-羟基丁酸的成骨诱导分化液(0μM,40μM,160μM,320μM),进行21天诱导分化,诱导21天后,利用激光共聚焦拉曼光谱检测胞外磷灰石的形成。
结果如图6所示。图6为激光共聚拉曼光谱分析hBMSCs胞外磷灰石的形成情况,骨髓间充质干细胞诱导分化21天后,胞外有大量的磷灰石形成且其含量随着3HB浓度的变大而增多,表明3-羟基丁酸能够促进骨髓间充质干细胞成骨分化磷灰石的形成。
(5)将人源骨髓间充质干细胞(hBMSCs),按照1×105细胞密度接种于6孔板,培养12小时后,向上述细胞中加入含有不同浓度的3-羟基丁酸(0μM,40μM,160μM,320μM)的成骨诱导分化液,进行7天诱导分化,诱导7天后,利用实时荧光定量PCR技术检测成骨分化相关基因(Runx相关转录因子2基因、骨钙素基因、骨保护素基因)表达情况。
结果如图7所示。图7为支架降解产物3-羟基丁酸对hBMSCs成骨分化相关基因表达的影响结果,与对照组(LA)相比,3HB刺激骨髓间充质干细胞诱导分化7天后促进了成骨分化相关基因的表达,而对照组则在一定程度上抑制了基因的表达,表明支架降解产物3-羟基丁酸能够促进hBMSCs成骨分化相关基因表达。
(6)将人源骨髓间充质干细胞(hBMSCs),按照1×104细胞密度接种于6孔板,培养12小时后,实验分为阳性对照组,阴性对照组以及实验组,阳性对照组用高糖培养基(HG)处理细胞6小时;阴性对照组用含有氧化磷酸化解偶联剂(CCCP)的高糖培养基处理细胞6小时;实验组用含有不同浓度(40μM,160μM,320μM)的3-羟基丁酸和乳酸(LA)的无糖培养基(GF)处理细胞6小时。
结果如图8所示。图8为支架降解产物3-羟基丁酸对细胞线粒体膜势能的影响结果,结果表明支架降解产物3-羟基丁酸能够为细胞提供ATP及维持线粒体膜势能(ΔΨm),且与LA组相比,3HB能为细胞提供较多的生物能量(ATP)并提高细胞膜势能。
Claims (10)
1.一种生物能量活性材料,其特征在于,所述生物能量活性材料为生物可降解聚合物;所述生物能量活性材料的降解产物为三羧酸循环和/或糖酵解途径中的代谢中间产物;
或者所述生物能量活性材料的降解产物为能够转换生成三羧酸循环和/或糖酵解途径中代谢中间产物的聚合物单体;
或者所述生物能量活性材料的降解产物为能够转换生成乙酰辅酶A的聚合物单体。
2.根据权利要求1所述的生物能量活性材料,其特征在于,所述三羧酸循环的代谢中间产物包括柠檬酸、顺乌头酸、异柠檬酸、草酰琥珀酸、a-酮戊二酸、琥珀酰辅酶A、琥珀酸、延胡索酸、苹果酸和三磷酸腺苷中的一种以上;
所述糖酵解途径的代谢中间产物包括葡萄糖-6-磷酸、果糖-6-磷酸、果糖-1,6-二磷酸、3-磷酸甘油醛、磷酸二羟丙酮、1,3-二磷酸甘油酸、3-磷酸甘油酸、2-磷酸甘油酸、磷酸烯醇式丙酮酸PEP和丙酮酸中的一种以上;
所述能够转换生成乙酰辅酶A的聚合物单体为3-羟基丁酸。
3.根据权利要求1所述的生物能量活性材料,其特征在于,所述生物能量活性材料为降解产物为3-羟基丁酸的聚羟基脂肪酸酯。
4.根据权利要求3所述的生物能量活性材料,其特征在于,所述聚羟基脂肪酸酯包括3-羟基丁酸-4-羟基丁酸共聚酯、聚-3-羟基丁酸酯、3-羟基丁酸-3-羟基戊酸共聚酯和3-羟基丁酸-3-羟基己酸共聚酯中的一种以上。
5.权利要求1-4任一项所述的生物能量活性材料在骨组织再生及修复领域中的应用。
6.权利要求1-4任一项所述的生物能量活性材料在制备骨组织修复多孔支架中的应用。
7.一种骨组织修复多孔支架,由权利要求1-4任一项所述的生物能量活性材料制备得到。
8.一种骨组织修复3D多孔支架的制备方法,其特征在于,包括如下步骤:
(1)合成权利要求1-4任一项所述的生物能量活性材料;
(2)结合3D打印技术制备骨组织修复3D多孔支架。
9.根据权利要求8所述的制备方法,其特征在于,所述生物能量活性材料通过微生物法或化学合成法合成。
10.权利要求8或9所述制备方法制备的骨组织修复3D多孔支架。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102936407A (zh) * | 2012-10-19 | 2013-02-20 | 华中科技大学 | 一种基于三羧酸循环的能量生物材料及其制备方法 |
CN103785059A (zh) * | 2014-01-03 | 2014-05-14 | 朱小龙 | 一种骨修复再生材料及其制备方法 |
WO2015117019A1 (en) * | 2014-01-30 | 2015-08-06 | Easel Biotechnologies, Llc | Improved carbon dioxide fixation via bypassing feedback regulation |
CN105771000A (zh) * | 2016-02-26 | 2016-07-20 | 华中科技大学 | 一种提供能量的骨修复可降解多孔支架及制备方法、应用 |
CN107375934A (zh) * | 2017-08-10 | 2017-11-24 | 浙江大学 | 含有果糖‑1,6‑二磷酸的组合物在制备抗肿瘤药物中的应用 |
CN112773940A (zh) * | 2021-01-26 | 2021-05-11 | 深圳市创想三维科技有限公司 | 一种3d打印用人工骨支架材料及其制备方法和应用 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
RU2708396C1 (ru) * | 2018-12-19 | 2019-12-06 | Федеральное государственное бюджетное учреждение науки Институт высокомолекулярных соединений Российской академии наук | Биосовместимый биоразлагаемый остеокондуктивный полимерный композиционный материал для регенерации костной ткани |
CN109676915B (zh) * | 2019-02-15 | 2022-02-15 | 复旦大学 | 一种车轮状多孔支架及其制备方法与应用 |
CN113546213A (zh) * | 2021-07-30 | 2021-10-26 | 深圳先进技术研究院 | 一种生物能量活性材料及其应用 |
-
2021
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-
2022
- 2022-08-01 US US18/574,727 patent/US20240299620A1/en active Pending
- 2022-08-01 CN CN202280004789.0A patent/CN115884799A/zh active Pending
- 2022-08-01 WO PCT/CN2022/109458 patent/WO2023006113A1/zh active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102936407A (zh) * | 2012-10-19 | 2013-02-20 | 华中科技大学 | 一种基于三羧酸循环的能量生物材料及其制备方法 |
CN103785059A (zh) * | 2014-01-03 | 2014-05-14 | 朱小龙 | 一种骨修复再生材料及其制备方法 |
WO2015117019A1 (en) * | 2014-01-30 | 2015-08-06 | Easel Biotechnologies, Llc | Improved carbon dioxide fixation via bypassing feedback regulation |
CN105771000A (zh) * | 2016-02-26 | 2016-07-20 | 华中科技大学 | 一种提供能量的骨修复可降解多孔支架及制备方法、应用 |
CN107375934A (zh) * | 2017-08-10 | 2017-11-24 | 浙江大学 | 含有果糖‑1,6‑二磷酸的组合物在制备抗肿瘤药物中的应用 |
CN112773940A (zh) * | 2021-01-26 | 2021-05-11 | 深圳市创想三维科技有限公司 | 一种3d打印用人工骨支架材料及其制备方法和应用 |
Non-Patent Citations (3)
Title |
---|
冯静等: "《几种高分子材料的生物降解研究进展》", 《塑料科技》 * |
张一鸣: "《生物化学与分子生物学》", 31 August 2018, 东南大学出版社 * |
李谦: "《生物化学》", 31 December 2019, 中国医药科技出版社 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023006113A1 (zh) * | 2021-07-30 | 2023-02-02 | 深圳先进技术研究院 | 一种生物能量活性材料及其应用 |
CN115444985A (zh) * | 2022-09-19 | 2022-12-09 | 中鼎凯瑞科技成都有限公司 | 三磷酸腺苷基高能骨修复材料及其制备方法 |
CN115444985B (zh) * | 2022-09-19 | 2023-09-26 | 中鼎凯瑞科技成都有限公司 | 三磷酸腺苷基高能骨修复材料及其制备方法 |
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