聚三亚甲基碳酸酯体外酶解性能的影响因素及其作用规律
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摘要
背景:聚三亚甲基碳酸酯具有良好的生物相容性和生物降解性能,具有较大的临床应用前景。目前,国内外关于聚三亚甲基碳酸酯体外酶解性能影响因素及其作用规律的研究较少。目的:研究聚三亚甲基碳酸酯的体外酶解性能,并探讨影响聚三亚甲基碳酸酯体外酶解性能的因素及其作用规律。方法:通过开环聚合反应制备聚三亚甲基碳酸酯均聚物及其共聚物,2种均聚物的分子质量分别为135,256 kDa,共聚物的分子质量为238 kDa,将分子质量为256 kDa的均聚物制成棒条与膜片2种形状,其余2种样品制成棒条。将均聚物、共聚物样品分别置于脂肪酶溶液中,均聚物样品于第1,2,4,8,10,12周取出,共聚物样品于3,6,9,12,15 d后取出样品,测量样品质量变化,计算失重率及降解速率常数。结果与结论:(1)分子质量、形状及共聚改性对聚三亚甲基碳酸酯的体外酶解性能有显著影响;(2)当分子质量由135kD a增加至256kD a时,聚三亚甲基碳酸酯均聚物棒条的酶解速率常数由每周1.46%增加到每周3.81%,说明分子质量越高,聚三亚甲基碳酸酯均聚物的体外酶解速率越快;(3)当形状由棒条转换为膜片时,分子质量为256 kDa聚三亚甲基碳酸酯均聚物的酶解速率常数由每周3.81%增加到每周9.16%,说明膜片形状样品降解速率更快;(4)在分子质量为256 kDa聚三亚甲基碳酸酯结构中引入等摩尔比己内脂成分后,其酶解速率常数由每周3.81%增加到每周14.49%,说明在结构中引入聚己内酯成分,可加速聚三亚甲基碳酸酯的降解速率;(5)各因素对聚三亚甲基碳酸酯体外酶解速率的影响程度顺序为:共聚组成>样品形状>分子质量。
BACKGROUND: Poly(trimethylene carbonate) has great potential in clinical applications due to the excellent biocompatibility and biodegradability. Little is reported on the factors influencing the in vitro enzymatic degradation of poly(trimethylene carbonate) and the underlying mechanism. OBJECTIVE: To investigate the in vitro enzymatic degradation of poly(trimethylene carbonate), and to explore the influencing factors and their effects on the poly(trimethylene carbonate) degradation. METHODS: poly(trimethylene carbonate) homopolymers and copolymers were prepared by ring-opening polymerization. The molecular mass of the two homopolymers was 135 and 256 kDa, respectively. The molecular mass of the copolymers was 238 kDa. The homopolymer of 256 kDa was made into two shapes(rods and films), and the other two samples were shaped into rods. The in vitro enzymatic degradation of poly(trimethylene carbonate) was conducted in lipase solutions, the homopolymer samples were taken out at 1, 2, 4, 8, 10, and 12 weeks, and the copolymer samples were taken at 3, 6, 9, 12, and 15 days. The mass loss and degradation rate constant were measured. RESULTS AND CONCLUSION: The molecular mass and shape as well as molar ratio played important roles on the in vitro enzymatic degradation behavior of poly(trimethylene carbonate). As the molecular mass increased from 135 to 256 kDa, the degradation rate constant of poly(trimethylene carbonate) homopolymer increased from 1.46% to 3.81%, indicating that the higher the molecular mass, the higher degradation rate of poly(trimethylene carbonate). The poly(trimethylene carbonate) film presented with higher degradation rate than the cylinder one with the same molecular weight, and the degradation rate constant increased from 3.81% to 9.16% as the shape of poly(trimethylene carbonate) with a molecular weight of 256 kDa changed from rods to films. The introduction of polycaprolactone segment accelerated the degradation rate of poly(trimethylene carbonate). The degradation rate constant increased from 3.81% to14.49% as the 50 mol% caprolactone content was introduced into the structure of poly(trimethylene carbonate)(256 kDa). In summary, the order of factors influencing the degradation rate of poly(trimethylene carbonate) is as follows: copolymer composition > shape > molecular mass.
BACKGROUND: Poly(trimethylene carbonate) has great potential in clinical applications due to the excellent biocompatibility and biodegradability. Little is reported on the factors influencing the in vitro enzymatic degradation of poly(trimethylene carbonate) and the underlying mechanism. OBJECTIVE: To investigate the in vitro enzymatic degradation of poly(trimethylene carbonate), and to explore the influencing factors and their effects on the poly(trimethylene carbonate) degradation. METHODS: poly(trimethylene carbonate) homopolymers and copolymers were prepared by ring-opening polymerization. The molecular mass of the two homopolymers was 135 and 256 kDa, respectively. The molecular mass of the copolymers was 238 kDa. The homopolymer of 256 kDa was made into two shapes(rods and films), and the other two samples were shaped into rods. The in vitro enzymatic degradation of poly(trimethylene carbonate) was conducted in lipase solutions, the homopolymer samples were taken out at 1, 2, 4, 8, 10, and 12 weeks, and the copolymer samples were taken at 3, 6, 9, 12, and 15 days. The mass loss and degradation rate constant were measured. RESULTS AND CONCLUSION: The molecular mass and shape as well as molar ratio played important roles on the in vitro enzymatic degradation behavior of poly(trimethylene carbonate). As the molecular mass increased from 135 to 256 kDa, the degradation rate constant of poly(trimethylene carbonate) homopolymer increased from 1.46% to 3.81%, indicating that the higher the molecular mass, the higher degradation rate of poly(trimethylene carbonate). The poly(trimethylene carbonate) film presented with higher degradation rate than the cylinder one with the same molecular weight, and the degradation rate constant increased from 3.81% to 9.16% as the shape of poly(trimethylene carbonate) with a molecular weight of 256 kDa changed from rods to films. The introduction of polycaprolactone segment accelerated the degradation rate of poly(trimethylene carbonate). The degradation rate constant increased from 3.81% to14.49% as the 50 mol% caprolactone content was introduced into the structure of poly(trimethylene carbonate)(256 kDa). In summary, the order of factors influencing the degradation rate of poly(trimethylene carbonate) is as follows: copolymer composition > shape > molecular mass.
引文
[1]Zhang Z,Kuijer R,Bulstra SK,et al.The in vivo and in vitro degradation behavior of poly(trimethylene carbonate).Biomaterials.2006;27(9):1741-1748.
[2]Zhu KJ,Hendren RW,Jensen K,et al.Synthesis,properties,and degradation of poly(1,3-trimethylene carbonate).Macromolecules.1991;24(8):1736-1740.
[3]杨立群,周群华,张巍,等.聚三亚甲基碳酸酯的研究进展[J].高分子通报,2016,29(8):29-43.
[4]赵宁,张现军,李伟,等.左旋聚乳酸羟基磷灰石生物材料与牙周膜细胞的生物相容性[J].中国组织工程研究,2016,20(12):1732-1737.
[5]马晓妍,石淑先,夏宇正,等.聚乳酸及其共聚物的制备和降解性能[J].北京化工大学学报(自然科学版),2004,31(1):51-56.
[6]胡露,魏坤,邹芬.聚乳酸-羟基乙酸共聚物/硅酸钙三维多孔骨组织工程支架的构建与性能[J].中国组织工程研究,2016,20(47):6997-7005.
[7]Albertsson AC,Eklund M.In vitro degradation of poly(trimethylene carbonate),poly(trimethylene carbonate-co-caprolactone),and poly(adipic anhydride).J Appl Polym Sci.1995;57(1):87-103.
[8]肖凯,郭颖志,顾忠伟.聚三亚甲基碳酸酯-丙交酯共聚物微球长效避孕体系的研究[J].中国计划生育学杂志,2008,16(4):219-222.
[9]杨立群,张金哲,张巍,等.聚三亚甲基碳酸酯的改性研究[J].高分子通报,2016,29(11):23-38.
[10]Bat E,Kothman BH,Higuera GA,et al.Ultraviolet light crosslinking of poly(trimethylene carbonate)for elastomeric tissue engineering scaffolds.Biomaterials.2010;31(33):8696-8705.
[11]韩雅如.完全生物可降解心血管支架用复合材料研究[D].上海:复旦大学,2011.
[12]Papenburg BJ,Schüller-Ravoo S,Bolhuis-Versteeg LA,et al.Designing porosity and topography of poly(1,3-trimethylene carbonate)scaffolds.Acta Biomaterialia.2009;5(9):3281-3294.
[13]Pego AP,Poot AA,Grijpma DW,et al.Copolymers of trimethylene carbonate andε-caprolactone for porous nerve guides:synthesis and properties.J Biomater Sci Polym Ed.2001;12(1):35-53.
[14]邵国喜,焦晶雪,刘钦毅,等.聚乳酸-聚三亚甲基碳酸酯/GDNF导管对大鼠脊髓损伤区GAP-43m RNA表达的影响[J].中国实验诊断学,2015,19(2):198-201.
[15]Fabre T,Schappacher M,Bareille R,et al.Study of a(trimethylenecarbonate-co-ε-caprolactone)polymer-Part 2:in vitro cytocompatibility analysis and in vivo ED1 cell response of a new nerve guide.Biomaterials.2001;22(22):2951-2958.
[16]裴香玲.恩诺沙星聚乳酸-聚三亚甲基碳酸酯的体内外降解和释药研究[D].兰州:甘肃农业大学,2007.
[17]吴雷刚,吴婧,侯瑞霞,等.他克莫司-聚三亚甲基碳酸酯药物洗脱支架涂层研究[J].功能材料,2011,42(11):1954-1957.
[18]Pego AP,Poot AA,Grijpma DW,et al.In vitro degradation of trimethylene carbonate based(co)polymers.Macromol Biosci.2002;2(9):411-419.
[19]周群华,杨立群,张巍,等.聚三亚甲基碳酸酯的生物相容性研究[J].高分子通报,2016,29(6):71-77.
[20]Yang LQ,Yang D,Guan YM,et al.Random copolymers based on trimethylene carbonate andε‐caprolactone for implant applications:Synthesis and properties.J Appl Polym Sci.2012;124(5):3714-3720.
[21]Yang L,Li J,Meng S,et al.The in vitro and in vivo degradation behavior of poly(trimethylene carbonate-co-ε-caprolactone)implants.Polymer.2014;55(20):5111-5124.
[22]Yang L,Li J,Jin Y,et al.In vitro enzymatic degradation of the cross-linked poly(ε-caprolactone)implants.Polym Degrad Stab.2015;112:10-19.
[23]Hou Z,Hu J,Li J,et al.The In Vitro Enzymatic Degradation of Cross-Linked Poly(trimethylene carbonate)Networks.Polymers.2017;9(11):605.
[24]Yang L,Li J,Zhang W,et al.The degradation of poly(trimethylene carbonate)implants:the role of molecular weight and enzymes.Polym Degrad Stab.2015;122:77-87.
[25]Chen B,Yin C,Cheng Y,et al.Using silk woven fabric as support for lipase immobilization:The effect of surface hydrophilicity/hydrophobicity on enzymatic activity and stability.Biomass Bioenergy.2012;39:59-66.
[26]Chen GJ,Kuo CH,Chen CI,et al.Effect of membranes with various hydrophobic/hydrophilic properties on lipase immobilized activity and stability.J Biosci Bioeng.2012;113(2):166-172.
[27]Sun H,Mei L,Song C,et al.The in vivo degradation,absorption and excretion of PCL-based implant.Biomaterials.2006;27(9):1735-1740.
[28]宋存先,王彭延.聚己内酯在体内的降解,吸收和排泄[J].生物医学工程学杂志,2000,17(1):25-28.
[29]於秋霞,朱光明,梁国正,等.聚ε-己内酯的合成、性能及应用进展[J].高分子材料科学与工程,2004,20(5):37-40.
[30]张金哲,周群华,杨立群,等.非生物降解型及生物降解型长效皮下埋植避孕剂的理论研究与应用进展[J].中国组织工程研究,2017,21(22):3595-3601.
[31]刘春海.可生物降解皮下埋植避孕剂Capronor的临床研究[J].中国计划生育学杂志,1996,4(3):187-189.
[32]Marten E,Muller RJ,Deckwer WD.Studies on the enzymatic hydrolysis of polyesters I.Low molecular mass model esters and aliphatic polyesters.Polym Degrad Stab.2003;80(3):485-501.
[2]Zhu KJ,Hendren RW,Jensen K,et al.Synthesis,properties,and degradation of poly(1,3-trimethylene carbonate).Macromolecules.1991;24(8):1736-1740.
[3]杨立群,周群华,张巍,等.聚三亚甲基碳酸酯的研究进展[J].高分子通报,2016,29(8):29-43.
[4]赵宁,张现军,李伟,等.左旋聚乳酸羟基磷灰石生物材料与牙周膜细胞的生物相容性[J].中国组织工程研究,2016,20(12):1732-1737.
[5]马晓妍,石淑先,夏宇正,等.聚乳酸及其共聚物的制备和降解性能[J].北京化工大学学报(自然科学版),2004,31(1):51-56.
[6]胡露,魏坤,邹芬.聚乳酸-羟基乙酸共聚物/硅酸钙三维多孔骨组织工程支架的构建与性能[J].中国组织工程研究,2016,20(47):6997-7005.
[7]Albertsson AC,Eklund M.In vitro degradation of poly(trimethylene carbonate),poly(trimethylene carbonate-co-caprolactone),and poly(adipic anhydride).J Appl Polym Sci.1995;57(1):87-103.
[8]肖凯,郭颖志,顾忠伟.聚三亚甲基碳酸酯-丙交酯共聚物微球长效避孕体系的研究[J].中国计划生育学杂志,2008,16(4):219-222.
[9]杨立群,张金哲,张巍,等.聚三亚甲基碳酸酯的改性研究[J].高分子通报,2016,29(11):23-38.
[10]Bat E,Kothman BH,Higuera GA,et al.Ultraviolet light crosslinking of poly(trimethylene carbonate)for elastomeric tissue engineering scaffolds.Biomaterials.2010;31(33):8696-8705.
[11]韩雅如.完全生物可降解心血管支架用复合材料研究[D].上海:复旦大学,2011.
[12]Papenburg BJ,Schüller-Ravoo S,Bolhuis-Versteeg LA,et al.Designing porosity and topography of poly(1,3-trimethylene carbonate)scaffolds.Acta Biomaterialia.2009;5(9):3281-3294.
[13]Pego AP,Poot AA,Grijpma DW,et al.Copolymers of trimethylene carbonate andε-caprolactone for porous nerve guides:synthesis and properties.J Biomater Sci Polym Ed.2001;12(1):35-53.
[14]邵国喜,焦晶雪,刘钦毅,等.聚乳酸-聚三亚甲基碳酸酯/GDNF导管对大鼠脊髓损伤区GAP-43m RNA表达的影响[J].中国实验诊断学,2015,19(2):198-201.
[15]Fabre T,Schappacher M,Bareille R,et al.Study of a(trimethylenecarbonate-co-ε-caprolactone)polymer-Part 2:in vitro cytocompatibility analysis and in vivo ED1 cell response of a new nerve guide.Biomaterials.2001;22(22):2951-2958.
[16]裴香玲.恩诺沙星聚乳酸-聚三亚甲基碳酸酯的体内外降解和释药研究[D].兰州:甘肃农业大学,2007.
[17]吴雷刚,吴婧,侯瑞霞,等.他克莫司-聚三亚甲基碳酸酯药物洗脱支架涂层研究[J].功能材料,2011,42(11):1954-1957.
[18]Pego AP,Poot AA,Grijpma DW,et al.In vitro degradation of trimethylene carbonate based(co)polymers.Macromol Biosci.2002;2(9):411-419.
[19]周群华,杨立群,张巍,等.聚三亚甲基碳酸酯的生物相容性研究[J].高分子通报,2016,29(6):71-77.
[20]Yang LQ,Yang D,Guan YM,et al.Random copolymers based on trimethylene carbonate andε‐caprolactone for implant applications:Synthesis and properties.J Appl Polym Sci.2012;124(5):3714-3720.
[21]Yang L,Li J,Meng S,et al.The in vitro and in vivo degradation behavior of poly(trimethylene carbonate-co-ε-caprolactone)implants.Polymer.2014;55(20):5111-5124.
[22]Yang L,Li J,Jin Y,et al.In vitro enzymatic degradation of the cross-linked poly(ε-caprolactone)implants.Polym Degrad Stab.2015;112:10-19.
[23]Hou Z,Hu J,Li J,et al.The In Vitro Enzymatic Degradation of Cross-Linked Poly(trimethylene carbonate)Networks.Polymers.2017;9(11):605.
[24]Yang L,Li J,Zhang W,et al.The degradation of poly(trimethylene carbonate)implants:the role of molecular weight and enzymes.Polym Degrad Stab.2015;122:77-87.
[25]Chen B,Yin C,Cheng Y,et al.Using silk woven fabric as support for lipase immobilization:The effect of surface hydrophilicity/hydrophobicity on enzymatic activity and stability.Biomass Bioenergy.2012;39:59-66.
[26]Chen GJ,Kuo CH,Chen CI,et al.Effect of membranes with various hydrophobic/hydrophilic properties on lipase immobilized activity and stability.J Biosci Bioeng.2012;113(2):166-172.
[27]Sun H,Mei L,Song C,et al.The in vivo degradation,absorption and excretion of PCL-based implant.Biomaterials.2006;27(9):1735-1740.
[28]宋存先,王彭延.聚己内酯在体内的降解,吸收和排泄[J].生物医学工程学杂志,2000,17(1):25-28.
[29]於秋霞,朱光明,梁国正,等.聚ε-己内酯的合成、性能及应用进展[J].高分子材料科学与工程,2004,20(5):37-40.
[30]张金哲,周群华,杨立群,等.非生物降解型及生物降解型长效皮下埋植避孕剂的理论研究与应用进展[J].中国组织工程研究,2017,21(22):3595-3601.
[31]刘春海.可生物降解皮下埋植避孕剂Capronor的临床研究[J].中国计划生育学杂志,1996,4(3):187-189.
[32]Marten E,Muller RJ,Deckwer WD.Studies on the enzymatic hydrolysis of polyesters I.Low molecular mass model esters and aliphatic polyesters.Polym Degrad Stab.2003;80(3):485-501.