锂掺杂聚癸二酸甘油酯复合支架的制备及其性能
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摘要
目的:利用锂(Li)离子的特异性作用及聚癸二酸甘油酯(PGS)优良的性能制备PGS-Li复合支架,为其作为牙骨质组织工程支架的应用提供依据。方法:将支架分为2组,PGS交联过程中加入磷酸锂后制备的PGS-Li组和等比例PGS与甘油重结晶纯化后合成的PGS组。凝胶渗透色谱法测定2组支架的相对分子质量,傅里叶红外光谱仪分析2组支架的结构,扫描电子显微镜观察2组支架的表面形态并测定2组支架的孔隙率及孔径,X射线光电子能谱(XPS)仪及电感耦合等离子体发射光谱仪测定2组支架的Li离子含量,热重分析仪分析2组支架的热稳定性,接触角测量仪比较2组支架的亲水性,体外失重实验测定2组支架的降解率。将OCCM-30细胞分为PGS-Li组(加入PGS-Li支架浸提液)、PGS组(加入PGS支架浸提液)及空白对照组(加入空白培养液),MTT法检测不同时间(24、48和72h)各组细胞的增殖活性,钙黄素-AM染色观察细胞形态。结果:凝胶渗透色谱,PGS-Li组支架的相对分子质量略大于PGS组支架。傅里叶红外光谱检测,PGS-Li组支架出现了磷酸根特异性吸收峰。扫描电子显微镜下观察,2组支架均呈无规则的三维网状结构,孔径为20～160μm,PGS组支架的孔隙率为(53.92±2.18)%,PGS-Li组支架的孔隙率为(53.58±1.73)%,2组支架孔隙率比较差异无统计学意义(P>0.05)。XPS检测,PGS-Li组支架的XPS在54.9eV处出现了与Li 1s相符的峰值,电感耦合等离子体发射光谱仪测试PGS-Li组支架中Li离子含量为0.084%。热重分析,PGS-Li组支架的起始失重温度高于PGS组支架,而停止失重的温度则低于PGS组支架。接触角测量,2组支架均为亲水性材料,PGS组支架材料的接触角为78.26°±2.00°,PGS-Li组支架材料的接触角为69.78°±1.15°,2组比较差异有统计学意义(P<0.05)。体外降解实验,PGS-Li组支架的降解速率快于PGS组。PGS-Li组OCCM-30细胞增殖活性与PGS组和空白对照组比较差异无统计学意义(P>0.05)。钙黄绿素-AM染色后PGS组和PGA-Li组OCCM-30细胞均呈现绿色荧光,细胞形态无明显差异。结论:PGS-Li支架与PGS支架有着相似的组成和结构,且在亲水性及热稳定性方面有着更优异的性能,对成牙骨质细胞增殖无影响,在牙骨质组织工程中有着广阔的应用前景。
Objective:To prepare the lithium-doped poly-glycerol sebacate(PGS-Li)scaffold using the specific effects of lithium ions and the excellent performance of PGS,and to provide the basis for its application prospects in cementation tissue engineering scaffold.Methods:The scaffolds were divided into two groups.The PGS-Li scaffolds prepared by adding lithium phosphate during the PGS cross-linking process were used as PGS-Li group,and the PGS scaffolds synthesized by the equal-purification of sebacic acid and glycerol were used as PGS group.The molecular weights of the scaffolds in two groups were determined by gel permeation chromatography.The structures of the scaffolds in two groups were analyzed by fourier transform infrared spectroscope.The surface morphology and the porosities and the pore sizes of the scaffolds in two groups were observed by scanning electron microscope.X-ray photoelectron(XPS)spectroscope and inductively coupled plasma optical emission spectrometer were used to determine the Li ion contents in the scaffolds in two groups.Thermogravimetric analyzer was used to analyze the thermal stabilities of the scaffolds in two groups.Contact angle measuring instrument was used to compare the hydrophilicities of the scaffolds in two groups.In vitro weight loss test was used to determine the degradation rates of the scaffolds in two groups.The OCCM-30 cells were divided into experimental group(added with PGS-Li scaffold extract),PGS group(added with PGS scaffold extract)and blank control group(added with DMEM culture medium).MTT assay was used to detect the proliferation activities of cells in various groups at different time(24,48 and 72 h);the cell morphology was observed by calcein-AM staining.Results:The gel permeation chromatography results showed that the molecular weight of the PGS-Li scaffold was slightly larger than that of the PGS scaffold.The specific absorption peak of phosphate was detected in the fourier infrared spectrum of the PGS-Li scaffold.The scaffolds in two groups had irregular three-dimensional network structures under scanning electron microscope,and the pore size was 20-160μm,the porosity of PGS scaffold was(53.92±2.18)%,and the porosity of PGS-Li scaffold was(53.58±1.73)%,there was no statistical difference between two groups(P>0.05).The XPS results showed that a peak appeared at 54.9 eV in PGS-Li group,which coincided with the Li 1 s binding energy,while the inductively coupled plasma emission spectrometer results showed that the Li ion content in the PGS-Li scaffold was 0.084%.The thermogravimetric analysis results showed that PGS-Li scaffolds began to degrade at a higher temperature and ceased at a lower temperature compared with PGS scaffolds.The contact angle measurement results indicated that both the materials were hydrophilic materials;the contact angle of PGS scaffold meterial was 78.26°±2.00°,and the contact angle of the PGS-Li scaffold material was 69.78°±1.15°;there was statistical difference between two groups(P <0.05).The in vitro degradation experiments showed that the degradation rate of PGS-Li scaffolds was faster than that of PGS scaffolds.The proliferation activity of OCCM-30 cells in PGS-Li group had no significant difference compared with PGS group and blank control group(P>0.05).The calcein-AM staining results showed the green fluorescence in the OCCM-30 cells in PGS and PGS-Li groups,and there were no significant changes in the morphology of cementoblasts.Conclusion:PGS-Li scaffolds have similar composition and structure to PGS scaffolds,and have better performance in hydrophilicity and thermal stability.PGS-Li scaffolds have no effect on the proliferation of cementoblasts and have broad application prospects in cementum tissue engineering.
Objective:To prepare the lithium-doped poly-glycerol sebacate(PGS-Li)scaffold using the specific effects of lithium ions and the excellent performance of PGS,and to provide the basis for its application prospects in cementation tissue engineering scaffold.Methods:The scaffolds were divided into two groups.The PGS-Li scaffolds prepared by adding lithium phosphate during the PGS cross-linking process were used as PGS-Li group,and the PGS scaffolds synthesized by the equal-purification of sebacic acid and glycerol were used as PGS group.The molecular weights of the scaffolds in two groups were determined by gel permeation chromatography.The structures of the scaffolds in two groups were analyzed by fourier transform infrared spectroscope.The surface morphology and the porosities and the pore sizes of the scaffolds in two groups were observed by scanning electron microscope.X-ray photoelectron(XPS)spectroscope and inductively coupled plasma optical emission spectrometer were used to determine the Li ion contents in the scaffolds in two groups.Thermogravimetric analyzer was used to analyze the thermal stabilities of the scaffolds in two groups.Contact angle measuring instrument was used to compare the hydrophilicities of the scaffolds in two groups.In vitro weight loss test was used to determine the degradation rates of the scaffolds in two groups.The OCCM-30 cells were divided into experimental group(added with PGS-Li scaffold extract),PGS group(added with PGS scaffold extract)and blank control group(added with DMEM culture medium).MTT assay was used to detect the proliferation activities of cells in various groups at different time(24,48 and 72 h);the cell morphology was observed by calcein-AM staining.Results:The gel permeation chromatography results showed that the molecular weight of the PGS-Li scaffold was slightly larger than that of the PGS scaffold.The specific absorption peak of phosphate was detected in the fourier infrared spectrum of the PGS-Li scaffold.The scaffolds in two groups had irregular three-dimensional network structures under scanning electron microscope,and the pore size was 20-160μm,the porosity of PGS scaffold was(53.92±2.18)%,and the porosity of PGS-Li scaffold was(53.58±1.73)%,there was no statistical difference between two groups(P>0.05).The XPS results showed that a peak appeared at 54.9 eV in PGS-Li group,which coincided with the Li 1 s binding energy,while the inductively coupled plasma emission spectrometer results showed that the Li ion content in the PGS-Li scaffold was 0.084%.The thermogravimetric analysis results showed that PGS-Li scaffolds began to degrade at a higher temperature and ceased at a lower temperature compared with PGS scaffolds.The contact angle measurement results indicated that both the materials were hydrophilic materials;the contact angle of PGS scaffold meterial was 78.26°±2.00°,and the contact angle of the PGS-Li scaffold material was 69.78°±1.15°;there was statistical difference between two groups(P <0.05).The in vitro degradation experiments showed that the degradation rate of PGS-Li scaffolds was faster than that of PGS scaffolds.The proliferation activity of OCCM-30 cells in PGS-Li group had no significant difference compared with PGS group and blank control group(P>0.05).The calcein-AM staining results showed the green fluorescence in the OCCM-30 cells in PGS and PGS-Li groups,and there were no significant changes in the morphology of cementoblasts.Conclusion:PGS-Li scaffolds have similar composition and structure to PGS scaffolds,and have better performance in hydrophilicity and thermal stability.PGS-Li scaffolds have no effect on the proliferation of cementoblasts and have broad application prospects in cementum tissue engineering.
引文
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[17]BODAKHE S,VENA SMGARJGAL K,et al.Injectable photocrosslinkable nanocomposite based on poly(glycerol sebacate)fumarate and hydroxyapatite:development,biocompatibility and bone regeneration in a rat calvarial bone defect model[J].Nanomedicine,2013,8(11):1777-1795.
[18]LIANG H J,LI R Y,LIU G C,et al.Design of the porous orthopedic implants:research and application status[J].Chin Tissue Eng Res,2017,21(15):2410-2417.
[19]常丽,张玉兰,袁源,等.45S5型生物活性玻璃制备及体外性能表征[J].中国医学物理学杂志,2017,34(5):521-526.
[20]黄术,吴江怡,杨君君,等.天冬氨酸修饰纳米羟基磷灰石/聚乳酸复合物制备软骨组织工程支架及其生物性能研究[J].中国医学物理学杂志,2018,35(5):616-620.
[2]WANG Y,AMEER G A,Sheppard B J,et al.A tough biodegradable elastomer[J].Nat Biotechnol,2002,20(6):602.
[3]HOU L,ZHANG X,MIKAEL P E,,et al.Biodegradable and bioactive PCL-PGS core-shell fibers for tissue engineering[J].ACS Omega,2017,2(10):6321-6328.
[4]SHIRAZAKI P,VARSHOSAZ J,KHARAZI A Z.Electrospun gelatin/poly(glycerol sebacate)membrane with controlled release of antibiotics for wound dressing[J].Adv Biomed Res,2017,6(1):105.
[5]ZHAO X,WU H,GUO B,et al.Antibacterial anti-oxidant electroactive injectable hydrogel as self-healing wound dressing with hemostasis and adhesiveness for cutaneous wound healing[J].Biomaterials,2017,122:34-47.
[6]WANG Z,MA Y,WANG Y X,et al.Urethane-based lowtemperature curing,highly-customized and multifunctional poly(glycerol sebacate)-co-poly(ethylene glycol)copolymers[J].Acta Biomaterialia,2018,71:279-292.
[7]DING X,WU Y L,GAO J,et al.Tyramine functionalization of poly(glycerol sebacate)increases the elasticity of the polymer[J].J Mater Chem B,2017,5(30):6097-6109.
[8]MASOUDI R M,NOURI K S,GHASEMI-MOBARAKEH L,et al.Fabrication and characterization of two-layered nanofibrous membrane for guided bone and tissue regeneration application[J].Mater Sci Eng C Mater Biol Appl,2017,80:75-87.
[9]SOUZA M T,TANSAZ S,ZANOTTO E D,et al.Bioactive glass fiber-reinforced PGS matrix composites for cartilage regeneration[J].Materials,2017,10(1):83.
[10]WANG M,LEI D,LIU Z,et al.A poly(glycerol sebacate)based photo/thermo dual curable biodegradable and biocompatible polymer for biomedical applications[J].J Biomater Sci Polym Ed,2017,28(15):1728-1739.
[11]NADIM A,KHORASANI S N,KHARAZIHA M,et al.Design and characterization of dexamethasone-loaded poly(glycerol sebacate)-poly caprolactone/gelatin scaffold by coaxial electro spinning for soft tissue engineering[J].Mater Sci Eng C Mater Biol Appl,2017,78:47-58.
[12]LIM W H,LIU B,MAN S J,et al.Alveolar bone turnover and periodontal ligament width are controlled by Wnt[J].J Periodontol,2015,86(2):319-326.
[13]REN Y,HAN X,HO S P,et al.Removal of SOST or blocking its product sclerostin rescues defects in the periodontitis mouse model[J].FASEB J,2015,29(7):2702-2711.
[14]WANG Y,GAO S,JIANG H,et al.Lithium chloride attenuates root resorption during orthodontic tooth movement in rats[J].Exp Ther Med,2014,7(2):468.
[15]CHEN X,H U C,WANG G,et al.Nuclear factor-κBmodulates osteogenesis of periodontal ligament stem cells through competition withβ-catenin signaling in inflammatory microenvironments[J].Cell Death Dis,2013,4(2):e510.
[16]POMERANTSEVA I,KREBS N,HART A,et al.Degradation behavior of poly(glycerol sebacate)[J].J Biomed Mater Res A,2009,91(4):1038-1047.
[17]BODAKHE S,VENA SMGARJGAL K,et al.Injectable photocrosslinkable nanocomposite based on poly(glycerol sebacate)fumarate and hydroxyapatite:development,biocompatibility and bone regeneration in a rat calvarial bone defect model[J].Nanomedicine,2013,8(11):1777-1795.
[18]LIANG H J,LI R Y,LIU G C,et al.Design of the porous orthopedic implants:research and application status[J].Chin Tissue Eng Res,2017,21(15):2410-2417.
[19]常丽,张玉兰,袁源,等.45S5型生物活性玻璃制备及体外性能表征[J].中国医学物理学杂志,2017,34(5):521-526.
[20]黄术,吴江怡,杨君君,等.天冬氨酸修饰纳米羟基磷灰石/聚乳酸复合物制备软骨组织工程支架及其生物性能研究[J].中国医学物理学杂志,2018,35(5):616-620.