羟基磷灰石/羧甲基壳聚糖/明胶复合支架的制备及表征
|
摘要
采用原位共沉淀结合冷冻干燥工艺制备羟基磷灰石/羧甲基壳聚糖-明胶复合支架材料,然后利用X射线衍射仪(XRD)、傅立叶红外光谱仪(FTIR)检测其组成,扫描电子显微镜(SEM)观察微观结构,模拟体液浸泡法测试其降解性能,直接接触法观察其细胞毒性。结果表明:羟基磷灰石通过化学键与羧甲基壳聚糖和明胶结合;支架为相互连通的多孔结构,孔径尺寸介于100μm到300μm之间;纳米羟基磷灰石呈针状,均匀分布在支架中。与纯羟基磷灰石相比,复合支架降解性更好,更利于细胞黏附和增殖。实验结果同时表明支架材料的微观结构、降解性和细胞毒性可以通过改变明胶的含量进行调节,以满足不同修复部位的需求,是一种理想的骨组织工程用支架材料。
Hydroxyapatite(HA)/carboxymethyl chitosan(CMCS)-gelatin(Gel) composite scaffold was designed and prepared by an in situ coprecipitation combined with lyophilization method. X-ray diffraction(XRD), fourier transform infrared spectroscopy(FTIR), scanning electron microscopy(SEM), bioresorbability study and direct contact test were used to investigated the composite scaffold. The results confirm the formation of HA in the composite and the chemical bonds between HA and organic phase. SEM images show macroporous internal morphology with pore size ranging from 100 to 300 μm and needle-like HA particles uniformly distributed in the composite scaffold. Compared with pure HA, the composite scaffold has better resorbability and provide a healthier environment for cell attachment and proliferation. It was inferred that the morphology, degradation and optical density can be altered by tuning the gelatin ratio. In conclusion, the developed composite scaffold is a potential candidate for bone tissue engineering application.
Hydroxyapatite(HA)/carboxymethyl chitosan(CMCS)-gelatin(Gel) composite scaffold was designed and prepared by an in situ coprecipitation combined with lyophilization method. X-ray diffraction(XRD), fourier transform infrared spectroscopy(FTIR), scanning electron microscopy(SEM), bioresorbability study and direct contact test were used to investigated the composite scaffold. The results confirm the formation of HA in the composite and the chemical bonds between HA and organic phase. SEM images show macroporous internal morphology with pore size ranging from 100 to 300 μm and needle-like HA particles uniformly distributed in the composite scaffold. Compared with pure HA, the composite scaffold has better resorbability and provide a healthier environment for cell attachment and proliferation. It was inferred that the morphology, degradation and optical density can be altered by tuning the gelatin ratio. In conclusion, the developed composite scaffold is a potential candidate for bone tissue engineering application.
引文
[1]李波.壳聚糖/胶原骨组织工程支架材料的仿生矿化及生物相容性研究[A].Healy K,Guldberg R.Bone tissue engineering[C].J.musculoskeletal and neuronal interaction,2007,7(4):328-330.
[2]Mathew Peter,Nitya Ganesh,N.Selvamurugan,et al.Preparation and characterization of chitosangelatin/nanohydroxyapatite composite scaffolds for tissue engineering applications[J].Carbohydrate Polymers,2010,80:687-694.
[3]Su Wen,Ma Xiaomin,Sun Zhe,et al.RhBMP-2 and concomitant rapid material degradation synergistically promote bone repair and regeneration with collagen-hydroxyapatite nanocomposites[J].Journal of materials chemistry.B,Materials for biology and medicine,2018,6(26):4338-4350.
[4]黄明华,陈寰贝,陈庆华,等.纳米羟基磷灰石/胶原/魔芋葡甘聚糖多孔骨支架研究[J].人工晶体学报,2015,44(7):1961-1967.
[5]Jiancang Wang,Chaozong Liu.Biomimetic collagen/hydroxyapatite composite scaffolds:fabrication and characterizations[J].Journal of Bionic Engineering,2014,11(4):600-609.
[6]Daniel G.T.Strange,Michelle L.Oyen.Biomimetic bone-like composites fabricated through an automated altermate soaking process[J].Acta Biomateri alia,2011,7(10):3586-3594.
[7]张晓庆,秦建华,刘慧.透明质酸-明胶-纳米羟基磷灰石仿生骨支架的制备及成骨活性研究[J].高校化学工程学报,2017,31(2):387-393.
[8]Raucci Maria Grazia,Demitri Christian,Soriente,Alessandra,et al.Gelatin/nano-hydroxyapatite hydrogel scaffold prepared by sol-gel technology as filler to repair bone defects[J].Journal of Biomedical Materials Research Part A,2018,106(7):2007-2019.
[9]Fayyazbakhsh Fateme,Solati-Hashjin Mehran,Keshtkar Abbas,et al.Release behavior and signaling effect of vitamin D3 in layered double hydroxides-hydroxyapatite/gelatin bone tissue engineering scaffold:An in vitro evaluation[J].Colloids and Surfaces B-biointerface,2017,158:697-708.
[10]Khaled R.Mohamed,Hanan H.Beherei,Zenab M.El-Rashidy.In vitro study of nano-hydroxyapatite/chitosan-gelatin composites for bio-applications[J].Journal of Advanced Research,2014,5:201-208.
[11]Lee Chang-Min,Yang Seong-Won,Jung Sang-Chul,et al.Oxygen Plasma Treatment on 3D-printed chitosan/gelatin/hydroxyapatite scaffolds for bone tissue engineering[J].Journal of Nanoscience and Nanotechnology,2017,17(4):2747-2750.
[12]Zahra Babaei,Mohsen Jahanshahi,Sayed Mahmood Rabiee.The fabrication of nanocomposites viac alcium p hosphate f ormation o n g elatinchitosan network and the g elatin in fluence on the properties of bi phasic composites[J].Material Science and Engineering C,2013,33:370-375.
[13]Teimouri Abbas,Azadi Mohammad.Beta-chitin/gelatin/nanohydroxyapatite composite scaffold prepared through freeze-drying method for tissue engineering applications[J].Polymer Bulletin,2016,73(12):3513-3529.
[14]K.C.Kavya,R.Jayakumar,Shantikumar Nair,et al.Fabrication and characterization of chitosan/gelatin/nSiO2 composite scaffold for bone tissue engineering[J].International Journal of Biological Macromolecules,2013,59:255-263.
[15]Linbin Jiang,Yuan Lu,Xingyun Liu,et al.Layer-by-layer immobilization of quaternized carboxymethyl chitosan/organic rectorite and alginate onto nanofibrous mats and their antibacterial application[J].Carbohydrate Polymers,2015,121:428-435.
[16]Nadia A.Mohamed,Riham R.Mohamed,Rania S.Seoudi.Synthesis and characterization of some novel antimicrobial thiosemicarbazone O-carboxymethyl chitosan derivatives[J].International Journal of Biological Macromolecules,2014,63:163-169.
[2]Mathew Peter,Nitya Ganesh,N.Selvamurugan,et al.Preparation and characterization of chitosangelatin/nanohydroxyapatite composite scaffolds for tissue engineering applications[J].Carbohydrate Polymers,2010,80:687-694.
[3]Su Wen,Ma Xiaomin,Sun Zhe,et al.RhBMP-2 and concomitant rapid material degradation synergistically promote bone repair and regeneration with collagen-hydroxyapatite nanocomposites[J].Journal of materials chemistry.B,Materials for biology and medicine,2018,6(26):4338-4350.
[4]黄明华,陈寰贝,陈庆华,等.纳米羟基磷灰石/胶原/魔芋葡甘聚糖多孔骨支架研究[J].人工晶体学报,2015,44(7):1961-1967.
[5]Jiancang Wang,Chaozong Liu.Biomimetic collagen/hydroxyapatite composite scaffolds:fabrication and characterizations[J].Journal of Bionic Engineering,2014,11(4):600-609.
[6]Daniel G.T.Strange,Michelle L.Oyen.Biomimetic bone-like composites fabricated through an automated altermate soaking process[J].Acta Biomateri alia,2011,7(10):3586-3594.
[7]张晓庆,秦建华,刘慧.透明质酸-明胶-纳米羟基磷灰石仿生骨支架的制备及成骨活性研究[J].高校化学工程学报,2017,31(2):387-393.
[8]Raucci Maria Grazia,Demitri Christian,Soriente,Alessandra,et al.Gelatin/nano-hydroxyapatite hydrogel scaffold prepared by sol-gel technology as filler to repair bone defects[J].Journal of Biomedical Materials Research Part A,2018,106(7):2007-2019.
[9]Fayyazbakhsh Fateme,Solati-Hashjin Mehran,Keshtkar Abbas,et al.Release behavior and signaling effect of vitamin D3 in layered double hydroxides-hydroxyapatite/gelatin bone tissue engineering scaffold:An in vitro evaluation[J].Colloids and Surfaces B-biointerface,2017,158:697-708.
[10]Khaled R.Mohamed,Hanan H.Beherei,Zenab M.El-Rashidy.In vitro study of nano-hydroxyapatite/chitosan-gelatin composites for bio-applications[J].Journal of Advanced Research,2014,5:201-208.
[11]Lee Chang-Min,Yang Seong-Won,Jung Sang-Chul,et al.Oxygen Plasma Treatment on 3D-printed chitosan/gelatin/hydroxyapatite scaffolds for bone tissue engineering[J].Journal of Nanoscience and Nanotechnology,2017,17(4):2747-2750.
[12]Zahra Babaei,Mohsen Jahanshahi,Sayed Mahmood Rabiee.The fabrication of nanocomposites viac alcium p hosphate f ormation o n g elatinchitosan network and the g elatin in fluence on the properties of bi phasic composites[J].Material Science and Engineering C,2013,33:370-375.
[13]Teimouri Abbas,Azadi Mohammad.Beta-chitin/gelatin/nanohydroxyapatite composite scaffold prepared through freeze-drying method for tissue engineering applications[J].Polymer Bulletin,2016,73(12):3513-3529.
[14]K.C.Kavya,R.Jayakumar,Shantikumar Nair,et al.Fabrication and characterization of chitosan/gelatin/nSiO2 composite scaffold for bone tissue engineering[J].International Journal of Biological Macromolecules,2013,59:255-263.
[15]Linbin Jiang,Yuan Lu,Xingyun Liu,et al.Layer-by-layer immobilization of quaternized carboxymethyl chitosan/organic rectorite and alginate onto nanofibrous mats and their antibacterial application[J].Carbohydrate Polymers,2015,121:428-435.
[16]Nadia A.Mohamed,Riham R.Mohamed,Rania S.Seoudi.Synthesis and characterization of some novel antimicrobial thiosemicarbazone O-carboxymethyl chitosan derivatives[J].International Journal of Biological Macromolecules,2014,63:163-169.