A plasma-modified polyurethane-collagen-chondroitin sulfate composite material: preparation and performance

doi:10.3969/j.issn.2095-4344.2016.43.010

Abstract

Abstract:

BACKGROUND: Polyurethane has excellent mechanical properties and perfect biocompatibility. But its hydrophobicity is not conductive to the adhesion and growth of cells, which limits its use as the scaffold of the skin.
OBJECTIVE: To modify a polyurethane scaffold by plasma processing and crosslinking then to test its hydrophilic, mechanical properties and cytotoxicity in vitro.
METHODS: Polyurethane porous materials were prepared into membranous materials by eletrospinning method and modified by CO2 and NH3 plasma. Then the plasma-modified polyurethane-collagen- chondroitin sulfate composite materials were prepared by crosslinking with collagen type I and chondroitin sulfate at the same time, which was named Group A. Polyurethane porous materials which were electrospun and modified by plasma were soaked into a mixed solution of collagen and chondroitin sulfate without crosslinking agent. After being air-dryed, the materials were prepared, named Group B. Polyurethane porous materials which were electrospun and modified by plasma were named Group C. Electrospun polyurethane porous materials without other treatments were named Group D. Then the morphology, hydrophilicity, mechanical property and in vitro cytotoxicity of the four groups were compared.
RESULTS AND CONCLUSION: The plasma-modified polyurethane-collagen-chondroitin sulfate composite materials enjoy excellent fiber morphology, hydrophilicity and mechanical properties. It can also promote cell adhesion, proliferation and have no cytotoxicity.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Collagen, Chondroitin Sulfates, Tissue Engineering

CLC Number:

R318

Xuan Guang-shan, Mu Lan-lan, Sun Tong, Li Qing.

A plasma-modified polyurethane-collagen-chondroitin sulfate composite material: preparation and performance

[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(43): 6451-6457.

Figures/Tables 4

2.2 扫描电镜观察支架材料表面形貌由图2扫描电镜照片可见，该静电纺丝条件制备的聚氨酯支架形貌良好，纤维平滑直径分布均匀，平均直径为1 μm左右，纤维较粗，这是由于与电压、溶液的流速、距离相比，溶剂是影响静电纺丝形貌的一个比较重要因素。实验选用的是N，N-二甲基甲酰胺和四氢呋喃摩尔比为2∶1的混合溶剂，四氢呋喃的表面张力低且易挥发，使射流纤维外层快速挥发，限制了纤维的进一步拉伸，使纤维直径较粗；N，N-二甲基甲酰胺沸点高，不易挥发，可使纤维在电场中有效拉伸。因此，适当增加N，N-二甲基甲酰胺比例，可一定程度减小纤维直径，但当N，N-二甲基甲酰胺量过多时，由于溶剂挥发过慢易形成连珠。等离子体处理聚氨酯表面稍有毛糙，这是由于等离子体处理对聚氨酯纤维有一定的刻蚀作用，但整体表观影响不大。等离子体处理聚氨酯-胶原-硫酸软骨素复合材料纤维表面被胶原包裹填充，经化学交联复合材料整体出现皱缩现象，纤维有弯曲收缩现象；未经等离子体处理聚氨酯支架材料表面物理涂布胶原硫酸软骨素涂层，纤维直径稍有变粗。 2.3 亲水性检测结果生物材料的亲疏水性是限制其应用的主要因素。就皮肤支架而言，适宜亲水性的皮肤修复材料可使伤口保持湿润，有利于细胞黏附生长，促进伤口愈合。对各种支架材料接触角进行分析发现，未经等离子体处理的聚氨酯支架材料接触角为110°，材料不发生浸润；经等离子体处理后，材料的亲水性显著增加，水滴滴至材料后被迅速吸收，无法测出接触角，涂布和接枝胶原硫酸软骨素的材料也迅速将水滴吸收。聚氨酯有大量非极性基团如酯基，有较高的疏水性，离子体处理可在表面接枝亲水基团，虽然等离子体只在接触面反应，但由于静电纺丝纤维之间空隙大，材料比表面积大，增加了处理效率，使亲水性显著增加；胶原蛋白中含有大量-NH2、-OH，硫酸软骨素含有-OH、-COOH、-SO3H亲水基团，也有很好的亲水性。故将聚氨酯进行等离子体处理并与胶原蛋白及硫酸软骨素偶联处理，可得亲水性良好的支架材料。"

References

[1]Mansbridge J. Skin tissue engineering.J Biomat Sci Polym E.2008;19(8):955-968.
[2]董丽,王旭昇,马绍英,等.组织工程皮肤的构建及组织形态学观察[J].中国组织工程研究,2011,15(41):7631-7634.
[3]Cao H,Chen MM,Liu Y,et al.Fish collagen-based scaffold containing PLGA microspheres for controlled growth factor delivery in skin tissue engineering. Colloids Surf B Biointerfaces. 2015;136:1098-1106.
[4]Wang X,You C,Hu X,et al.The roles of knitted mesh-reinforced collagen–chitosan hybrid scaffold in the one-step repair of full-thickness skin defects in rats.Acta Biomaterialia.2013; 9(8):7822-7832.
[5]Lee CH,Chang SH,Chen WJ,et al.Augmentation of diabetic wound healing and enhancement of collagen content using nanofibrous glucophage-loaded collagen/PLGA scaffold membranes.J Colloid Interface Sci.2015;439:88-97.
[6]Sundaramurthi D,Vasanthan KS,Kuppan P,et al.Electrospun nanostructured chitosan–poly (vinyl alcohol) scaffolds: a biomimetic extracellular matrix as dermal substitute.Biomed Mater.2012; 7(4):045005.
[7]Gholipour-Kanani A,Bahrami SH, Samadi-Kochaksaraie A,et al.Effect of tissue-engineered chitosan-poly (vinyl alcohol) nanofibrous scaffolds on healing of burn wounds of rat skin.IET Nanobiotechnol.2012;6(4):129-135.
[8]Prasad T,Shabeena EA,Vinod D,et al.Characterization and in vitro evaluation of electrospun chitosan/polycaprolactone blend fibrous mat for skin tissue engineering. J Mater Sci Mater Med.2015;26(1):1-13.
[9]Zhang J,Li G,Gao S,et al.Monocyte chemoattractant protein-1 released from polycaprolactone/chitosan hybrid membrane to promote angiogenesis in vivo.J Bioact Compat Pol.2014:0883911514554146.
[10]Shalumon KT,Anulekha KH,Chennazhi KP,et al. Fabrication of chitosan/poly (caprolactone) nanofibrous scaffold for bone and skin tissue engineering.Int J Biol Macromol.2011;48(4):571-576.
[11]Por?ba R,Špírková M,Bro?ová L,et al.Aliphatic polycarbonate‐based polyurethane elastomers and nanocomposites. II. Mechanical, thermal, and gas transport properties.J Appl Polym Sci.2013;127(1): 329-341.
[12]Kojio K,Furukawa M,Shimada M,et al.Improvement of the Low-Temperature Property of Aliphatic Polycarbonate Glycols-Based Polyurethane Elastomers. SciAdv Mater.2015;7(5):934-939.
[13]Zhou H,Wang H,Tian X,et al.Effect of 3-Aminopropyltriethoxysilane on polycarbonate based waterborne polyurethane transparent coating.Prog Org Coat.2014;77(6):1073-1078.
[14]Por?ba R,Špírková M,Bro?ová L,et al.Aliphatic polycarbonate‐based polyurethane elastomers and nanocomposites. II.Mechanical, thermal, and gas transport properties.J Appl Polym Sci.2013;127(1): 329-341.
[15]Kucinska-Lipka J,Gubanska I,Janik H,et al.Fabrication of polyurethane and polyurethane based composite fibres by the electrospinning technique for soft tissue engineering of cardiovascular system.Mater Sci Eng C.2015;46:166-176.
[16]Sheikh FA,Macossay J,Cantu T,et al.Imaging, spectroscopy, mechanical, alignment and biocompatibility studies of electrospun medical grade polyurethane (Carbothane 3575A) nanofibers and composite nanofibers containing multiwalled carbon nanotubes.J Mech Behav Biomed Mater.2015;41: 189-198.
[17]Shi C,Yuan W,Khan M,et al.Hydrophilic PCU scaffolds prepared by grafting PEGMA and immobilizing gelatin to enhance cell adhesion and proliferation.Mater Sci Eng C.2015;50:201-209.
[18]Gauvin R,Berthod F.Collagen-based biomaterials for tissue engineering applications. Materials. 2010;3(3): 1863-1887.
[19]Huang R,Li W,Lv X,et al.Biomimetic LBL structured nanofibrous matrices assembled by chitosan/collagen for promoting wound healing.Biomaterials. 2015;53: 58-75.
[20]Wang W,Lin S,Xiao Y,et al.Acceleration of diabetic wound healing with chitosan-crosslinked collagen sponge containing recombinant human acidic fibroblast growth factor in healing-impaired STZ diabetic rats.Life Sci.2008;82(3):190-204.
[21]Mikami T,Kitagawa H.Biosynthesis and function of chondroitin sulfate.Biochimica et Biophysica Acta. 2013;1830(10):4719-4733.
[22]GB/T 16886.5-2003,医疗器械生物学评价第十二部分:样品制备与参照样品[S].中国标准出版社,2005.
[23]GB/T 16886.5-2003,医疗器械生物学评价第五部分:体外细胞毒性试验[S].中国标准出版社,2005.
[24]Yang J,Wan Y,Yang J,et al.Plasma-treated, collagen-anchored polylactone: Its cell affinity evaluation under shear or shear-free conditions.J Biomed Mater Res Part A.2003; 67(4):1139-1147.
[25]Desmet T,Morent R,Geyter ND,et al.Nonthermal plasma technology as a versatile strategy for polymeric biomaterials surface modification: a review.Biomacromolecules.2009;10(9):2351-2378.
[26]张燕搏,姜明,胡平,等.聚氨酯不同低温等离子体改性及生物相容性评价[J].中华实用诊断与治疗杂志, 2011, 25(10): 944-946.
[27]张燕搏,王强.低温等离子体技术在生物医用材料改性中的应用[J].中华实用诊断与治疗杂志, 2012, 26(1): 1-3.
[28]陈璐,杨庆,邵梅玲.PHBV/PCL静电纺纤维膜的低温等离子处理研究[J].合成纤维工业,2013,36(4): 30-33.
[29]Sugiyama K,Okamura A,Kawazoe N,et al.Coating of collagen on a poly (l-lactic acid) sponge surface for tissue engineering.Mater Sci Eng C. 2012;32(2): 290-295.
[30]Chen XX,Zhang XR,Li QS.Research advance of collagen protein in eye diseases.Guoji Yanke Zazhi (Int Eye Sci).2015;15(10):1745-1748.
[31]Müller R,Abke J,Schnell E,et al.Surface engineering of stainless steel materials by covalent collagen immobilization to improve implant biocompatibility. Biomaterials.2005; 26(34): 6962-6972.
[32]陈锐.静电纺制备胶原蛋白/聚氨酯心脏瓣膜组织工程支架材料的研究[D].东华大学,2010.