壳聚糖-胶原角膜修复材料的制备及生物相容性
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摘要
背景:将胶原和壳聚糖按一定比例制成复合膜后,能够降低壳聚糖的正电荷,改善壳聚糖的吸附力,促进细胞的黏附生长、迁移和增殖,增强壳聚糖的生物学性能,成为十分优良的生物材料。目的:制备壳聚糖-胶原复合膜,观察其与角膜基质层的组织相容性。方法:制作壳聚糖-胶原复合膜材料。将16只新西兰兔随机分为两组,实验组于右眼角膜基质袋内植入壳聚糖-胶原复合膜,对照组右眼角膜基质袋内植入壳聚糖膜。移植后进行裂隙灯显微镜、前节-光学相干断层成像及组织学观察。结果与结论:(1)裂隙灯显微镜:移植后第8周,实验组膜片中央出现降解浸润,皱褶屈曲不明显;对照组膜片全部浸润,皱褶屈曲明显。(2)前节-光学相干断层成像:移植后第6周,实验组的复合膜边界模糊,密度与正常角膜组织很接近,角膜形态恢复正常。(3)组织学观察:移植后第8周,实验组膜片表层少量降解,降解材料与角膜基质融合,膜片周围角膜基质有少量炎性细胞浸润;对照组膜片表层降解程度大于实验组,降解物质与角膜基质交织融合,膜片周围角膜基质有较多炎性细胞浸润。表明壳聚糖-胶原复合膜具有可降解性和良好的组织相容性。
BACKGROUND: The collagen and chitosan composite membrane made at a certain percentage can reduce the positive charge of chitosan, improve the adsorption force of chitosan, promote the cell adhesion and growth, migration and proliferation, and enhance the biological properties of chitosan, which has become a very excellent biological material. OBJECTIVE: To prepare chitosan-collagen composite membrane and observe its histocompatibility with corneal stroma layer. METHODS: Chitosan-collagen composite membrane was prepared. Totally 16 New Zealand rabbits were randomly divided into experimental and control groups. The chitosan-collagen composite membrane was implanted into right eye cornea stroma bag in the experimental group, and the chitosan membrane was implanted into right eye cornea stroma bag in the control group. The slit lamp microscope, anterior segment optical coherence tomography and histological observation were conducted after transplantation. RESULTS AND CONCLUSION: Slit lamp microscope observation results showed that after 8 weeks of transplantation, degradation infiltration was visible in the center of the diaphragm, crease buckling was not obvious in the experimental group; the diaphragm was infiltrated completely and crease buckling was notable in the control group. The anterior segment optical coherence tomography results showed that at the 6th week aftertransplantation, the composite membrane boundary was fuzzy, its density was very close to the normal corneal tissue, and the corneal shape recovered to normal in the experimental group. The histological findings showed that after 8 weeks of transplantation, in the experimental group, the membrane surface was degraded a little; biodegradable materials were fused with the corneal stroma; there were a small amount of inflammatory cells infiltrated on the corneal stroma around the iris. In the control group, the membrane degradation was greater than that of the experimental group; degradable substances intertwined and fused with corneal stroma; there were more inflammatory cells infiltrated on the corneal stroma around the iris. These results demonstrate that chitosan-collagen composite membrane has degradability and good histocompatibility.
BACKGROUND: The collagen and chitosan composite membrane made at a certain percentage can reduce the positive charge of chitosan, improve the adsorption force of chitosan, promote the cell adhesion and growth, migration and proliferation, and enhance the biological properties of chitosan, which has become a very excellent biological material. OBJECTIVE: To prepare chitosan-collagen composite membrane and observe its histocompatibility with corneal stroma layer. METHODS: Chitosan-collagen composite membrane was prepared. Totally 16 New Zealand rabbits were randomly divided into experimental and control groups. The chitosan-collagen composite membrane was implanted into right eye cornea stroma bag in the experimental group, and the chitosan membrane was implanted into right eye cornea stroma bag in the control group. The slit lamp microscope, anterior segment optical coherence tomography and histological observation were conducted after transplantation. RESULTS AND CONCLUSION: Slit lamp microscope observation results showed that after 8 weeks of transplantation, degradation infiltration was visible in the center of the diaphragm, crease buckling was not obvious in the experimental group; the diaphragm was infiltrated completely and crease buckling was notable in the control group. The anterior segment optical coherence tomography results showed that at the 6th week aftertransplantation, the composite membrane boundary was fuzzy, its density was very close to the normal corneal tissue, and the corneal shape recovered to normal in the experimental group. The histological findings showed that after 8 weeks of transplantation, in the experimental group, the membrane surface was degraded a little; biodegradable materials were fused with the corneal stroma; there were a small amount of inflammatory cells infiltrated on the corneal stroma around the iris. In the control group, the membrane degradation was greater than that of the experimental group; degradable substances intertwined and fused with corneal stroma; there were more inflammatory cells infiltrated on the corneal stroma around the iris. These results demonstrate that chitosan-collagen composite membrane has degradability and good histocompatibility.
引文
[1]黄飞华,孔繁智,朱婉萍,等.不同相对分子质量壳聚糖乳酸盐体外抑菌实验研究[J].中国海洋药物,2009,28(2):36-39.
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[12]Melton JT,Wilson AJ,Chapman-Sheath P,et al.Tru Fit CB bone plug:chondral repair,scaffold design,surgical technique and early experiences.Expert Rev Med Devices.2010;7(3):333-341.
[13]龙玉宇,任力,王家鸣,等.角膜修复用改性明胶交联膜的生物相容性评价[J].生物医学工程学杂志,2013,20(1):170-175.
[14]Badawi AA,El-Laithy HM,El Qidra RK,et al.Chitosan based nanocarriers for indomethacin ocular delivery.Arch Pharm Res.2008;31(8):1040-1049.
[15]Shah A,Brugnano J,Sun S,et al.The development of a tissue-engineered cornea:biomaterials and culture methods.Pediatr Res.2008;63(5):535-544.
[16]王雪,颜华.组织工程角膜上皮支架材料研究进展[J].眼科研究,2010,28(10):998-1002.
[17]Bu P,Vin AP,Sethupathi P,et al.Effects of activated omental cells on rat limbal corneal alkali injury.Exp Eye Res.2014;121:143-146.
[18]Murphy CM,O'Brien FJ.Understanding the effect of mean pore size on cell activity in collagen-glycosaminoglycan scaffolds.Cell Adh Migr.2010;4(3):377-381.
[19]侯江平,李国星,李玉莉,等.角膜内不同部位植入壳聚糖-胶原复合膜的生物相容性[J].中国组织工程研究与临床康复,2010,14(34):6319-6322.
[20]Hongyok T,Chae JJ,Shin YJ,et al.Effect of chitosan-Nacetylcysteine conjugate in a mouse model of botulinum toxin B-induced dry eye.Arch Ophthal mol.2009;127(4):525-532.
[21]郭丽,王迎军,任力,等.明胶固定化壳聚糖膜的细胞相容性评价[J].功能材料,2009,40(9):1525-1528.
[22]Qu C,Xiong Y,Mahmood A,et al.Treatment of traumatic brain injury in mice with bone marrow stromal cell-impregnated collagen scaffolds.J Neurosurg.2009;111(4):658-665.
[23]Notara M,Hernandez D,Mason C,et al.Characterization of the phenotype and functionality of corneal epithelial cells derived from mouse embryonic stem cells.Regen Med.2012;7(2):167-178.
[24]Hu K,Shi H,Zhu J,et al.Compressed collagen gel as the scaffold for skin engineering.Biomed Microdevices.2010;12(4):627-635.
[25]周晓伟,李国星,朱显丰,等.壳聚糖胶原复合膜在兔角膜基质层间的组织相容性[J].眼视光学杂志,2009,11(6):419-426.
[26]Wang F,Li Y,Shen Y,et al.The functions and applications of RGD in tumor therapy and tissue engineering.Int J Mol Sci.2013;14(7):13447-13462.
[27]李娜,周伟,孙恒.人工角膜的研究进展[J].医学综述,2009,15(4):575-579.
[2]李玉文,张梁栋.甲壳素及其衍生物在医药领域的应用[J].潍坊学院学报,2011,11(4):80-84.
[3]Pillai CKS,Paul W,Sharma CP.Chitin and chitosan polymers:chemistry,solubility and fiber formation.Prog Polym Sci.2009;34(7):641-678.
[4]Cheba BA.Chitin and chitosan:marine biopolymers with unique properties and versatile applications.Global J Biotech Bio-Chem.2011;(3):149-153.
[5]马群,刘万顺,韩宝芹,等.角膜组织工程支架壳聚糖基共混膜的制备及性能评价[J].功能材料,2010,41(9):1547-1551.
[6]Liang Y,Liu W,Han B,et al.Fabrication and characters of a corneal endothelial cells scaffold based on chitasan.J Mater Sci:Mater Med.2011;22(1):175-183.
[7]Keogh MB,O'Brien FJ,Daly JS.A novel collagen scaffold supports human osteogenesis--applications for bone tissue engineering.Cell Tissue Res.2010;340(1):169-177.
[8]王家鸣,任力,龙玉宇,等.壳聚糖-胶原复合膜渗透性能的研究[J].功能材料,2012,43(20):2747-2750.
[9]Wang L,Stegemann JP.Thermogelling chitosan and collagen composite hydrogels initiated with beta-glycerophosphate for bone tissue engineering.Biomaterials.2010;31(14):3976-3985.
[10]马群,刘万顺,韩宝芹,等.角膜组织工程支架壳聚糖基共混膜的制备及性能评价[J].功能材料,2010,41(9):1547-1551.
[11]Venugopal J,Prabhakaran MP,Zhang Y,et al.Biomimetic hydroxyapatite-containing composite nanofibrous substrates for bone tissue engineering.Philos Trans A Math Phys Eng Sci.2010;368(1917):2065-2081.
[12]Melton JT,Wilson AJ,Chapman-Sheath P,et al.Tru Fit CB bone plug:chondral repair,scaffold design,surgical technique and early experiences.Expert Rev Med Devices.2010;7(3):333-341.
[13]龙玉宇,任力,王家鸣,等.角膜修复用改性明胶交联膜的生物相容性评价[J].生物医学工程学杂志,2013,20(1):170-175.
[14]Badawi AA,El-Laithy HM,El Qidra RK,et al.Chitosan based nanocarriers for indomethacin ocular delivery.Arch Pharm Res.2008;31(8):1040-1049.
[15]Shah A,Brugnano J,Sun S,et al.The development of a tissue-engineered cornea:biomaterials and culture methods.Pediatr Res.2008;63(5):535-544.
[16]王雪,颜华.组织工程角膜上皮支架材料研究进展[J].眼科研究,2010,28(10):998-1002.
[17]Bu P,Vin AP,Sethupathi P,et al.Effects of activated omental cells on rat limbal corneal alkali injury.Exp Eye Res.2014;121:143-146.
[18]Murphy CM,O'Brien FJ.Understanding the effect of mean pore size on cell activity in collagen-glycosaminoglycan scaffolds.Cell Adh Migr.2010;4(3):377-381.
[19]侯江平,李国星,李玉莉,等.角膜内不同部位植入壳聚糖-胶原复合膜的生物相容性[J].中国组织工程研究与临床康复,2010,14(34):6319-6322.
[20]Hongyok T,Chae JJ,Shin YJ,et al.Effect of chitosan-Nacetylcysteine conjugate in a mouse model of botulinum toxin B-induced dry eye.Arch Ophthal mol.2009;127(4):525-532.
[21]郭丽,王迎军,任力,等.明胶固定化壳聚糖膜的细胞相容性评价[J].功能材料,2009,40(9):1525-1528.
[22]Qu C,Xiong Y,Mahmood A,et al.Treatment of traumatic brain injury in mice with bone marrow stromal cell-impregnated collagen scaffolds.J Neurosurg.2009;111(4):658-665.
[23]Notara M,Hernandez D,Mason C,et al.Characterization of the phenotype and functionality of corneal epithelial cells derived from mouse embryonic stem cells.Regen Med.2012;7(2):167-178.
[24]Hu K,Shi H,Zhu J,et al.Compressed collagen gel as the scaffold for skin engineering.Biomed Microdevices.2010;12(4):627-635.
[25]周晓伟,李国星,朱显丰,等.壳聚糖胶原复合膜在兔角膜基质层间的组织相容性[J].眼视光学杂志,2009,11(6):419-426.
[26]Wang F,Li Y,Shen Y,et al.The functions and applications of RGD in tumor therapy and tissue engineering.Int J Mol Sci.2013;14(7):13447-13462.
[27]李娜,周伟,孙恒.人工角膜的研究进展[J].医学综述,2009,15(4):575-579.