CN114306750B - H-CNC multi-orientation coaxial artificial blood vessel and preparation method thereof - Google Patents
H-CNC multi-orientation coaxial artificial blood vessel and preparation method thereofDownload PDFInfo
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Abstract
Description
Translated fromChinese技术领域technical field
本发明属于生物制造技术领域，具体涉及到一种H-CNC多取向同轴人工血管及其制备方法。The invention belongs to the technical field of biological manufacturing, and particularly relates to an H-CNC multi-orientation coaxial artificial blood vessel and a preparation method thereof.
背景技术Background technique
目前，人工血管成为血管替代物得到快速发展。小口径人工血管（直径小于6 mm）因其生物相容性不佳、后期炎症、血栓和堵塞等不良反应，限制了小口径人工血管的应用。原位组织工程人工血管，是具有生物活性功能的非细胞血管移植物，可利用其取向性、生物相容性募集宿主细胞、诱导其原位再生出新生血管，成为治疗小口径血管疾病、促进损伤小口径血管原位再生最有潜力的治疗策略。At present, artificial blood vessels have been rapidly developed as blood vessel substitutes. Small-diameter artificial blood vessels (less than 6 mm in diameter) limit the application of small-diameter artificial blood vessels due to their poor biocompatibility, late-stage inflammation, thrombosis, and blockage. In situ tissue-engineered artificial blood vessels are acellular vascular grafts with bioactive functions. They can recruit host cells and induce them to regenerate new blood vessels in situ by using their orientation and biocompatibility. The most promising therapeutic strategy for in situ regeneration of injured small-caliber vessels.
然而现有原位组织工程人工血管制备技术中，沿血流方向取向排列的纳米纤维常采用平面取向薄膜卷曲分体制备工艺，会使人工血管内层产生卷曲沟痕，使血液产生紊流现象，血小板凝集，引发血栓，同时非一体成形，易产生分层现象；沿周向取向排列的纳米纤维常采用高速旋转制备工艺，因高速离心作用使纤维收集粘结松散，同时增加了危险系数；无序取向排列的纳米纤维常采用接收转轴接地，使其纳米纤维直接电纺在接收转轴上，随着电纺层厚的增加，纤维层的电荷累积使电纺纤维较难持续累积，影响纳米纤维层的制备；常采用的COL作为人工血管的生物材料可以增加生物相容性，然而COL的力学性能较差。However, in the existing in-situ tissue engineering artificial blood vessel preparation technology, the nanofibers oriented along the blood flow direction are often prepared by a plane-oriented film curling split preparation process, which will cause curling groove marks in the inner layer of the artificial blood vessel and cause turbulent blood flow. , platelet aggregation, causing thrombosis, and non-integrated forming, prone to delamination; nanofibers arranged in the circumferential direction are often prepared by high-speed rotation, due to the high-speed centrifugation, the fibers are loosely collected and bonded, and the risk factor is increased at the same time; The nanofibers with disordered orientation are often grounded by the receiving shaft, so that the nanofibers are directly electrospun on the receiving shaft. Preparation of fiber layer; COL is often used as a biomaterial for artificial blood vessels to increase biocompatibility, but COL has poor mechanical properties.
发明内容SUMMARY OF THE INVENTION
发明目的：本发明的目的在于提供一种可模拟血管三层组织结构，具有较好的生物力学性能，具有较强弹性模量和断裂韧性的H-CNC多取向同轴人工血管及其制备方法。Purpose of the invention: The purpose of the present invention is to provide a H-CNC multi-oriented coaxial artificial blood vessel that can simulate the three-layer tissue structure of blood vessels, has good biomechanical properties, and has strong elastic modulus and fracture toughness and its preparation method. .
技术方案：本发明的H-CNC多取向同轴人工血管，包括同轴且从内往外紧密结合在一起的水凝胶内层、MOCT和水凝胶外层，所述水凝胶内层和外层为H-CNC、COL和PDGF制成，所述MOCT从内往外依次由AOCNs层、COCNs层和DOCNs层，所述AOCNs层的纤维方向与接收转轴的轴向方向平行，采用平行电极工艺制备，COCNs层的纤维方向与AOCNs层纤维方向垂直，采用直角电极工艺制备，DOCNs层为无序取向纤维，采用平板电极工艺制备，所述同轴纳米纤维依次由纳米纤维芯层和纳米纤维壳层紧密结合而成，形成由纳米纤维壳层包裹纳米纤维芯层的复合结构，所述纳米纤维芯层由RAPA和第一PLCL复合材料制成，所述纳米纤维壳层由TPU和第二PLCL复合材料制成。Technical solution: The H-CNC multi-orientation coaxial artificial blood vessel of the present invention comprises an inner hydrogel layer, a MOCT and an outer hydrogel layer that are coaxially and tightly bonded together from the inside to the outside. The outer layer is made of H-CNC, COL and PDGF. The MOCT is composed of AOCNs layer, COCNs layer and DOCNs layer from the inside to the outside. The fiber direction of the AOCNs layer is parallel to the axial direction of the receiving shaft, and the parallel electrode process is adopted. Preparation, the fiber direction of the COCNs layer is perpendicular to the fiber direction of the AOCNs layer, and is prepared by a right-angle electrode process. The DOCNs layer is a disordered oriented fiber, which is prepared by a flat electrode process. The coaxial nanofibers are sequentially composed of nanofiber core layer and nanofiber shell. The layers are tightly combined to form a composite structure in which the nanofiber core layer is wrapped by the nanofiber shell layer, the nanofiber core layer is made of RAPA and the first PLCL composite material, and the nanofiber shell layer is made of TPU and the second PLCL Made of composite material.
其中，术语“H-CNC”是指：高长径比的纤维素纳米晶须，长径比为50~70；Wherein, the term "H-CNC" refers to: cellulose nanowhiskers with a high aspect ratio, with an aspect ratio of 50 to 70;
术语“H-MOCV”是指：H-CNC多取向同轴人工血管；The term "H-MOCV" refers to: H-CNC multi-orientation coaxial artificial blood vessel;
术语“MOCNs”是指：多取向同轴纳米纤维；The term "MOCNs" refers to: multi-oriented coaxial nanofibers;
术语“MOCT”是指：多取向同轴纳米纤维管；The term "MOCT" refers to: multi-oriented coaxial nanofibrous tubes;
术语“AOCNs”是指：轴向取向同轴纳米纤维；The term "AOCNs" refers to: axially oriented coaxial nanofibers;
术语“COCNs”是指：周向取向同轴纳米纤维；The term "COCNs" refers to: circumferentially oriented coaxial nanofibers;
术语“DOCNs”是指：无序取向同轴纳米纤维；The term "DOCNs" refers to: disorderly oriented coaxial nanofibers;
术语“PLCL”是指：聚乳酸-己内酯共聚物，其中PLA:PCL=50:50；The term "PLCL" refers to: polylactic acid-caprolactone copolymer, wherein PLA:PCL=50:50;
术语“HFIP”是指：六氟异丙醇The term "HFIP" refers to: Hexafluoroisopropanol
术语“RAPA”是指：雷帕霉素；The term "RAPA" refers to: rapamycin;
术语“TPU”是指：热塑性聚氨酯；The term "TPU" means: thermoplastic polyurethane;
术语“COL”是指：胶原蛋白；The term "COL" refers to: collagen;
术语“PDGF”是指：血小板衍生生长因子。The term "PDGF" refers to: Platelet-Derived Growth Factor.
进一步地，所述第一PLCL复合材料和第二PLCL复合材料由PLCL和基体材料共混制成，所述基体材料为HFIP，PLCL和HFIP的质量体积比为（5-15）g：100 mL。Further, the first PLCL composite material and the second PLCL composite material are made by blending PLCL and a matrix material, the matrix material is HFIP, and the mass-volume ratio of PLCL and HFIP is (5-15) g: 100 mL .
进一步地，所述RAPA和第一PLCL复合材料中的PLCL的组分质量配比为（1-100）：1000，所述TPU和第二PLCL复合材料中的PLCL的组分质量配比为（1-10）：10。Further, the component mass ratio of PLCL in the RAPA and the first PLCL composite material is (1-100): 1000, and the component mass ratio of the PLCL in the TPU and the second PLCL composite material is ( 1-10): 10.
进一步地，所述PDGF、H-CNC和COL的组分质量配比为（1-100）：（1-500）：1000，水凝胶复合材料由PDGF、H-CNC和凝胶基体材料共混制成，其中PDGF、H-CNC（长径比为50~70），凝胶基体材料为去离子水，COL与去离子水的质量体积比为（1-20）g：100 mL。Further, the mass ratio of the components of PDGF, H-CNC and COL is (1-100): (1-500): 1000, and the hydrogel composite material is composed of PDGF, H-CNC and gel matrix material. It is mixed into PDGF, H-CNC (length-diameter ratio is 50-70), the gel matrix material is deionized water, and the mass-volume ratio of COL to deionized water is (1-20) g: 100 mL.
本发明还公开了H-CNC多取向同轴人工血管的制备方法，包括如下步骤：The invention also discloses a preparation method of the H-CNC multi-orientation coaxial artificial blood vessel, comprising the following steps:
（1）将PLCL溶于HFIP中，磁力搅拌直至PLCL完全溶解，配制成质量体积比浓度为5-15 wt%的PLCL溶液；(1) Dissolve PLCL in HFIP, stir magnetically until PLCL is completely dissolved, and prepare a PLCL solution with a mass-volume ratio of 5-15 wt%;
（2）将TPU溶于PLCL溶液，配制成质量体积比浓度为5-15 wt%的同轴静电纺丝壳层溶液；(2) Dissolving TPU in PLCL solution to prepare a coaxial electrospinning shell solution with a mass-volume ratio of 5-15 wt%;
（3）将RAPA溶于PLCL溶液，配制成质量体积比浓度为5-15 wt%的同轴静电纺丝芯层溶液；(3) Dissolve RAPA in PLCL solution to prepare a coaxial electrospinning core layer solution with a mass-volume ratio concentration of 5-15 wt%;
（4）将接收转轴表面均匀涂覆普朗尼科F127水凝胶;(4) Evenly coat the surface of the receiving shaft with Pluronic F127 hydrogel;
（5）分别采用同轴静电纺丝芯层溶液和同轴静电纺丝壳层溶液，通过同轴静电纺丝工艺电纺在接收转轴上，得到三层结构的MOCT，从内到外依次为AOCNs层、COCNs层、DOCNs层；(5) Using the coaxial electrospinning core layer solution and the coaxial electrospinning shell solution respectively, and electrospinning on the receiving shaft through the coaxial electrospinning process, the MOCT with a three-layer structure is obtained. AOCNs layer, COCNs layer, DOCNs layer;
（6）将MOCT连同接收转轴一起取下竖直放于0~4 ℃的环境中，普朗尼科F127液化后MOCT可从接收转轴上轻松滑落，放于无水乙醇中清洗掉残余的普朗尼科F127水凝胶；(6) Remove the MOCT together with the receiving shaft and place it vertically in the environment of 0~4 ℃. After the liquefaction of Pluronic F127, the MOCT can easily slide off the receiving shaft, and put it in anhydrous ethanol to wash off the residual common Lannico F127 hydrogel;
（7）将清洗后的MOCT放于T型杆上并置于浸渍液中，浸渍结束将MOCT拿出直至MOCT上无液体滴落；(7) Put the cleaned MOCT on the T-bar and place it in the immersion solution, and after the immersion, take out the MOCT until there is no liquid dripping on the MOCT;
（8）将装在T型杆上且浸渍完成的MOCT置于交联剂溶液中交联，直至MOCT上的浸渍液完成交联，取出静置，直至无液体滴落，放于无水乙醇中清洗掉残余的交联剂；(8) Place the dipped MOCT mounted on the T-bar in the cross-linking agent solution for cross-linking until the dipping solution on the MOCT is cross-linked, take it out and stand until no liquid drips, and place it in anhydrous ethanol. Wash off the residual cross-linking agent;
（9）将MOCT从T型杆上取出，冷冻干燥后得到成形的H-MOCV产物。(9) The MOCT was taken out from the T-bar, and the formed H-MOCV product was obtained after freeze-drying.
进一步地，所述浸渍液的制备步骤为，Further, the preparation step of the dipping solution is,
（11）将COL溶于37 ℃的去离子水中，配制成质量体积比为1-20 wt%的COL溶液；(11) Dissolve COL in deionized water at 37 °C to prepare a COL solution with a mass-to-volume ratio of 1-20 wt%;
（12）将H-CNC溶于COL溶液，配制成质量体积比为1-20 wt%的H-CNC/COL溶液；(12) Dissolving H-CNC in COL solution to prepare H-CNC/COL solution with a mass-volume ratio of 1-20 wt%;
（13）将PDGF溶于H-CNC/COL溶液，配制成质量体积比为1-20 wt%的水凝胶溶液。(13) Dissolve PDGF in H-CNC/COL solution to prepare a hydrogel solution with a mass-to-volume ratio of 1-20 wt%.
进一步地，制备所述MOCT的AOCNs层时，接收转轴的转速为1~50 r/min；平行电极板的A板与B板的垂直距离为50~200 mm，平行电极板为尺寸相同的两个导电型圆形金属板，各自接地，与接收转轴之间为间隙配合，即平行电极板固定，接收转轴自转，平行电极板外径与接收转轴外径的比值为（1.1~3）：1，平行电极板中心与接收转轴的轴心重合；同轴喷头与接收转轴之间的水平距离为5~20 cm，纺丝时间为0.5~8h；Further, when preparing the AOCNs layer of the MOCT, the rotating speed of the receiving shaft is 1-50 r/min; the vertical distance between the A plate and the B plate of the parallel electrode plate is 50-200 mm, and the parallel electrode plate is two with the same size. Each conductive type circular metal plate is grounded, and the receiving shaft is a clearance fit, that is, the parallel electrode plate is fixed, the receiving shaft rotates, and the ratio of the outer diameter of the parallel electrode plate to the outer diameter of the receiving shaft is (1.1~3): 1 , the center of the parallel electrode plate coincides with the axis of the receiving shaft; the horizontal distance between the coaxial nozzle and the receiving shaft is 5~20 cm, and the spinning time is 0.5~8h;
制备所述MOCT的COCNs层时，When preparing the COCNs layer of the MOCT,
接收转轴的转速为50~1000 r/min，距离导电型直角架A面的距离为1~10 cm，距离B面的距离为1~10 cm，A面和B面垂直，直角架与接收转轴分别接地，纺丝时间为0.5~8 h；同轴喷头与接收转轴之间的水平距离为5~20 cm；The rotating speed of the receiving shaft is 50~1000 r/min, the distance from the conductive right angle frame A surface is 1~10 cm, and the distance from the B face is 1~10 cm, the A and B faces are perpendicular, and the right angle frame is perpendicular to the receiving shaft Grounded separately, the spinning time is 0.5~8 h; the horizontal distance between the coaxial nozzle and the receiving shaft is 5~20 cm;
制备所述MOCT的DOCNs层时，When preparing the DOCNs layer of the MOCT,
接收转轴的转速为50~200 r/min，距离导电型平板的垂直距离为1~10 cm，轴线通过平板中心线垂直平面，平板接地，纺丝时间0.5~8 h；同轴喷头与接收转轴之间的水平距离为5~20 cm。The rotating speed of the receiving shaft is 50~200 r/min, the vertical distance from the conductive plate is 1~10 cm, the axis passes through the vertical plane of the center line of the flat plate, the flat plate is grounded, and the spinning time is 0.5~8 h; the coaxial nozzle and the receiving shaft are The horizontal distance between them is 5~20 cm.
进一步地，制备所述MOCT时，通过两个微量泵同时给同轴喷头的芯层供料和壳层供料，芯层供料速度为10~100 μL/min，壳层供料速度为10~100 μL/min，纺丝电压为7-18kV，接收转轴采用可导电的金属材质，接收转轴的轴长为50~200 mm，外径为1~30 mm；同轴喷头的内径为0.1~0.5 mm，外径为0.5~2 mm。Further, when preparing the MOCT, the core layer and the shell layer of the coaxial nozzle are simultaneously fed by two micro-pumps, the core layer feeding speed is 10~100 μL/min, and the shell layer feeding speed is 10. ~100 μL/min, the spinning voltage is 7-18kV, the receiving shaft is made of conductive metal material, the shaft length of the receiving shaft is 50~200 mm, and the outer diameter is 1~30 mm; the inner diameter of the coaxial nozzle is 0.1~ 0.5 mm, the outer diameter is 0.5~2 mm.
进一步地，所述T型杆的小端面圆柱直径为0.5~30 mm，小端面高度为50~250 mm，大端面圆柱直径为1~40 mm，大端面圆柱高度为0.1~2 mm，MOCT套装在小端面圆柱上，MOCT浸在浸渍液内的一端与大端面圆柱接触。Further, the diameter of the small end face cylinder of the T-shaped rod is 0.5~30 mm, the height of the small end face is 50~250 mm, the diameter of the large end face cylinder is 1~40 mm, the height of the large end face cylinder is 0.1~2 mm, MOCT suit On the small-end cylinder, one end of the MOCT immersed in the immersion liquid is in contact with the large-end cylinder.
进一步地，所述浸渍时间为30~100 min，所述交联剂为京尼平的无水乙醇溶液，浓度为3~6 mM，交联时间为12~72 h，所述冷冻干燥时间为4~10 h。Further, the immersion time is 30-100 min, the cross-linking agent is an anhydrous ethanol solution of genipin, the concentration is 3-6 mM, the cross-linking time is 12-72 h, and the freeze-drying time is 4 to 10 hours.
进一步地，普朗尼科F127水凝胶的制备步骤为，将普朗尼科F127溶于0~4 ℃的去离子水中，配制成质量体积比为30-50 wt%的普朗尼科F127水凝胶。Further, the preparation step of Pluronic F127 hydrogel is as follows: Pluronic F127 is dissolved in deionized water at 0-4 °C to prepare Pluronic F127 with a mass-volume ratio of 30-50 wt%. Hydrogels.
有益效果：与现有技术相比，本发明具有如下显著优点：本发明制备的原位血管组织工程H-MOCV，由MOCT和生物水凝胶构成，MOCT的内层由AOCNs组成，纤维取向与血流方向相同，可诱导ECs轴向取向生长，形成仿生天然内皮层结构的取向形态，降低血流阻力，抑制血栓；中间层由COCNs组成，促进SMCs周向取向生长，诱导其向收缩表型转变，可改善径向受力，增强爆破压；外层由DOCNs组成，可增加孔隙率，促进细胞的粘附增殖和铺展；H-MOCV的生物水凝胶层通过载有PDGF/H-CNC/COL的浸渍交联工艺制成，其PDGF可通过分子化学作用促细胞的增殖，加速血管再生，H-CNC可增加水凝胶的弹性模量和断裂韧性90%以上，COL具有细胞粘附信号肽序列，可介导细胞特异性识别，促进细胞的粘附增殖，且无免疫源性，提高人工血管的生物学性能；采用普朗尼科F127涂覆工艺，可使MOCT从接收转轴上轻松取下，提高H-MOCV的制备成功率90%以上。Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the in-situ vascular tissue engineering H-MOCV prepared by the present invention is composed of MOCT and biological hydrogel, the inner layer of MOCT is composed of AOCNs, and the fiber orientation is the same as that of AOCNs. The same blood flow direction can induce the axial oriented growth of ECs, form the oriented morphology of the biomimetic natural endothelial layer structure, reduce the blood flow resistance, and inhibit the thrombus; the middle layer is composed of COCNs, which promotes the circumferential oriented growth of SMCs and induces its contraction phenotype. transformation, which can improve the radial force and enhance the burst pressure; the outer layer is composed of DOCNs, which can increase the porosity and promote the adhesion, proliferation and spreading of cells; the biohydrogel layer of H-MOCV is composed of PDGF/H-CNC loaded with PDGF/H-CNC. /COL is made by dipping and cross-linking process. Its PDGF can promote cell proliferation and accelerate angiogenesis through molecular chemistry. H-CNC can increase the elastic modulus and fracture toughness of hydrogels by more than 90%. COL has cell adhesion. The signal peptide sequence can mediate the specific recognition of cells, promote the adhesion and proliferation of cells, and has no immunogenicity to improve the biological properties of artificial blood vessels; the use of Pluronic F127 coating process can make MOCT from the receiving shaft It can be easily removed, which improves the preparation success rate of H-MOCV by more than 90%.
附图说明Description of drawings
图1 为本发明的制备流程示意图；Fig. 1 is the preparation flow schematic diagram of the present invention;
图2 为本发明中H-MOCV及MOCT的结构示意图；Fig. 2 is the structural representation of H-MOCV and MOCT in the present invention;
图3为本发明中同轴纳米纤维及MOCNs的微观结构示意图；3 is a schematic view of the microstructure of coaxial nanofibers and MOCNs in the present invention;
图中，1芯层微量泵，2壳层微量泵，3同轴喷头，4平行电极板，401平行电极A板，402平行电极B板，5接收转轴，7高压电源，8直角架，801直角架A面，802直角架B面，10导电型平板，11T型杆，111T型杆大端面圆柱，112T型杆小端面圆柱，12MOCT，13浸渍液，14冷冻干燥机，15-H-MOCV，16水凝胶内层，17MOCT，18水凝胶外层，19AOCNs层，20COCNs层，21DOCNs层。In the figure, 1 core micro pump, 2 shell micro pump, 3 coaxial nozzle, 4 parallel electrode plates, 401 parallel electrode A plate, 402 parallel electrode B plate, 5 receiving shaft, 7 high voltage power supply, 8 right angle frame, 801 Right angle frame A side, 802 right angle frame B side, 10 conductive type flat plate, 11T type rod, 111T type rod large end face cylinder, 112T type rod small end face cylinder, 12MOCT, 13 immersion liquid, 14 freeze dryer, 15-H-MOCV , 16 hydrogel inner layer, 17MOCT, 18 hydrogel outer layer, 19AOCNs layer, 20COCNs layer, 21DOCNs layer.
具体实施方式Detailed ways
下面结合附图对本发明的技术方案作进一步说明。The technical solutions of the present invention will be further described below with reference to the accompanying drawings.
术语“H-CNC”是指：高长径比的纤维素纳米晶须，长径比为50~70；The term "H-CNC" refers to: cellulose nanowhiskers with a high aspect ratio, with an aspect ratio of 50 to 70;
术语“H-MOCV”是指：H-CNC多取向同轴人工血管；The term "H-MOCV" refers to: H-CNC multi-orientation coaxial artificial blood vessel;
术语“MOCNs”是指：多取向同轴纳米纤维；The term "MOCNs" refers to: multi-oriented coaxial nanofibers;
术语“MOCT”是指：多取向同轴纳米纤维管；The term "MOCT" refers to: multi-oriented coaxial nanofibrous tubes;
术语“AOCNs”是指：轴向取向同轴纳米纤维；The term "AOCNs" refers to: axially oriented coaxial nanofibers;
术语“COCNs”是指：周向取向同轴纳米纤维；The term "COCNs" refers to: circumferentially oriented coaxial nanofibers;
术语“DOCNs”是指：无序取向同轴纳米纤维；The term "DOCNs" refers to: disorderly oriented coaxial nanofibers;
术语“PLCL”是指：聚乳酸-己内酯共聚物，其中PLA:PCL=50:50； The term "PLCL" refers to: polylactic acid-caprolactone copolymer, wherein PLA:PCL=50:50;
术语“HFIP”是指：六氟异丙醇The term "HFIP" refers to: Hexafluoroisopropanol
术语“RAPA”是指：雷帕霉素；The term "RAPA" refers to: rapamycin;
术语“TPU”是指：热塑性聚氨酯；The term "TPU" means: thermoplastic polyurethane;
术语“COL”是指：胶原蛋白；The term "COL" refers to: collagen;
术语“PDGF”是指：血小板衍生生长因子。The term "PDGF" refers to: Platelet-Derived Growth Factor.
实施例1Example 1
如图1所示，本发明提供一种H-CNC多取向同轴人工血管，包括同轴且从内往外紧密结合在一起的水凝胶内层、MOCT和水凝胶外层，所述水凝胶内层和外层为H-CNC、COL和PDGF制成，所述MOCT从内往外依次由AOCNs层、COCNs层和DOCNs层，所述AOCNs层的纤维方向与接收转轴的轴向方向平行，COCNs层的纤维方向与AOCNs层纤维方向垂直，DOCNs层为无序取向纤维，所述同轴纳米纤维依次由纳米纤维芯层和纳米纤维壳层紧密结合而成，形成由纳米纤维壳层包裹纳米纤维芯层的复合结构，所述纳米纤维芯层由RAPA和第一PLCL复合材料制成，所述纳米纤维壳层由TPU和第二PLCL复合材料制成。As shown in FIG. 1 , the present invention provides an H-CNC multi-orientation coaxial artificial blood vessel, comprising an inner hydrogel layer, a MOCT and an outer hydrogel layer that are coaxially and tightly bonded together from the inside to the outside. The inner and outer layers of the gel are made of H-CNC, COL and PDGF. The MOCT is composed of an AOCNs layer, a COCNs layer and a DOCNs layer in turn from the inside to the outside. The fiber direction of the AOCNs layer is parallel to the axial direction of the receiving shaft. , the fiber direction of the COCNs layer is perpendicular to the fiber direction of the AOCNs layer, and the DOCNs layer is disordered oriented fibers. The composite structure of the nanofiber core layer, the nanofiber core layer is made of RAPA and the first PLCL composite material, and the nanofiber shell layer is made of TPU and the second PLCL composite material.
进一步的，第一PLCL复合材料和第二PLCL复合材料由PLCL和基体材料共混制成，所述基体材料为HFIP，PLCL和HFIP的质量体积比为（5-15）g：100 mL；RAPA和第一PLCL复合材料中的PLCL的组分质量配比为（1-100）：1000,所述TPU和第二PLCL复合材料中的PLCL的组分质量配比为（1-10）：10。Further, the first PLCL composite material and the second PLCL composite material are made by blending PLCL and a matrix material, and the matrix material is HFIP, and the mass-to-volume ratio of PLCL and HFIP is (5-15) g: 100 mL; RAPA The component mass ratio with the PLCL in the first PLCL composite material is (1-100): 1000, and the component mass ratio of the PLCL in the TPU and the second PLCL composite material is (1-10): 10 .
进一步的，PDGF、H-CNC和COL的组分质量配比为（1-100）：（1-500）：1000，水凝胶复合材料由PDGF、H-CNC（长径比为50~70）和凝胶基体材料共混制成，凝胶基体材料为去离子水，COL与去离子水的质量体积比为（1-20）g：100 mL。Further, the mass ratio of components of PDGF, H-CNC and COL is (1-100): (1-500): 1000, and the hydrogel composite material is composed of PDGF, H-CNC (length-diameter ratio of 50~70). ) and gel matrix material, the gel matrix material is deionized water, and the mass-volume ratio of COL to deionized water is (1-20) g: 100 mL.
实施例2Example 2
参考图1，与实施例1的不同之处在于，本实施例提供一种H-CNC多取向同轴人工血管的制备方法，包括以下步骤，Referring to FIG. 1 , the difference from
将PLCL溶于HFIP中，磁力搅拌直至PLCL完全溶解，配制成质量体积比浓度为7 wt%的PLCL溶液50 g；Dissolve PLCL in HFIP, stir magnetically until PLCL is completely dissolved, and prepare 50 g of PLCL solution with a mass-volume ratio concentration of 7 wt%;
将TPU溶于PLCL溶液，TPU与PLCL的质量比为3:7，配制成质量体积比浓度为10 wt%的同轴静电纺丝壳层溶液；Dissolve TPU in PLCL solution, the mass ratio of TPU and PLCL is 3:7, and prepare the coaxial electrospinning shell solution with a mass-volume ratio concentration of 10 wt%;
将RAPA溶于PLCL溶液，RAPA与PLCL的质量比为1:70，配制成质量体积比浓度为7.1wt%的同轴静电纺丝芯层溶液；Dissolve RAPA in PLCL solution, the mass ratio of RAPA and PLCL is 1:70, and prepare a coaxial electrospinning core layer solution with a mass-volume ratio concentration of 7.1 wt%;
将COL溶于37 ℃的去离子水中，配制成质量体积比为10 wt%的COL溶液50 g；Dissolve COL in deionized water at 37 °C to prepare 50 g of COL solution with a mass-to-volume ratio of 10 wt%;
将PDGF和H-CNC溶于COL溶液，PDGF、H-CNC与COL的质量比为1:10：100，配制成质量体积比为11.1 wt%的水凝胶溶液；Dissolve PDGF and H-CNC in COL solution, the mass ratio of PDGF, H-CNC and COL is 1:10:100, and prepare a hydrogel solution with a mass-volume ratio of 11.1 wt%;
将接收转轴表面均匀涂覆35 wt%的普朗尼科F127水凝胶；The surface of the receiving shaft was evenly coated with 35 wt% Pluronic F127 hydrogel;
分别采用同轴静电纺丝芯层溶液和同轴静电纺丝壳层溶液，通过同轴静电纺丝工艺电纺在接收转轴上，得到三层结构的MOCT，从内到外依次为AOCNs层、COCNs层、DOCNs层，相应的纳米纤维结构如图3所示；The coaxial electrospinning core layer solution and the coaxial electrospinning shell layer solution were respectively used, and electrospun on the receiving shaft through the coaxial electrospinning process to obtain a MOCT with a three-layer structure. From the inside to the outside, the AOCNs layer, The COCNs layer, DOCNs layer, and the corresponding nanofiber structure are shown in Figure 3;
将MOCT连同接收转轴一起取下竖直放于0~4 ℃的环境中，普朗尼科F127液化后MOCT可从接收转轴上轻松滑落，放于无水乙醇中清洗掉残余的普朗尼科F127水凝胶；Remove the MOCT together with the receiving shaft and place it vertically at 0~4 ℃. After the Pluronic F127 is liquefied, the MOCT can easily slide off the receiving shaft, and then put it in anhydrous ethanol to wash off the residual Pluronic. F127 hydrogel;
将清洗后的MOCT放于T型杆上并置于浸渍液中，浸渍结束将MOCT拿出直至MOCT上无液体滴落；Put the cleaned MOCT on the T-bar and place it in the dipping solution, and after the dipping, take out the MOCT until there is no liquid dripping on the MOCT;
将装在T型杆上且浸渍完成的MOCT置于交联剂溶液中交联，直至MOCT上的浸渍液完成交联，取出静置，直至无液体滴落，放于无水乙醇中清洗掉残余的交联剂；Place the dipped MOCT mounted on the T-bar in the cross-linking agent solution for cross-linking, until the dipping solution on the MOCT is cross-linked, take it out and stand until no liquid drips, and wash it off in absolute ethanol. residual crosslinking agent;
将MOCT从T型杆上取出，放于冷冻干燥机中冷冻干燥，冷冻干燥时间为6h，冷冻干燥后得到内层水凝胶，中间层MOCT及外层水凝胶组成的H-MOCV。The MOCT was taken out from the T-bar and placed in a freeze-drying machine for freeze-drying. The freeze-drying time was 6 h. After freeze-drying, an H-MOCV composed of an inner layer of hydrogel, a middle layer of MOCT and an outer layer of hydrogel was obtained.
其中，在制备MOCT的AOCNs层时，接收转轴的转速为10 r/min，平行电极板的A板与B板的垂直距离为100 mm，外径为5 mm，分别接地，同轴喷头与接收转轴之间的水平距离为10 cm，纺丝时间为1 h；纺丝结束，制备COCNs层，调节接收转轴的转速，接收转轴的转速为500 r/min，距离导电型直角架A面的距离为5 cm，距离B面的距离为5 cm，直角架接地，纺丝时间为1 h；同轴喷头与接收转轴之间的水平距离为10 cm；纺丝结束，制备DOCNs层，接收转轴的转速为70 r/min，距离导电型平板的垂直距离为5 cm，平板接地，纺丝时间1 h；同轴喷头与接收转轴之间的水平距离为10 cm。Among them, when preparing the AOCNs layer of MOCT, the rotating speed of the receiving shaft is 10 r/min, the vertical distance between the A plate and the B plate of the parallel electrode plate is 100 mm, and the outer diameter is 5 mm, which are grounded respectively, and the coaxial nozzle is connected to the receiving shaft. The horizontal distance between the rotating shafts was 10 cm, and the spinning time was 1 h; after spinning, the COCNs layer was prepared, and the rotational speed of the receiving shaft was adjusted. It is 5 cm, the distance from the B surface is 5 cm, the right angle frame is grounded, and the spinning time is 1 h; the horizontal distance between the coaxial nozzle and the receiving shaft is 10 cm; after the spinning, the DOCNs layer is prepared, and the receiving shaft is The rotation speed was 70 r/min, the vertical distance from the conductive plate was 5 cm, the plate was grounded, and the spinning time was 1 h; the horizontal distance between the coaxial nozzle and the receiving shaft was 10 cm.
供料时，为了描述的方便，将给壳层供料的称为壳层微量泵，给芯层供料的称为芯层微量泵，本实施例中，在制备中间层时，芯层微量泵的供料速度为为50 μL/min，壳层供料速度为50 μL/min，纺丝电压为12 kV，接收转轴材质为不锈钢，接收转轴的轴长为100 mm，外径为4 mm；同轴喷头的内径为0.3 mm，外径为1 mm；制备水凝胶内层和外层时，浸渍时间为30 min，所述交联剂为京尼平的无水乙醇溶液，浓度为4 mM，交联时间为12 h，所用的T型杆的小端面圆柱直径为3 mm，小端面高度为150 mm，大端面圆柱直径为8 mm，大端面圆柱高度为1 mm。When feeding, for the convenience of description, the shell layer is called the shell micro pump, and the core layer is called the core micro pump. In this embodiment, when the intermediate layer is prepared, the core layer micro pump The feeding speed of the pump is 50 μL/min, the feeding speed of the shell layer is 50 μL/min, the spinning voltage is 12 kV, the receiving shaft is made of stainless steel, the shaft length of the receiving shaft is 100 mm, and the outer diameter is 4 mm. ; the inner diameter of the coaxial nozzle is 0.3 mm, and the outer diameter is 1 mm; when preparing the inner and outer layers of the hydrogel, the immersion time is 30 min, and the cross-linking agent is anhydrous ethanol solution of genipin with a concentration of 4 mM, the cross-linking time was 12 h, the diameter of the small end face cylinder of the T-bar used was 3 mm, the height of the small end face was 150 mm, the diameter of the large end face cylinder was 8 mm, and the height of the large end face cylinder was 1 mm.
在本实施例中，制备出的H-MOCV的结构如图2所示，各个纳米纤维的结构如图3所示，从图3中可以看出，同轴纳米纤维具有清晰的芯/壳结构，DOCNs具有高的孔隙率且纤维平滑，AOCNs、COCNs具有高的取向度，验证了该工艺的有效性。In this example, the structure of the prepared H-MOCV is shown in Figure 2, and the structure of each nanofiber is shown in Figure 3. It can be seen from Figure 3 that the coaxial nanofibers have a clear core/shell structure , DOCNs have high porosity and smooth fibers, and AOCNs and COCNs have high degree of orientation, verifying the effectiveness of this process.
实施例3Example 3
将PLCL溶于HFIP中，磁力搅拌直至PLCL完全溶解，配制成质量体积比浓度为7 wt%的PLCL溶液50 g；Dissolve PLCL in HFIP, stir magnetically until PLCL is completely dissolved, and prepare 50 g of PLCL solution with a mass-volume ratio concentration of 7 wt%;
将TPU溶于PLCL溶液，TPU与PLCL的质量比为4:7，配制成质量体积比浓度为11 wt%的同轴静电纺丝壳层溶液；Dissolve TPU in PLCL solution, the mass ratio of TPU and PLCL is 4:7, and prepare the coaxial electrospinning shell solution with a mass-volume ratio concentration of 11 wt%;
将RAPA溶于PLCL溶液，RAPA与PLCL的质量比为2:70，配制成质量体积比浓度为7.2wt%的同轴静电纺丝芯层溶液；Dissolve RAPA in PLCL solution, the mass ratio of RAPA and PLCL is 2:70, and prepare a coaxial electrospinning core layer solution with a mass-volume ratio concentration of 7.2wt%;
将COL溶于37 ℃的去离子水中，配制成质量体积比为10 wt%的COL溶液50 g；Dissolve COL in deionized water at 37 °C to prepare 50 g of COL solution with a mass-to-volume ratio of 10 wt%;
将PDGF和H-CNC溶于COL溶液，PDGF、H-CNC与COL的质量比为1:20：100，配制成质量体积比为12.1 wt%的水凝胶溶液；Dissolve PDGF and H-CNC in COL solution, the mass ratio of PDGF, H-CNC and COL is 1:20:100, and prepare a hydrogel solution with a mass-volume ratio of 12.1 wt%;
将接收转轴表面均匀涂覆35 wt%的普朗尼科F127水凝胶；The surface of the receiving shaft was evenly coated with 35 wt% Pluronic F127 hydrogel;
分别采用同轴静电纺丝芯层溶液和同轴静电纺丝壳层溶液，通过同轴静电纺丝工艺电纺在接收转轴上，得到三层结构的MOCT，从内到外依次为AOCNs层、COCNs层、DOCNs层；The coaxial electrospinning core layer solution and the coaxial electrospinning shell layer solution were respectively used, and electrospun on the receiving shaft through the coaxial electrospinning process to obtain a MOCT with a three-layer structure. From the inside to the outside, the AOCNs layer, COCNs layer, DOCNs layer;
将MOCT连同接收转轴一起取下竖直放于0~4 ℃的环境中，普朗尼科F127液化后MOCT可从接收转轴上轻松滑落，放于无水乙醇中清洗掉残余的普朗尼科F127水凝胶；Remove the MOCT together with the receiving shaft and place it vertically at 0~4 ℃. After the Pluronic F127 is liquefied, the MOCT can easily slide off the receiving shaft, and then put it in anhydrous ethanol to wash off the residual Pluronic. F127 hydrogel;
将清洗后的MOCT放于T型杆上并置于浸渍液中，浸渍结束将MOCT拿出直至MOCT上无液体滴落；Put the cleaned MOCT on the T-bar and place it in the dipping solution, and after the dipping, take out the MOCT until there is no liquid dripping on the MOCT;
将装在T型杆上且浸渍完成的MOCT置于交联剂溶液中交联，直至MOCT上的浸渍液完成交联，取出静置，直至无液体滴落，放于无水乙醇中清洗掉残余的交联剂；Place the dipped MOCT mounted on the T-bar in the cross-linking agent solution for cross-linking, until the dipping solution on the MOCT is cross-linked, take it out and stand until no liquid drips, and wash it off in absolute ethanol. residual crosslinking agent;
将MOCT从T型杆上取出，放于冷冻干燥机中冷冻干燥，冷冻干燥时间为8h，冷冻干燥后得到内层水凝胶，中间层MOCT及外层水凝胶组成的H-MOCV。The MOCT was taken out from the T-bar and placed in a freeze-drying machine for freeze-drying. The freeze-drying time was 8 h. After freeze-drying, an H-MOCV composed of an inner layer of hydrogel, a middle layer of MOCT and an outer layer of hydrogel was obtained.
其中，在制备MOCT的AOCNs层时，接收转轴的转速为20 r/min，平行电极板的A板与B板的垂直距离为100 mm，外径为7 mm，分别接地，同轴喷头与接收转轴之间的水平距离为10 cm，纺丝时间为1.5 h；纺丝结束，制备COCNs层，调节接收转轴的转速，接收转轴的转速为700 r/min，距离导电型直角架A面的距离为5 cm，距离B面的距离为5 cm，直角架接地，纺丝时间为1.5 h；同轴喷头与接收转轴之间的水平距离为10 cm；纺丝结束，制备DOCNs层，接收转轴的转速为80 r/min，距离导电型平板的垂直距离为5 cm，平板接地，纺丝时间1.5 h；同轴喷头与接收转轴之间的水平距离为10 cm。Among them, when preparing the AOCNs layer of MOCT, the rotating speed of the receiving shaft is 20 r/min, the vertical distance between the A plate and the B plate of the parallel electrode plate is 100 mm, and the outer diameter is 7 mm, which are grounded respectively, and the coaxial nozzle is connected to the receiving shaft. The horizontal distance between the rotating shafts was 10 cm, and the spinning time was 1.5 h; after spinning, the COCNs layer was prepared, and the rotational speed of the receiving shaft was adjusted. is 5 cm, the distance from the B surface is 5 cm, the right angle frame is grounded, and the spinning time is 1.5 h; the horizontal distance between the coaxial nozzle and the receiving shaft is 10 cm; after the spinning, the DOCNs layer is prepared, and the receiving shaft is The rotation speed was 80 r/min, the vertical distance from the conductive plate was 5 cm, the plate was grounded, and the spinning time was 1.5 h; the horizontal distance between the coaxial nozzle and the receiving shaft was 10 cm.
供料时，为了描述的方便，将给壳层供料的称为壳层微量泵，给芯层供料的称为芯层微量泵，本实施例中，在制备中间层时，芯层微量泵的供料速度为60 μL/min，壳层供料速度为60 μL/min，纺丝电压为12 kV，接收转轴材质为不锈钢，接收转轴的轴长为100 mm，外径为5 mm；同轴喷头的内径为0.3 mm，外径为1 mm；制备水凝胶内层和外层时，浸渍时间为45 min，所述交联剂为京尼平的无水乙醇溶液，浓度为4 mM，交联时间为24 h，所用的T型杆的小端面圆柱直径为4 mm，小端面高度为150 mm，大端面圆柱直径为8 mm，大端面圆柱高度为1 mm。When feeding, for the convenience of description, the shell layer is called the shell micro pump, and the core layer is called the core micro pump. In this embodiment, when the intermediate layer is prepared, the core layer micro pump The feeding speed of the pump is 60 μL/min, the feeding speed of the shell layer is 60 μL/min, the spinning voltage is 12 kV, the receiving shaft is made of stainless steel, the shaft length of the receiving shaft is 100 mm, and the outer diameter is 5 mm; The inner diameter of the coaxial nozzle is 0.3 mm, and the outer diameter is 1 mm; when preparing the inner and outer layers of the hydrogel, the immersion time is 45 min, and the cross-linking agent is an anhydrous ethanol solution of genipin with a concentration of 4 mM, the cross-linking time was 24 h, the diameter of the small end face cylinder of the T-bar used was 4 mm, the height of the small end face was 150 mm, the diameter of the large end face cylinder was 8 mm, and the height of the large end face cylinder was 1 mm.
在本实施例中，与实施例1和实施例2相比：增加了纤维的芯/壳层浓度，同轴纳米纤维直径增大；增加了RAPA浓度，抗血栓性增强；增加了TPU的浓度，MOCT弹性增加；增加了AOCNs纺丝时间，AOCNs层的层厚增加；增加了COCNs层的转速、纺丝时间，COCNs层的取向度增加，层厚增加；增加了DOCNs层的转速、纺丝时间，DOCNs层的层厚增加；增加了H-CNC的浓度，浸渍时间、交联时间，水凝胶层的弹性模量和断裂韧性增加；增加了接收转轴的直径，H-MOCV的内径增加；综上，其生物力学性能将会增加。In this example, compared with Example 1 and Example 2: the core/shell concentration of the fiber was increased, and the diameter of the coaxial nanofibers was increased; the concentration of RAPA was increased, and the antithrombotic property was enhanced; the concentration of TPU was increased , MOCT elasticity increases; increasing the spinning time of AOCNs, the layer thickness of the AOCNs layer increases; increasing the rotational speed and spinning time of the COCNs layer, the orientation degree of the COCNs layer increases, and the layer thickness increases; increasing the rotational speed and spinning of the DOCNs layer time, the layer thickness of the DOCNs layer increased; increasing the concentration of H-CNC, immersion time, cross-linking time, the elastic modulus and fracture toughness of the hydrogel layer increased; increasing the diameter of the receiving shaft, the inner diameter of H-MOCV increased In conclusion, its biomechanical properties will increase.
实施例4Example 4
将PLCL溶于HFIP中，磁力搅拌直至PLCL完全溶解，配制成质量体积比浓度为7 wt%的PLCL溶液50 g；Dissolve PLCL in HFIP, stir magnetically until PLCL is completely dissolved, and prepare 50 g of PLCL solution with a mass-volume ratio concentration of 7 wt%;
将TPU溶于PLCL溶液，TPU与PLCL的质量比为5:7，配制成质量体积比浓度为12 wt%的同轴静电纺丝壳层溶液；Dissolve TPU in PLCL solution, the mass ratio of TPU and PLCL is 5:7, and prepare the coaxial electrospinning shell solution with a mass-volume ratio concentration of 12 wt%;
将RAPA溶于PLCL溶液，RAPA与PLCL的质量比为2:70，配制成质量体积比浓度为7.2wt%的同轴静电纺丝芯层溶液；Dissolve RAPA in PLCL solution, the mass ratio of RAPA and PLCL is 2:70, and prepare a coaxial electrospinning core layer solution with a mass-volume ratio concentration of 7.2wt%;
将COL溶于37 ℃的去离子水中，配制成质量体积比为10 wt%的COL溶液50 g；Dissolve COL in deionized water at 37 °C to prepare 50 g of COL solution with a mass-to-volume ratio of 10 wt%;
将PDGF和H-CNC溶于COL溶液，PDGF、H-CNC与COL的质量比为1:30：100，配制成质量体积比为13.1 wt%的水凝胶溶液；Dissolve PDGF and H-CNC in COL solution, the mass ratio of PDGF, H-CNC and COL is 1:30:100, and prepare a hydrogel solution with a mass-volume ratio of 13.1 wt%;
将接收转轴表面均匀涂覆35 wt%的普朗尼科F127水凝胶；The surface of the receiving shaft was evenly coated with 35 wt% Pluronic F127 hydrogel;
分别采用同轴静电纺丝芯层溶液和同轴静电纺丝壳层溶液，通过同轴静电纺丝工艺电纺在接收转轴上，得到三层结构的MOCT，从内到外依次为AOCNs层、COCNs层、DOCNs层；The coaxial electrospinning core layer solution and the coaxial electrospinning shell layer solution were respectively used, and electrospun on the receiving shaft through the coaxial electrospinning process to obtain a MOCT with a three-layer structure. From the inside to the outside, the AOCNs layer, COCNs layer, DOCNs layer;
将MOCT连同接收转轴一起取下竖直放于0~4 ℃的环境中，普朗尼科F127液化后MOCT可从接收转轴上轻松滑落，放于无水乙醇中清洗掉残余的普朗尼科F127水凝胶；Remove the MOCT together with the receiving shaft and place it vertically at 0~4 ℃. After the Pluronic F127 is liquefied, the MOCT can easily slide off the receiving shaft, and then put it in anhydrous ethanol to wash off the residual Pluronic. F127 hydrogel;
将清洗后的MOCT放于T型杆上并置于浸渍液中，浸渍结束将MOCT拿出直至MOCT上无液体滴落；Put the cleaned MOCT on the T-bar and place it in the dipping solution, and after the dipping, take out the MOCT until there is no liquid dripping on the MOCT;
将装在T型杆上且浸渍完成的MOCT置于交联剂溶液中交联，直至MOCT上的浸渍液完成交联，取出静置，直至无液体滴落，放于无水乙醇中清洗掉残余的交联剂；Place the dipped MOCT mounted on the T-bar in the cross-linking agent solution for cross-linking, until the dipping solution on the MOCT is cross-linked, take it out and stand until no liquid drips, and wash it off in absolute ethanol. residual crosslinking agent;
将MOCT从T型杆上取出，放于冷冻干燥机中冷冻干燥，冷冻干燥时间为10 h，冷冻干燥后得到内层水凝胶，中间层MOCT及外层水凝胶组成的H-MOCV。The MOCT was taken out from the T-bar and placed in a freeze-drying machine for freeze-drying. The freeze-drying time was 10 h. After freeze-drying, the H-MOCV composed of the inner layer of hydrogel, the middle layer of MOCT and the outer layer of hydrogel was obtained.
其中，在制备MOCT的AOCNs层时，接收转轴的转速为20 r/min，平行电极板的A板与B板的垂直距离为100 mm，外径为7 mm，分别接地，同轴喷头与接收转轴之间的水平距离为15 cm，纺丝时间为1.5 h；纺丝结束，制备COCNs层，调节接收转轴的转速，接收转轴的转速为800 r/min，距离导电型直角架A面的距离为5 cm，距离B面的距离为5 cm，直角架接地，纺丝时间为1.5 h；同轴喷头与接收转轴之间的水平距离为15 cm；纺丝结束，制备DOCNs层，接收转轴的转速为80 r/min，距离导电型平板的垂直距离为5 cm，平板接地，纺丝时间1.5 h；同轴喷头与接收转轴之间的水平距离为15 cm。Among them, when preparing the AOCNs layer of MOCT, the rotating speed of the receiving shaft is 20 r/min, the vertical distance between the A plate and the B plate of the parallel electrode plate is 100 mm, and the outer diameter is 7 mm, which are grounded respectively, and the coaxial nozzle is connected to the receiving shaft. The horizontal distance between the rotating shafts was 15 cm, and the spinning time was 1.5 h; after spinning, the COCNs layer was prepared, and the rotational speed of the receiving shaft was adjusted. is 5 cm, the distance from the B surface is 5 cm, the right angle frame is grounded, and the spinning time is 1.5 h; the horizontal distance between the coaxial nozzle and the receiving shaft is 15 cm; after spinning, the DOCNs layer is prepared, and the receiving shaft is The rotating speed was 80 r/min, the vertical distance from the conductive plate was 5 cm, the plate was grounded, and the spinning time was 1.5 h; the horizontal distance between the coaxial nozzle and the receiving shaft was 15 cm.
供料时，为了描述的方便，将给壳层供料的称为壳层微量泵，给芯层供料的称为芯层微量泵，本实施例中，在制备中间层时，芯层微量泵的供料速度为65 μL/min，壳层供料速度为65 μL/min，纺丝电压为14 kV，接收转轴材质为不锈钢，接收转轴的轴长为100 mm，外径为5 mm；同轴喷头的内径为0.3 mm，外径为1 mm；制备水凝胶内层和外层时，浸渍时间为45 min，所述交联剂为京尼平的无水乙醇溶液，浓度为4 mM，交联时间为24 h，所用的T型杆的小端面圆柱直径为4 mm，小端面高度为150 mm，大端面圆柱直径为8 mm，大端面圆柱高度为1 mm。When feeding, for the convenience of description, the shell layer is called the shell micro pump, and the core layer is called the core micro pump. In this embodiment, when the intermediate layer is prepared, the core layer micro pump The feeding speed of the pump is 65 μL/min, the feeding speed of the shell layer is 65 μL/min, the spinning voltage is 14 kV, the receiving shaft is made of stainless steel, the shaft length of the receiving shaft is 100 mm, and the outer diameter is 5 mm; The inner diameter of the coaxial nozzle is 0.3 mm, and the outer diameter is 1 mm; when preparing the inner and outer layers of the hydrogel, the immersion time is 45 min, and the cross-linking agent is an anhydrous ethanol solution of genipin with a concentration of 4 mM, the cross-linking time was 24 h, the diameter of the small end face cylinder of the T-bar used was 4 mm, the height of the small end face was 150 mm, the diameter of the large end face cylinder was 8 mm, and the height of the large end face cylinder was 1 mm.
本实施例与实施例2和实施例3相比：增加了TPU的浓度，MOCT弹性增加；增加了H-CNC的浓度，水凝胶层的弹性模量和断裂韧性增加；增加了芯/壳供料速度及纺丝电压，单位时间内收集到的纳米纤维数将会增加；延长了冷冻干燥时间，挥发性有害气体的残留将会降低；增加了COCNs层的转速，周向取向度增加；相对于实施例2和实施例3，本实施例制备的H-MOCV在纳米纤维平行度、生物力学方面将会增加优秀，使其在原位组织工程人工血管的应用中具有更广阔的使用前景及发展空间。Compared with Example 2 and Example 3, this example: increasing the concentration of TPU, the MOCT elasticity increases; increasing the concentration of H-CNC, increasing the elastic modulus and fracture toughness of the hydrogel layer; increasing the core/shell Feeding speed and spinning voltage, the number of nanofibers collected per unit time will increase; prolong the freeze-drying time, the residual volatile harmful gas will decrease; increase the rotation speed of the COCNs layer, the degree of circumferential orientation will increase; Compared with Example 2 and Example 3, the H-MOCV prepared in this example will be more excellent in nanofiber parallelism and biomechanics, making it more promising in the application of in situ tissue engineering artificial blood vessels. and development space.
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