丝素蛋白涂层人工血管的研制
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摘要
丝素蛋白是一种具生物相容性,可降解和无毒性的天然蛋白。本研究运用丝素蛋白浸渍涂层多孔针织涤纶血管(InterVascular~((?)),InterVascular有限公司,法国)研制一种新的涂层人工血管,并通过体外和体内试验来评价该血管假体是否具备植入时既不漏血而又保留足够孔隙便于受体组织侵入生长并包裹愈合的性能。
在体外试验中,丝素蛋白涂层人工血管的涂层材料占重,血管壁渗水率,形态学以及力学性能将与未涂层的采用相同材料编织的人工血管(InterVascular~((?)))及商业化的胶原涂层人工血管(InterGard~(TM),InterVascular有限公司,法国)进行比较。对照组的未涂层人工血管管壁的渗水率(1846ml/min*cm~2在120mmHg压力下)通过丝素蛋白浸渍涂层降低了99%,达到了手术植入时不漏血的目的。丝素蛋白涂层的占重(117±22mg/g)较InterGard~(TN)血管胶原涂层占重(302±23mg/g)要低的多。
在体内试验中,6例丝素蛋白涂层人工血管植入杂种犬的腹主动脉,分别在植入后4h,3d,2w,1m,3m和6m取材。对照组分未涂层组(InterVascular~((?)))和胶原涂层组(InterGard~(TM)),分别在植入后1m、3m和6m取材。通过对取材血管假体的大体观察和病理学检查来评价移植物在体内的愈合情况。
结果显示丝素蛋白涂层组人工血管在炎症和异物巨细胞反应较对照组轻。在植入1m后,丝素蛋白涂层大部分被吸收降解。丝素蛋白涂层人工血管组在植入后1、3和6m后血管内壁内皮样细胞覆盖率较胶原涂层组要高(48%vs 34%,85%vs 46%和97%vs 90%)。
本研究初步证实使用丝素蛋白浸渍涂层的人工血管可操控性好,手术植入时不漏血。同时丝素蛋白浸渍涂层人工血管植入后异物反应和炎症反应较轻,而且似能促进内皮细胞贴附和生长。
Silk fibroin (SF) is well known to be biocompatible, degradable, and nontoxic. In this study, SF was impregnated into a porous polyester graft (InterVascular~(?) external velour, InterVascular Inc, La Ciotat, France). The SF-impregnated graft was investigated in vitro and in vivo to evaluate its potential for use as a new vascular graft impervious to blood, while retaining high porosity for tissue ingrowth and biological healing.
For in vitro investigation, the water permeability, coating weight, morphology, and mechanical properties of the SF-impregnated grafts were compared to collagen coated grafts (InterGard~(TN) grafts, InterVascular Inc, La Ciotat, France). The water permeability of the controls (1388±30.5mL/cm~2·min at 120 mmHg) was reduced > 99% by SF impregnation, rendering the graft impervious to blood. The coating weight of the collagen was 117±22 mg/g of graft, producing a slightly lower value than the InterGard~(TN) prosthesis (302±23 mg/g).
For in vivo experiment, six SF-sealed vascular grafts were implanted in the abdominal aorta of dogs for scheduled periods ranging from 4 hours to 6 months. Commercial collagen-impregnated grafts (InterGard~(TN)) and untreated external velour grafts (InterVascular~(?)) were also implanted for scheduled periods ranging from 1 to 6 months for comparison. Gross observation of the explanted grafts and histological examination of the representative sections were conducted for two types of grafts using a light microscope after hematoxylin-eosin staining.
These SF-impregnated grafts showed less foreign body and inflammation reactions, and the SF layer was almost completely absorbed. The average of the values in each period for the SF-grafts was 48% neointima at 1 month, 85% at 3 months, and 97% at 6 months, whereas those of the InterGard~(TN) prostheses was 34%, 46%, and 90% respectively.
This study demonstrated that the use of a biodegradable SF as biological sealant can be a feasible approach to prepare impervious textile arterial prostheses. The SF-impregnated graft showed less thrombogenesis and induced host cell migration along the matrix without foreign body or inflammatory reactions. Moreover, it appears to facilitate the development of endothelial-like cells.
在体外试验中,丝素蛋白涂层人工血管的涂层材料占重,血管壁渗水率,形态学以及力学性能将与未涂层的采用相同材料编织的人工血管(InterVascular~((?)))及商业化的胶原涂层人工血管(InterGard~(TM),InterVascular有限公司,法国)进行比较。对照组的未涂层人工血管管壁的渗水率(1846ml/min*cm~2在120mmHg压力下)通过丝素蛋白浸渍涂层降低了99%,达到了手术植入时不漏血的目的。丝素蛋白涂层的占重(117±22mg/g)较InterGard~(TN)血管胶原涂层占重(302±23mg/g)要低的多。
在体内试验中,6例丝素蛋白涂层人工血管植入杂种犬的腹主动脉,分别在植入后4h,3d,2w,1m,3m和6m取材。对照组分未涂层组(InterVascular~((?)))和胶原涂层组(InterGard~(TM)),分别在植入后1m、3m和6m取材。通过对取材血管假体的大体观察和病理学检查来评价移植物在体内的愈合情况。
结果显示丝素蛋白涂层组人工血管在炎症和异物巨细胞反应较对照组轻。在植入1m后,丝素蛋白涂层大部分被吸收降解。丝素蛋白涂层人工血管组在植入后1、3和6m后血管内壁内皮样细胞覆盖率较胶原涂层组要高(48%vs 34%,85%vs 46%和97%vs 90%)。
本研究初步证实使用丝素蛋白浸渍涂层的人工血管可操控性好,手术植入时不漏血。同时丝素蛋白浸渍涂层人工血管植入后异物反应和炎症反应较轻,而且似能促进内皮细胞贴附和生长。
Silk fibroin (SF) is well known to be biocompatible, degradable, and nontoxic. In this study, SF was impregnated into a porous polyester graft (InterVascular~(?) external velour, InterVascular Inc, La Ciotat, France). The SF-impregnated graft was investigated in vitro and in vivo to evaluate its potential for use as a new vascular graft impervious to blood, while retaining high porosity for tissue ingrowth and biological healing.
For in vitro investigation, the water permeability, coating weight, morphology, and mechanical properties of the SF-impregnated grafts were compared to collagen coated grafts (InterGard~(TN) grafts, InterVascular Inc, La Ciotat, France). The water permeability of the controls (1388±30.5mL/cm~2·min at 120 mmHg) was reduced > 99% by SF impregnation, rendering the graft impervious to blood. The coating weight of the collagen was 117±22 mg/g of graft, producing a slightly lower value than the InterGard~(TN) prosthesis (302±23 mg/g).
For in vivo experiment, six SF-sealed vascular grafts were implanted in the abdominal aorta of dogs for scheduled periods ranging from 4 hours to 6 months. Commercial collagen-impregnated grafts (InterGard~(TN)) and untreated external velour grafts (InterVascular~(?)) were also implanted for scheduled periods ranging from 1 to 6 months for comparison. Gross observation of the explanted grafts and histological examination of the representative sections were conducted for two types of grafts using a light microscope after hematoxylin-eosin staining.
These SF-impregnated grafts showed less foreign body and inflammation reactions, and the SF layer was almost completely absorbed. The average of the values in each period for the SF-grafts was 48% neointima at 1 month, 85% at 3 months, and 97% at 6 months, whereas those of the InterGard~(TN) prostheses was 34%, 46%, and 90% respectively.
This study demonstrated that the use of a biodegradable SF as biological sealant can be a feasible approach to prepare impervious textile arterial prostheses. The SF-impregnated graft showed less thrombogenesis and induced host cell migration along the matrix without foreign body or inflammatory reactions. Moreover, it appears to facilitate the development of endothelial-like cells.
引文
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