Evaluation of small-diameter silk vascular grafts implanted in dogs.

Objectives: Vascular replacement is one treatment for cardiovascular disease. However, in blood vessels with a diameter less than 6 mm, the existing artificial vascular grafts may be occluded by thrombus formation or intimal hyperplasia. Thus, new artificial vascular grafts need to be developed. We have developed a small-diameter artificial vascular graft made of silk fibroin. The implantation of such graft has been evaluated mainly in rats. However, only a few reports describe long-term implantation in large animal models. Therefore, modified silk fibroin artificial vascular grafts were implanted in the femoral arteries of dogs, and their patency and remodeling ability were investigated. Methods: Six beagles weighing 10 to 12 kg were used for the in vivo study. Grafts (4 cm length × 3.5 mm inner diameter) were implanted in the femoral artery of 6 dogs. The patency of the graft was monitored using vascular ultrasound apparatus. At 3 months', 5 months', and 1-year postimplantation, the graft was retrieved and conducted histopathologic examination. Results: No side effects, such as ischemia, paralysis, and edema of the hind legs, were observed postimplantation. Five of the 6 grafts exhibited a high patency rate, and the lumen was covered with vascular endothelial cells in the central part of the graft 3 months' postimplantation. Conclusions: Based on these results, artificial silk fibroin vascular grafts implanted in the femoral arteries of dogs exhibit high patency and remodeling ability. Silk fibroin grafts may be clinically applicable as an artificial vascular graft in small-diameter <6 mm.

Development of Small-diameter Polyester Vascular Grafts Coated with Silk Fibroin Sponge.

In recent years, the demand for functional small-diameter (< 6 mm) artificial vascular grafts has greatly increased due to an increase in the number of patients with vascular heart disease. However, currently, there are no available commercial small-diameter grafts. The objective of this research was to develop a porous silk fibroin (SF)-coated poly(ethylene terephthalate) (PET) graft with a diameter < 6 mm. The graft was compared with a gelatin-coated PET graft because the latter PET graft with a diameter ~ 6 mm was widely used as a commercial vascular graft. Initially, porous SF was prepared using Glyc as the porogen [termed SF(Glyc)] and the PET grafts were prepared through the double-Raschel knitting method. Subsequently, the degradation of the SF coating was monitored using protease XIV in vitro and was compared with that observed in gelatin-coated PET grafts. Finally, these grafts were also implanted into rats for an in vivo comparison. In degradation experiments, after 7 days, the SF was clearly digested by protease XIV, but the gelatin on the graft was still remained at the outer surface. In implantation experiments in rats, the SF(Glyc)-coated PET graft was rapidly degraded in vivo and remodeling to self-tissues was promoted compared with the gelatin-coated PET graft. Thrombus formation and intimal hyperplasia were observed in the gelatin-coated PET graft; however, such side reactions were not observed in the SF(Glyc)-coated PET graft. Thus, the porous SF(Glyc)-coated PET graft with a small diameter < 6 mm may be useful as a commercial vascular graft.

Rapid endothelialization and thin luminal layers in vascular grafts using silk fibroin.

The histological effects of silk fibroin (SF) in vascular grafts have not been clarified comprehensively in a large-animal model. This study aimed to observe the histological changes in vascular grafts by using Bombyx mori SF in a dog model. A splice graft consisting of SF and control grafts were implanted in the abdominal aorta of dogs, and the histological characteristics of the 2 types of grafts in each splice graft were compared. Five splice grafts consisted of one graft made of polyester (PE) fibers coated with SF and another coated with gelatin, 4 splice grafts consisted of one graft made of SF fibers coated with gelatin and another made of PE fibers coated with gelatin, and 2 splice grafts consisted of one graft made of SF fibers coated with SF and another made of PE fibers coated with SF. The graft made of PE fibers coated with SF showed more endothelial cells than the graft made of PE fibers coated with gelatin. The grafts using SF as a coating material or graft fibers showed a thinner luminal layer than the grafts made of PE fibers coated with gelatin. This study suggests that SF use for vascular grafts has advantages of rapid endothelialization and tendency to form thin luminal layers.

Small-diameter silk vascular grafts (3 mm diameter) with a double-raschel knitted silk tube coated with silk fibroin sponge.

Small-diameter (less than 6 mm in diameter) vascular grafts are highly desirable due to the large demand for surgical revascularization; however, there are no available artificial grafts. Vascular grafts of 1.5 mm diameter prepared by our group with silk fibroin fiber have been proved to be excellent grafts with remarkably high patency and remodeling, based on rat implantation experiment (Enomoto et al., 2010). In this study, a silk fibroin vascular graft with 3 mm diameter which can be used for the coronary arteries or lower extremity arteries is prepared with a double-raschel knitted Bombyx mori silk fiber tube coated with B. mori silk fibroin sponge. Here the silk sponge is prepared from an aqueous solution of the silk fibroin and poly(ethylene) glycol diglycidyl ether as porogen. Sufficient strength, proper elasticity, and protection from loose ends in the implantation process are obtained for the silk fibroin graft; low water permeability and relatively large compliance are also attained. These excellent physical properties make silk fibroin grafts suitable to be implanted in a canine model.

Preparation of double-raschel knitted silk vascular grafts and evaluation of short-term function in a rat abdominal aorta.

Silk fibroin fiber has a long history of use in sutures because of its high strength and toughness. In the work reported in this paper, small-diameter vascular grafts 1.5 mm in diameter and 10 mm in length were prepared by coating a double-raschel knitted silk fiber graft with silk fibroin aqueous solution containing poly(ethylene glycol diglycidyl ether) as a cross-linking agent. The most important character of silk fibroin graft is remodeling, which is never observed for polyester fiber or expanded polytetrafluoroethylene grafts. The double-raschel knitted silk fiber graft with coating has sufficient physical strength and protects the ladder from the end in the implantation process. The coating also gives protection against leakage of blood from the graft, and elasticity to the graft. Eight weeks after implantation of the grafts in rat abdominal aorta, early formation of thrombosis was avoided.