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组织工程人工韧带的研制:用弹性蛋白-胶原蛋白与韧带细胞复合人工韧带重建兔内侧副韧带损伤

Development of a Tissue-Engineered Artificial Ligament: Reconstruction of Injured Rabbit Medial Collateral Ligament With Elastin-Collagen and Ligament Cell Composite Artificial Ligament.

作者信息

Hirukawa Masaki, Katayama Shingo, Sato Tatsuya, Inoue Kota, Niwa Kosuke, Ito Naoya, Hattori Tetsuya, Hosoi Takashi, Unno Hironori, Suzuki Yoshiaki, Hasegawa Masahiro, Miyamoto Keiichi, Horiuchi Takashi

机构信息

Department of Chemistry for Materials, Mie University Graduate School of Medicine, Tsu, Mie, Japan.

Department of Orthopedic Surgery, Mie University Graduate School of Medicine, Tsu, Mie, Japan.

出版信息

Artif Organs. 2018 Jul;42(7):736-745. doi: 10.1111/aor.13124. Epub 2018 Apr 16.

Abstract

Ligament reconstruction using a tissue-engineered artificial ligament (TEAL) requires regeneration of the ligament-bone junction such that fixation devices such as screws and end buttons do not have to be used. The objective of this study was to develop a TEAL consisting of elastin-coated polydioxanone (PDS) sutures covered with elastin and collagen fibers preseeded with ligament cells. In a pilot study, a ring-type PDS suture with a 2.5 mm (width) bone insertion was constructed with/without elastin coating (Ela-coat and Non-coat) and implanted into two bone tunnels, diameter 2.4 mm, in the rabbit tibia (6 cases each) to access the effect of elastin on the bond strength. PDS specimens taken together with the tibia at 6 weeks after implantation indicated growth of bone-like hard tissues around bone tunnels accompanied with narrowing of the tunnels in the Ela-coat group and not in the Non-coat group. The drawout load of the Ela-coat group was significantly higher (28.0 ± 15.1 N, n = 4) than that of the Non-coat group (7.6 ± 4.6 N, n = 5). These data can improve the mechanical bulk property of TEAL through extracellular matrix formation. To achieve this TEAL model, 4.5 × 10 ligament cells were seeded on elastin and collagen fibers (2.5 cm × 2.5 cm × 80 µm) prior to coil formation around the elastin-coated PDS core sutures having ball-shape ends with a diameter of 2.5 mm. Cell-seeded and cell-free TEALs were implanted across the femur and the tibia through bone tunnels with a diameter of 2.4 mm (6 cases each). There was no incidence of TEAL being pulled in 6 weeks. Regardless of the remarkable degradation of PDS observed in the cell-seeded group, both the elastic modulus and breaking load of the cell-seeded group (n = 3) were comparable to those of the sham-operation group (n = 8) (elastic modulus: 15.4 ± 1.3 MPa and 18.5 ± 5.7 MPa; breaking load: 73.0 ± 23.4 N and 104.8 ± 21.8 N, respectively) and higher than those of the cell-free group (n = 5) (elastic modulus: 5.7 ± 3.6 MPa; breaking load: 48.1 ± 11.3 N) accompanied with narrowed bone tunnels and cartilage matrix formation. These data suggest that elastin increased the bond strength of TEAL and bone. Furthermore, our newly developed TEAL from elastin, collagen, and ligament cells maintained the strength of the TEAL even if PDS was degraded.

摘要

使用组织工程人工韧带(TEAL)进行韧带重建需要韧带-骨结合部再生,这样就无需使用螺钉和端钮等固定装置。本研究的目的是开发一种TEAL,它由涂有弹性蛋白的聚二氧六环酮(PDS)缝线组成,表面覆盖有弹性蛋白和预种韧带细胞的胶原纤维。在一项初步研究中,构建了带有/不带有弹性蛋白涂层(弹性蛋白涂层组和无涂层组)、骨插入宽度为2.5 mm的环形PDS缝线,并将其植入兔胫骨中直径2.4 mm的两个骨隧道(每组6例),以研究弹性蛋白对结合强度的影响。植入6周后与胫骨一起取出的PDS标本显示,弹性蛋白涂层组骨隧道周围有类骨硬组织生长,同时隧道变窄,而无涂层组则没有。弹性蛋白涂层组的拔出载荷(28.0±15.1 N,n = 4)显著高于无涂层组(7.6±4.6 N,n = 5)。这些数据可通过细胞外基质形成改善TEAL的机械性能。为实现这种TEAL模型,在围绕直径为2.5 mm的球形末端的涂有弹性蛋白的PDS核心缝线形成线圈之前,将4.5×10个韧带细胞接种在弹性蛋白和胶原纤维(2.5 cm×2.5 cm×80 µm)上。将接种细胞的和未接种细胞的TEAL通过直径2.4 mm的骨隧道植入股骨和胫骨之间(每组6例)。6周内没有TEAL被拉出的情况。尽管在接种细胞组中观察到PDS有明显降解,但接种细胞组(n = 3)的弹性模量和断裂载荷与假手术组(n = 8)相当(弹性模量:分别为15.4±1.3 MPa和18.5±5.7 MPa;断裂载荷:分别为73.0±23.4 N和104.8±21.8 N),且高于未接种细胞组(n = 5)(弹性模量:5.7±3.6 MPa;断裂载荷:48.1±11.3 N),同时伴有骨隧道变窄和软骨基质形成。这些数据表明弹性蛋白增加了TEAL与骨的结合强度。此外,我们新开发的由弹性蛋白、胶原和韧带细胞组成的TEAL即使在PDS降解的情况下仍能保持TEAL的强度。

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