A micro-experimental study on tendon defect rapair with xenogeneic tendon as a growth scaffold

doi:10.3969/j.issn.2095-4344.2015.08.007

Abstract

Abstract:

BACKGROUND: Autologous tendons for repair of tendon defects are limited due to the supply of available and can induce tendon dysfunction. Limited sources and high cost of tendon allograft are also difficult to meet the clinical needs.

OBJECTIVE: To provide n theoretical basis for xenogeneic tendon as a clinical tissue-engineered tendon scaffold by observing microscopic changes of xenogeneic tendon in repair of tendon defects at different periods.

METHODS: Flexor tendons from Leghorn chickens (6 months old) were treated chemically as xenogeneic tendon donors. Thirty-six Japanese white rabbits were used to make 2 cm defect models of the intermediate beam in the Achilles tendon of bilateral hind limbs and then divided into two groups randomly, 18 in each group. The experimental group was treated with acellular xenogeneic tendon transplantation and the control group was treated with autologous tendon transplantation. Tendon suture was done using 4-0 non-invasive tendon suture line in double “8” way and the grafts were straightened and immobilized by plaster for 2 weeks. The donor tendons were conducted by general observation, biomechanical test, and engineering observation by optical and electron microscope before and after decellularization. At 2, 4, 9 weeks after transplantation, six rabbits in each group were evaluated by means of optical and electron microscope observation.

RESULTS AND CONCLUSION: (1) The color of tendon cells treated chemically became white and the texture was softer than that before decellularization. The cells and collagen fibers arranged closely and alternately before decellularization, but the arrangement of the collagen fibers was loose relatively and there were no cells and cell debris after decellularization. Moreover, the mechanical strength of the acellular tendon was reduced than that before decellularization. (2) Electron microscope observation: With the increase of transplantation time, the collagen fibers of the transplanted thick chicken tendons were gradually replaced by the ones of the regenerated rabbit tendons. The newly generated collagen fibers were transferred to same-thickness fibers and arrange parallelly, and their structure and function were close to the normal tendons. The maximum tensile resistance after decellularization was 83.44% as before, which could meet the needs of tendon mechanical strength. Ultimately tendon repair is the result of the collagen fiber regeneration, and the xenogeneic tendon treated by chemical methods can be used as the growth scaffold for clinical tendon repair.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: Tendons, Transplantation, Heterologous, Biomechanics

CLC Number:

R318

Nie Ji-ping, Wen Shu-zheng, Bai Zhi-gang. A micro-experimental study on tendon defect rapair with xenogeneic tendon as a growth scaffold[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(8): 1184-1189.

Figures/Tables 3

2.1 肌腱去细胞前后肌腱大体观察未去细胞的肌腱呈黄白色或淡白色，腱膜完整，表面光泽较弱，质地较硬；去细胞肌腱呈瓷白色，腱膜完整，表面光泽较强，质地柔软、韧性好。 2.2 供体肌腱去细胞前后生物力学测定结果肌腱经脱细胞处理后，其力学强度有一定降低，去细胞前与去细胞后对比有显著性意义[(116.98±23.36)，(97.61±11.38) N，t=2.36，P=0.030]；去细胞后肌腱最大抗拉力是去细胞前的83.44%，能满足肌腱移植的生物力学要求。 2.3 供体肌腱去细胞前后光镜及电镜检测结果苏木精-伊红染色光镜100倍下见去细胞前胶原纤维平行紧密排列，束间可见细胞呈串平行排列(见图1A)；去细胞后胶原纤维结构松散，呈波浪状排列，纤维束间无细胞及细胞碎片残留(见图1B)。去细胞前电镜下见粗大的胶原纤维平行排列、结构致密，可见明显周期横纹，细胞核呈梭形、不规则形，细胞器很不发达，数量少，界限不清；去细胞后粗大的胶原纤维结构变的松散，走行基本一致，仍可见周期横纹，但无细胞及细胞碎片残留(见图2)。 2.4 移植后2，4，9周组织学光镜观察结果天狼星红对胶原纤维染色具有其与胶原分子吸附紧密，反应稳定，染色后不褪色且具有特异性，是目前胶原染色的最佳染料。染色后在偏振光显微镜下可见Ⅰ、Ⅲ型胶原显示不同的颜色，Ⅰ型胶原为鲜红色或橙黄色较粗的纤维，成强双折光；Ⅲ型胶原为绿色较细的纤维，呈弱双折光，对比鲜明，极易区别。移植后2周时，两组胶原纤维染色都以Ⅲ型胶原为主，以绿色纤维为主，红色纤维散在其间(见图3A，B)；移植后9周时，两组胶原都较成熟，以Ⅰ型胶原纤维为主，染色呈红色，绿色纤维很少，接近正常兔纤维染色(见图3E，F，G)。移植后4周处于2周与9周的过渡期，红绿纤维交替排列较为明显，但以红色为主(见图3C，D)。 2.5 移植后2，4，9周组织学电镜观察结果移植后2周：异种肌腱移植组见未被替代的粗大鸡纤维与新生细小兔纤维交错排列，新生纤维呈短节状，不连续，排列稀疏，靠近细胞处新生兔纤维较多，可见未被替代完全的粗大鸡纤维阴影，细胞核大，呈椭圆形，细胞器发达，呈球囊状扩张(见图4A-C)；自体肌腱移植组见胶原纤维纤细，排列稀疏，细胞核大，细胞器发达，呈球囊状扩张(见图4D，E；以上结果提示，2周时两组都处于肌腱修复的早期。移植后4周：异种肌腱移植组见新生兔胶原纤维较2周时增粗，排列整齐有序，可见残存的粗大鸡纤维，细胞核呈长椭圆形，胞质少，细胞器不发达，胞质内可见其模糊轮廓(见图4F，G)；自体肌腱移植组见胶原纤维较2周时增粗，纤维数量较多，排列整齐有序，细胞核及细胞器形态同异种肌腱移植组相似(见图4H，I)。移植后9周：异种肌腱移植组见新生兔胶原纤维经改造塑形后变得更加粗大，致密有序排列，可见明显周期横纹，偶而模糊可见残存鸡纤维替代阴影，细胞核呈长梭形，细胞器极不发达，模糊不清(见图4J，K)；自体肌腱移植组见胶原纤维粗大致密，整齐有序排列，横纹明显，细胞核呈长梭形，细胞器极不发达，模糊不清(见图4L，M)；以上结果显示，9周时两组基本都处于肌腱修复的晚期。"

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