Repair of Achilles tendon defect with autologous ASCs engineered tendon in a rabbit model
Repair of Achilles tendon defect with autologous ASCs engineered tendon in a rabbit model
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自体 ASC 工程肌腱修复兔模型跟腱缺损
DOI：
10.1016/j.biomaterials.2014.06.058
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发表时间：
2014-10-01
期刊：
影响因子：
14
通讯作者：
Liu, Wei
中科院分区：
文献类型：
--
作者：
Deng, Dan;Wang, Wenbo;Liu, Wei
脂肪干细胞（ASC）是组织再生的重要细胞来源，并已被证明具有体外肌腱分化的潜力。本研究探讨了在兔跟腱模型中使用 ASC 进行体内工程肌腱修复的可行性。共有 30 只兔子参与了这项研究。复合肌腱支架由内部聚乙醇酸（PGA）无纺纤维和外部由PGA/PLA（聚乳酸）纤维编织的网组成，用于提供机械强度。从颈项皮下脂肪组织中收获自体ASC并进行体外扩增。实验组收获扩增的ASC，重悬于培养基中并均匀接种到支架上，而无细胞支架作为对照组。两组构建体均在生物反应器内动态拉伸下培养 5 周。在每只 30 只兔子的跟腱右侧制作 2 cm 的缺损，然后在实验组（15 只兔子）中移植 3 cm 的细胞接种支架，或在对照组（15 只兔子）中移植 3 cm 的无细胞支架。术后12、21和45周处死动物进行大体观察、组织学和机械分析。结果表明，短期体外培养使 ASC 能够在 PGA 纤维上产生基质，并且两组结构的拉伸强度均约为 50 MPa（p > 0.05）。随着植入时间的增加，细胞接种的构建体逐渐形成新肌腱，并在45周时变得更加成熟，其组织学结构与天然肌腱相似，并且存在形成的胶原纤维的双极模式和D周期结构。此外，胶原纤维直径和拉伸强度均持续增加，不同时间点之间差异显着（p < 0.05）。相比之下，无细胞构建体未能形成高质量的肌腱组织，仅在 45 周时才能观察到纤维结构。在所有检查时间点，两组之间的胶原原纤维直径和拉伸强度均存在显着差异（p < 0.05）。这项研究的结果表明，ASC 可能是体内肌腱工程和再生的潜在细胞来源。 (C) 2014 Elsevier Ltd. 保留所有权利。
Adipose derived stem cells (ASCs) are an important cell source for tissue regeneration and have been demonstrated the potential of tenogenic differentiation in vitro. This study explored the feasibility of using ASCs for engineered tendon repair in vivo in a rabbit Achilles tendon model. Total 30 rabbits were involved in this study. A composite tendon scaffold composed of an inner part of polyglycolic acid (PGA) unwoven fibers and an outer part of a net knitted with PGA/PLA (polylactic acid) fibers was used to provide mechanical strength. Autologous ASCs were harvested from nuchal subcutaneous adipose tissues and in vitro expanded. The expanded ASCs were harvested and resuspended in culture medium and evenly seeded onto the scaffold in the experimental group, whereas cell-free scaffolds served as the control group. The constructs of both groups were cultured inside a bioreactor under dynamic stretch for 5 weeks. In each of 30 rabbits, a 2 cm defect was created on right side of Achilles tendon followed by the transplantation of a 3 cm cell-seeded scaffold in the experimental group of 15 rabbits, or by the transplantation of a 3 cm cell-free scaffold in the control group of 15 rabbits. Animals were sacrificed at 12, 21 and 45 weeks post-surgery for gross view, histology, and mechanical analysis. The results showed that short term in vitro culture enabled ASCs to produce matrix on the PGA fibers and the constructs showed tensile strength around 50 MPa in both groups (p > 0.05). With the increase of implantation time, cell-seeded constructs gradually form neo-tendon and became more mature at 45 weeks with histological structure similar to that of native tendon and with the presence of bipolar pattern and D-periodic structure of formed collagen fibrils. Additionally, both collagen fibril diameters and tensile strength increased continuously with significant difference among different time points (p < 0.05). In contrast, cell-free constructs failed to form good quality tendon tissue with fibril structure observable only at 45 weeks. There were significant differences in both collagen fibril diameter and tensile strength between two groups at all examined time points (p < 0.05). The results of this study support that ASCs are likely to be a potential cell source for in vivo tendon engineering and regeneration. (C) 2014 Elsevier Ltd. All rights reserved.