Application of Autogenous Urine-Derived Stem Cell Sheet Enhances Rotator Cuff Healing in a Canine Model
Application of Autogenous Urine-Derived Stem Cell Sheet Enhances Rotator Cuff Healing in a Canine Model
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应用自体尿液干细胞片增强犬模型的肩袖愈合
DOI：
10.1177/0363546520962774
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发表时间：
2020-12-01
影响因子：
4.8
通讯作者：
Chen, Can
中科院分区：
文献类型：
--
作者：
Chen, Yang;Xu, Yan;Chen, Can
背景：修复后的肩袖（RC）通常愈合时伴有瘢痕组织，使得修复容易失败。尿源性干细胞（USCs）具有强大的增殖能力和多谱系分化，可以从尿液中分离，避免了用于收获细胞的侵入性和痛苦的外科手术。这些优点使其成为自体移植促进RC愈合的新细胞来源。假设：将自体USC片植入损伤部位将促进RC愈合。研究设计：对照实验室研究。方法：采用抗坏血酸和转化生长因子β3培养人尿USCs。16只成年雄性比格犬接受了双侧肩关节手术。右侧肩关节仅行冈下肌腱止点分离修复，另一侧肩关节行冈下肌腱止点分离修复，然后植入自体USC片。其中3只动物在右肩植入Dil（1，1 ′-双十八烷基-3，3，3 ′，3 ′-四甲基吲哚羰花青高氯酸盐）标记的USC片植入物，并在术后6周处死进行细胞追踪。其余动物于术后12周处死，取IT-肱骨复合体进行大体观察、显微计算机断层扫描评价和组织学分析（n = 5）以及力学测试（n = 8）。此外，还包括13个未配对的犬尸体肩关节作为自体对照。结果如下：显微计算机断层扫描分析显示，与对照组相比，USC片材组RC愈合部位的骨体积/总体积和小梁厚度显著增加（所有P <0.05）。组织学上，在RC愈合部位仍可见Dil标记的USC片，这表明植入的USC在术后6周仍有活力。同时，USC片组愈合界面再生的附着点样组织明显多于对照组（P <0.05）。此外，与对照组相比，USC片材组的愈合界面呈现出更大的纤维软骨面积、更多的蛋白多糖沉积和更高的胶原双折射（P <0.05）。生物力学方面，USC片材组的失效载荷和刚度显著高于对照组（P <0.05）。结论：USC片材能够增强犬模型中的RC愈合。临床相关性：研究结果表明，USC片材植入可作为RC愈合的实际应用。
Background: A repaired rotator cuff (RC) often heals with interposed scar tissue, making repairs prone to failure. Urine-derived stem cells (USCs), with robust proliferation ability and multilineage differentiation, can be isolated from urine, avoiding invasive and painful surgical procedures for harvesting the cells. These advantages make it a novel cell source for autologous transplantation to enhance RC healing. Hypothesis: Implantation of an autogenous USC sheet to the injury site will enhance RC healing. Study Design: Controlled laboratory study. Methods: USCs isolated from urine were cultured using ascorbic acid and transforming growth factor β3 to form a cell sheet. Sixteen male mature beagles underwent bilateral shoulder surgery. The right shoulder underwent infraspinatus tendon (IT) insertion detachment and repair only, and the other was subjected to IT insertion detachment and repair, followed by autogenous USC sheet implantation. Among the animals, 3 received a Dil (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate)- labeled USC sheet implant in the right shoulder and were sacrificed at postoperative 6 weeks for cell tracking. The other animals were sacrificed at postoperative 12 weeks, and the IT-humerus complexes were harvested for gross observation, micro–computed tomography evaluation and histological analysis (n = 5), and mechanical testing (n = 8). Additionally, 13 unpaired canine cadaveric shoulders were included as native controls. Results: Micro–computed tomography analysis showed that the USC sheet group had a significant increase in bone volume/total volume and trabecular thickness at the RC healing site when compared with the control group (P < .05 for all). Histologically, the Dil-labeled USC sheet was still visible at the RC healing site, which suggested that the implanted USCs remained viable at postoperative 6 weeks. Meanwhile, the healing interface in the USC sheet group regenerated significantly more enthesis-like tissue than did that of the control group (P < .05). Additionally, the healing interface in the USC sheet group presented a larger fibrocartilage area, more proteoglycan deposition, and higher collagen birefringence than did that of the control group (P < .05 for all). Biomechanically, the USC sheet group showed significantly higher failure load and stiffness versus the control group (P < .05 for all). Conclusion: A USC sheet was able to enhance RC healing in a canine model. Clinical Relevance: The findings of the study showed that USC sheet implantation could serve as a practical application for RC healing.