自体尿源性干细胞片在犬模型中增强肩袖愈合的应用。
Application of Autogenous Urine-Derived Stem Cell Sheet Enhances Rotator Cuff Healing in a Canine Model.
机构信息
Department of Sport Medicine, Xiangya Hospital, Central South University, Changsha, China.
Department of Orthopedics, Xiangya Hospital, Central South University, Changsha, China.
出版信息
Am J Sports Med. 2020 Dec;48(14):3454-3466. doi: 10.1177/0363546520962774. Epub 2020 Nov 2.
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 ( < .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 ( < .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 ( < .05 for all). Biomechanically, the USC sheet group showed significantly higher failure load and stiffness versus the control group ( < .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.
背景
修复后的肩袖(RC)常因瘢痕组织的介入而愈合,使修复更容易失败。尿源性干细胞(USC)具有强大的增殖能力和多能分化能力,可从尿液中分离出来,避免了采集细胞的侵袭性和痛苦的手术。这些优势使其成为增强 RC 愈合的自体移植的新型细胞来源。
假设
将自体 USC 片植入损伤部位将增强 RC 愈合。
研究设计
对照实验室研究。
方法
使用抗坏血酸和转化生长因子β3培养从尿液中分离的 USC 以形成细胞片。16 只雄性成熟比格犬进行双侧肩部手术。右肩仅进行肩胛下肌腱(IT)插入部脱离和修复,另一只进行 IT 插入部脱离和修复,然后植入自体 USC 片。在这些动物中,3 只接受 Dil(1,1'-二辛基-3,3,3',3'-四甲基吲哚碳菁高氯酸盐)标记的 USC 片植入右肩,并在术后 6 周进行细胞追踪后处死。其余动物在术后 12 周处死,收获 IT-肱骨复合体进行大体观察、微计算机断层扫描评估和组织学分析(n = 5)和机械测试(n = 8)。此外,还纳入了 13 个未配对的犬尸体肩部作为天然对照。
结果
微计算机断层扫描分析显示,与对照组相比,USC 片组在 RC 愈合部位的骨体积/总体积和小梁厚度均有显著增加(所有 P <.05)。组织学上,在 RC 愈合部位仍可见 Dil 标记的 USC 片,表明植入的 USC 在术后 6 周时仍存活。同时,USC 片组的愈合界面再生出明显更多的类骨骺组织,而对照组则没有(所有 P <.05)。此外,USC 片组的愈合界面表现出比对照组更大的纤维软骨区、更多的糖胺聚糖沉积和更高的胶原双折射(所有 P <.05)。生物力学上,与对照组相比,USC 片组的失效负荷和刚度显著更高(所有 P <.05)。
结论
在犬模型中,USC 片能够增强 RC 的愈合。
临床相关性
该研究结果表明,USC 片植入可作为 RC 愈合的一种实用方法。