Application of Autogenous Urine-Derived Stem Cell Sheet Enhances Rotator Cuff Healing in a Canine Model

Abstract

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.

Evaluation of patches for rotator cuff repair: A systematic review and meta-analysis based on animal studies

“…Sun, Y [ 125 ] demonstrated that the bone-tendon composite autograft group had a higher tendon maturity score than simple suture. Chen, Y [ 126 ] discussed the stem cell sheets derived from urine of the patient, and the comparison with the tendon-bone suture showed that the cell sheet group presented more collagen fibers at the tendon-bone interface. Sener, M [ 127 ] compared the results between patellar tendon-bone autograft with free flexor tendon autograft, and found that the patellar tendon-bone autograft was completely integrated with the tibia.…”

Infrapatellar Fat Pad Mesenchymal Stromal Cell–Derived Exosomes Accelerate Tendon-Bone Healing and Intra-articular Graft Remodeling After Anterior Cruciate Ligament Reconstruction

“…12,44 The multilineage differentiation potential of the expanded IPFP MSCs toward bone, cartilage, and fat was characterized, and the positive induction of osteogenesis, chondrogenesis, and adipogenesis was determined using alkaline phosphatase staining, safranin O staining, and oil red O staining, respectively. 6,7,13 Isolation and Identification of IPFP MSC-Derived Exosomes (IMEs) Exosome isolation from IPFP MSCs was conducted in accordance with a previously described protocol. 54,58 When the IPFP MSCs reached 70% to 80% confluency on a 10-cm dish (EasYDish; Thermo Fisher Scientific), the cells (~10 6 /10-cm dish) were washed twice using PBS, and the medium was replaced by medium containing exosome-depleted fetal bovine serum (Invitrogen).…”

Application of antibody-conjugated small intestine submucosa to capture urine-derived stem cells for bladder repair in a rabbit model

“…As progenitor cells derived from the urinary system, the USCs can be isolated readily from urine and expand extensively through culturing [ 54 ]. Such cells possess the desired regenerative properties including robust proliferative capacity [ 55 ], multipotential for differentiation [ [56] , [57] , [58] , [59] , [60] ], paracrine effect [ [61] , [62] , [63] ], and immune-modulatory property [ 29 ], and have been tested in animal models for a multitude of human diseases [ 29 , [64] , [65] , [66] , [67] , [68] ]. Previous studies have also demonstrated that the USCs could differentiate into multiple bladder cell lineages identifiable by particular gene and/or protein markers, and provide an ideal cell source for the reconstruction of urinary tract [ 12 , 57 , 69 , 70 ].…”