PMID- 38180714 OWN - NLM STAT- MEDLINE DCOM- 20240322 LR - 20240422 IS - 2092-9293 (Electronic) IS - 1976-9571 (Linking) VI - 46 IP - 4 DP - 2024 Apr TI - Human umbilical cord mesenchymal stem cells overexpressing RUNX1 promote tendon-bone healing by inhibiting osteolysis, enhancing osteogenesis and promoting angiogenesis. PG - 461-473 LID - 10.1007/s13258-023-01478-3 [doi] AB - BACKGROUND: Rotator cuff injury (RCI) is a common shoulder injury, which is difficult to be completely repaired by surgery. Hence, new strategies are needed to promote the healing of tendon-bone. OBJECTIVE: We aimed to investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) overexpressing RUNX1 on the tendon-bone healing after RCI, and to further explore its mechanism. METHODS: Lentiviral vector was used to mediate the overexpression of RUNX1. RUNX1-overexpressed UCB-MSCs (referred to as MSC-RUNX1) were co-cultured with osteoclasts, and TRAP staining was performed to observe the formation of osteoclasts. Then MSC-RUNX1 was cultured in osteogenic differentiation medium, Alizarin red staining was conducted to detect osteogenic differentiation. The expression of markers of osteogenesis and osteoclast was detected by RT-qPCR. EA. hy926 cells were co-cultured with MSC-RUNX1. Transwell assay was used to detect the migration, and the expression of angiogenesis related-genes VEGF and TGF-beta was detected by RT-qPCR. The rat rotator cuff reconstruction model was established and MSCs were injected at the tendon-bone junction. Biomechanical test and micro-CT scanning were performed, and HE, Masson and Alcian Blue staining were used for histological evaluation of tendon-bone healing. TUNEL and PCNA immunofluorescence (IF) staining were performed to evaluate apoptosis and proliferation at the tendon-bone healing site. The levels of TNF-alpha, IL-6 and IL-8 in serum were detected by ELISA. The expression of CD31 and Endomucin that related to angiogenesis was detected by IF. Safranin O-fast and TRAP/CD40L immunohistochemical staining were used to assess the levels of osteoclasts and osteoblasts at the tendon-bone healing site. RESULTS: hUC-MSCs overexpressing RUNX1 inhibited osteoclast formation and promoted osteogenic differentiation. MSC-RUNX1 could promote the migration and tube formation of EA. hy926 cells, and up-regulate the levels of VEGF and TGF-beta. Model mice treated with MSC-RUNX1 partially restored the biomechanical indexes. Treatment of MSC-RUNX1 obviously increased the bone density, accompanied by the formation of new bone. In vivo experiments showed that MSC-RUNX1 treatment could promote tendon-bone healing and inhibit inflammatory response in rats. MSC-RUNX1 treatment also promoted angiogenesis at the tendon-bone healing site, while inhibiting osteoclast formation and promoting osteogenic differentiation. CONCLUSION: hUC-MSCs overexpressing RUNX1 can inhibit the formation of osteoclasts and differentiation of osteoblasts, promote angiogenesis and inhibit inflammation, thereby promoting tendon-bone healing after RCI. CI - (c) 2024. The Author(s) under exclusive licence to The Genetics Society of Korea. FAU - Guo, Dan AU - Guo D AD - Department of Orthopedic, Yangzhou Clinical Medical College of Nanjing Medical University, Yangzhou, 225001, Jiangsu, China. FAU - Yang, Jian AU - Yang J AD - Department of Orthopedic, Yangzhou Clinical Medical College of Nanjing Medical University, Yangzhou, 225001, Jiangsu, China. FAU - Liu, Dianwei AU - Liu D AD - Department of Orthopedic, Yangzhou Clinical Medical College of Nanjing Medical University, Yangzhou, 225001, Jiangsu, China. FAU - Zhang, Pei AU - Zhang P AD - Department of Orthopedic, Yangzhou Clinical Medical College of Nanjing Medical University, Yangzhou, 225001, Jiangsu, China. FAU - Sun, Hao AU - Sun H AD - Department of Orthopedic, Yangzhou Clinical Medical College of Nanjing Medical University, Yangzhou, 225001, Jiangsu, China. FAU - Wang, Jingcheng AU - Wang J AD - Department of Orthopedic, Yangzhou Clinical Medical College of Nanjing Medical University, Yangzhou, 225001, Jiangsu, China. wangjcyangzhou@163.com. LA - eng PT - Journal Article DEP - 20240105 PL - Korea (South) TA - Genes Genomics JT - Genes & genomics JID - 101481027 RN - 0 (Vascular Endothelial Growth Factor A) RN - 0 (Core Binding Factor Alpha 2 Subunit) RN - 0 (Transforming Growth Factor beta) RN - 0 (RUNX1 protein, human) SB - IM MH - Humans MH - Rats MH - Mice MH - Animals MH - Osteogenesis MH - Vascular Endothelial Growth Factor A/genetics MH - Angiogenesis MH - Core Binding Factor Alpha 2 Subunit/metabolism MH - *Osteolysis MH - Tendons MH - Umbilical Cord MH - *Mesenchymal Stem Cells/metabolism MH - Transforming Growth Factor beta/metabolism OTO - NOTNLM OT - Human umbilical cord mesenchymal stem cells OT - RUNX1 OT - Rotator cuff injury OT - Tendon-bone healing EDAT- 2024/01/05 12:44 MHDA- 2024/03/22 06:44 CRDT- 2024/01/05 11:15 PHST- 2023/09/28 00:00 [received] PHST- 2023/11/23 00:00 [accepted] PHST- 2024/03/22 06:44 [medline] PHST- 2024/01/05 12:44 [pubmed] PHST- 2024/01/05 11:15 [entrez] AID - 10.1007/s13258-023-01478-3 [pii] AID - 10.1007/s13258-023-01478-3 [doi] PST - ppublish SO - Genes Genomics. 2024 Apr;46(4):461-473. doi: 10.1007/s13258-023-01478-3. Epub 2024 Jan 5.