PMID- 37051812 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20230415 IS - 2046-3758 (Print) IS - 2046-3758 (Electronic) IS - 2046-3758 (Linking) VI - 12 IP - 3 DP - 2023 Mar 10 TI - TGF-beta1 derived from macrophages contributes to load-induced tendon-bone healing in the murine rotator cuff repair model by promoting chondrogenesis. PG - 219-230 LID - 10.1302/2046-3758.123.BJR-2022-0368.R1 [doi] AB - It has been established that mechanical stimulation benefits tendon-bone (T-B) healing, and macrophage phenotype can be regulated by mechanical cues; moreover, the interaction between macrophages and mesenchymal stem cells (MSCs) plays a fundamental role in tissue repair. This study aimed to investigate the role of macrophage-mediated MSC chondrogenesis in load-induced T-B healing in depth. C57BL/6 mice rotator cuff (RC) repair model was established to explore the effects of mechanical stimulation on macrophage polarization, transforming growth factor (TGF)-beta1 generation, and MSC chondrogenesis within T-B enthesis by immunofluorescence and enzyme-linked immunosorbent assay (ELISA). Macrophage depletion was performed by clodronate liposomes, and T-B healing quality was evaluated by histology and biomechanics. In vitro, bone marrow-derived macrophages (BMDMs) were stretched with CELLOAD-300 load system and macrophage polarization was identified by flow cytometry and quantitative real-time polymerase chain reaction (qRT-PCR). MSC chondrogenic differentiation was measured by histochemical analysis and qRT-PCR. ELISA and qRT-PCR were performed to screen the candidate molecules that mediated the pro-chondrogenic function of mechanical stimulated BMDMs. Mechanical stimulation promoted macrophage M2 polarization in vivo and in vitro. The conditioned media from mechanically stimulated BMDMs (MS-CM) enhanced MSC chondrogenic differentiation, and mechanically stimulated BMDMs generated more TGF-beta1. Further, neutralizing TGF-beta1 in MS-CM can attenuate its pro-chondrogenic effect. In vivo, mechanical stimulation promoted TGF-beta1 generation, MSC chondrogenesis, and T-B healing, which were abolished following macrophage depletion. Macrophages subjected to appropriate mechanical stimulation could polarize toward the M2 phenotype and secrete TGF-beta1 to promote MSC chondrogenesis, which subsequently augments T-B healing. CI - (c) 2023 Author(s) et al. FAU - Wang, Linfeng AU - Wang L AUID- ORCID: 0000-0001-6269-8289 AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. FAU - Li, Shengcan AU - Li S AUID- ORCID: 0000-0002-6131-4770 AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. AD - Department of Orthopedics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China. FAU - Xiao, Han AU - Xiao H AUID- ORCID: 0000-0003-4283-4559 AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. FAU - Zhang, Tao AU - Zhang T AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. FAU - Liu, Yuqian AU - Liu Y AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. FAU - Hu, Jianzhong AU - Hu J AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. AD - Department of Spine Surgery and Orthopaedics, Xiangya Hospital, Central South University, Changsha, China. FAU - Xu, Daqi AU - Xu D AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. FAU - Lu, Hongbin AU - Lu H AUID- ORCID: 0000-0001-7749-3593 AD - Department of Sports Medicine, Xiangya Hospital, Central South University, Changsha, China. AD - Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, China. AD - National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China. LA - eng PT - Journal Article DEP - 20230310 PL - England TA - Bone Joint Res JT - Bone & joint research JID - 101586057 PMC - PMC10032229 COIS- All authors declare no conflicts of interest. EDAT- 2023/04/14 06:00 MHDA- 2023/04/14 06:01 PMCR- 2023/03/10 CRDT- 2023/04/13 04:23 PHST- 2023/04/14 06:01 [medline] PHST- 2023/04/13 04:23 [entrez] PHST- 2023/04/14 06:00 [pubmed] PHST- 2023/03/10 00:00 [pmc-release] AID - BJR-2022-0368.R1 [pii] AID - 10.1302/2046-3758.123.BJR-2022-0368.R1 [doi] PST - epublish SO - Bone Joint Res. 2023 Mar 10;12(3):219-230. doi: 10.1302/2046-3758.123.BJR-2022-0368.R1.