title: Low intensity pulsed ultrasound enhanced mesenchymal stem cell recruitment through stromal derived factor-1 signaling in fracture healing.
abstract: Low intensity pulsed ultrasound (LIPUS) has been proven effective in promoting fracture healing but the underlying mechanisms are not fully depicted. We examined the effect of LIPUS on the recruitment of mesenchymal stem cells (MSCs) and the pivotal role of stromal cell-derived factor-1/C-X-C chemokine receptor type 4 (SDF-1/CXCR4) pathway in response to LIPUS stimulation, which are essential factors in bone fracture healing. For in vitro study, isolated rat MSCs were divided into control or LIPUS group. LIPUS treatment was given 20 minutes/day at 37 °C for 3 days. Control group received sham LIPUS treatment. After treatment, intracellular CXCR4 mRNA, SDF-1 mRNA and secreted SDF-1 protein levels were quantified, and MSCs migration was evaluated with or without blocking SDF-1/CXCR4 pathway by AMD3100. For in vivo study, fractured 8-week-old young rats received intracardiac administration of MSCs were assigned to LIPUS treatment, LIPUS+AMD3100 treatment or vehicle control group. The migration of transplanted MSC to the fracture site was investigated by ex vivo fluorescent imaging. SDF-1 protein levels at fracture site and in serum were examined. Fracture healing parameters, including callus morphology, micro-architecture of the callus and biomechanical properties of the healing bone were investigated. The in vitro results showed that LIPUS upregulated SDF-1 and CXCR4 expressions in MSCs, and elevated SDF-1 protein level in the conditioned medium. MSCs migration was promoted by LIPUS and partially inhibited by AMD3100. In vivo study demonstrated that LIPUS promoted MSCs migration to the fracture site, which was associated with an increase of local and serum SDF-1 level, the changes in callus formation, and the improvement of callus microarchitecture and mechanical properties; whereas the blockade of SDF-1/CXCR4 signaling attenuated the LIPUS effects on the fractured bones. These results suggested SDF-1 mediated MSCs migration might be one of the crucial mechanisms through which LIPUS exerted influence on fracture healing.
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Fang-Yuan
authors.initials: FY
authors.lastname: Wei
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China; Translational Medicine Research & Development Center, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
authors.forename: Kwok-Sui
authors.initials: KS
authors.lastname: Leung
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Jianghui
authors.initials: J
authors.lastname: Qin
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Simon Kwoon-Ho
authors.initials: SK
authors.lastname: Chow
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Huang
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Ming-Hui
authors.initials: MH
authors.lastname: Sun
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China; Translational Medicine Research & Development Center, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Qin
authors.affiliation: Department of Orthopaedics and Traumatology, Clinical Sciences Building, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China; Translational Medicine Research & Development Center, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
authors.forename: Wing-Hoi
authors.initials: WH
authors.lastname: Cheung
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D002465:Cell Movement; D054377:Chemokine CXCL12 / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005269:Femur / Q000166:cytology / Q000000981:diagnostic imaging / Q000293:injuries / Q000502:physiology; D017102:Fracture Healing; D005786:Gene Expression Regulation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019718:Receptors, CXCR4 / Q000378:metabolism; D015398:Signal Transduction; D013977:Tibia / Q000166:cytology / Q000000981:diagnostic imaging / Q000293:injuries / Q000502:physiology; D014465:Ultrasonics; D055114:X-Ray Microtomography
chemical_list: C575935:CXCL12 protein, rat; D054377:Chemokine CXCL12; C486264:Cxcr4 protein, rat; D019718:Receptors, CXCR4