title: Spatiotemporal Analyses of Osteogenesis and Angiogenesis via Intravital Imaging in Cranial Bone Defect Repair.
abstract: Osteogenesis and angiogenesis are two integrated components in bone repair and regeneration. A deeper understanding of osteogenesis and angiogenesis has been hampered by technical difficulties of analyzing bone and neovasculature simultaneously in spatiotemporal scales and in 3D formats. To overcome these barriers, a cranial defect window chamber model was established that enabled high-resolution, longitudinal, and real-time tracking of angiogenesis and bone defect healing via multiphoton laser scanning microscopy (MPLSM). By simultaneously probing new bone matrix via second harmonic generation (SHG), neovascular networks via intravenous perfusion of fluorophore, and osteoblast differentiation via 2.3-kb collagen type I promoter-driven GFP (Col2.3GFP), we examined the morphogenetic sequence of cranial bone defect healing and further established the spatiotemporal analyses of osteogenesis and angiogenesis coupling in repair and regeneration. We showed that bone defect closure was initiated in the residual bone around the edge of the defect. The expansion and migration of osteoprogenitors into the bone defect occurred during the first 3 weeks of healing, coupled with vigorous microvessel angiogenesis at the leading edge of the defect. Subsequent bone repair was marked by matrix deposition and active vascular network remodeling within new bone. Implantation of bone marrow stromal cells (BMSCs) isolated from Col2.3GFP mice further showed that donor-dependent bone formation occurred rapidly within the first 3 weeks of implantation, in concert with early angiogenesis. The subsequent bone wound closure was largely host-dependent, associated with localized modest induction of angiogenesis. The establishment of a live imaging platform via cranial window provides a unique tool to understand osteogenesis and angiogenesis in repair and regeneration, enabling further elucidation of the spatiotemporal regulatory mechanisms of osteoprogenitor cell interactions with host bone healing microenvironment.
authors: Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY, USA.
authors: Chunlan
authors: C
authors: Huang
authors: Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY, USA.
authors: Vincent P
authors: VP
authors: Ness
authors: Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY, USA.
authors: Xiaochuan
authors: X
authors: Yang
authors: Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY, USA.
authors: Hongli
authors: H
authors: Chen
authors: Department of Computer Science, University of Rochester, Rochester, NY, USA.
authors: Jiebo
authors: J
authors: Luo
authors: Department of Biomedical Engineering, University of Rochester, Rochester, NY, USA.
authors: Edward B
authors: EB
authors: Brown
authors: Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY, USA.
authors: Xinping
authors: X
authors: Zhang
mesh_terms: D000818:Animals; D002449:Cell Aggregation; D003199:Computer Systems; D004195:Disease Models, Animal; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D018919:Neovascularization, Physiologic; D064231:Nestin / Q000378:metabolism; D010006:Osteoblasts / Q000378:metabolism; D010012:Osteogenesis; D012038:Regeneration; D012886:Skull / Q000473:pathology*; D062211:Spatio-Temporal Analysis; D014945:Wound Healing
chemical_list: D064231:Nestin; D049452:Green Fluorescent Proteins
keywords: ANGIOGENESIS; BMSC; CRANIAL DEFECT REPAIR; INTRAVITAL IMAGING; OSTEOGENESIS
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