title: Co-Seeding Human Endothelial Cells with Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells on Calcium Phosphate Scaffold Enhances Osteogenesis and Vascularization in Rats.
abstract: A major challenge in repairing large bone defects with tissue-engineered constructs is the poor vascularization in the defect. The lack of vascular networks leads to insufficient oxygen and nutrients supply, which compromises the survival of seeded cells. To achieve favorable regenerative effects, prevascularization of tissue-engineered constructs by co-culturing of endothelial cells and bone cells is a promising strategy. The aim of this study was to investigate the effects of human-induced pluripotent stem cell-derived mesenchymal stem cells (hiPSC-MSCs) co-cultured with human umbilical vein endothelial cells (HUVECs) for prevascularization of calcium phosphate cement (CPC) scaffold on bone regeneration in vivo for the first time. HUVECs co-cultured with hiPSC-MSCs formed microcapillary-like structures in vitro. HUVECs promoted mineralization of hiPSC-MSCs on CPC scaffolds. Four groups were tested in a cranial bone defect model in nude rats: (1) CPC scaffold alone (CPC control); (2) HUVEC-seeded CPC (CPC-HUVEC); (3) hiPSC-MSC-seeded CPC (CPC-hiPSC-MSC); and (4) HUVECs co-cultured with hiPSC-MSCs on CPC scaffolds (co-culture group). After 12 weeks, the co-culture group achieved the greatest new bone area percentage of 46.38% ± 3.8% among all groups (p < 0.05), which was more than four folds of the 10.61% ± 1.43% of CPC control. In conclusion, HUVECs co-cultured with hiPSC-MSCs substantially promoted bone regeneration. The novel construct of HUVECs co-cultured with hiPSC-MSCs delivered via CPC scaffolds is promising to enhance bone and vascular regeneration in orthopedic applications.
authors.affiliation: 1 State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University , Chengdu, Sichuan, China .|2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Xian
authors.initials: X
authors.lastname: Liu
authors.affiliation: 1 State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University , Chengdu, Sichuan, China .|2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Wenchuan
authors.initials: W
authors.lastname: Chen
authors.affiliation: 1 State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University , Chengdu, Sichuan, China .|2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Chi
authors.initials: C
authors.lastname: Zhang
authors.affiliation: 2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Wahwah
authors.initials: W
authors.lastname: Thein-Han
authors.affiliation: 2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Kevin
authors.initials: K
authors.lastname: Hu
authors.affiliation: 2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Reynolds
authors.affiliation: 1 State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University , Chengdu, Sichuan, China .
authors.forename: Chongyun
authors.initials: C
authors.lastname: Bao
authors.affiliation: 2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.affiliation: 2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.|3 Department of Orthopedic Surgery, Nanfang Hospital, Southern Medical University , Guangzhou, Guangdong, China .
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: 2 Department of Endodontics, Periodontics and Prosthodontics, University of Maryland School of Dentistry , Baltimore, Maryland.|4 Center for Stem Cell Biology and Regenerative Medicine, University of Maryland School of Medicine , Baltimore, Maryland.|5 Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine , Baltimore, Maryland.|6 Mechanical Engineering Department, University of Maryland , Baltimore County, Maryland.
authors.forename: Hockin H K
authors.initials: HHK
authors.lastname: Xu
mesh_terms: D000818:Animals; D001861:Bone Regeneration; D002130:Calcium Phosphates / Q000737:chemistry*; D018920:Coculture Techniques; D064593:Heterografts; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism / Q000473:pathology / Q000637:transplantation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology / Q000637:transplantation; D008297:Male; D008819:Mice, Nude; D018919:Neovascularization, Physiologic; D010012:Osteogenesis; D051381:Rats; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D002130:Calcium Phosphates; C020243:calcium phosphate
keywords: animal model; bone tissue engineering; endothelial cells; induced pluripotent stem cells; mesenchymal stem cells; vascularization