title: Prevascularization of biofunctional calcium phosphate cement for dental and craniofacial repairs.
abstract: OBJECTIVE
  Calcium phosphate cement (CPC) is promising for dental and craniofacial repairs. Vascularization in bone tissue engineering constructs is currently a major challenge. The objectives of this study were to investigate the prevascularization of macroporous CPC via coculturing human umbilical vein endothelial cells (HUVEC) and human osteoblasts (HOB), and determine the effect of RGD in CPC on microcapillary formation for the first time.
  
  
  METHODS
  Macroporous CPC scaffold was prepared using CPC powder, chitosan liquid and gas-foaming porogen. Chitosan was grafted with Arg-Gly-Asp (RGD) to biofunctionalize the CPC. HUVEC and HOB were cocultured on macroporous CPC-RGD and CPC control without RGD for up to 42d. The osteogenic and angiogenic differentiation, bone matrix mineral synthesis, and formation of microcapillary-like structures were measured.
  
  
  RESULTS
  RGD-grafting in CPC increased the gene expressions of osteogenic and angiogenic differentiation markers than those of CPC control without RGD. Cell-synthesized bone mineral content also increased on CPC-RGD, compared to CPC control (p<0.05). Immunostaining with endothelial marker showed that the amount of microcapillary-like structures on CPC scaffolds increased with time. At 42d, the cumulative vessel length for CPC-RGD scaffold was 1.69-fold that of CPC control. SEM examination confirmed the morphology of self-assembled microcapillary-like structures on CPC scaffolds.
  
  
  CONCLUSIONS
  HUVEC+HOB coculture on macroporous CPC scaffold successfully achieved prevascularization. RGD incorporation in CPC enhanced osteogenic differentiation, bone mineral synthesis, and microcapillary-like structure formation. The novel prevascularized CPC-RGD constructs are promising for dental, craniofacial and orthopedic applications.
authors.affiliation: State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, China; Biomaterials & Tissue Engineering Division, Department of Endodontics, Prosthodontics and Operative Dentistry, University of Maryland Dental School, Baltimore, MD 21201, USA.
authors.forename: Wenchuan
authors.initials: W
authors.lastname: Chen
authors.affiliation: Biomaterials & Tissue Engineering Division, Department of Endodontics, Prosthodontics and Operative Dentistry, University of Maryland Dental School, Baltimore, MD 21201, USA.
authors.forename: WahWah
authors.initials: W
authors.lastname: Thein-Han
authors.affiliation: Biomaterials & Tissue Engineering Division, Department of Endodontics, Prosthodontics and Operative Dentistry, University of Maryland Dental School, Baltimore, MD 21201, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Weir
authors.affiliation: State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, China.
authors.forename: Qianming
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Biomaterials & Tissue Engineering Division, Department of Endodontics, Prosthodontics and Operative Dentistry, University of Maryland Dental School, Baltimore, MD 21201, USA; Center for Stem Cell Biology and Regenerative Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA; University of Maryland Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201, USA; Department of Mechanical Engineering, University of Maryland, Baltimore County, MD 21250, USA. Electronic address: hxu@umaryland.edu.
authors.forename: Hockin H K
authors.initials: HH
authors.lastname: Xu
mesh_terms: D001861:Bone Regeneration; D001842:Bone and Bones / Q000098:blood supply; D002130:Calcium Phosphates; D018920:Coculture Techniques; D003738:Dental Cements; D003793:Dental Restoration, Permanent; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D010006:Osteoblasts / Q000166:cytology; D023822:Tissue Engineering
chemical_list: D002130:Calcium Phosphates; D003738:Dental Cements; C020243:calcium phosphate
keywords: Bone regeneration; Endothelial cells; Macroporous calcium phosphate cement; Osteoblasts; Prevascularization; RGD biofunctionalization
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