title: Repair of segmental bone defect using Totally Vitalized tissue engineered bone graft by a combined perfusion seeding and culture system.
abstract: BACKGROUND
  The basic strategy to construct tissue engineered bone graft (TEBG) is to combine osteoblastic cells with three dimensional (3D) scaffold. Based on this strategy, we proposed the "Totally Vitalized TEBG" (TV-TEBG) which was characterized by abundant and homogenously distributed cells with enhanced cell proliferation and differentiation and further investigated its biological performance in repairing segmental bone defect.
  
  
  METHODS
  In this study, we constructed the TV-TEBG with the combination of customized flow perfusion seeding/culture system and β-tricalcium phosphate (β-TCP) scaffold fabricated by Rapid Prototyping (RP) technique. We systemically compared three kinds of TEBG constructed by perfusion seeding and perfusion culture (PSPC) method, static seeding and perfusion culture (SSPC) method, and static seeding and static culture (SSSC) method for their in vitro performance and bone defect healing efficacy with a rabbit model.
  
  
  RESULTS
  Our study has demonstrated that TEBG constructed by PSPC method exhibited better biological properties with higher daily D-glucose consumption, increased cell proliferation and differentiation, and better cell distribution, indicating the successful construction of TV-TEBG. After implanted into rabbit radius defects for 12 weeks, PSPC group exerted higher X-ray score close to autograft, much greater mechanical property evidenced by the biomechanical testing and significantly higher new bone formation as shown by histological analysis compared with the other two groups, and eventually obtained favorable healing efficacy of the segmental bone defect that was the closest to autograft transplantation.
  
  
  CONCLUSIONS
  This study demonstrated the feasibility of TV-TEBG construction with combination of perfusion seeding, perfusion culture and RP technique which exerted excellent biological properties. The application of TV-TEBG may become a preferred candidate for segmental bone defect repair in orthopedic and maxillofacial fields.
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Xiang-Yu
authors.initials: XY
authors.lastname: Ma
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Ya-Fei
authors.initials: YF
authors.lastname: Feng
authors.affiliation: School of Mechanical Engineering, Shanghai Jiao Tong University, State Key Laboratory of Mechanical System and Vibration, Shanghai, People's Republic of China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Yun-Yu
authors.initials: YY
authors.lastname: Hu
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Zhen-Sheng
authors.initials: ZS
authors.lastname: Ma
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Lei
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001696:Biomechanical Phenomena / Q000187:drug effects; D019149:Bioreactors; D001861:Bone Regeneration / Q000187:drug effects; D016025:Bone Transplantation; D002130:Calcium Phosphates / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D019245:Compressive Strength / Q000187:drug effects; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D020007:Implants, Experimental; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D010477:Perfusion; D011817:Rabbits; D011884:Radius / Q000000981:diagnostic imaging / Q000187:drug effects / Q000473:pathology* / Q000601:surgery; D013194:Staining and Labeling; D013997:Time Factors; D046509:Tissue Culture Techniques; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D014945:Wound Healing / Q000187:drug effects; D055114:X-Ray Microtomography
chemical_list: D002130:Calcium Phosphates; D005456:Fluorescent Dyes; C485817:beta-tricalcium phosphate; D000469:Alkaline Phosphatase; D005947:Glucose
references.citation: Trends Biotechnol. 2008 Aug;26(8):434-41
references.pmid: 18585808
references.citation: Bone. 1992;13(1):81-8
references.pmid: 1581112
references.citation: Ann Biomed Eng. 2007 Mar;35(3):429-42
references.pmid: 17216348
references.citation: Proc Natl Acad Sci U S A. 2003 Dec 9;100(25):14683-8
references.pmid: 14657343
references.citation: Biomaterials. 2004 Mar;25(6):1049-58
references.pmid: 14615170
references.citation: Biomaterials. 2003 Jun;24(13):2363-78
references.pmid: 12699674
references.citation: J Mater Sci Mater Med. 2010 Nov;21(11):3039-48
references.pmid: 20857322
references.citation: Tissue Eng. 2001 Oct;7(5):585-98
references.pmid: 11694192
references.citation: Biotechnol Bioeng. 1994 Mar 25;43(7):573-81
references.pmid: 11540959
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11203-8
references.pmid: 15277663
references.citation: Macromol Biosci. 2007 May 10;7(5):579-88
references.pmid: 17457938
references.citation: Biomaterials. 2010 Dec;31(36):9452-61
references.pmid: 20869769
references.citation: Biomaterials. 2009 Oct;30(30):6142-9
references.pmid: 19674779
references.citation: Clin Orthop Relat Res. 1986 Apr;(205):299-308
references.pmid: 3084153
references.citation: Biomech Model Mechanobiol. 2002 Jun;1(1):69-82
references.pmid: 14586708
references.citation: Cell Stress Chaperones. 2013 Mar;18(2):193-201
references.pmid: 23054889
references.citation: J Biomed Mater Res A. 2009 Dec;91(3):855-65
references.pmid: 19065569
references.citation: Tissue Eng. 2003 Apr;9(2):315-25
references.pmid: 12740094
references.citation: Ann Biomed Eng. 2005 Jan;33(1):63-70
references.pmid: 15709706
references.citation: Biotechnol Prog. 1998 Mar-Apr;14(2):193-202
references.pmid: 9548769
references.citation: J Biomed Mater Res A. 2006 Nov;79(2):338-48
references.pmid: 16878315
references.citation: Spine (Phila Pa 1976). 2005 May 15;30(10):1134-8
references.pmid: 15897826
references.citation: J Biomed Mater Res. 2002 Sep 5;61(3):380-90
references.pmid: 12115463
references.citation: Biomaterials. 2006 Feb;27(4):586-95
references.pmid: 16026825
references.citation: Biomaterials. 2001 Oct;22(19):2581-93
references.pmid: 11519777
references.citation: Biomaterials. 2005 Sep;26(27):5474-91
references.pmid: 15860204
references.citation: Tissue Eng. 2003 Apr;9(2):327-36
references.pmid: 12740095
references.citation: J Mater Sci Mater Med. 2005 Dec;16(12):1111-9
references.pmid: 16362209
references.citation: J Mater Sci Mater Med. 2001 Jun;12(6):471-8
references.pmid: 15348260
references.citation: Biomaterials. 2004 Feb;25(5):757-68
references.pmid: 14609664
references.citation: J Biomed Mater Res A. 2003 Dec 15;67(4):1228-37
references.pmid: 14624509
references.citation: FASEB J. 2005 Mar;19(3):482-4
references.pmid: 15625080
references.citation: J Biomed Mater Res A. 2003 Feb 1;64(2):235-41
references.pmid: 12522809
references.citation: J Bone Joint Surg Am. 2008 Feb;90 Suppl 1:36-42
references.pmid: 18292355
references.citation: J Bone Joint Surg Br. 2006 Apr;88(4):421-6
references.pmid: 16567773
references.citation: J Biomed Mater Res A. 2003 Aug 1;66(2):275-82
references.pmid: 12888997
references.citation: Biomaterials. 2001 Jun;22(11):1279-88
references.pmid: 11336300
references.citation: J Biomed Mater Res A. 2005 Aug 1;74(2):171-80
references.pmid: 15965910
references.citation: Clin Orthop Relat Res. 1994 Jan;(298):286-95
references.pmid: 8118989
references.citation: Tissue Eng. 2000 Dec;6(6):605-17
references.pmid: 11103082
references.citation: Biomaterials. 2009 Jul;30(21):3560-6
references.pmid: 19359037
references.citation: J Biomed Mater Res A. 2009 Oct;91(1):102-13
references.pmid: 18767058
references.citation: Tissue Eng. 2006 Apr;12(4):801-9
references.pmid: 16674293
references.citation: Tissue Eng. 2001 Feb;7(1):23-33
references.pmid: 11224921
references.citation: Biotechnol Bioeng. 2005 Aug 20;91(4):482-93
references.pmid: 15895382
references.citation: Biomaterials. 2012 Mar;33(9):2656-72
references.pmid: 22217806
references.citation: Biomaterials. 2010 May;31(13):3527-35
references.pmid: 20170950
references.citation: Ann Biomed Eng. 2005 Sep;33(9):1238-48
references.pmid: 16133930
references.citation: Tissue Eng. 2002 Feb;8(1):43-52
references.pmid: 11886653