Co-culture of bone marrow fibroblasts and endothelial cells on modified polycaprolactone substrates for enhanced potentials in bone tissue engineering.

The creation of a vascularized bed makes the survival of seeded cells on 3-dimensional scaffolds much more likely. However, relying purely on random capillary ingrowth into the porous scaffolds from the host may compromise vascularization of a scaffold. One solution is to transplant cells capable of differentiating into new blood vessels into the scaffolds to accelerate the creation of a vascularized scaffold. Because endothelial cells are the key cells involved in blood vessel formation, the present study was designed to investigate the development of a biomaterial surface that supports endothelial cell attachment and proliferation. The subsequent effects of the material surface modifications on the differentiation and proliferation of human bone marrow-derived fibroblasts (HBMFs) when grown in co-culture with a human bone marrow endothelial cell line (HBMEC-60) were studied. Endothelialization studies showed that the gelatin-coated and hydroxyapatite-coated substrates were superior for HBMEC-60 attachment and proliferation to hydrolyzed-only or untreated polycaprolactone substrates. Co-culture studies showed that the presence of the HBMEC-60 specifically enhanced HBMF cell proliferation and differentiation and that this effect was not observed with co-culture with skin fibroblasts. It is concluded that the co-culture of endothelial cells with HBMFs could be a promising culture system for bone tissue- engineering applications.

[1]  D. Ingber,et al.  Prevascularization of porous biodegradable polymers , 1993, Biotechnology and bioengineering.

[2]  Early Osteoblast Attachment, Spreading, and Focal Adhesions on RGD Coated Surfaces , 1998 .

[3]  D H Kohn,et al.  Sustained release of vascular endothelial growth factor from mineralized poly(lactide-co-glycolide) scaffolds for tissue engineering. , 2000, Biomaterials.

[4]  J. Vacanti,et al.  Tissue engineering : Frontiers in biotechnology , 1993 .

[5]  A. Shuttleworth,et al.  Direct cell contact influences bone marrow mesenchymal stem cell fate. , 2004, The international journal of biochemistry & cell biology.

[6]  P. Kelly.  Anatomy, physiology, and pathology of the blood supply of bones. , 1968, The Journal of bone and joint surgery. American volume.

[7]  J. Folkman,et al.  Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis. , 1977, Microvascular research.

[8]  J Amédée,et al.  Effect of human endothelial cells on Human Bone Marrow Stromal Cell phenotype: Role of VEGF? , 2000, Journal of cellular biochemistry.

[9]  H. Augustin,et al.  Bi-directional cell contact-dependent regulation of gene expression between endothelial cells and osteoblasts in a three-dimensional spheroidal coculture model. , 2004, Biochemical and biophysical research communications.

[10]  Anthony Atala,et al.  Principals of neovascularization for tissue engineering. , 2002, Molecular aspects of medicine.

[11]  J. Kohn,et al.  Physico-mechanical properties of degradable polymers used in medical applications: a comparative study. , 1991, Biomaterials.

[12]  C. Choong,et al.  Simple surface modification of poly(ε-caprolactone) for apatite deposition from simulated body fluid , 2005 .

[13]  S. Monroe,et al.  Clinical evaluation of the Capronor contraceptive implant: preliminary report. , 1989, American journal of obstetrics and gynecology.

[14]  R Langer,et al.  Long-term engraftment of hepatocytes transplanted on biodegradable polymer sponges. , 1997, Journal of biomedical materials research.

[15]  M. Nimni,et al.  Promotion of calvarial cell osteogenesis by endothelial cells , 1990, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[16]  J. Feijen,et al.  Interaction of cultured human endothelial cells with polymeric surfaces of different wettabilities. , 1985, Biomaterials.

[17]  B. Decker,et al.  Relationships between endothelial cells, pericytes, and osteoblasts during bone formation in the sheep femur following implantation of tricalciumphosphate‐ceramic , 1995, The Anatomical record.

[18]  P. Rood,et al.  Immortalisation of human bone marrow endothelial cells: characterisation of new cell lines , 2000, European journal of clinical investigation.

[19]  U Kneser,et al.  Modulation of in vitro angiogenesis in a three-dimensional spheroidal coculture model for bone tissue engineering. , 2004, Tissue engineering.

[20]  P. Collin‐Osdoby.  Role of vascular endothelial cells in bone biology , 1994, Journal of cellular biochemistry.

[21]  J. Vacanti,et al.  Human hepatocyte isolation and transplantation into an athymic rat, using prevascularized cell polymer constructs. , 1995, Journal of pediatric surgery.

[22]  B. Guillotin,et al.  Human Primary Endothelial Cells Stimulate Human Osteoprogenitor Cell Differentiation , 2004, Cellular Physiology and Biochemistry.

[23]  D. Darland,et al.  Cell-cell interactions in vascular development. , 2001, Current topics in developmental biology.

[24]  David J Mooney,et al.  Engineering vascular networks in porous polymer matrices. , 2002, Journal of biomedical materials research.

[25]  Y. Ikada,et al.  Bioabsorbable thread for tight tying of bones , 2000, Journal of orthopaedic science : official journal of the Japanese Orthopaedic Association.

[26]  Denys N. Wheatley,et al.  An in vitro model of angiogenesis: Basic features , 2004, Angiogenesis.

[27]  David J. Mooney,et al.  Increased Vascularization and Heterogeneity of Vascular Structures Occurring in Polyglycolide Matrices Containing Aortic Endothelial Cells Implanted in the Rat , 1997 .

[28]  G. Bowlin,et al.  Electrostatic endothelial cell seeding technique for small-diameter (<6 mm) vascular prostheses: feasibility testing. , 1997, Cell transplantation.

[29]  Kanji Sato,et al.  Anabolic effects of 1,25-dihydroxyvitamin D3 on osteoblasts are enhanced by vascular endothelial growth factor produced by osteoblasts and by growth factors produced by endothelial cells. , 1997, Endocrinology.

[30]  A. Göpferich,et al.  Biomimetic polymers in pharmaceutical and biomedical sciences. , 2004, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[31]  J. Folkman,et al.  Vasculogenesis, Angiogenesis, and Growth Factors: Ephrins Enter the Fray at the Border , 1998, Cell.

[32]  C. G. Pitt.  Poly-ε-caprolactone and its copolymers , 1990 .

[33]  G. Nicolardi,et al.  Plasma-treated PET surfaces improve the biocompatibility of human endothelial cells. , 2000, Journal of biomedical materials research.

[34]  C. Little,et al.  Morphogenesis of the First Blood Vessels , 1998, Annals of the New York Academy of Sciences.

[35]  J. Vacanti,et al.  Transplantation of hepatocytes using porous, biodegradable sponges. , 1994, Transplantation proceedings.

[36]  B. Christ,et al.  Embryonic angiogenesis: A review , 1996, Naturwissenschaften.

[37]  C. Colton,et al.  Implantable biohybrid artificial organs. , 1995, Cell transplantation.

[38]  J. Hubbell,et al.  Human endothelial cell interactions with surface-coupled adhesion peptides on a nonadhesive glass substrate and two polymeric biomaterials. , 1991, Journal of biomedical materials research.

[39]  I Zein,et al.  Mechanical properties and cell cultural response of polycaprolactone scaffolds designed and fabricated via fused deposition modeling. , 2001, Journal of biomedical materials research.

[40]  K. Boström,et al.  Endothelial Cells Modulate Osteogenesis in Calcifying Vascular Cells , 2004, Journal of Vascular Research.

[41]  G. Johnson,et al.  Cell responses to biomaterials. I: Adhesion and growth of vascular endothelial cells on poly(hydroxyethyl methacrylate) following surface modification by hydrolytic etching. , 1987, Journal of biomedical materials research.

[42]  I. Zein,et al.  Fused deposition modeling of novel scaffold architectures for tissue engineering applications. , 2002, Biomaterials.

[43]  C. Choong,et al.  Polycaprolactone scaffolds for bone tissue engineering: Effects of a calcium phosphate coating layer on osteogenic cells , 2004 .

[44]  K. Anselme,et al.  Osteoblast adhesion on biomaterials. , 2000, Biomaterials.

[45]  D. Vittet,et al.  In Vitro Models of Vasculogenesis and Angiogenesis , 2001, Laboratory Investigation.

[46]  K J Gooch,et al.  Biomaterial-microvasculature interactions. , 2000, Biomaterials.

[47]  C. Kirkpatrick,et al.  Endothelialization of a non-woven silk fibroin net for use in tissue engineering: growth and gene regulation of human endothelial cells. , 2004, Biomaterials.

[48]  I. A. Jones,et al.  Physical and biocompatibility properties of poly-epsilon-caprolactone produced using in situ polymerisation: a novel manufacturing technique for long-fibre composite materials. , 2000, Biomaterials.

[49]  C. Lewis,et al.  Comparison of three in vitro human ‘angiogenesis’ assays with capillaries formed in vivo , 2004, Angiogenesis.

[50]  Makoto Kodama,et al.  Pore size, tissue ingrowth, and endothelialization of small-diameter microporous polyurethane vascular prostheses. , 2004, Biomaterials.

[51]  J. Feijen,et al.  Adhesion of cultured human endothelial cells onto methacrylate polymers with varying surface wettability and charge. , 1987, Biomaterials.