Abstract
Human mesenchymal stem cells (hMSCs) are an attractive tissue engineering avenue for the repair and regeneration of bone. In this study we detail the in vivo performance of a novel electrospun polycaprolactone scaffold incorporating the glycosaminoglycan heparan sulfate (HS) as a carrier for hMSC. HS is a multifunctional regulator of many key growth factors expressed endogenously during bone wound repair, and we have found it to be a potent stimulator of proliferation in hMSCs. To assess the potential of the scaffolds to support hMSC function in vivo, hMSCs pre-committed to the osteogenic lineage (human osteoprogenitor cells) were seeded onto the scaffolds and implanted subcutaneously into the dorsum of nude rats. After 6 weeks the scaffolds were retrieved and examined by histological methods. Implanted human cells were identified using a human nuclei-specific antibody. The host response to the implants was characterized by ED1 and ED2 antibody staining for monocytes/macrophages and mature tissue macrophages, respectively. It was found that the survival of the implanted human cells was affected by the host response to the implant regardless of the presence of HS, highlighting the importance of controlling the host response to tissue engineering devices.
Similar content being viewed by others
References
Ali SA, Doherty PJ, Williams DF (1994) Molecular biointeractions of biomedical polymers with extracellular exudate and inflammatory cells and their effects on the biocompatibility, in vivo. Biomaterials 15:779–785
Blanquaert F, Barritault D, Caruelle JP (1999) Effects of heparan-like polymers associated with growth factors on osteoblast proliferation and phenotype expression. J Biomed Mater Res 44:63–72
Brickman YG, Ford MD, Gallagher JT, Nurcombe V, Bartlett PF, Turnbull JE (1998) Structural modification of fibroblast growth factor-binding heparan sulfate at a determinative stage of neural development. J Biol Chem 273:4350–4359
Burger D, Dayer JM (2002) Cytokines, acute-phase proteins, and hormones: IL-1 and TNF-alpha production in contact-mediated activation of monocytes by T lymphocytes. Ann NY Acad Sci 966:464–473
Dayoub H, Dumont RJ, Li JZ, Dumont AS, Hankins GR, Kallmes DF, Helm GA (2003) Human mesenchymal stem cells transduced with recombinant bone morphogenetic protein-9 adenovirus promote osteogenesis in rodents. Tissue Eng 9:347–356
De Jong WH, Bergsma JE, Robinson JE, Bos RRM (2005) Tissue response to partially in vitro predegraded poly-L-lactide implants. Biomaterials 26:1781–1791
Dijkstra CD, Döpp EA, Joling P, Kraal G (1985) The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3. Immunology 54:589–599
Hagerty RD, Salzmann DL, Kleinert LB, Williams SK (2000) Cellular proliferation and macrophage populations associated with implanted expanded polytetrafluoroethylene and polyethyleneterephthalate. J Biomed Mater Res 49:489–497
Hausser HJ, Brenner RE (2004) Low doses and high doses of heparin have different effects on osteoblast-like Saos-2 cells in vitro. J Cell Biochem 91:1062–1073
Hernandez-Pando R, Bornstein QL, Aguilar Leon D, Orozco EH, Madrigal VK, Martinez Cordero E (2000) Inflammatory cytokine production by immunological and foreign body multinucleated giant cells. Immunology 100:352–358
Hu WJ, Eaton JW, Ugarova TP, Tang L (2001) Molecular basis of biomaterial-mediated foreign body reactions. Blood 98:1231–1238
Hutmacher DW, Schantz T, Zein I, Ng KW, Teoh SH, Tan KC (2001) Mechanical properties and cell cultural response of polycaprolactone scaffolds designed and fabricated via fused deposition modeling. J Biomed Mater Res 55:203–216
Irie A, Habuchi H, Kimata K, Sanai Y (2003) Heparan sulfate is required for bone morphogenetic protein-7 signaling. Biochem Biophys Res Commun 308:858–865
Jackson RA, McDonald MM, Nurcombe V, Little DG, Cool SM (2006) The use of heparan sulfate to augment fracture repair in a rat fracture model. J Orthop Res 24:636–644
Jackson RA, Murali S, van Wijnen AJ, Stein GS, Nurcombe V, Cool SM (2007) Heparan sulfate regulates the anabolic activity of MC3T3-E1 preosteoblast cells by induction of Runx2. J Cell Physiol 210:38–50
Kasten P, Vogel J, Luginbuhl R, Niemeyer P, Tonak M, Lorenz H, Helbig L, Weiss S, Fellenberg J, Leo A et al (2005) Ectopic bone formation associated with mesenchymal stem cells in a resorbable calcium deficient hydroxyapatite carrier. Biomaterials 26:5879–5889
Kim KH, Jeong L, Park HN, Shin SY, Park WH, Lee SC, Kim TI, Park YJ, Seol YJ, Lee YM et al (2005) Biological efficacy of silk fibroin nanofiber membranes for guided bone regeneration. J Biotechnol 120:327–339
Kresse H, Schönherr E (2001) Proteoglycans of the extracellular matrix and growth control. J Cell Physiol 198:266–274
Lafont J, Blanquaert F, Colombier ML, Barritault D, Carueelle JP, Saffar JL (2004) Kinetic study of early regenerative effects of RGTA11, a heparan sulfate mimetic, in rat craniotomy defects. Calcif Tissue Int 75:517–525
Li C, Vepari C, Jin HJ, Kim HJ, Kaplan DL (2006) Electrospun silk-BMP-2 scaffolds for bone tissue engineering. Biomaterials 27:3115–3124
Li WJ, Laurencin CT, Caterson EJ, Tuan RS, Ko FK (2002) Electrospun nanofibrous structure: a novel scaffold for tissue engineering. J Biomed Mater Res 60:613–621
Li WJ, Tuli R, Okafor C, Derfoul A, Danielson KG, Hall DJ, Tuan RS (2005) A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells. Biomaterials 26:599–609
Lindahl U, Lidholt K, Spillmann D, Kjellen L (1994) More to “heparin” than anticoagulation. Thromb Res 75:1–32
Luong-Van E, Grondahl L, Chua KN, Leong KW, Nurcombe V, Cool SM (2006) Controlled release of heparin from poly(epsilon-caprolactone) electrospun fibers. Biomaterials 27:2042–2050
Luong-Van E, Grondahl L, Nurcombe V, Cool S (2007) In vitro biocompatibility and bioactivity of microencapsulated heparan sulfate. Biomaterials 28:2127–2136
Marques AP, Reis RL, Hunt JA (2005) An in vivo study of the host response to starch-based polymers and composites subcutaneously implanted in rats. Macromol Biosci 5:775–785
Ng KW, Hutmacher DW, Schantz JT, Ng CS, Too HP, Lim TC, Phan TT, Teoh SH (2001) Evaluation of ultra-thin poly(epsilon-caprolactone) films for tissue-engineered skin. Tissue Eng 7:441–455
Nurcombe V, Ford MD, Wildschut JA, Bartlett PF (1993) Developmental regulation of neural response to FGF-1 and FGF-2 by heparan sulfate proteoglycan. Science 260:103–106
Ornitz DM (2000) FGFs, heparan sulfate and FGFRs: complex interactions essential for development. Bioessays 22:108–112
Pitt CG, Gratzl MM, Kimmel GL, Surles J, Schindler A (1981) Aliphatic polyesters II. The degradation of poly (DL-lactide), poly (epsilon-caprolactone), and their copolymers in vivo. Biomaterials 2:215–220
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147
Rai B, Teoh SH, Hutmacher DW, Cao T, Ho KH (2005) Novel PCL-based honeycomb scaffolds as drug delivery systems for rhBMP-2. Biomaterials 26:3739–3748
Rapraeger AC, Krufka A, Olwin BB (1991) Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science 252:1705–1708
Rhodes NP, Hunt JA, Williams DF (1997) Macrophage subpopulation differentiation by stimulation with biomaterials. J Biomed Mater Res 37:481–488
Rosengren A, Danielsen N, Bjursten LM (1997) Inflammatory reaction dependence on implant localization in rat soft tissue models. Biomaterials 18:979–987
Sarkar S, Lee GY, Wong JY, Desai TA (2006) Development and characterization of a porous micro-patterned scaffold for vascular tissue engineering applications. Biomaterials 27:4775–4782
Shin M, Ishii O, Sueda T, Vacanti JP (2004a) Contractile cardiac grafts using a novel nanofibrous mesh. Biomaterials 25:3717–3723
Shin M, Yoshimoto H, Vacanti JP (2004b) In vivo bone tissue engineering using mesenchymal stem cells on a novel electrospun nanofibrous scaffold. Tissue Eng 10:33–41
Spivak-Kroizman T, Lemmon MA, Dikic I, Ladbury JE, Pinchasi D, Huang J, Jaye M, Crumley G, Schlessinger J, Lax I (1994) Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation. Cell 79:1015–1024
Sun TC, Mori S, Roper J, Brown C, Hooser T, Burr DB (1992) Do different fluorochrome labels give equivalent histomorphometric information? Bone 13:443–446
Sung HJ, Meredith C, Johnson C, Galis ZS (2004) The effect of scaffold degradation rate on three-dimensional cell growth and angiogenesis. Biomaterials 25:5735–5742
Takada T, Katagiri T, Ifuku M, Morimura N, Kobayashi M, Hasegawa K, Ogamo A, Kamijo R (2003) Sulfated polysaccharides enhance the biological activities of bone morphogenetic proteins. J Biol Chem 278:43229–43235
Tang L, Eaton JW (1993) Fibrin(ogen) mediates acute inflammatory responses to biomaterials. J Exp Med 178:2147–2156
van Luyn MJ, Khouw IM, van Wachem PB, Blaauw EH, Werkmeister JA (1998) Modulation of the tissue reaction to biomaterials. II. The function of T cells in the inflammatory reaction to crosslinked collagen implanted in T-cell-deficient rats. J Biomed Mater Res 39:398–406
Vos JG, Berkvens JM, Kruijt BC (1980) The athymic nude rat. I. Morphology of lymphoid and endocrine organs. Clin Immunol Immunopathol 15:213–228
Xia Z, Taylor PR, Locklin RM, Gordon S, Cui Z, Triffitt JT (2006) Innate immune response to human bone marrow fibroblastic cell implantation in CB17 scid/beige mice. J Cell Biochem 98:966–980
Yang Q, Chen L, Shen XY, Tan ZQ (2006) Preparation of polycaprolactone tissue engineering scaffolds by improved solvent casting/particulate leaching method. J Macromol Sci B Phys 45:1171–1181
Yayon A, Klagsbrun M, Esko JD, Leder P, Ornitz DM (1991) Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell 64:841–848
Yoshimoto H, Shin YM, Terai H, Vacanti JP (2003) A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. Biomaterials 24:2077–2082
Zhou Y, Chen F, Ho ST, Woodruff MA, Lim TM, Hutmacher DW (2007) Combined marrow stromal cell-sheet techniques and high-strength biodegradable composite scaffolds for engineered functional bone grafts. Biomaterials 28:814–824
Acknowledgments
The authors would like to acknowledge the grant support from Singapore’s Agency for Science Technology and Research (A-STAR), the Biomedical Research Council (BMRC) of Singapore and the Institute of Molecular and Cell Biology (IMCB) Singapore. This study was also supported by research grants from the Australian Research Council (DP0209873) and the Wesley Research Institute, Australia (2000100294).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Luong-Van, E., Grøndahl, L., Song, S. et al. The in vivo assessment of a novel scaffold containing heparan sulfate for tissue engineering with human mesenchymal stem cells. J Mol Hist 38, 459–468 (2007). https://doi.org/10.1007/s10735-007-9129-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-007-9129-y