Interactions of osteoblasts and macrophages with biodegradable and highly porous polyesterurethane foam and its degradation products.

The macrophage cell line J774, primary rat osteoblasts, and the osteoblast cell line MC3T3-E1 were used to examine the biocompatibility of a newly developed polyesterurethane foam and the possible use of this structure as bone-repair materials. The newly developed, biodegradable, and highly porous (pore size 100-150 microns) DegraPol/btc polyesterurethane foam was found to exhibit good cell compatibility; the cell-to-substrate interactions induced neither cytotoxic effects nor activation of macrophages. Osteoblasts and macrophages exhibited normal cell morphology. No signs of cell damage were detected using scanning electron microscopy (SEM). No significant increase in the production of tumor necrosis factor-alpha (TNF-alpha) or nitric oxide (NO) was detected in macrophages. Compared with cells cultured on tissue culture polystyrene (TCPS), macrophages exhibited relatively high cell attachment (150% of TCPS) but significantly high doubling time (about 8 days) compared with TCPS (4.6 days). Primary rat osteoblasts and the osteoblast cell line exhibited relatively high attachment (140% and 180% of TCPS, respectively) and a doubling time of about 5 days, compared with TCPS (6 days and 8.8 days, respectively). Eight days after cell seeding, osteoblasts exhibited a confluent cell multilayer and migrated into the pores of the polymer. In addition they produced high concentrations of collagen type I, the main protein of the bone, and expressed increasing alkaline phosphatase activity and osteocalcin production throughout the 12 days of the experiment. During degradation of these polymers, small crystalline particles of short-chain poly[(R)-3-hydroxybutyric acid] (M(n) approximately 2300) (PHB-P) are released. Therefore PHB-P (diameter, 2-20 microns), as possible degradation products of the polymer, are investigated here for their effects on macrophages and osteoblasts. Results obtained in the present study clearly indicate that macrophages and, to a lesser degree, osteoblasts have the ability to take up (phagocytose) PHB-P. At low concentrations particles of PHB failed to induce cytotoxic effects or to activate macrophages. Osteoblasts showed only limited PHB-P phagocytosis and no signs of cellular damage. At high concentrations of PHB-P, this process was accompanied by cytotoxic effects in macrophages (> 200 pg PHB-P/cell) and to a lesser extent in osteoblasts (> 400 pg PHB-P/cell).

[1]  G. Stein,et al.  Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells , 1990, Journal of cellular physiology.

[2]  L. Quarles,et al.  Distinct proliferative and differentiated stages of murine MC3T3‐E1 cells in culture: An in vitro model of osteoblast development , 1992, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[3]  C. Laurencin,et al.  Osteoblast-like cell (MC3T3-E1) proliferation on bioerodible polymers: an approach towards the development of a bone-bioerodible polymer composite material. , 1993, Biomaterials.

[4]  H. Busscher,et al.  Interaction of fibroblasts and polymer surfaces: relationship between surface free energy and fibroblast spreading. , 1983, Journal of biomedical materials research.

[5]  H. Sudo,et al.  Development of a new system for evaluating the biocompatibility of implant materials using an osteogenic cell line (MC3T3-E1). , 1988, Journal of biomedical materials research.

[6]  James M. Anderson,et al.  Chapter 4 Mechanisms of inflammation and infection with implanted devices , 1993 .

[7]  W. Jeter,et al.  Immunological Responses to Bone , 1972, Clinical orthopaedics and related research.

[8]  F. A. Scholl,et al.  Crude liver membrane fractions and extracellular matrix components as substrata regulate differentially the preservation and inducibility of cytochrome P-450 isoenzymes in cultured rat hepatocytes. , 1993, European journal of biochemistry.

[9]  A. Raz,et al.  Cell-substrate interaction. A method for evaluating the possible correlation between metastatic phenotype and cell surface energy. , 1984, Experimental cell research.

[10]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[11]  C. G. Groot,et al.  Behaviour of fetal rat osteoblasts cultured in vitro on bioactive glass and nonreactive glasses. , 1992, Biomaterials.

[12]  G. Francis,et al.  A micromechanical technique for monitoring cell-substrate adhesiveness: measurements of the strength of red blood cell adhesion to glass and polymer test surfaces. , 1989, Journal of biomedical materials research.

[13]  D. Puleo,et al.  Osteoblasts on hydroxyapatite, alumina and bone surfaces in vitro: morphology during the first 2 h of attachment. , 1992, Biomaterials.

[14]  D. Puleo,et al.  Osteoblast responses to orthopedic implant materials in vitro. , 1991, Journal of biomedical materials research.

[15]  G. Ciardelli,et al.  Cell response of cultured macrophages, fibroblasts, and co-cultures of Kupffer cells and hepatocytes to particles of short-chain poly[(R)-3-hydroxybutyric acid] , 1996 .

[16]  M. Marinkovich,et al.  Collagen synthesis and deposition during mammary epithelial cell spreading on collagen gels , 1986, Journal of cellular physiology.

[17]  R Langer,et al.  Switching from differentiation to growth in hepatocytes: Control by extracellular matrix , 1992, Journal of cellular physiology.

[18]  T. Espevik,et al.  A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis factor from human monocytes. , 1986, Journal of immunological methods.

[19]  G. Ciardelli,et al.  Phagocytosis and biodegradation of short-chain poly [(R)-3-hydroxybutyric acid] particles in macrophage cell line , 1995 .

[20]  K. Langford,et al.  Hydroxylapatite: an adjunct to cranial bone grafting. , 1988, Journal of neurosurgery.

[21]  Y. Amagai,et al.  In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria , 1983, The Journal of cell biology.

[22]  R. W. Bussian,et al.  Short-term cell-attachment rates: a surface-sensitive test of cell-substrate compatibility. , 1987, Journal of biomedical materials research.

[23]  J. Turnay,et al.  Cell morphology, proliferation and collagen synthesis of human fibroblasts cultured on sepiolite-collagen complexes. , 1988, Journal of biomedical materials research.

[24]  G. Friedlaender Bone grafts. The basic science rationale for clinical applications. , 1987, The Journal of bone and joint surgery. American volume.

[25]  O. H. Lowry,et al.  The quantitative histochemistry of brain. II. Enzyme measurements. , 1954, The Journal of biological chemistry.

[26]  B. Tighe,et al.  Cellular interactions with synthetic polymer surfaces in culture. , 1985, Biomaterials.

[27]  M. Kodama,et al.  Porous polyurethane vascular prostheses with variable compliances. , 1992, Journal of biomedical materials research.

[28]  A. Mikos,et al.  Osteoblast function on synthetic biodegradable polymers. , 1994, Journal of biomedical materials research.

[29]  A. Albini,et al.  Osteoblastic cells from rat long bone. I. Characterization of their differentiation in culture. , 1995, Bone.

[30]  Kevin E. Healy,et al.  A novel method to fabricate bioabsorbable scaffolds , 1995 .

[31]  S. Pal,et al.  Mechanical properties of bone cement: a review. , 1984, Journal of biomedical materials research.

[32]  G. Ciardelli,et al.  Characterization of the cell response of cultured macrophages and fibroblasts to particles of short-chain poly[(R)-3-hydroxybutyric acid]. , 1996, Journal of biomedical materials research.

[33]  K. Heiple,et al.  Immune responses of rats to frozen bone allografts. , 1983, The Journal of bone and joint surgery. American volume.