Human fibroblast reactions to standard and electropolished titanium and Ti-6Al-7Nb, and electropolished stainless steel.

Stainless steel (SS), titanium (cpTi), and Ti-6Al-7Nb (TAN) are frequently used metals in orthopedic internal fracture fixation. Although reactivity to SS and cpTi are noted in reference, the soft tissue compatibility of TAN has not been comprehensively studied. This study focuses on the in vitro soft tissue compatibility of TAN in comparison to SS and cpTi using a human fibroblast model. The industrial standard surface finishes of these three materials vary considerably in view of their use in similar applications. To distinguish between material parameters of topography and chemistry, we have included electropolished (e.p) counterparts of the standard preparations of cpTi and TAN in the study (standard SS is e.p). All materials were characterized using atomic force microscopy, profilometry, and scanning electron microscopy. Our findings demonstrate that cell morphology and growth rate was similar for SS, and e.p. cpTi and TAN, with cells well spread and forming a confluent monolayer by 10 days. Cell growth on standard cpTi was similar to the electropolished samples; however, they showed a less spread morphology with more filopodia and surface ruffling present. Cell morphology on standard TAN was rounded or elongated and proliferation was inhibited at all time points, with possible cell necrosis by day 10. We found evidence of endocytosis of beta-phase particles originating from the standard TAN surface. We believe that the particle uptake coupled with the characteristic surface topography contribute to the noncytocompatibility of fibroblasts on standard TAN.

[1]  J. Campisi.  The biology of replicative senescence. , 1997, European journal of cancer.

[2]  W. Kolch,et al.  Comparison of anthracycline-induced death of human leukemia cells: Programmed cell death versus necrosis , 2002, Apoptosis.

[3]  A S G Curtis,et al.  Investigating the limits of filopodial sensing: a brief report using SEM to image the interaction between 10 nm high nano‐topography and fibroblast filopodia , 2004, Cell biology international.

[4]  A. Gristina,et al.  Implant failure and the immuno-incompetent fibro-inflammatory zone. , 1994, Clinical orthopaedics and related research.

[5]  O. Pohler,et al.  Unalloyed titanium for implants in bone surgery. , 2000, Injury.

[6]  M. Lanotte,et al.  Multiple apoptotic death types triggered through activation of separate pathways by cAMP and inhibitors of protein phosphatases in one (IPC leukemia) cell line. , 1994, Journal of cell science.

[7]  E. Osano,et al.  Phagocytosis of titanium particles and necrosis in TNF-α-resistant mouse sarcoma L929 cells , 2003 .

[8]  S. Gilbert,et al.  The development of the distal femur Less Invasive Stabilization System (LISS). , 2001, Injury.

[9]  L. Eschbach,et al.  Stainless steel in bone surgery. , 2000, Injury.

[10]  David L. Cochran,et al.  Osteoblasts generate an osteogenic microenvironment when grown on surfaces with rough microtopographies. , 2003, European cells & materials.

[11]  Nikolaj Gadegaard,et al.  Investigating filopodia sensing using arrays of defined nano-pits down to 35 nm diameter in size. , 2004, The international journal of biochemistry & cell biology.

[12]  E. Zamir,et al.  Molecular complexity and dynamics of cell-matrix adhesions. , 2001, Journal of cell science.

[13]  R. Mostardi,et al.  In vitro response of human fibroblasts to commercially pure titanium. , 1999, Journal of biomedical materials research.

[14]  David A. Puleo,et al.  An Introduction To Tissue-Biomaterial Interactions: Tissue-Biomaterial , 2003 .

[15]  M. Textor,et al.  Surface characterization , 1999, Journal of materials science. Materials in medicine.

[16]  T Zislis,et al.  A scanning electron microscopic study of in vitro toxicity of ethylene-oxide-sterilized bone repair materials. , 1989, The Journal of oral implantology.

[17]  J. Folkman,et al.  Role of cell shape in growth control , 1978, Nature.

[18]  Implants for Surgery – A Survey on Metallic Materials , 2005 .

[19]  S. Perren,et al.  Evaluation of soft tissue reactions at the interface of titanium limited contact dynamic compression plate implants with different surface treatments: an experimental sheep study. , 1996, Biomaterials.

[20]  W. Maloney,et al.  In vitro activation of human fibroblasts by retrieved titanium alloy wear debris , 1996, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  R. Rubin,et al.  THE EFFECT OF CELL-TO-CELL CONTACT ON THE SURFACE MORPHOLOGY OF CHINESE HAMSTER OVARY CELLS , 1973, The Journal of cell biology.

[22]  Cell-substratum interactions and the cytoskeleton in cell shape-mediated growth regulation of lens epithelial cells. , 1991, Lens and eye toxicity research.

[23]  M. Iwig,et al.  Growth regulation by cell shape alteration and organization of the cytoskeleton. , 1995, European journal of cell biology.

[24]  C. O'neill,et al.  Evidence for two distinct mechanisms of anchorage stimulation in freshly explanted and 3T3 Swiss mouse fibroblasts , 1986, Cell.

[25]  M. A. Wood,et al.  Steps toward a model nanotopography. , 2002, IEEE transactions on nanobioscience.

[26]  A. Pardee.  G1 events and regulation of cell proliferation. , 1989, Science.

[27]  G. Voggenreiter,et al.  Immuno-inflammatory tissue reaction to stainless-steel and titanium plates used for internal fixation of long bones. , 2003, Biomaterials.

[28]  M. Braun,et al.  Bone remodelling along a titanium hip arthroplasty stem after resection of a chondrosarcoma , 2002, Archives of Orthopaedic and Trauma Surgery.

[29]  B. Kasemo,et al.  Biomaterial and implant surfaces: a surface science approach. , 1988, The International journal of oral & maxillofacial implants.

[30]  C. Lohmann,et al.  Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition. , 1998, Biomaterials.

[31]  L. Spångberg,et al.  Morphologic effects on L929 fibroblasts of titanium tetrafluoride application. , 1998, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[32]  R. Mostardi,et al.  Prosthetic metals have a variable necrotic threshold in human fibroblasts: an in vitro study. , 2002, Journal of biomedical materials research.

[33]  B. Kasemo.  Biocompatibility of titanium implants: surface science aspects. , 1983, The Journal of prosthetic dentistry.

[34]  M. Zlowodzki,et al.  Less Invasive Stabilization System (LISS) for fractures of the proximal tibia: indications, surgical technique and preliminary results of the UMC Clinical Trial. , 2003, Injury.

[35]  Paul Martin,et al.  Structures in focus--filopodia. , 2002, The international journal of biochemistry & cell biology.

[36]  Marcus Textor,et al.  Titanium in Medicine : material science, surface science, engineering, biological responses and medical applications , 2001 .

[37]  M. Textor,et al.  The implant material, Ti6Al7Nb: surface microstructure, composition and properties , 1999, Journal of materials science. Materials in medicine.

[38]  S. S. Davis,et al.  An introduction to tissue-biomaterial interactions , 2002 .

[39]  C. Wilkinson,et al.  Topographical control of cell behaviour: II. Multiple grooved substrata. , 1990, Development.

[40]  G. Stein,et al.  Progressive development of the rat osteoblast phenotype in vitro: Reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix , 1990, Journal of cellular physiology.

[41]  Donald E. Ingber,et al.  The structural and mechanical complexity of cell-growth control , 1999, Nature Cell Biology.

[42]  C. Leyens,et al.  Titanium and titanium alloys : fundamentals and applications , 2005 .

[43]  B. König,et al.  In vitro and in vivo biocompatibility testing of Ti-6Al-7Nb alloy with and without plasma-sprayed hydroxyapatite coating. , 2001, Journal of biomedical materials research.

[44]  S. Perren,et al.  Evaluation of the soft tissue interface at titanium implants with different surface treatments: experimental study on rabbits. , 1994, Bio-medical materials and engineering.

[45]  A. F. Recum.  Handbook of biomaterials evaluation: Scientific, technical, and clinical testing of implant materials , 1986 .

[46]  A. Bershadsky,et al.  Focal contacts of normal and RSV-transformed quail cells. Hypothesis of the transformation-induced deficient maturation of focal contacts. , 1985, Experimental cell research.

[47]  J. Disegi.  Anodizing treatments for titanium implants , 1997, Proceedings of the 1997 16 Southern Biomedical Engineering Conference.

[48]  D E Ingber,et al.  Cell shape, cytoskeletal mechanics, and cell cycle control in angiogenesis. , 1995, Journal of biomechanics.

[49]  L. Claes,et al.  Proliferation and differentiation parameters of human osteoblasts on titanium and steel surfaces. , 2001, Journal of biomedical materials research.

[50]  B D Boyan,et al.  Effect of titanium surface roughness on proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63). , 1995, Journal of biomedical materials research.

[51]  D. Brune.  Surface characterization : a user's sourcebook , 1997 .

[52]  J A Disegi,et al.  Implant materials for fracture fixation: a clinical perspective. , 1989, Orthopedics.

[53]  P. Giannoudis,et al.  Distal femoral fractures: long-term outcome following stabilisation with the LISS. , 2004, Injury.

[54]  L. Claes,et al.  A scanning electron microscopy study of human osteoblast morphology on five orthopedic metals. , 2002, Journal of biomedical materials research.