Novel hydrophilic nanostructured microtexture on direct metal laser sintered Ti-6Al-4V surfaces enhances osteoblast response in vitro and osseointegration in a rabbit model.

The purpose of this study was to compare the biological effects in vivo of hierarchical surface roughness on laser sintered titanium-aluminum-vanadium (Ti-6Al-4V) implants to those of conventionally machined implants on osteoblast response in vitro and osseointegration. Laser sintered disks were fabricated to have micro-/nano-roughness and wettability. Control disks were computer numerical control (CNC) milled and then polished to be smooth (CNC-M). Laser sintered disks were polished smooth (LST-M), grit blasted (LST-B), or blasted and acid etched (LST-BE). LST-BE implants or implants manufactured by CNC milling and grit blasted (CNC-B) were implanted in the femurs of male New Zealand white rabbits. Most osteoblast differentiation markers and local factors were enhanced on rough LST-B and LST-BE surfaces in comparison to smooth CNC-M or LST-M surfaces for MG63 and normal human osteoblast cells. To determine if LST-BE implants were osteogenic in vivo, we compared them to implant surfaces used clinically. LST-BE implants had a unique surface with combined micro-/nano-roughness and higher wettability than conventional CNC-B implants. Histomorphometric analysis demonstrated a significant improvement in cortical bone-implant contact of LST-BE implants compared to CNC-B implants after 3 and 6 weeks. However, mechanical testing revealed no differences between implant pullout forces at those time points. LST surfaces enhanced osteoblast differentiation and production of local factors in vitro and improved the osseointegration process in vivo. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2086-2098, 2016.

[1]  Ichiro Nishimura,et al.  Three-dimensional bone-implant integration profiling using micro-computed tomography. , 2006, The International journal of oral & maxillofacial implants.

[2]  M. Zwahlen,et al.  Improvements in implant dentistry over the last decade: comparison of survival and complication rates in older and newer publications. , 2014, The International journal of oral & maxillofacial implants.

[3]  R. Tannenbaum,et al.  The roles of titanium surface micro/nanotopography and wettability on the differential response of human osteoblast lineage cells. , 2013, Acta biomaterialia.

[4]  David L Cochran,et al.  Osteoblast maturation and new bone formation in response to titanium implant surface features are reduced with age , 2012, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[5]  C. Vanhove,et al.  Utilizing micro-computed tomography to evaluate bone structure surrounding dental implants: a comparison with histomorphometry. , 2013, Journal of biomedical materials research. Part B, Applied biomaterials.

[6]  S. Cho,et al.  Oxygen effects on the mechanical properties and lattice strain of Ti and Ti-6Al-4V , 2011 .

[7]  K. Suzuki,et al.  Effects of surface roughness of titanium implants on bone remodeling activity of femur in rabbits. , 1997, Bone.

[8]  D. Landolt,et al.  Differential regulation of osteoblasts by substrate microstructural features. , 2005, Biomaterials.

[9]  B. Boyan,et al.  Implant Materials Generate Different Peri-implant Inflammatory Factors , 2015, Spine.

[10]  Barbara D Boyan,et al.  Additively manufactured 3D porous Ti-6Al-4V constructs mimic trabecular bone structure and regulate osteoblast proliferation, differentiation and local factor production in a porosity and surface roughness dependent manner , 2014, Biofabrication.

[11]  F Rupp,et al.  High surface energy enhances cell response to titanium substrate microstructure. , 2005, Journal of biomedical materials research. Part A.

[12]  N. Spencer,et al.  The role of nanostructures and hydrophilicity in osseointegration: In-vitro protein-adsorption and blood-interaction studies. , 2015, Journal of biomedical materials research. Part A.

[13]  Lars Sennerby,et al.  Early tissue response to titanium implants inserted in rabbit cortical bone , 1993 .

[14]  R. Mowry Biological Stains. A Handbook on the Nature and Uses of the Dyes Employed in the Biological Laboratory , 1963 .

[15]  Michiel Mulier,et al.  Bone regeneration performance of surface-treated porous titanium. , 2014, Biomaterials.

[16]  A Piattelli,et al.  Direct laser metal sintering as a new approach to fabrication of an isoelastic functionally graded material for manufacture of porous titanium dental implants. , 2008, Dental materials : official publication of the Academy of Dental Materials.

[17]  A. Hoffmann,et al.  Osseointegration by bone morphogenetic protein-2 and transforming growth factor beta2 coated titanium implants in femora of New Zealand white rabbits , 2011, Indian journal of orthopaedics.

[18]  F. Schwarz,et al.  Effects of surface hydrophilicity and microtopography on early stages of soft and hard tissue integration at non-submerged titanium implants: an immunohistochemical study in dogs. , 2007, Journal of periodontology.

[19]  L. Breschi,et al.  Effects of acid-etching solutions on human enamel and dentin. , 1995, Quintessence international.

[20]  Takashi Nakamura,et al.  Bioactive Ti metal analogous to human cancellous bone: Fabrication by selective laser melting and chemical treatments. , 2011, Acta biomaterialia.

[21]  B. Boyan,et al.  Titanium surface characteristics, including topography and wettability, alter macrophage activation. , 2016, Acta biomaterialia.

[22]  A. Virdi,et al.  Limitations of using micro‐computed tomography to predict bone–implant contact and mechanical fixation , 2012, Journal of microscopy.

[23]  S. Ferguson,et al.  Biomechanical comparison of the sandblasted and acid-etched and the machined and acid-etched titanium surface for dental implants. , 2002, Journal of biomedical materials research.

[24]  Ralf Schumacher,et al.  Bone regeneration by the osteoconductivity of porous titanium implants manufactured by selective laser melting: a histological and micro computed tomography study in the rabbit. , 2013, Tissue engineering. Part A.

[25]  R. G. Richards,et al.  Animal models for implant biomaterial research in bone: a review. , 2007, European cells & materials.

[26]  B. Boyan,et al.  A review on the wettability of dental implant surfaces II: Biological and clinical aspects. , 2014, Acta biomaterialia.

[27]  Manjeet Mapara,et al.  Rabbit as an animal model for experimental research , 2012, Dental research journal.

[28]  M. Heiland,et al.  Cytocompatibility of Direct Laser Interference-patterned Titanium Surfaces for Implants. , 2017, In Vivo.

[29]  M. D. del Cerro,et al.  Stevenel's Blue, an excellent stain for optical microscopical study of plastic embedded tissues. , 1980, Microscopica acta.

[30]  K. Sandhage,et al.  Differential responses of osteoblast lineage cells to nanotopographically-modified, microroughened titanium-aluminum-vanadium alloy surfaces. , 2012, Biomaterials.

[31]  P. Layrolle,et al.  Enhanced osseointegration of titanium implants with nanostructured surfaces: an experimental study in rabbits. , 2015, Acta biomaterialia.

[32]  F. Pezzetti,et al.  Zirconium oxide coating improves implant osseointegration in vivo. , 2008, Dental materials : official publication of the Academy of Dental Materials.

[33]  Jae-ho Yang,et al.  A histomorphometric study of dental implants with different surface characteristics , 2010, The journal of advanced prosthodontics.

[34]  J. Katz,et al.  Population-Based Rates of Revision of Primary Total Hip Arthroplasty: A Systematic Review , 2010, PloS one.

[35]  B. Boyan,et al.  Imaging analysis of the interface between osteoblasts and microrough surfaces of laser‐sintered titanium alloy constructs , 2018, Journal of microscopy.

[36]  B. Boyan,et al.  Novel Osteogenic Ti-6Al-4V Device For Restoration Of Dental Function In Patients With Large Bone Deficiencies: Design, Development And Implementation , 2016, Scientific Reports.

[37]  P. Thomsen,et al.  Early tissue response to titanium implants inserted in rabbit cortical bone , 1993 .

[38]  P. Ullrich,et al.  Osteoblasts exhibit a more differentiated phenotype and increased bone morphogenetic protein production on titanium alloy substrates than on poly-ether-ether-ketone. , 2012, The spine journal : official journal of the North American Spine Society.

[39]  A. Wennerberg,et al.  Spontaneously formed nanostructures on titanium surfaces. , 2013, Clinical oral implants research.

[40]  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.

[41]  Yang Yang,et al.  Comparison of conventional reconstruction plate versus direct metal laser sintering plate: an in vitro mechanical characteristics study , 2017, Journal of Orthopaedic Surgery and Research.

[42]  M. McCracken,et al.  Dental implant materials: commercially pure titanium and titanium alloys. , 1999, Journal of prosthodontics : official journal of the American College of Prosthodontists.

[43]  P. Layrolle,et al.  Histomorphometric analysis of the osseointegration of four different implant surfaces in the femoral epiphyses of rabbits. , 2008, Clinical oral implants research.

[44]  B. Boyan,et al.  Integrin α2β1 plays a critical role in osteoblast response to micron-scale surface structure and surface energy of titanium substrates , 2008, Proceedings of the National Academy of Sciences.

[45]  B. Boyan,et al.  Direct and indirect effects of microstructured titanium substrates on the induction of mesenchymal stem cell differentiation towards the osteoblast lineage. , 2010, Biomaterials.

[46]  Kozo Osakada,et al.  Microstructure and mechanical properties of pure titanium models fabricated by selective laser melting , 2004 .

[47]  L. Murr,et al.  Microstructure and mechanical behavior of Ti-6Al-4V produced by rapid-layer manufacturing, for biomedical applications. , 2009, Journal of the mechanical behavior of biomedical materials.

[48]  R. Ladda,et al.  Smoking and dental implants , 2012, Journal of International Society of Preventive & Community Dentistry.

[49]  C. Maniatopoulos,et al.  An improved method for preparing histological sections of metallic implants. , 1986, The International journal of oral & maxillofacial implants.

[50]  J. Buckwalter,et al.  Use of animal models in musculoskeletal research. , 1998, The Iowa orthopaedic journal.

[51]  P Nilsson,et al.  Biomechanical characterization of osseointegration during healing: an experimental in vivo study in the rat. , 1997, Biomaterials.

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

[53]  B. Boyan,et al.  Implant osseointegration and the role of microroughness and nanostructures: lessons for spine implants. , 2014, Acta biomaterialia.

[54]  J. Granjeiro,et al.  Basic research methods and current trends of dental implant surfaces. , 2009, Journal of biomedical materials research. Part B, Applied biomaterials.

[55]  P. Ullrich,et al.  Rough titanium alloys regulate osteoblast production of angiogenic factors. , 2013, The spine journal.

[56]  H. Aro,et al.  Low BMD affects initial stability and delays stem osseointegration in cementless total hip arthroplasty in women , 2012, Acta orthopaedica.

[57]  U. Holzwarth,et al.  Effect of surface finish on the osseointegration of laser-treated titanium alloy implants. , 2004, Biomaterials.

[58]  M. Meyers,et al.  Biomedical applications of titanium and its alloys , 2008 .

[59]  Michel Dard,et al.  Nanostructures and hydrophilicity influence osseointegration: a biomechanical study in the rabbit tibia. , 2014, Clinical oral implants research.

[60]  F. He,et al.  Mechanical and histomorphometric evaluations of rough titanium implants treated with hydrofluoric acid/nitric acid solution in rabbit tibia. , 2011, The International journal of oral & maxillofacial implants.

[61]  R. van Noort,et al.  Direct Metal Laser Sintering Titanium Dental Implants: A Review of the Current Literature , 2014, International journal of biomaterials.

[62]  F. He,et al.  Bone responses to titanium implants surface-roughened by sandblasted and double etched treatments in a rabbit model. , 2008, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[63]  Abraham Marmur,et al.  A review on the wettability of dental implant surfaces I: theoretical and experimental aspects. , 2014, Acta biomaterialia.

[64]  B. Boyan,et al.  Osteoblast Lineage Cells Can Discriminate Microscale Topographic Features on Titanium–Aluminum–Vanadium Surfaces , 2014, Annals of Biomedical Engineering.

[65]  Ana Mellado-Valero,et al.  Effects of diabetes on the osseointegration of dental implants. , 2007, Medicina oral, patologia oral y cirugia bucal.

[66]  M. McGuire,et al.  Commentary: from normal scientific progress to game changers: the impact on periodontal clinical practice. , 2014, Journal of periodontology.

[67]  R. Noort Titanium: The implant material of today , 1987 .

[68]  R. Tannenbaum,et al.  The effects of combined micron-/submicron-scale surface roughness and nanoscale features on cell proliferation and differentiation. , 2011, Biomaterials.