Garvin, Kevin L.

Kim, Bongok

Koepsell, Laura

Miralami, Raheleh

Namavar, Fereydoon

Premaraj, Thyagaseely Sheela

Sharp, J. Graham

Thiele, Geoffrey M.

English

University of Nebraska Medical Center Research: DigitalCommons@UNMC

University of Nebraska Medical Center DigitalCommons@UNMC Posters and Presentations: Orthopaedic Surgery & Rehab Orthopaedic Surgery & Rehab 2013 Comparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline Titanium Raheleh Miralami University of Nebraska Medical Center, raheleh.miralami@unmc.edu Laura Koepsell University of Nebraska Medical Center Thyagaseely Sheela Premaraj University of Nebraska Medical Center, sheelapremaraj@unmc.edu Bongok Kim University of Nebraska - Lincoln Geoffrey M. Thiele University of Nebraska Medical Center, gthiele@unmc.edu See next page for additional authors Follow this and additional works at: https://digitalcommons.unmc.edu/com_ortho_pres  Part of the Orthopedics Commons Recommended Citation Miralami, Raheleh; Koepsell, Laura; Premaraj, Thyagaseely Sheela; Kim, Bongok; Thiele, Geoffrey M.; Sharp, J. Graham; Garvin, Kevin L.; and Namavar, Fereydoon, "Comparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline Titanium" (2013). Posters and Presentations: Orthopaedic Surgery & Rehab. 2. https://digitalcommons.unmc.edu/com_ortho_pres/2 This Conference Proceeding is brought to you for free and open access by the Orthopaedic Surgery & Rehab at DigitalCommons@UNMC. It has been accepted for inclusion in Posters and Presentations: Orthopaedic Surgery & Rehab by an authorized administrator of DigitalCommons@UNMC. For more information, please contact digitalcommons@unmc.edu. Authors Raheleh Miralami, Laura Koepsell, Thyagaseely Sheela Premaraj, Bongok Kim, Geoffrey M. Thiele, J. Graham Sharp, Kevin L. Garvin, and Fereydoon Namavar This conference proceeding is available at DigitalCommons@UNMC: https://digitalcommons.unmc.edu/com_ortho_pres/2 DISCUSSIONMETHODS (con’t)INTRODUCTIONMETHODSComparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline TitaniumR. Miralami1, L. Koepsell1, T. Premaraj2, B. Kim3, G.M. Thiele4, J.G. Sharp5, K.L. Garvin1, F. Namavar11Department of Orthopaedic Surgery and Rehabilitation, UNMC, Omaha, NE;  2Department of Growth & Development, University of Nebraska-Lincoln, Lincoln, NE;  3Department of Adult Restorative Dentistry, University of Nebraska - Lincoln, Lincoln, NE; 4Department of Internal Medicine – Rheumatology, UNMC, Omaha, NE; 5Department of Genetics, Cell Biology & Anatomy, UNMC, Omaha, NE Kurtz, et. al.[1], predict that by 2010 hip and knee replacements will number over one million, and by 2030 we will see 4.5 million annually.Primary concerns of orthopaedic devices are wear failure of biointegration [2,3].Thus, there is a need to accommodate such overload by the development of biocompatible surfaces with superior cell adhesion, survival, growth and proliferation, which is the overall objective of our lab.It was shown that the biocompatibility of titanium (Ti) metal is due to the presence of a thin native sub-stoichiometric titanium-oxide (TiO2) layer that enhances the adsorption of mediating proteins on the surface [4].The present study evaluates the adhesion, survival and growth of cells on the engineered nanocrystalline TiO2 and Ti produced by ion-beam-assisted-deposition (IBAD) technique, and compares them with microcrystalline Ti (orthopaedic grade). Nanocrystalline surfaces (TiO2 and  Ti) are superior in supporting growth, adhesion, and proliferation as compared to microcrystalline Ti due to the nano-crystal film characteristics that affect the wettability and mechanical properties of the coatings. Nano-engineered TiO2 is superior to both nano and biomedical grade Ti because of the improved quality of the oxide layer by employing an IBAD technique.  Osteoblastic cells deposit calcium in order to support bone construction. The cells cultured on the nano surfaces showed more calcium deposition (brighter red color in alizarin red staining), which implies a higher degree of the differentiation of the cells. Therefore, enhanced bone formation ability can be expected from the engineered nano structured surfaces. Method of Nano-fabrication (IBAD Technique):The process combines physical vapor deposition with concurrent ion beam bombardment in a high vacuum environment. The nano-structure films produced by IBAD (with 1 to 70nm grain size) possess combined properties of super hardness and complete wetting behavior.Cell cultureThe SAOS-2 cell line was used, which is a human osteosarcoma cell line, and maintains the cellular features of the osteoblasts with the ability to deposit extracellular matrix and mineralization.Comparing cell adhesion and growthCell counting was done using DAPI (6-diamidino-2-phenylindole) fluorescent staining and ImageJ software.Comparing cell morphologyImmunofluorescence staining was done to monitor actin stress fiber shapes.Comparing calcium depositionAlizarin red assay was used to detect calcium compounds on surfaces using a standard plate reader.REFERENCES1. S. Kurtz, et. al, American Academy of Orthopaedic Surgeons,  Chicago, IL, 2006.2. L.G. Harris, et. al. Injury. Int. J. Care Injured. 37,S3-S14, 2006.3. M. Viceconti, et. al. Med Biol Eng Comput. 42, 222-229, 2004.4. F. Namavar, R. Sabirianov; et. al. Orthopaedic Research Society, San Francisco, Feb 2012. Fig. 3: Atomic Force Microscopy Images of Nano-structure TiO2 Films Deposited on Silicon Substrates Using IBAD with Different BeamsFig. 6: Comparing Calcium Deposition on Different Substrates Using Alizarin Red Assay for 7 and 14  DaysFig. 5: Number of Nuclei by DAPIFigure 5 shows a greater number of DAPI-stained cells on nanocrystalline TiO2 and Ti compared to microcrystalline Ti, which indicates more adhesion and growth on nano surfaces.Figure 6 indicates the difference in calcium deposition on nano-engineered TiO2 compared to nanocrystalline Ti and orthopaedic grade Ti. Besides DAPI staining, actin stress fiber shapes were also monitored (Figure 7) because by only observing nuclei, it is impossible to know if cells are healthy and prolific on the surface or not.Figure 8 shows cell morphology on different substrates. Cells look more extended and healthier on nano-surfaces (TiO2 and Ti) compared to microcrystalline Ti.Fig. 1:IBAD SystemFig. 2:Schematic of IBAD ProcessorFig. 4: Atomic Force Microscopy Images(5µm Scan Size) of Ion Beam Deposited Nanocrystalline Ti and TiO2 Deposited at Room TemperatureFig. 7: Merged Actin and DAPI Stained Cells on Nanocrystalline TiO2 and Microcrystalline Ti, Respectively   7 Days 14 Days Fig. 8: Comparing Cell Adhesion on Nanocrystalline TiO2, Ti, and Biomedical Grade of Ti by Fluorescence Images Microscopy. 50000 Cells Were Incubated for 48 Hours. Images (a), (b) and (c) are DAPI and (d), (e) and (f) are Actin Stain Experiments  RESULTSCONCLUSIONACKNOWLEDGMENTThis work was supported by department of energy, material science smart coating (DE-SC0005318).  Argon RMS 64.4 nm Nitrogen RMS 19.4 nm Oxygen RMS 20.4 nm    NanoCrystalline Titanium  MicroCrystalline Titanium  

Comparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline Titanium

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Comparing Biocompatibility of Nanocrystalline Titanium and Titanium-Oxide with Microcrystalline Titanium

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