Dhaliwal, J.S., Marulanda, J., Li, J. et al. In vitro comparison of two titanium dental implant surface treatments: 3M™ESPE™ MDIs versus Ankylos®.
Int J Implant Dent 3, 27 (2017). https://doi.org/10.1186/s40729-017-0083-5
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Received: 19 January 2017
Accepted: 22 May 2017
Published: 27 June 2017
DOI: https://doi.org/10.1186/s40729-017-0083-5
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
Faculty of Dentistry, McGill University, Montreal, Quebec, Canada
Jagjit Singh Dhaliwal, Juliana Marulanda, Sharifa Alebrahim, Jocelyne Sheila Feine & Monzur Murshed
PAPRSB, Institute of Health Sciences, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong, BE1410, Brunei Darussalam
Jagjit Singh Dhaliwal
Faculty of Medicine, McGill University, Montreal, Quebec, Canada
Jingjing Li & ...
The authors would like to thank Prof. Georgios Romanos, School of Dental Medicine, Dept. of Periodontology, Stony Brook University, Stony Brook, NY, USA, for his contribution to the study. We would also like to thank 3M ESPE and Dentsply Friadent for providing the implant disks. This work is a part of the thesis submitted by JD for attaining a PhD degree at the Faculty of Dentistry, McGill Univers...
Wennerberg A, Albrektsson T. Effects of titanium surface topography on bone integration: a systematic review. Clin Oral Implants Res. 2009;20 Suppl 4:172–84.
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Our results demonstrate that both implant surfaces are conducive for osteoblastic cell attachment, proliferation, and mineralization. However, 3M™ESPE™ MDI surface shows more pronounced effects on cell proliferation, which may in turn facilitate better osseointegration by enhancing ECM mineralization. Our ongoing research will provide further information on how implant surfaces may affect cell...
ALPL is a late osteogenic marker, which is essential for normal bone mineralization. ALPL-deficient osteoblasts fail to mineralize in culture. Considering that there was no significant difference in the relative ALPL activity in cells grown on two surfaces, it is unlikely that the surface property of the disks affected cell differentiation. This observation does not support the findings of Davies ...
MC3T3-E1 cells have been extensively used in numerous cell culture experiments as a model for osteoblasts [36]. Under differentiating conditions, e.g., in the presence of ascorbic acid and β-glycerol phosphate, these cells upregulate the osteogenic markers and, more importantly, promote the deposition of calcium phosphate minerals within and around the collagen-rich extracellular matrix (ECM). In...
In the current study, we used an in vitro cell culture system to evaluate the biocompatibility of two implant materials with different surface topography. Our objective was to establish the osseointegration potential of MDIs versus an established regular implant. Disks prepared from the implant material were coated with gelatin to grow cells, and proliferation and osteogenic differentiation parame...
Pre-osteoblastic MC3T3-E1 cells were plated on each implant disk (40,000 cells/disk) and were differentiated with mineralization medium for 12 days. Quantification of cells after nuclear staining by H33258 revealed an increased number of cells on the 3M™ESPE™ MDI disks (Fig. 5a). Measurement of cell viability by the reduction of Alamar blue® after 3 days of culture of MC3T3-E1 cells furthe...
The variable sizes of the implant disks obtained from two different manufacturers demanded an innovative culture system to ensure equal surface areas on both disks for the cell culture experiments. We achieved this by attaching constant diameter (5 mm) plastic cylinders to the disk surface. Disks were sterilized with absolute alcohol, and polystyrene cloning cylinders (Sigma) were attached onto t...
While performing the experiments, JM (first co-author) was not aware of the sources/manufacturers of the disks, which were identified by their size (small and large) only. At the end of the analyses, each disk’s manufacturer was revealed to her by JSD (first co-author).
Protein samples from the transfected cells were prepared in 1× SDS gel-loading buffer (Laemmli buffer) without adding β-mercaptoethanol and quantified using the Pierce™ Coomassie Plus Assay kit (Thermo Scientific, Rockford, IL, USA). Without heat denaturation, equal amount of protein samples (50 μg) were loaded on a 10% SDS-polyacrylamide gel. After electrophoresis, the gel was incubated in N...
Titanium disks made up with the same materials and surface characteristics as those with the original implants were obtained from the respective manufacturers. Two types of disks were used; the small disks represented 3M™ESPE™ MDI implants, while the large disks represented Ankylos®, Dentsply Friadent implants. A total of 10 disks of each brand were used for the study.
Disks were sterilized ...
Despite the advantages of the MDI, evidence on their potential for osseointegration and long-term success is lacking. [15,16,17,18]. Newer implant systems entering the market must be studied first in vitro and then in vivo with animal models followed by human studies to demonstrate their osseointegration capability.
Modifications of implant surface properties have been shown to have a positive in...
Prosthetic devices are often used as surrogates for missing skeletal and dental elements. These devices are in close contact with the surrounding tissues, and their functionality and stability are critically dependent on the successful integration within the tissue’s extracellular matrix (ECM). The surface of the implanted device directly interacts with cell and extracellular milieu and influenc...
An ideal implant should have a surface that is conducive to osseointegration. In vitro cell culture studies using disks made of same materials and surface as of implants may provide useful information on the events occurring at the implant-tissue interface. In the current study, we tested the hypothesis that there is no difference in the proliferation and differentiation capacities of osteoblastic...
Figure 5. a Florescence microscopy showing H33258-stained MC3T3-E1 cells on Ankylos® and 3M™ESPE™ MDI disks. Although equal numbers of cells were plated, after 12 days of culture, more cells were detected on the 3M™ESPE™ MDI disks. b Increased Alamar blue® reduction in MC3T3-E1 cells seeded on 3M™ESPE™ MDI disks when compared to cells cultured on Ankylos®. c Increase...
Figure 4. a C2C12 cells (control) and pBMP-2-transfected C2C12 cells were seeded on a 24-well plate (50,000 cell/well) and cultured in DMEM medium for 48 h. ALPL assay showing upregulated ALPL activity in the BMP-2-transfected C2C12 cells. b Cell extracts of C2C12 cells and pBMP-2-transfected cells were run on a 10% SDS-PAGE under non-denaturing conditions. The gel was then stained with ...
Figure 3. Increased proliferation of C2C12 cells grown on 3M™ESPE™ MDI disks in comparison to the cells grown on the Ankylos® disks untreated and treated with bone morphogenetic protein-BMP-2
Figure 2. Implant surface characterization under SEM. Increased surface roughness in the 3M™ESPE™ MDI dental implants when compared to Ankylos® implants
Figure 1. Preparation of specimens. Small disks represent 3M™ESPE™ MDI implants, and large disks represent Ankylos® implants. Note that the attachment of polystyrene rings ensures the area of culture remains constant regardless of the disk size
MacDonald et al. have shown that wettability, i.e., hydrophilic surfaces support cell interactions and biological fluids better than the hydrophobic surfaces. It has also been shown that roughening the titanium surface improves hydrophilicity. In addition, many authors have stated that rougher surfaces promote differentiation, growth and attachment of bone cells, and higher production of gro...
Because of their pluripotency, these cells have been considered as a type of mesenchymal stem cells. It has been shown that when treated with BMPs, these cells readily upregulate many key osteoblast markers including RUNX2, OSX, osteocalcin, and alkaline phosphatase (ALPL). In the current study, we used C2C12 cells that were treated with BMP-2 or stably transfected with a BMP-2 expression v...
Discussion
In the current study, we used an in vitro cell culture system to evaluate the biocompatibility of two implant materials with different surface topography. Our objective was to establish the osseointegration potential of MDIs versus an established regular implant. Disks prepared from the implant material were coated with gelatin to grow cells, and proliferation and osteogenic diff...
Alkaline phosphatase immunostaining and assay
BMP-2-transfected C2C12 cells were fixed in 4% PFA for 15 min, and then blocked with 5% bovine serum albumin (Fisher, Pittsburgh, PA, USA) in Tris buffered saline-0.025%Triton for 30 min at room temperature, followed by overnight incubation with an anti-mouse alkaline phosphatase antibody (R&D systems, Minneapolis, MN, USA). Detection was done by ...
The reduction of Alamar blue was measured at 560 nm (reference wavelength 610 nm) after 5-h incubation at 37 °C using a microplate reader (Infinite 200, Tecan).
Generation of BMP2 expressing C2C12 cells
C2C12 cells were electroporated together with 0.4 μg of a BMP-2 expression vector (a kind gift from Dr. Katagiri) and 0.1 μg of pCMV-Tag, which expresses a neomycin-resistance gene. Culture...
Results
Ring culture technique
The variable sizes of the implant disks obtained from two different manufacturers demanded an innovative culture system to ensure equal surface areas on both disks for the cell culture experiments. We achieved this by attaching constant diameter (5 mm) plastic cylinders to the disk surface. Disks were sterilized with absolute alcohol, and polystyrene cloning cyli...
Methods
Implant disks
Titanium disks made up with the same materials and surface characteristics as those with the original implants were obtained from the respective manufacturers. Two types of disks were used; the small disks represented 3M™ESPE™ MDI implants, while the large disks represented Ankylos®, Dentsply Friadent implants. A total of 10 disks of each brand were used for the stud...
Background
Prosthetic devices are often used as surrogates for missing skeletal and dental elements. These devices are in close contact with the surrounding tissues, and their functionality and stability are critically dependent on the successful integration within the tissue’s extracellular matrix (ECM). The surface of the implanted device directly interacts with cell and extracellular milie...
In vitro comparison of two titanium dental implant surface treatments: 3M™ESPE™ MDIs versus Ankylos®
Abstract
Background
An ideal implant should have a surface that is conducive to osseointegration. In vitro cell culture studies using disks made of same materials and surface as of implants may provide useful information on the events occurring at the implant-tissue interface. In the curr...