Fig. 5. Osteoblasts with an orientation tendency after 24 h of rotation (phallacidin fluorescence staining). On the left side with 200× and on the right side with 400× magnification. The yellow arrows show the orientation of the cells. The red arched arrow within the coloured circle shows the direction of rotation. The dashed white line oriented to the right stands for the resulting centrifug...
Fig. 4. Randomly orientated osteoblasts without influence of rotation (phallacidin fluorescence staining). On the left side with 200× and on the right side with 400× magnification. The white X on the coloured circle marks the location upon the plate where the osteoblasts were located. The red X marks the centre of the plate
Fig. 4. Randomly orientated osteoblasts without influence of rotati...
centrifugal force and the glass plates’ dimensions. For example, at a distance of 25 mm from the centre of the upper plate, the shear forces’ value is 8.33 dyn/cm2, together with an additional centrifugal force that has a value of 0.55 dyn/cm2
Fig. 3. Diagram for visualisation of the calculation of shear stress rates taking into account the centrifugal force and the glass plates’ dim...
earing gap and bottom plate are shown on the left side; rotation speed = 200 rpm; colour code bar (left edge) showing shear force values [Pa] [1 Pa = 10 dyn/cm2]; flow direction presented by arrows
Fig. 2. Side view of a computerized simulation, showing the flow chambers’ lower compartment and the flow profile in between the two plates; shearing gap and bottom plate are shown on ...
Fig. 1. Three-dimensional illustration (a–e) and photography (f) of the experimental setup with the components marked numerical. a as the lower plate); 2 Rotating glass panel [60 mm diameter (cell bearing)]; 3 Titanium axis. b4 Liquid medium (red). cGearwheel with set screw. eng ring with additional set screw
Fig. 1. Three-dimensional illustration (a–e) and photography (f) of the experim...
Culture medium/additives
Manufacturer
Order no.
Concentration
Dulbecco’s modified Eagle medium (DMEM) with l-glutamine, plus 4.5 g glucose,...
Component
Manufacturer
Order no.
Large petri dish
Becton Dickinson, Franklin Lakes, NJ, USA
...
Kämmerer, P.W., Thiem, D.G.E., Alshihri, A. et al. Cellular fluid shear stress on implant surfaces—establishment of a novel experimental set up.
Int J Implant Dent 3, 22 (2017). https://doi.org/10.1186/s40729-017-0085-3
Download citation
Received: 27 February 2017
Accepted: 22 May 2017
Published: 31 May 2017
DOI: https://doi.org/10.1186/s40729-017-0085-3
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...
Correspondence to
D. G. E. Thiem.
Department of Oral and Maxillofacial Surgery, Facial Plastic Surgery, University Medical Centre Rostock, Schillingallee 35, 18057, Rostock, Germany
P. W. Kämmerer & D. G. E. Thiem
Department of Prosthetic and Biomaterial Sciences, King Saud University, Riyadh, Saudi Arabia
A. Alshihri
Harvard School of Dental Medicine, Boston, MA, USA
A. Alshihri
Department of Oral and Maxillofacial Surge...
The authors thank the Department of Hydraulic Machines, Faculty of Mechanical Engineering, Technical University of Munich, Germany, for helping with the computerised simulations.
Nothing to declare
The datasets supporting the conclusions of this article are available at the repository of the University Medical Centre Mainz, Germany, and can be provided on request.
PK had substantial contributio...
Papadaki M, Eskin SG. Effects of fluid shear stress on gene regulation of vascular cells. Biotechnol Prog. 1997;13(3):209–21.
James NL, Harrison DG, Nerem RM. Effects of shear on endothelial cell calcium in the presence and absence of ATP. FASEB J. 1995;9(10):968–73.
Kämmerer PW, Lehnert M, Al-Nawas B, Kumar VV, Hagmann S, Alshihri A, et al. Osseoconductivity of a specific streptavidin-biot...
Hughes-Fulford M. Signal transduction and mechanical stress. Sci STKE. 2004;2004(249):RE12.
Ruel J, Lemay J, Dumas G, Doillon C, Charara J. Development of a parallel plate flow chamber for studying cell behavior under pulsatile flow. ASAIO J. 1995;41(4):876–83.
Kazakidi A, Sherwin SJ, Weinberg PD. Effect of Reynolds number and flow division on patterns of haemodynamic wall shear stress near br...
Becker J, Kirsch A, Schwarz F, Chatzinikolaidou M, Rothamel D, Lekovic V, et al. Bone apposition to titanium implants biocoated with recombinant human bone morphogenetic protein-2 (rhBMP-2). A pilot study in dogs. Clin Oral Investig. 2006;10(3):217–24.
Hung CT, Allen FD, Pollack SR, Brighton CT. What is the role of the convective current density in the real-time calcium response of cultured bon...
Ehrlich PJ, Lanyon LE. Mechanical strain and bone cell function: a review. Osteoporos Int. 2002;13(9):688–700.
Vaughan TJ, Haugh MG, Mcnamara LM. A fluid-structure interaction model to characterize bone cell stimulation in parallel-plate flow chamber systems. J R Soc Interface. 2013;10(81):20120900.
Weinbaum S, Cowin SC, Zeng Y. A model for the excitation of osteocytes by mechanical loading-in...
To create fluid shear stress under in vitro conditions, several flow chambers have been developed in the past. The experimental setup of the flow chamber in the centre of this study offers advantages such as simplicity to assemble and ease of use as well as the creation of reproducible fluid shear forces on cells. Due to the new design, different cell types could be simultaneously analysed under r...
Besides, in the model reported in this study, microscopic examinations are possible after completing the experiment only. Nevertheless, an advantage of the new flow chamber is the possibility of testing different cell colonies simultaneously in one single experiment by placing cells in different radial locations on the spinning disc. Due to the current flow gradient from the centre to the peripher...
Due to the fact that constant flows were generated within the parallel flow chamber only, the situations of in vitro experiments differ from in vivo setting where dynamic flow profiles are particular [33]. As the constant laminar flow profile is not physiological in bones [34], vessels and other tissues [35], the informative value of the experimental setting is limited but it could be used for var...
The aim of this study was to establish a new FSS model that is easy to use as well as simple to assemble in order to create reproducible fluid shear forces on cells close to implant material surfaces. Todays’ commonly used commercial flow devices differ in geometry and function, which makes comparisons between experiments difficult [4, 10, 26, 27]. The benefits of this novel testing device are r...
in which ρ = density, h = height, ω = angular velocity and r = radius.
Figure 3 shows the respective physical force and its dependence on a bigger radius and higher rotational speed. The results of this study indicate that the centrifugal force represents only a little proportion of effective forces. Hence, the centrifugal forces’ impacts on the tested cells are considered to be insignifican...
Our analysis was focused on two main aspects:
Simulation of the fluid flow characteristics as well as quantification of the arising shear forces at the plate/plate flow chamber with reliable reproducibility
Assessment of the impact of fluid shear stress on osteoblast cells in terms of altered cell morphology and intracellular structural changes
The computational fluid dynamic analysis and the q...
For constant and fully developed laminar flow between the two parallel plates, the magnitude of the wall shear stress (τ) in between was calculated by formula 1:
in which η is the dynamic fluid viscosity (dyn/cm2), r is the radius of the plate (cm), ω stands for angular velocity and H for height (vertical distance in between the two plates).
To get information whether the flow is laminar or t...
A three-dimensional illustration and photography of the plate/plate flow chamber model is shown in Fig. 1. A detailed list of used parts can be found in Appendix 1.
The circulation within the flow chamber was generated by an externally attached electric motor, which rotates up to 500 rounds per minute (rpm). A commercial grade 4 pure medical titanium gear shaft (length = 40 mm, diameter =...
Therefore, the aim of the present study was to establish a new cell chamber model for FSS simulation and stimulation. In addition to its ease of use, the reported model in this study should meet the requirements of a simple design, generating reproducible flow characteristics next to laminar flows and clearly defined flow gradients on implant surfaces.
Cells can be influenced by different mechanostimuli, which lead to an activation of cellular and inter-cellular responses. These reactions may be caused by either a direct stimulation of the cell body (mechanoreception) or indirect cellular stimulation (response) [1,2,3]. Extracellular fluid movement induces fluid shear stress (FSS) that can result in different cellular processes including prolife...
Mechanostimuli of different cells can affect a wide array of cellular and inter-cellular biological processes responsible for dental implant healing. The purpose of this in vitro study was to establish a new test model to create a reproducible flow-induced fluid shear stress (FSS) of osteoblast cells on implant surfaces.
As FSS effects on osteoblasts are detectable at 10 dyn/cm2, a custom-made f...
Fig. 5. Osteoblasts with an orientation tendency after 24 h of rotation (phallacidin fluorescence staining). On the left side with 200× and on the right side with 400× magnification. The yellow arrows show the orientation of the cells. The red arched arrow within the coloured circle shows the direction of rotation. The dashed white line oriented to the right stands for the resulting centrifug...
Fig. 4. Randomly orientated osteoblasts without influence of rotation (phallacidin fluorescence staining). On the left side with 200× and on the right side with 400× magnification. The white X on the coloured circle marks the location upon the plate where the osteoblasts were located. The red X marks the centre of the plate
Fig. 4. Randomly orientated osteoblasts without influence of rotati...
to account the centrifugal force and the glass plates’ dimensions. For example, at a distance of 25 mm from the centre of the upper plate, the shear forces’ value is 8.33 dyn/cm2, together with an additional centrifugal force that has a value of 0.55 dyn/cm2
Fig. 3. Diagram for visualisation of the calculation of shear stress rates taking into account the centrifugal force and the glas...
e in between the two plates; shearing gap and bottom plate are shown on the left side; rotation speed = 200 rpm; colour code bar (left edge) showing shear force values [Pa] [1 Pa = 10 dyn/cm2]; flow direction presented by arrows
Fig. 2. Side view of a computerized simulation, showing the flow chambers’ lower compartment and the flow profile in between the two plates; shearing gap...
Fig. 1. Three-dimensional illustration (a–e) and photography (f) of the experimental setup with the components marked numerical. a lower plate); 2 Rotating glass panel [60 mm diameter (cell bearing)]; 3 Titanium axis. b4 Liquid medium (red). cwheel with set screw. e ring with additional set screw
Fig. 1. Three-dimensional illustration (a–e) and photography (f) of the experimental setup w...
Culture medium/additives
Manufacturer
Order no.
Concentration
Dulbecco’s modified Eagle medium (DMEM) with l-glutamine, plus 4.5 g glucose,...
Component
Manufacturer
Order no.
Large petri dish
Becton Dickinson, Franklin Lakes, NJ, USA
...
Kämmerer, P.W., Thiem, D.G.E., Alshihri, A. et al. Cellular fluid shear stress on implant surfaces—establishment of a novel experimental set up.
Int J Implant Dent 3, 22 (2017). https://doi.org/10.1186/s40729-017-0085-3
Download citation
Received: 27 February 2017
Accepted: 22 May 2017
Published: 31 May 2017
DOI: https://doi.org/10.1186/s40729-017-0085-3
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...
Correspondence to
D. G. E. Thiem.
Department of Oral and Maxillofacial Surgery, Facial Plastic Surgery, University Medical Centre Rostock, Schillingallee 35, 18057, Rostock, Germany
P. W. Kämmerer & D. G. E. Thiem
Department of Prosthetic and Biomaterial Sciences, King Saud University, Riyadh, Saudi Arabia
A. Alshihri
Harvard School of Dental Medicine, Boston, MA, USA
A. Alshihri
Department of Oral and Maxillofacial Surge...
The authors thank the Department of Hydraulic Machines, Faculty of Mechanical Engineering, Technical University of Munich, Germany, for helping with the computerised simulations.
Nothing to declare
The datasets supporting the conclusions of this article are available at the repository of the University Medical Centre Mainz, Germany, and can be provided on request.
PK had substantial contributio...
Papadaki M, Eskin SG. Effects of fluid shear stress on gene regulation of vascular cells. Biotechnol Prog. 1997;13(3):209–21.
James NL, Harrison DG, Nerem RM. Effects of shear on endothelial cell calcium in the presence and absence of ATP. FASEB J. 1995;9(10):968–73.
Kämmerer PW, Lehnert M, Al-Nawas B, Kumar VV, Hagmann S, Alshihri A, et al. Osseoconductivity of a specific streptavidin-biot...
Hughes-Fulford M. Signal transduction and mechanical stress. Sci STKE. 2004;2004(249):RE12.
Ruel J, Lemay J, Dumas G, Doillon C, Charara J. Development of a parallel plate flow chamber for studying cell behavior under pulsatile flow. ASAIO J. 1995;41(4):876–83.
Kazakidi A, Sherwin SJ, Weinberg PD. Effect of Reynolds number and flow division on patterns of haemodynamic wall shear stress near br...
Becker J, Kirsch A, Schwarz F, Chatzinikolaidou M, Rothamel D, Lekovic V, et al. Bone apposition to titanium implants biocoated with recombinant human bone morphogenetic protein-2 (rhBMP-2). A pilot study in dogs. Clin Oral Investig. 2006;10(3):217–24.
Hung CT, Allen FD, Pollack SR, Brighton CT. What is the role of the convective current density in the real-time calcium response of cultured bon...
Ehrlich PJ, Lanyon LE. Mechanical strain and bone cell function: a review. Osteoporos Int. 2002;13(9):688–700.
Vaughan TJ, Haugh MG, Mcnamara LM. A fluid-structure interaction model to characterize bone cell stimulation in parallel-plate flow chamber systems. J R Soc Interface. 2013;10(81):20120900.
Weinbaum S, Cowin SC, Zeng Y. A model for the excitation of osteocytes by mechanical loading-in...
To create fluid shear stress under in vitro conditions, several flow chambers have been developed in the past. The experimental setup of the flow chamber in the centre of this study offers advantages such as simplicity to assemble and ease of use as well as the creation of reproducible fluid shear forces on cells. Due to the new design, different cell types could be simultaneously analysed under r...
Besides, in the model reported in this study, microscopic examinations are possible after completing the experiment only. Nevertheless, an advantage of the new flow chamber is the possibility of testing different cell colonies simultaneously in one single experiment by placing cells in different radial locations on the spinning disc. Due to the current flow gradient from the centre to the peripher...
Due to the fact that constant flows were generated within the parallel flow chamber only, the situations of in vitro experiments differ from in vivo setting where dynamic flow profiles are particular [33]. As the constant laminar flow profile is not physiological in bones [34], vessels and other tissues [35], the informative value of the experimental setting is limited but it could be used for var...
The aim of this study was to establish a new FSS model that is easy to use as well as simple to assemble in order to create reproducible fluid shear forces on cells close to implant material surfaces. Todays’ commonly used commercial flow devices differ in geometry and function, which makes comparisons between experiments difficult [4, 10, 26, 27]. The benefits of this novel testing device are r...
in which ρ = density, h = height, ω = angular velocity and r = radius.
Figure 3 shows the respective physical force and its dependence on a bigger radius and higher rotational speed. The results of this study indicate that the centrifugal force represents only a little proportion of effective forces. Hence, the centrifugal forces’ impacts on the tested cells are considered to be insignifican...
Our analysis was focused on two main aspects:
Simulation of the fluid flow characteristics as well as quantification of the arising shear forces at the plate/plate flow chamber with reliable reproducibility
Assessment of the impact of fluid shear stress on osteoblast cells in terms of altered cell morphology and intracellular structural changes
The computational fluid dynamic analysis and the q...
For constant and fully developed laminar flow between the two parallel plates, the magnitude of the wall shear stress (τ) in between was calculated by formula 1:
in which η is the dynamic fluid viscosity (dyn/cm2), r is the radius of the plate (cm), ω stands for angular velocity and H for height (vertical distance in between the two plates).
To get information whether the flow is laminar or t...
A three-dimensional illustration and photography of the plate/plate flow chamber model is shown in Fig. 1. A detailed list of used parts can be found in Appendix 1.
The circulation within the flow chamber was generated by an externally attached electric motor, which rotates up to 500 rounds per minute (rpm). A commercial grade 4 pure medical titanium gear shaft (length = 40 mm, diameter =...
Therefore, the aim of the present study was to establish a new cell chamber model for FSS simulation and stimulation. In addition to its ease of use, the reported model in this study should meet the requirements of a simple design, generating reproducible flow characteristics next to laminar flows and clearly defined flow gradients on implant surfaces.
Cells can be influenced by different mechanostimuli, which lead to an activation of cellular and inter-cellular responses. These reactions may be caused by either a direct stimulation of the cell body (mechanoreception) or indirect cellular stimulation (response) [1,2,3]. Extracellular fluid movement induces fluid shear stress (FSS) that can result in different cellular processes including prolife...
Mechanostimuli of different cells can affect a wide array of cellular and inter-cellular biological processes responsible for dental implant healing. The purpose of this in vitro study was to establish a new test model to create a reproducible flow-induced fluid shear stress (FSS) of osteoblast cells on implant surfaces.
As FSS effects on osteoblasts are detectable at 10 dyn/cm2, a custom-made f...
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
Download citation
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.
Download references
Sudo H, Kodama HA, Amagai Y, Yamamoto S, Kasai S. In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol. 1983;96(1):191–8.
Novaes Jr AB, de Souza SL, de Barros RR, Pereira KK, Iezzi G, Piattelli A. Influence of implant surfaces on osseointegration. Braz Dent J. 2010;21(6):471–81.
MacDonald DE, Markovic B, Allen M, Som...
Buser D, Nydegger T, Oxland T, Cochran DL, Schenk RK, Hirt HP, et al. Interface shear strength of titanium implants with a sandblasted and acid-etched surface: a biomechanical study in the maxilla of miniature pigs. J Biomed Mater Res. 1999;45(2):75–83.
Li D, Ferguson SJ, Beutler T, Cochran DL, Sittig C, Hirt HP, et al. Biomechanical comparison of the sandblasted and acid-etched and the machine...
Griffitts TM, Collins CP, Collins PC. Mini dental implants: an adjunct for retention, stability, and comfort for the edentulous patient. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2005;100(5):e81–4.
Cooper LF, Zhou Y, Takebe J, Guo J, Abron A, Holmen A, et al. Fluoride modification effects on osteoblast behavior and bone formation at TiO2 grit-blasted c.p. titanium endosseous implants. B...
Branemark PI, Hansson BO, Adell R, Breine U, Lindstrom J, Hallen O, et al. Osseointegrated implants in the treatment of the edentulous jaw. Experience from a 10-year period. Scand J Plast Reconstr Surg Suppl. 1977;16:1–132.
Bulard RA, Vance JB. Multi-clinic evaluation using mini-dental implants for long-term denture stabilization: a preliminary biometric evaluation. Compend Contin Educ Dent. 20...
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...
Fig. 1. PRISMA flow diagram of literature search
Fig. 1. PRISMA flow diagram of literature search
Study (year)
Sample (size)
Treatment group (size)
Methodology
Parameter
...
Study (year)
Sample (size)
Treatment group (size)
Methodology
Parameter
...
Study (year)
Sample
Treatment group
Methodology
Parameter
Outco...
Item
Domain
5
Ethical statement
6
Study design
...
Study
Random sequence generation
Allocation concealment
Blinding of participants/personnel
Blinding of outcome assessment
...
Item
Domain
1
Abstract: structured summary of trial design, methods, results, and conclusions
Introduction
...
Luo, J., Miller, C., Jirjis, T. et al. The effect of non-steroidal anti-inflammatory drugs on the osteogenic activity in osseointegration: a systematic review.
Int J Implant Dent 4, 30 (2018). https://doi.org/10.1186/s40729-018-0141-7
Download citation
Received: 22 May 2018
Accepted: 13 July 2018
Published: 09 October 2018
DOI: https://doi.org/10.1186/s40729-018-0141-7
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 m...
Not applicable
Not applicable
Jie Denny Luo, Catherine Miller, Tamara Jirjis, Masoud Nasir, and Dileep Sharma declare that they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
College of Medicine & Dentistry, James Cook University, 14-88 McGregor Road, Smithfield, QLD, 4878, Australia
Jie Denny Luo, Tamara Jirjis, Masoud Nasir & Dileep Sharma
College of Public Health, Medical and Veterinary Sciences, James Cook University, 14-88 McGregor Road, Smithfield, QLD, 4878, Australia
Catherine Miller
You can also search for this author in
PubMed...
The authors acknowledge and are grateful for the help and preparation of manuscript by the supporting research supervisors: Dr. Ernest Jennings and Prof. Alan Nimmo.
The systematic review is funded by James Cook University College of Medicine and Dentistry as part of a Dentistry Honours Research Project.
A meta-analysis was not conducted for this systematic review. The critical analysis tables t...
Winnett B, Tenenbaum HC, Ganss B, Jokstad A. Perioperative use of non-steroidal anti-inflammatory drugs might impair dental implant osseointegration. Clin Oral Implants Res. 2016;27(2):E1–7.
Goodman SB, Ma T, Mitsunaga L, Miyanishi K, Genovese MC, Smith RL. Temporal effects of a COX-2-selective NSAID on bone ingrowth. J Biomed Mater Res A. 2005;72((3):279–87.
Ribeiro FV, Cesar-Neto JB, Nocit...
Marquez-Lara A, Hutchinson ID, Nuñez F, Smith TL, Miller AN. Nonsteroidal anti-inflammatory drugs and bone-healing: a systematic review of research quality. JBJS Rev. 2016;4(3). https://doi.org/10.2106/JBJS.RVW.O.00055.
Liberati A, Altman DG, Tetzlaff J, Mulrow C, Gøtzsche PC, Ioannidis JPA, et al. The PRISMA statement for reporting systematic reviews and meta-analyses of studies that evaluate...
Salari P, Abdollahi M. Controversial effects of non-steroidal anti-inflammatory drugs on bone: a review. Inflamm Allergy Drug Targets. 2009;8(3):169–75.
Boursinos LA, Karachalios T, Poultsides L, Malizos KN. Do steroids, conventional non-steroidal anti-inflammatory drugs and selective Cox-2 inhibitors adversely affect fracture healing? J Musculoskelet Neuronal Interact. 2009;9(1):44–52.
Kaly...
Cyclooxygenase
Non-steroidal anti-inflammatory drug/s
Prostaglandin
Prostaglandin E2
The analgesic and therapeutic effects of NSAIDs are achieved by COX-2 inhibition [4]. It is likely that COX inhibition by NSAIDs is detrimental to the bone healing process, given the favourable actions of PG on this process [4]. Osteoblasts have the capacity to produce PGs, where PGE2 is most abundant, through the COX pathway though the evidence asserting that PGs have a direct role in bone healin...
The majority of the included studies revealed a high risk of bias, and conclusions from studies that have a high risk of bias are sufficient to affect interpretation of data [16,17,18]. Publication and selection bias is apparent in several included studies, as the negative effects of NSAIDs on osseointegration can be expected in the studies that administered NSAID at a high concentration and/or fo...
The influence of NSAIDS on bone healing in animal models has been shown to be related to the duration of treatment and drug selectivity [5]. A total of seven studies were identified that investigated the effect of NSAIDs on the osseointegration of titanium implants in animals: mice, rabbits, and rats (Table 8).
The duration of treatment is a factor to consider when using NSAIDs, and a study cond...
The effects of NSAIDs on the osteogenic activity of osteoblasts have been extensively studied at the molecular pharmacological level [23]. However, only two studies have been identified that investigated the effect of NSAIDs on osteoblasts attached to titanium surfaces (Table 6). In the study conducted by Boyan et al., their results demonstrated that a non-selective COX inhibitor (indomethacin, 0...
The eligibility and study selection criteria as mentioned above were applied to the 79 full-text articles. A total of 66 studies were excluded after a full-text assessment for the following reasons:
The study did not explore the role of COX pathway in osseointegration (n = 26).
The effects of NSAIDs on osteoblasts were not investigated on titanium (n = 24).
The study was a systematic re...
The full-text manuscripts of included studies were catalogued into in vitro, clinical, and in vivo studies. The data from the included studies were independently extracted by the primary (JDL) and the second reviewer (TJ) according to the “Data items” section as listed below. Disagreements or uncertainties were discussed with the third reviewer (MN) until an agreement was reached.
The data co...
An electronic search into four databases: Ovid, Pubmed, Scopus, and Web of Science was performed to systematically identify the available literature. Articles published between January 1, 1999, and July 7, 2018, were considered.
The focus question, used to guide the search strategy, according to the PICO schema is “Will variables such as the dosage, duration of administration, and selectivity o...
Cyclooxygenases have an important role in the production of PGs where these enzymes in bone tissues show increased activity under the influence of hypoxia-inducible factors [6, 11]. Therefore, local activity of COX enzymes promotes bone formation and resorption through the production of PGs [12]. Non-selective NSAIDs are reported to inhibit the activity of COX-1 equally, if not more than COX-2 [2]...
Non-steroidal anti-inflammatory drugs (NSAIDs) are a group of drugs with anti-inflammatory, analgesic, and antipyretic effects. They are commonly used in dentistry for management of dental pain associated with inflammation. NSAIDs exert their effects through the inhibition of the cyclooxygenase (COX) enzyme, therefore interfering with the synthesis of prostaglandins (PG) and thromboxanes; PGs and ...
Non-steroidal anti-inflammatory drugs are commonly used in implant dentistry for management of post-operative pain. The objective of this systematic review was to analyse the effect of non-steroidal anti-inflammatory drugs on the osteogenic activity of osteoblasts with an emphasis on its effect on osseointegration. A systematic literature search for in vitro, animal models, and clinical trials was...
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...
Figure 5. Osteoblasts with an orientation tendency after 24 h of rotation (phallacidin fluorescence staining). On the left side with 200× and on the right side with 400× magnification. The yellow arrows show the orientation of the cells. The red arched arrow within the coloured circle shows the direction of rotation. The dashed white line oriented to the right stands for the r...
Figure 4. Randomly orientated osteoblasts without influence of rotation (phallacidin fluorescence staining). On the left side with 200× and on the right side with 400× magnification. The white X on the coloured circle marks the location upon the plate where the osteoblasts were located. The red X marks the centre of the plate
Figure 3. Diagram for visualisation of the calculation of shear stress rates taking into account the centrifugal force and the glass plates’ dimensions. For example, at a distance of 25 mm from the centre of the upper plate, the shear forces’ value is 8.33 dyn/cm2, together with an additional centrifugal force that has a value of 0.55 dyn/cm2
Figure 2. Side view of a computerized simulation, showing the flow chambers’ lower compartment and the flow profile in between the two plates; shearing gap and bottom plate are shown on the left side; rotation speed = 200 rpm; colour code bar (left edge) showing shear force values [Pa] [1 Pa = 10 dyn/cm2]; flow direction presented by arrows
Figure 1. Three-dimensional illustration (a–e) and photography (f) of the experimental setup with the components marked numerical. a 1 Lower petri dish (s’ bottom serving as the lower plate); 2 Rotating glass panel [60 mm diameter (cell bearing)]; 3 Titanium axis. b 4 Liquid medium (red). c 5 Reversed upper petri dish. d 6 Gearwheel with set screw. e 7 Closing; 8 Electronic motor device and ...
References
Ehrlich PJ, Lanyon LE. Mechanical strain and bone cell function: a review. Osteoporos Int. 2002;13(9):688–700.
Vaughan TJ, Haugh MG, Mcnamara LM. A fluid-structure interaction model to characterize bone cell stimulation in parallel-plate flow chamber systems. J R Soc Interface. 2013;10(81):20120900.
Weinbaum S, Cowin SC, Zeng Y. A model for the excitation of osteocytes by mec...
Besides, in the model reported in this study, microscopic examinations are possible after completing the experiment only. Nevertheless, an advantage of the new flow chamber is the possibility of testing different cell colonies simultaneously in one single experiment by placing cells in different radial locations on the spinning disc. Due to the current flow gradient from the centre to the pe...
Due to the fact that constant flows were generated within the parallel flow chamber only, the situations of in vitro experiments differ from in vivo setting where dynamic flow profiles are particular. As the constant laminar flow profile is not physiological in bones, vessels and other tissues, the informative value of the experimental setting is limited but it could be used for various cell...
Further on, the simulations indicated that the flow profile in between the two plates was not influenced by peripheral turbulences alongside the peripheral regions. To verify a cellular realignment towards the shear direction, cells were microscopically examined prior and after exposure to shear forces for 24 h upon a spinning disc at a speed level of 200 rpm. Even if not sufficiently ...
Discussion
The aim of this study was to establish a new FSS model that is easy to use as well as simple to assemble in order to create reproducible fluid shear forces on cells close to implant material surfaces. Todays’ commonly used commercial flow devices differ in geometry and function, which makes comparisons between experiments difficult. The benefits of this novel testing device are...
Regarding the upper compartment, peripheral turbulent flow along the outer edges was similar to the fluid movements within the area in between the plates. At the top, the turbulent flow directed from the centre to the periphery whereas the turbulences at the bottom were orientated in reverse to that. Moreover, the effect of the shear forces on the osteoblast cells was also influenced by the centri...
Results
Our analysis was focused on two main aspects:
Simulation of the fluid flow characteristics as well as quantification of the arising shear forces at the plate/plate flow chamber with reliable reproducibility
Assessment of the impact of fluid shear stress on osteoblast cells in terms of altered cell morphology and intracellular structural changes
Evaluation of the f...
Test procedure
The experimental process involved three steps. First, a count of n = 50.000 commercially available osteoblasts (PromoCell, Heidelberg, Germany) per millilitre of culture medium were cultured on the bottom of the cell-bearing surface (glass panel). Therefore, cells were seeded in a culture medium (cf. Appendix 2 for a detailed composition) at 37 °C. Prior to the test proce...
Analytical formula for evaluating the flow characteristics
Frequently used flow chambers are characterised by an internal fluid flow along a stationary cell-bearing surface, whereas the osteoblast test cells of this newly developed model are circulating within a resting culture medium.
For constant and fully developed laminar flow between the two parallel plates, the magnitude of the wall sh...
Methods
Experimental setup
A three-dimensional illustration and photography of the plate/plate flow chamber model is shown in Fig. 1. A detailed list of used parts can be found in Appendix 1.
The circulation within the flow chamber was generated by an externally attached electric motor, which rotates up to 500 rounds per minute (rpm). A commercial grade 4 pure medical titanium gear shaft (len...
Abstract
Background
Mechanostimuli of different cells can affect a wide array of cellular and inter-cellular biological processes responsible for dental implant healing. The purpose of this in vitro study was to establish a new test model to create a reproducible flow-induced fluid shear stress (FSS) of osteoblast cells on implant surfaces.
Methods
As FSS effects on osteoblasts are detectabl...
Kesuksesan implan gigi ditentukan oleh pembentukan tulang baru. Pembentukan tulang baru ditentukan oleh pembentukan osteoblas. Pembentukan osteoblas ditentukan oleh gen. Gen adalah salah satu faktor penentu apakah implan gigi akan jadi.Sel induk mesenkim atau sel prekursor dapat berdiferensiasi jadi osteoblas, kondrosit, mioblas, adiposit, dan sel tendon. Faktor-faktor transkripsi menentukan se...
Osteoblas adalah salah satu macem sel tulang yang bertanggungjawab atas pembentukan tulang baru. Kehadiran osteoblas itu penting karena osteoblas memungkinkan pembentukan tulang, pemodelan ulang tulang, dan perbaikan tulang.
Osteoblas hanya mempunyai 1 nukleus dan berukuran lebih kecil daripada osteoklas. Satu osteoblas tidak menghasilkan tulang baru. Saat sedang aktif, badan Golgi (Golgi ap...