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1.  Comparing the TiOblast and Osseospeed surfaces. Histomorphometric and histological analysis in humans 
Oral & Implantology  2008;1(1):34-42.
The aim of the present study was to compare two implant surfaces, the TiOblast (Astra Tech) surface, manufactured by blasting the surface and already present in literature and the Osseospeed (Astra Tech) surface, manufactured by blasting and treating the surface with fluoride ions and recently launched onto the market with the modified surfaces of the latest generation. This study is part of a more extensive research project whose protocol required the insertion of 10 couples of implants; thus in the present discussion partial data are being taken into consideration, with an eye at collecting more data in the future, regarding both microscopy and histomorphometric histological analysis on 5 couples of implants. The purpose of the study is to investigate how the modified surfaces of the latest generation can guarantee a greater osseointegration both from a qualitative and quantitative level compared to the surfaces presently used and that they may represent the first example of “bioactivity”, that is, an active interaction with the processes of new bone formation and tissue healing.
PMCID: PMC3476499  PMID: 23285334
sandblasted surface; fluoride; histology; histomorphometry; microthreads; macrothreads
2.  A comparative study on the initial stability of different implants placed above the bone level using resonance frequency analysis 
This study evaluated the initial stability of different implants placed above the bone level in different types of bone.
As described by Lekholm and Zarb, cortical layers of bovine bone specimens were trimmed to a thickness of 2 mm, 1 mm or totally removed to reproduce bone types II, III, and IV respectively. Three Implant system (Brånemark System® Mk III TiUnite™, Straumann Standard Implant SLA®, and Astra Tech Microthread™-OsseoSpeed™) were tested. Control group implants were placed in level with the bone, while test group implants were placed 1, 2, 3, and 4 mm above the bone level. Initial stability was evaluated by resonance frequency analysis. Data was statistically analyzed by one-way analysis of variance in confidence level of 95%. The effective implant length and the Implant Stability Quotient (ISQ) were compared using simple linear regression analysis.
In the control group, there was a significant difference in the ISQ values of the 3 implants in bone types III and IV (P<.05). The ISQ values of each implant decreased with increased effective implant length in all types of bone. In type II bone, the decrease in ISQ value per 1-mm increase in effective implant length of the Brånemark and Astra implants was less than that of the Straumann implant. In bone types III and IV, this value in the Astra implant was less than that in the other 2 implants.
The initial stability was much affected by the implant design in bone types III, IV and the implant design such as the short pitch interval was beneficial to the initial stability of implants placed above the bone level.
PMCID: PMC3259444  PMID: 22259702
Effective implant length; Initial stability; Implant design; Resonance frequency analysis
3.  A Focus on Soft Tissue in Dental Implantology 
Most of the focus in the early dental implant literature is on the bone to titanium interface because a successful Osseo integrated implant requires direct bone contact to the implant surface. The importance of soft tissue in the ability of dental implants to restore function and esthetics has often been underestimated. This paper reviews the pertinent literature on soft tissue healing and management in partially edentulous dental implant patients. Patients seek treatment to replace missing teeth and to improve comfort, function and/or esthetics. Healing around dental implants is affected by the patient’s health, soft and hard tissue contours, and the use and care of the prosthesis, surgical augmentation and placement, and the design of the definitive prosthesis. Several surgical and non-surgical procedures have been proposed to treat the soft tissue deformities in the interproximal areas. This review also discusses the interdental papilla and various approaches to preserve and restore the same. Most of the research was based on scientifically legitimate sources of information obtained from primary literature, other appropriate technical references and searching using various online resources.
PMCID: PMC3416938  PMID: 23997462
Implants; Soft tissue; Surgical management; Non-surgical management
4.  Enhanced Cell Integration to Titanium Alloy by Surface Treatment with Microarc Oxidation: A Pilot Study 
Microarc oxidation (MAO) is a surface treatment that provides nanoporous pits, and thick oxide layers, and incorporates calcium and phosphorus into the coating layer of titanium alloy. We presumed such modification on the surface of titanium alloy by MAO would improve the ability of cementless stems to osseointegrate. We therefore compared the in vitro ability of cells to adhere to MAOed titanium alloy to that of two different types of surface modifications: machined and grit-blasted. We performed energy-dispersive x-ray spectroscopy and scanned electron microscopy investigations to assess the structure and morphology of the surfaces. Biologic and morphologic responses to osteoblast cell lines (SaOS-2) were then examined by measuring cell proliferation, cell differentiation (alkaline phosphatase activity), and αvβ3 integrin. The cell proliferation rate, alkaline phosphatase activity, and cell adhesion in the MAO group increased in comparison to those in the machined and grit-blasted groups. The osteoblast cell lines of the MAO group were also homogeneously spread on the surface, strongly adhered, and well differentiated when compared to the other groups. This method could be a reasonable option for treating the surfaces of titanium alloy for better osseointegration.
PMCID: PMC2866922  PMID: 19434468
5.  Behavior of osteoblastic cells cultured on titanium and structured zirconia surfaces 
Head & Face Medicine  2008;4:29.
Osseointegration is crucial for the long-term success of dental implants and depends on the tissue reaction at the tissue-implant interface. Mechanical properties and biocompatibility make zirconia a suitable material for dental implants, although surface processings are still problematic. The aim of the present study was to compare osteoblast behavior on structured zirconia and titanium surfaces under standardized conditions.
The surface characteristics were determined by scanning electron microscopy (SEM). In primary bovine osteoblasts attachment kinetics, proliferation rate and synthesis of bone-associated proteins were tested on different surfaces.
The results demonstrated that the proliferation rate of cells was significantly higher on zirconia surfaces than on titanium surfaces (p < 0.05; Student's t-test). In contrast, attachment and adhesion strength of the primary cells was significant higher on titanium surfaces (p < 0.05; U test). No significant differences were found in the synthesis of bone-specific proteins. Ultrastructural analysis revealed phenotypic features of osteoblast-like cells on both zirconia and titanium surfaces.
The study demonstrates distinct effects of the surface composition on osteoblasts in culture. Zirconia improves cell proliferation significantly during the first days of culture, but it does not improve attachment and adhesion strength. Both materials do not differ with respect to protein synthesis or ultrastructural appearance of osteoblasts. Zirconium oxide may therefore be a suitable material for dental implants.
PMCID: PMC2614982  PMID: 19063728
6.  Osteoblasts exhibit a more differentiated phenotype and increased bone morphogenetic protein production on titanium alloy substrates than on poly-ether-ether-ketone 
Background Context
Multiple biomaterials are clinically available to spine surgeons for performing interbody fusion. Poly-ether-ether-ketone (PEEK) is used frequently for lumbar spine interbody fusion, but alternative materials are also used, including titanium (Ti) alloys. Previously, we showed that osteoblasts exhibit a more differentiated phenotype when grown on machined or grit-blasted titanium aluminum vanadium (Ti6Al4V) alloys with micron-scale roughened surfaces than when grown on smoother Ti6Al4V surfaces or on tissue culture polystyrene (TCPS). We hypothesized that osteoblasts cultured on rough Ti alloy substrates would present a more mature osteoblast phenotype than cells cultured on PEEK, suggesting that textured Ti6Al4V implants may provide a more osteogenic surface for interbody fusion devices.
The aim of the present study was to compare osteoblast response to smooth Ti6Al4V (sTiAlV) and roughened Ti6Al4V (rTiAlV) with their response to PEEK with respect to differentiation and production of factors associated with osteogenesis.
Study Design
This in vitro study compared the phenotype of human MG63 osteoblast-like cells cultured on PEEK, sTiAlV, or rTiAlV surfaces and their production of bone morphogenetic proteins (BMPs).
Surface properties of PEEK, sTiAlV, and rTiAlV discs were determined. Human MG63 cells were grown on TCPS and the discs. Confluent cultures were harvested, and cell number, alkaline phosphatase–specific activity, and osteocalcin were measured as indicators of osteoblast maturation. Expression of messenger RNA (mRNA) for BMP2 and BMP4 was measured by real-time polymerase chain reaction. Levels of BMP2, BMP4, and BMP7 proteins were also measured in the conditioned media of the cell cultures.
Although roughness measurements for sTiAlV (Sa=0.09±0.01), PEEK (Sa=0.43±0.07), and rTiAlV (Sa= 1.81±0.51) varied, substrates had similar contact angles, indicating comparable wettability. Cell morphology differed depending on the surface. Cells cultured on Ti6Al4V had lower cell number and increased alkaline phosphatase specific activity, osteocalcin, BMP2, BMP4, and BMP7 levels in comparison to PEEK. In particular, roughness significantly increased the mRNA levels of BMP2 and BMP4 and secreted levels of BMP4.
These data demonstrate that rTiAlV substrates increase osteoblast maturation and produce an osteogenic environment that contains BMP2, BMP4, and BMP7. The results show that modifying surface structure is sufficient to create an osteogenic environment without addition of exogenous factors, which may induce better and faster bone during interbody fusion.
PMCID: PMC3618467  PMID: 22424980
Ti6Al4V; PEEK; Osteoblast; BMP; Roughness
7.  In vitro assay for osteoinductive activity of different demineralized freeze-dried bone allograft 
Various bone graft materials have been used for periodontal tissue regeneration. Demineralized freeze-dried bone allograft (DFDBA) is a widely used bone substitute. The current widespread use of DFDBA is based on its potential osteoinductive ability. Due to the lack of verifiable data, the purpose of this study was to assess the osteoinductive activity of different DFDBAs in vitro.
Sarcoma osteogenic (SaOS-2) cells (human osteoblast-like cells) were exposed to 8 mg/mL and 16 mg/mL concentrations of three commercial types of DFDBA: Osseo+, AlloOss, and Cenobone. The effect of these materials on cell proliferation was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The osteoinductive ability was evaluated using alizarin red staining, and the results were confirmed by evaluating osteogenic gene expression using reverse transcription polymerase chain reaction (RT-PCR).
In the SaOS-2 cells, an 8 mg/mL concentration of Osseo+ and Cenobone significantly increased cell proliferation in 48 hours after exposure (P<0.001); however, in these two bone materials, the proliferation of cells was significantly decreased after 48 hours of exposure with a 16 mg/mL concentration (P<0.001). The alizarin red staining results demonstrated that the 16 mg/mL concentration of all three tested DFDBA induced complete morphologic differentiation and mineralized nodule production of the SaOS-2 cells. The RT-PCR results revealed osteopontin gene expression at a 16 mg/mL concentration of all three test groups, but not at an 8 mg/mL concentration.
These commercial types of DFDBA are capable of decreasing proliferation and increasing osteogenic differentiation of the SaOS-2 cell line and have osteoinductive activity in vitro.
PMCID: PMC3543938  PMID: 23346466
Alizarin red; Bone graft; Cell differentiation; Cell proliferation; Regeneration
8.  In vitro antimicrobial and biological properties of laser assisted tricalcium phosphate coating 
Implant related infections are of great concern in modern surgery. In order to improve the implant performance and to reduce implant related infections, titanium (Ti) surface was modified to simultaneously improve cell- materials interactions and antimicrobial activity. Ti surface was first coated with tricalcium phosphate (TCP) using Laser Engineered Net Shaping (LENS™) to improve biocompatibility. Silver (Ag) was then electrodeposited from different concentrations of silver nitrate (AgNO3) solutions to improve the antimicrobial activity. The Ag-TCP coatings were tested for cytotoxicy with human osteoblast cells. The antimicrobial activities of the Ag-TCP coatings were evaluated using Pseudomonas aeruginosa and Staphylococcus aureus bacteria. In vitro bacterial adhesion study indicated a significant reduction in bacterial colony on Ag-TCP coated surfaces when compared to TCP coated surface.
PMCID: PMC3065785  PMID: 21461340
Tricalcium phosphate coating; Silver; cytotoxicity; antimicrobial activity
9.  Influence of bone density on implant stability parameters and implant success: a retrospective clinical study 
BMC Oral Health  2008;8:32.
The aim of the present clinical study was to determine the local bone density in dental implant recipient sites using computerized tomography (CT) and to investigate the influence of local bone density on implant stability parameters and implant success.
A total of 300 implants were placed in 111 patients between 2003 and 2005. The bone density in each implant recipient site was determined using CT. Insertion torque and resonance frequency analysis were used as implant stability parameters. The peak insertion torque values were recorded with OsseoCare machine. The resonance frequency analysis measurements were performed with Osstell instrument immediately after implant placement, 6, and 12 months later.
Of 300 implants placed, 20 were lost, meaning a survival rate of %. 93.3 after three years (average 3.7 ± 0.7 years). The mean bone density, insertion torque and RFA recordings of all 300 implants were 620 ± 251 HU, 36.1 ± 8 Ncm, and 65.7 ± 9 ISQ at implant placement respectively; which indicated statistically significant correlations between bone density and insertion torque values (p < 0.001), bone density and ISQ values (p < 0.001), and insertion torque and ISQ values (p < 0.001). The mean bone density, insertion torque and RFA values were 645 ± 240 HU, 37.2 ± 7 Ncm, and 67.1 ± 7 ISQ for 280 successful implants at implant placement, while corresponding values were 267 ± 47 HU, 21.8 ± 4 Ncm, and 46.5 ± 4 ISQ for 20 failed implants; which indicated statistically significant differences for each parameter (p < 0.001).
CT is a useful tool to determine the bone density in the implant recipient sites, and the local bone density has a prevailing influence on primary implant stability, which is an important determinant for implant success.
PMCID: PMC2614413  PMID: 19025637
10.  Immediate implants in anterior maxillary arch 
The aim of the study was to evaluate the osseo-integration and soft tissue status of the endosseous implants placed in immediate extraction socket.
Seven patients (4 males and 3 females) aged 20-30 years were selected for the study. Nine implants were placed in seven patients in the maxillary arch. All the patients were clinically αnd thoroughly examined. Under local anesthesia, the indicated tooth was extracted. The extracted socket was prepared using standard drills with palatal wall as guide. The longest and widest implants were placed (Hi-Tec Implants). All implants showed good primary stability. The implants used in the study were tapered design endosseous implants with Threaded implants (TI) unit plasma-sprayed surface. Surgical re-entry (secondary surgery) was performed to remove the healing cap after 6 months for supra crestal fabrication. All patients were reviewed periodically at 3rd and 6th month interval and the following clinical parameters including modified plaque index (mPlI), modified bleeding index (mBI), probing depth (PD), attachment level (AL), and distance between the implant shoulder and mucosal margin (DIM), distance between the implant shoulder and first bone-implant contact, and Clinical Mobility Index were recorded. The results were computed and subjected to statistical evaluation.
The mPlI, mBI, PD, AL, and DIM were evaluated around the implants at baseline, 3rd and 6th month intervals and analyzed statistically by Friedman T-test. The results of the above were shown to be statistically non-significant. The distance between the implant shoulder and first bone implant contact was evaluated around the implants at base line, 3rd and 6th month intervals. The results proved to be statistically significant (0.01) implying that there was a bone apposition around the implants.
During the course of the study, soft tissue status around implants was found to be healthy. Osseointegration as assessed by clinical and radiographic findings was found to be sound.
PMCID: PMC3961959  PMID: 24678203
Bone borne implants; dental implants; endosseous implants; immediate implants; osseointegration
11.  The Osteogenic Study of Tissue Engineering Bone with BMP2 and BMP7 Gene-Modified Rat Adipose-Derived Stem Cell 
To evaluate the feasibility and advantages of constructing a novel tissue engineering bone, using β-tricalcium phosphate (β-TCP) and rat adipose-derived stem cells (ADSCs), modified with BMP2 and BMP7 by lentivirus. In the present study, ADSCs transfected with Lv-BMP2 and Lv-BMP7, alone or together, were seeded on β-TCP scaffold and cultured in vitro. Based on the results of DNA assay, alkaline phosphatase (ALP) activity, alizarin red staining and osteogenic marker genes expression analysis, the BMP2 and BMP7 genes cotransfection group exhibited a higher degree of osteogenic differentiation in vitro. To investigate the in vivo osteogenesis of the tissue engineering bone, the ADSCs/β-TCP constructs were implanted in rat femurs defects for 6 weeks and studied histomorphology and radiography. The results showed that BMP2 and BMP7 genes cotransfection group dramatically enhanced the efficiency of new bone formation than BMP2 group and BMP7 group in vivo. These results demonstrated that it was advantageous to construct tissue engineering bone using ADSCs cotransfected with BMP2 and BMP7 on β-TCP, providing a potential way for treating bone defects.
PMCID: PMC3388521  PMID: 22778550
12.  In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics 
In order to understand the mechanisms of poor osseointegration following dental implants in type 2 diabetics, it is important to study the biological properties of alveolar bone osteoblasts isolated from these patients. We collected alveolar bone chips under aseptic conditions and cultured them in vitro using the tissue explants adherent method. The biological properties of these cells were characterized using the following methods: alkaline phosphatase (ALP) chemical staining for cell viability, Alizarin red staining for osteogenic characteristics, MTT test for cell proliferation, enzyme dynamics for ALP contents, radio-immunoassay for bone gla protein (BGP) concentration, and ELISA for the concentration of type I collagen (COL-I) in the supernatant. Furthermore, we detected the adhesion ability of two types of cells from titanium slices using non-specific immunofluorescence staining and cell count. The two cell forms showed no significant difference in morphology under the same culture conditions. However, the alveolar bone osteoblasts received from type 2 diabetic patients had slower growth, lower cell activity and calcium nodule formation than the normal ones. The concentration of ALP, BGP and COL-I was lower in the supernatant of alveolar bone osteoblasts received from type 2 diabetic patients than in that received from normal subjects (P < 0.05). The alveolar bone osteoblasts obtained from type 2 diabetic patients can be successfully cultured in vitro with the same morphology and biological characteristics as those from normal patients, but with slower growth and lower concentration of specific secretion and lower combining ability with titanium than normal ones.
PMCID: PMC3854304  PMID: 22473318
Alveolar bone; Cell culture; Implant; Osteoblast; Type 2 diabetics
13.  Effect of etched microgrooves on hydrophilicity of titanium and osteoblast responses: A pilot study 
The aim of this pilot study was to investigate the effect of etched microgrooves on the hydrophilicity of Ti and osteoblast responses.
Microgrooves were applied on Ti to have 15 and 60 µm width, and 3.5 and 10 µm depth by photolithography, respectively. Further acid etching was applied to create Ti surfaces with etched microgrooves. Both smooth- and acid-etched Ti were used as the controls. The hydrophilicity of Ti was analyzed by determining contact angles. Cell proliferation and osteogenic activity of MC3T3 mouse preosteoblasts were analyzed by bromodeoxyuridine assay and alkaline phosphatase (ALP) activity test, respectively. One-way ANOVA, Pearson's correlation analysis and multiple regression analysis were used for statistics.
Etched microgrooves significantly increased the hydrophilicity of Ti compared to the smooth Ti. 60 µm-wide etched microgrooves significantly enhanced cell proliferation, whereas the osteogenic activity showed statistically non-significant differences between groups. Result of the osteogenic activity significantly correlated with those of hydrophilicity and cell proliferation. Hydrophilicity was determined to be an influential factor on osteogenic activity.
This study indicates that increase in hydrophilicity of Ti caused by etched microgrooves acts as an influential factor on osteogenic activity. However, statistically non-significant increase in the ALP activity suggests further investigation.
PMCID: PMC2984515  PMID: 21165183
Titanium; Etched microgrooves; Hydrophilicity; Osteoblast response
14.  Surface Properties and Osteoblastic Cytocompatibility of Two Blasted and Acid-Etched Titanium Implant Systems with Distinct Microtopography 
The aim of this study is to compare two commercially available screw-type sandblasted and acid-etched (SLA) Ti implant systems from Eckermann Laboratorium S.L., with similar geometry and distinct microtopography, regarding surface properties and osteoblastic cytocompatibility.
Material and Methods
Implant I (referred as a conventional SLA system) and Implant II (a system patented as Eckcyte®) were characterized for macro and microtopograpphy, surface roughness and chemical composition. For the cytocompatibility studies, human bone marrow osteoblastic cells were seeded over the implants' surface, and the cell response was assessed for cell adhesion and proliferation, alkaline phosphatase (ALP) activity and matrix mineralization.
Implant I presented a rough surface with irregularly shaped and sized cavities among flatter-appearing areas, whereas Implant II exhibited a homogeneous rough microporous surface. Compared to Implant I, Implant II presented higher Ra values (0.8 [SD 0.008] μm and 1.21 [SD 0.15] μm, respectively, P < 0.05) and also increased values of Rz, Rt and Rsm, a more negative value of Rsk, and similar RKu values. XPS showed the expected presence of Ti, O, C and N; Al, Si, F, P and Ca were detected in low concentrations. Implant II exhibited significantly lower Al levels. Both implants supported the adhesion, proliferation and differentiation of osteoblastic cells. Implant II showed a thicker fibrilar cell layer and an earlier onset and more abundant matrix mineralization.
The homogeneous rough and microporous surface of Implant II is most probably a main contributor for its improved cell response.
PMCID: PMC3886090  PMID: 24422006
dental implants; surface properties; bone marrow; osteoblasts; differentiation cell; cell culture.
15.  Treatment of protruding osseo integrated dental implant 
Titanium dental implants have been used in the treatment of partial or complete edentulism. The height and width of the residual alveolus and surrounding anatomical structures can determine the proper position and path of insertion of dental implants. The following case report describes the treatment of a malpositioned osseo integrated dental implant with an apex perforating the buccal cortex of alveolar bone. A 61-year-old male was referred by his local dentist for the chief complaint of a swelling at site of tooth 14 where an implant was present. Intraoral clinical examination revealed an implant supported porcelain fused to metal crown replacing the maxillary right first premolar. A peri-apical radiograph of the implant revealed no signs of peri-implant bone loss or radiolucency. Surgical exploration and modification of the protruding implant. The area healed uneventfully without the need of explantation of the implant in site of tooth 14. We felt that the conservative treatment provided was prudent and treatment of choice and anticipate that the implant will most likely continue to function for a lifetime.
PMCID: PMC2933530  PMID: 20922080
Dental implant; osseointegration; perforation
16.  Promotion of osteoblast proliferation on complex coacervation-based hyaluronic acid – recombinant mussel adhesive protein coatings on titanium 
Biomaterials  2009;31(6):1080-1084.
Many biological polyelectrolytes are capable of undergoing a fluid–fluid phase separation known as complex coacervation. Coacervates were prepared using hyaluronic acid (HA) and a recombinant fusion protein consisting of mussel adhesive motifs and the RGD peptide (fp-151-RGD). The low interfacial energy of the coacervate was exploited to coat titanium (Ti), a metal widely used in implant materials. The coacervate effectively distributed both HA and fp-151-RGD over the Ti surfaces and enhanced osteoblast proliferation. Approximately half of total fp-151-RGD and HA in the solution transferred to the titanium surface within 2 h. Titanium coated with coacervates having high residual negative surface charge showed the highest cell proliferation of preosteoblast cells (MC-3T3) compared to the treatments tested. Indeed, MC-3T3 cells on complex coacervate coated titanium foils exhibited over 5 times greater cell proliferation than bare, HA coated or fp-151-RGD coated titanium.
PMCID: PMC2835630  PMID: 19892396
Complex coacervate; Hyaluronic acid; Mussel adhesive protein; fp-151-RGD; MC-3T3; Biocoating
17.  Survival rate of Astra Tech implants with maxillary sinus lift 
The aim of this study was to retrospectively evaluate the clinical survival rate of Astra Tech implants in the maxillary molar region performed with sinus lift and bone graft.
Materials and Methods
Ninety-nine Astra Tech implants (Osseospeed) placed in the maxillary molar region using sinus lift from September 2009 to February 2012 were selected with a minimum follow-up period of 1 year. The height of alveolar bone, sinus approach technique, bone material and implant survival rate were evaluated.
Of the 99 implants, the survival rate was 90.9%; 8 implants failed within 1 year after implant placement, and 1 implant failed 1 year after implant loading. All failed implants were placed with sinus lift simultaneously. The average height of alveolar bone before implant placement was 6.9 mm, while the height of alveolar bone of failed implants was 2.1 mm, on average.
Astra Tech implants placed in the maxillary molar region had generally good survival rates, but the relationship between reduced pre-implant alveolar bone height and implant failure requires further attention.
PMCID: PMC3949495  PMID: 24627838
Sinus floor augmentation; Dental implants
18.  Stress Distribution in Implant Retained Finger Prosthesis: A Finite Element Study 
Background: Finger amputation may result from congenital cause, trauma, infection and tumours. The finger amputation may be rehabilitated with dental implant-retained finger prosthesis. The success of implant-retained finger prosthesis is determined by the implant loading. The type of the force is a determining factor in implant loading.
Objective: To evaluate stress distributions in finger bone when the loading force is applied along the long axis of the implant using finite element analysis.
Method: The finite element models were created. The finger bone model containing cortical bone and cancellous bone was constructed by using radiograph. Astra Tech Osseo Speed bone level implant of 4.5 mm diameter and 14 mm length was selected. The force was applied to the top of the abutment along the long axis of the implant.
Results: Finite element analysis indicated that the maximum stress was located at the head of abutment screw. The minimum stress was located in the apical third of the implant fixture. The weakest point was calculated by safety factor which is located in the spongy bone at apical third of the fixtures. Finally, 4.9 times yield stress of spongy bone was needed for the deformation of the spongy bone.
Conclusion: Finite element study showed that when the force was applied along the long axis of the implant, the maximum stress was located around the neck of the implant and the cortex bone received more stress than cancellous bone. So, to achieve long term success, the designers of implant systems must confront biomaterial and biomechanical problems including in vivo forces on implants, load transmission to the interface and interfacial tissue response.
PMCID: PMC3919359  PMID: 24551656
Dental implant; Finger prosthesis; Stress; Finite element analysis
19.  Cellular Performance Comparison of Biomimetic Calcium Phosphate Coating and Alkaline-Treated Titanium Surface 
BioMed Research International  2013;2013:832790.
The influence of biomimetic calcium phosphate coating on osteoblasts behavior in vitro is not well established yet. In this study, we investigated the behavior of osteoblastic rat osteosarcoma 17/2.8 cells (ROS17/2.8) on two groups of biomaterial surfaces: alkaline-treated titanium surface (ATT) and biomimetic calcium phosphate coated ATT (CaP). The cell attachment, proliferation, differentiation, and morphology on these surfaces were extensively evaluated to reveal the impact of substrate surface on osteoblastic cell responses. It was found that the ROS17/2.8 cells cultured on the ATT surface had higher attachment and proliferation rates compared to those on the CaP surface. Our results also showed that the calcium phosphate coatings generated in this work have an inhibiting effect on osteoblast adhesion and further influenced the proliferation and differentiation of osteoblast compared to the ATT surface in vitro. Cells on the ATT surface also exhibited a higher alkaline phosphatase activity than on the CaP surface after two weeks of culture. Immunofluorescence staining and scanning electron microscopy results showed that the cells adhered and spread faster on the ATT surface than on the CaP surface. These results collectively suggested that substrate surface properties directly influence cell adhesion on different biomaterials, which would result in further influence on the cell proliferation and differentiation.
PMCID: PMC3884630  PMID: 24455730
20.  Implant adaptation of stock abutments versus CAD/CAM abutments: a radiographic and Scanning Electron Microscopy study 
Annali di Stomatologia  2011;1(3-4):9-13.
The study evaluated a null-hypothesis of no differences of fit between stock abutments and CAD/CAM titanium, gold sputtered and zirconia abutments when examined for radiographic adaptation and Scanning Electron Microcopy (SEM) at their inner aspect. The agreement between microscopic and radiographic fit was also assessed.
Implants (Osseospeed, Astra Tech, Mölndal, Sweden) were connected to titanium abutments (Ti-design, Astra Tech, Mölndal, Sweden) (control group n=12), to stock zirconia abutments (Zir-design, Astra Tech) (group 1 n=12) and to third party zirconia abutments (Aadva Zr abutment, GC, Tokyo, Japan) as observed under SEM (JEOL JSM-6060LV, Tokyo, Japan). Two independent operators blindly evaluated the images, according to a three-score scale: perfect adaptation, no complete adaptation, and clear evidence of no adaptation. A Kruskal-Wallis test was applied to assess significant differences in adaptation scores between the groups.
All specimens showed precise SEM adaptation at all tested interfaces and no radiographically apparent gaps. No significant differences were found and therefore the null-hypothesis tested was accepted. Radiographic and SEM scores were in agreement.
CAD/CAM titanium, gold sputtered and zirconia abutments and third-part CAD/CAM zirconia abutments show an adaptation to Astra Tech implants that is comparable to that of stock titanium and zirconia abutments. Clinicians might be able to verify such adaptation with an x-ray. In-vivo studies would be needed to evaluate the clinical outcome of CAD/CAM abutments.
PMCID: PMC3254378  PMID: 22238709
titanium; zirconia; interface; abutment
21.  Biomolecular surface coating to enhance orthopaedic tissue healing and integration 
Biomaterials  2007;28(21):3228-3235.
Implant osseointegration is a prerequisite for clinical success in orthopaedic and dental applications, many of which are restricted by loosening. Biomaterial surface modification approaches, including calcium-phosphate ceramic coatings and macro/microporosity, have had limited success in promoting integration. To improve osseointegration, titanium surfaces were coated with the GFOGER collagen-mimetic peptide, selectively promoting α2β1 integrin binding, a crucial event for osteoblastic differentiation. Titanium surfaces presenting GFOGER triggered osteoblastic differentiation and mineral deposition in bone marrow stromal cells, leading to enhanced osteoblastic function compared to unmodified titanium. Furthermore, this integrin-targeted coating significantly improved in vivo peri-implant bone regeneration and osseointegration, as characterized by bone-implant contact and mechanical fixation, compared to untreated titanium in a rat cortical bone-implant model. GFOGER-modified implants also significantly enhanced osseointegration compared to surfaces modified with full-length type I collagen, highlighting the importance of presenting specific biofunctional domains within the native ligand. In addition, this biomimetic implant coating is generated using a simple, single-step procedure that readily translates to a clinical environment with minimal processing and cytotoxicity concerns. Therefore, this study establishes a biologically active and clinically relevant implant coating strategy that enhances bone repair and orthopaedic implant integration.
PMCID: PMC2034748  PMID: 17448533
biomimetic material; cell adhesion; collagen; osseointegration; integrin
22.  Exogenous heparin binds and inhibits bone morphogenetic protein 6 biological activity 
International Orthopaedics  2013;37(3):529-541.
The purpose of this study was to explore the effect of heparin on bone morphogenetic protein 6 (BMP6) osteogenic activity.
Western blot analysis was used to confirm the binding of BMP6 to heparin and to observe its effect on BMP6 signaling in C2C12-BRE-Luc myoblasts. Real-time RT-PCR was performed for the expression analysis of alkaline phosphatase (ALP) and osteocalcin (OC) in C2C12 myoblasts treated with BMP6 and heparin for 72 hours. Rat ectopic bone formation assay was performed to explore the effect of heparin on BMP6 osteogenic activity. Two weeks following implantation the implants were analysed morphologically and histologically. A mouse osteoporotic model was used to test the ability of BMP6 to improve the bone quality in vivo in the presence of heparin, followed by DEXA and μCT analyses. Blood coagulation was tested in rats previously treated with BMP6.
BMP6 specifically bound to heparin and induced Smad1/5/8 phosphorylation which was inhibited by heparin. After 48 and 72 hours of treatment, heparin inhibited BMP6-induced ALP and OC expression in C2C12 cells. Heparin dose dependently inhibited BMP6-induced new bone and cartilage formation in the rat ectopic bone formation assay, while in osteoporotic mice heparin inhibited the BMP6 potential to improve the bone quality as evidenced by decreased bone mineral density and trabecular bone parameters. Interestingly, BMP6 prevented the effect of heparin on the blood coagulation parameters.
The interaction of BMP6 with heparin might contribute to the heparin-induced osteoporosis and blood coagulation.
PMCID: PMC3580086  PMID: 23307015
23.  The combined effect of parathyroid hormone and bone graft on implant fixation 
Impaction allograft is an established method of securing initial stability of an implant in arthroplasty. Subsequent bone integration can be prolonged, and the volume of allograft may not be maintained. Intermittent administration of parathyroid hormone has an anabolic effect on bone and may therefore improve integration of an implant.
Using a canine implant model we tested the hypothesis that administration of parathyroid hormone may improve osseo-integration of implants surrounded by bone graft. In 20 dogs a cylindrical porous-coated titanium alloy implant was inserted into normal cancellous bone in the proximal humerus and surrounded by a circumferential gap of 2.5 mm. Morsellised allograft was impacted around the implant. Half of the animals were given daily injections of human parathyroid hormone (1-34) 5 μg/kg for four weeks and half received control injections. The two groups were compared by mechanical testing and histomorphometry. We observed a significant increase in new bone formation within the bone graft in the parathyroid hormone group. There were no significant differences in the volume of allograft, bone-implant contact or in the mechanical parameters.
These findings suggest that parathyroid hormone improves new bone formation in impacted morsellised allograft around an implant and retains the graft volume without significant resorption. Fixation of the implant was neither improved nor compromised at the final follow-up of four weeks.
PMCID: PMC3681808  PMID: 21196558
24.  Effects of TGF-β1 and VEGF-A transgenes on the osteogenic potential of bone marrow stromal cells in vitro and in vivo 
Journal of Tissue Engineering  2012;3(1):2041731412459745.
An exogenous supply of growth factors and bioreplaceable scaffolds may help bone regeneration. The aim of this study was to examine the effects of TGF-β1 and VEGF-A transgenes on the osteogenic potential of bone marrow stromal cells. Rat bone marrow stromal cells were transfected with plasmids encoding mouse TGF-β1 and/or VEGF-A complementary DNAs and cultured for up to 28 days. Furthermore, collagen scaffolds carrying combinations of the plasmids-transfected cells were implanted subcutaneously in rats. The transgenes increased alkaline phosphatase activity, enhanced mineralized nodule formation, and elevated osteogenic gene expressions in vitro. In vivo, messenger RNA expression of osteogenic genes such as BMPs and Runx2 elevated higher by the transgenes. The data indicate that exogenous TGF-β1 and VEGF-A acted synergistically and could induce osteoblastic differentiation of bone marrow stromal cells in both cell culture and an animal model. The results may provide valuable information to optimize protocols for transgene-and-cell-based tissue engineering.
PMCID: PMC3434762  PMID: 22962632
Gene transfer; bone marrow; TGF-β1; VEGF-A; tissue engineering
25.  Response of osteoblast-like cells cultured on zirconia to bone morphogenetic protein-2 
The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2.
MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days.
At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7.
The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.
PMCID: PMC3213233  PMID: 22087413
Bone morphogenetic protein-2; Cell differentiation; Cell proliferation; Zirconium oxide

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