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.
sandblasted surface; fluoride; histology; histomorphometry; microthreads; macrothreads
This study evaluated the initial stability of different implants placed above the bone level in different types of bone.
MATERIALS AND METHODS
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.
Effective implant length; Initial stability; Implant design; Resonance frequency analysis
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.
Implants; Soft tissue; Surgical management; Non-surgical management
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.
Alizarin red; Bone graft; Cell differentiation; Cell proliferation; Regeneration
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.
Alveolar bone; Cell culture; Implant; Osteoblast; Type 2 diabetics
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.
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.
Ti6Al4V; PEEK; Osteoblast; BMP; Roughness
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.
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.
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.
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.
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.
Dental implant; osseointegration; perforation
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.
Tricalcium phosphate coating; Silver; cytotoxicity; antimicrobial activity
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.
Dental implant; Finger prosthesis; Stress; Finite element analysis
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.
titanium; zirconia; interface; abutment
The aim of this pilot study was to investigate the effect of etched microgrooves on the hydrophilicity of Ti and osteoblast responses.
MATERIAL AND METHODS
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.
Titanium; Etched microgrooves; Hydrophilicity; Osteoblast response
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
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
The homogeneous rough and microporous surface of Implant II is most
probably a main contributor for its improved cell response.
dental implants; surface properties; bone marrow; osteoblasts; differentiation cell; cell culture.
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.
Gene transfer; bone marrow; TGF-β1; VEGF-A; tissue engineering
Functional tissue engineering for bone augmentation requires the appropriate combination of biomaterials, mesenchymal stem cells, and specific differentiation factors. Therefore, we investigated the morphology, attachment, viability, and proliferation of human dental pulp stem cells cultured in xeno-free conditions in human serum medium seeded on β-tricalcium phosphate/poly(l-lactic acid/caprolactone) three-dimensional biomaterial scaffold. Additionally, osteogenic inducers dexamethasone and vitamin D3 were compared to achieve osteogenic differentiation. Dental pulp stem cells cultured in human serum medium maintained their morphology; furthermore, cells attached, remained viable, and increased in cell number within the scaffold. Alkaline phosphatase staining showed the osteogenic potential of dental pulp stem cells under the influence of osteogenic medium containing vitamin D3 or dexamethasone within the scaffolds. Maintenance of dental pulp stem cells for 14 days in osteogenic medium containing vitamin D3 resulted in significant increase in osteogenic markers as shown at mRNA level in comparison to osteogenic medium containing dexamethasone. The results of this study show that osteogenic medium containing vitamin D3 osteo-induced dental pulp stem cells cultured in human serum medium within β-tricalcium phosphate/poly(l-lactic acid/caprolactone) three-dimensional biomaterial, which could be directly translated clinically.
dental pulp stem cells; human serum; β-tricalcium phosphate/poly(l-lactic acid/caprolactone); vitamin D3; osteogenic differentiation
Experimental animal study to evaluate the osseo integration, inflammatory response, dislocation and the reabsorption timing of the reasorbeable Mitek® mini anchor, in comparison with the non reasorbeable titanium based.
Twenty rabbits were used: divided into two groups of ten. Each animal underwent sectioning and reinsertion of the Achilles tendon bilaterally, using a reasorbeable mini anchor on one side and a controlateral non reasorbeable mini anchor. The first group was sacrificed after 40 days and the other after 120 days. The bone tendon complex was subjected to histological study.
No histological and statistical significative difference were observed in each group, with a lower inflammation response in the reasorbeable implant. Electronic microscopy evaluation demonstrates good stability of the implant in each group. At day 120 the reasorbeable anchors were not yet disappeared.
Reasorbeable Mitek mini anchors are to be considered to guarantee a similar response and similar bone stability than non reasorbeable ones.
anchor; reabsorption; tendon; osseointegration; rabbit
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.
Complex coacervate; Hyaluronic acid; Mussel adhesive protein; fp-151-RGD; MC-3T3; Biocoating
Many challenges exist in improving early osseointegration, one of the most critical factors in the long-term clinical success of dental implants. Recently, ultraviolet (UV) light-mediated photofunctionalization of titanium as a new potential surface treatment has aroused great interest. This study examines the bioactivity of titanium surfaces treated with UV light of different wavelengths and the underlying associated mechanism. Micro-arc oxidation (MAO) titanium samples were pretreated with UVA light (peak wavelength of 360 nm) or UVC light (peak wavelength of 250 nm) for up to 24 h. UVC treatment promoted the attachment, spread, proliferation and differentiation of MG-63 osteoblast-like cells on the titanium surface, as well as the capacity for apatite formation in simulated body fluid (SBF). These biological influences were not observed after UVA treatment, apart from a weaker effect on apatite formation. The enhanced bioactivity was substantially correlated with the amount of Ti-OH groups, which play an important role in improving the hydrophilicity, along with the removal of hydrocarbons on the titanium surface. Our results showed that both UVA and UVC irradiation altered the chemical properties of the titanium surface without sacrificing its excellent physical characteristics, suggesting that this technology has extensive potential applications and merits further investigation.
Microstructured and high surface energy titanium substrates increase osseointegration in vivo. In vitro, osteoblast differentiation is increased, but effects of the surface directly on multipotent mesenchymal stem cells (MSCs) and consequences for MSCs in the peri-implant environment are not known. We evaluated responses of human MSCs to substrate surface properties and examined the underlying mechanisms involved. MSCs exhibited osteoblast characteristics (alkaline phosphatase, RUNX2, and osteocalcin) when grown on microstructured Ti; this effect was more robust with increased hydrophilicity. Factors produced by osteoblasts grown on microstructured Ti were sufficient to induce co-cultured MSC differentiation to osteoblasts. Silencing studies showed that this was due to signaling via α2β1 integrins in osteoblasts on the substrate surface and paracrine action of secreted Dkk2. Thus, human MSCs are sensitive to substrate properties that induce osteoblastic differentiation; osteoblasts interact with these surface properties via α2β1 and secrete Dkk2, which acts on distal MSCs.
Several bone implants are applied in clinical practice, but none meets the requirements of an ideal implant. Platelet-rich plasma (PRP) is an easy and inexpensive way to obtain growth factors in physiologic proportions that might favour the regenerative process. The aim of this review is to analyse clinical studies in order to investigate the role of PRP in favouring bone integration of graft, graft substitutes, or implants, and to identify the materials for which the additional use of PRP might be associated with superior osseo- and soft tissues integration.
A search on PubMed database was performed considering the literature from 2000 to 2012, using the following string: ("Bone Substitutes"[Mesh] OR "Bone Transplantation"[Mesh] OR "Bone Regeneration"[Mesh] OR "Osseointegration"[Mesh]) AND ("Blood Platelets"[Mesh] OR "Platelet-Rich Plasma"[Mesh]). After abstracts screening, the full-texts of selected papers were analyzed and the papers found from the reference lists were also considered. The search focused on clinical applications documented in studies in the English language: levels of evidence included in the literature analysis were I, II and III.
Literature analysis showed 83 papers that fulfilled the inclusion criteria: 26 randomized controlled trials (RCT), 14 comparative studies, 29 case series, and 14 case reports. Several implant materials were identified: 24 papers on autologous bone, 6 on freeze-dried bone allograft (FDBA), 16 on bovine porous bone mineral (BPBM), 9 on β-tricalcium phosphate (β-TCP), 4 on hydroxyapatite (HA), 2 on titanium (Ti), 1 on natural coral, 1 on collagen sponge, 1 on medical-grade calcium sulphate hemihydrate (MGCSH), 1 on bioactive glass (BG) and 18 on a combination of biomaterials. Only 4 papers were related to the orthopaedic field, whereas the majority belonged to clinical applications in oral/maxillofacial surgery.
The systematic research showed a growing interest in this approach for bone implant integration, with an increasing number of studies published over time. However, knowledge on this topic is still preliminary, with the presence mainly of low quality studies. Many aspects still have to be understood, such as the biomaterials that can benefit most from PRP and the best protocol for PRP both for production and application.
PRP; Implant integration; Platelets; Bone; Regenerative medicine
The aim of the study was to control the in vivo localisation of implanted cells in cell-based therapies. Labelling cells with 111indium-oxine is one of the most interesting methods proposed. We evaluated this method in the setting of autologous osteoblast implantation in nonunion fractures.
An in vitro study of osteoblasts was conducted after 111indium-oxine labelling. Radioactivity retention and viability, proliferation and the ability to produce alkaline phosphatase were evaluated in a seven-day culture. In vivo labelling of implanted osteoblastic cells was conducted during a therapeutic trial of atrophic nonunion fractures, with the leakage outside the nonunion site and local uptake evolution at four, 24 and 48 hour being studied.
The mean labelling efficiency for osteoprogenitors was 78.8 ± 4.6 %. The intracellular retention was 89.4 ± 2.1 % at three hours and 67.3 ± 4.7 % at 18 hours. The viability assessed at three hours was 93.7 ± 0.6 %. After seven days of culture, morphology and alkaline phosphatase staining were similar for both labelled and unlabelled control cells, although the proliferation rate was decreased in the labelled cells. Some local intraosseous leakage was observed in four of 17 cases. All patients showed uptake at the injection site, with four having no other uptake. Four patients showed additional uptake in the bladder, liver and spleen, while 11 patients had additional uptake in the lungs in addition to the bladder, liver and spleen. The activity ratios (injection site/body) were 48 ± 28 % at four hours, 40 ± 25 % at 24 hours and 35 ± 25 % at 48 hours. After correcting for decay, the activity within the injection site was 82 ± 15 % at 24 hours and 69 ± 11 % at 48 hours compared with the activity measured at four hours. No relationship was found between uptake and radiological bone repair.
The 111indium-oxine labelling appears to be a good method for monitoring the behaviour of the osteoblastic cells after their implantation in atrophic nonunion fractures.
The surface properties of materials contribute to host cellular response and play a significant role in determining the overall success or failure of an implanted biomaterial. Rough titanium (Ti) surface microtopography and high surface free energy have been shown to enhance osteoblast maturation in vitro and increase bone formation in vivo. While the surface properties of Ti are known to affect osteoblast response, host bone quality also plays a significant role in determining successful osseointegration. One factor affecting host bone quality is patient age. We examined both in vitro and in vivo whether response to Ti surface features was affected by animal age. Calvarial osteoblasts isolated from 1-, 3-, and 11-month-old rats all displayed a reduction in cell number and increases in alkaline phosphatase specific activity and osteocalcin in response to increasing Ti surface microtopography and surface energy. Further, osteoblasts from the three ages examined displayed increased production of osteocalcin and local factors osteoprotegerin, VEGF-A, and active TGF-β1 in response to increasing Ti surface roughness and surface energy. Latent TGF-β1 only increased in cultures of osteoblasts from 1- and 3-month-old rats. Treatment with the systemic osteotropic hormone 1α,25(OH)2D3 further enhanced the response of osteoblasts to Ti surface features for all three age groups. However, osteoblasts derived from 11-month-old animals had a reduced response to 1α,25(OH)2D3 as compared to osteoblasts derived from 1-or 3-month-old animals. These results were confirmed in vivo. Ti implants placed in the femoral intramedullary canal of old (9-month) mice yielded lower bone-to-implant contract and neovascularization in response to Ti surface roughness and energy compared to younger (2-month) mice. These results show that rodent osteoblast maturation in vitro as well as new bone formation in vivo is reduced with age. Whether comparable age differences exist in humans needs to be determined.