Immunolocalization of fibronectin and vitronectin receptors in human gingival fibroblasts spreading on titanium surfaces

The adhesion and spreading of human gingival fibroblasls on glass and differently processed titanium surfaces was studied by immunolocalization of vinculin and the alpha and beta subunits of the fibronectin (α₅β₁) and vitronectin (α_vβ3) receptors. Vinculin-containing focal contacts were present both at 4 and 24 h of spreading in cells grown on glass or electropolished or etched titanium surfaces but not in cells spreading on sandblasted titanium surfaces. Immunostaining for the α5 and β1 subunits of the fibronectin receptor showed only a diffuse membrane fluorescence after 4 h of cell spreading irrespective of the growth surface. The α_v and β₃ subunits of the vitronectin receptor were at this stage detected in focal contacts in cells spreading on glass or electropolished or etched titanium surfaces. In cells spreading on sandblasted titanium surfaces, however, the vitronectin receptor had only a diffuse distribution. In cells that had been allowed to spread for 24 h on glass or electropolished or etched titanium surfaces the α₅ and β₁ integrin subunits were either diffusely distributed or showed a localization typical of extracellular matrix contacts. The α_v and β₃ integrin subunits were, as earlier, localized to typical focal contacts in cells grown on glass or electropolished or etched titanium surfaces. Cells attached to sandblasted titanium surfaces still expressed all the integrin subunits only diffusely. The results show that the surface texture of the substratum can affect the expression of integrin subunits in human gingival fibroblasts. As evidenced by the recruitment of integrin subunits to focal and extracellular matrix contacts, smooth or finely grooved titanium surfaces appear to be optimal in supporting the attachment of human gingival fibroblasts.     

Altered cell motility and attachment with titanium surface modifications

Background: Titanium implants are widely used in dentistry to replace lost teeth. Various surface modifications have been used to improve implant retention and osseointegration. This study is designed to compare the ability of three titanium surfaces to promote cell attachment and cell motility of cells relevant to periodontal tissues. Methods: Three clinically relevant surfaces were tested: 1) machined titanium; 2) a titanium surface roughened through acid etching (dual thermal‐etched titanium [DTET]); and 3) a titanium surface roughened with nanometer‐scale calcium phosphate deposition (nanoscale calcium phosphate–impregnated titanium [NCPIT]). Cell attachment and migration were examined for four cell types: rat osteosarcoma cells, human osteoblasts, and gingival and periodontal ligament (PDL) fibroblasts. Results: All four cell types attached to each of the three titanium surfaces equally by 2 hours, and the PDL and gingival fibroblasts generally displayed less attachment than the osteosarcoma cells and osteoblasts. The cells displayed differential motility and long‐term attachment to each of the titanium surfaces. Osteosarcoma cells displayed preferential motility on NCPIT, whereas PDL fibroblasts were more motile on machined titanium, and gingival fibroblasts moved more rapidly on both DTET and NCPIT. Osteoblasts displayed little motility on any of the titanium surfaces and lost viability on NCPIT after 24 hours. Gingival fibroblasts lost attachment to machined titanium. Conclusions: Periodontal cells displayed differential motility and long‐term attachment to titanium surfaces. Selective modification of titanium surface properties in various regions of an implant may be useful in guiding specific cell populations to specific locations where they might best aid in osseointegration and soft tissue remodeling.     

Early cell response of osteogenic cells on differently modified implant surfaces: Sequences of cell proliferation,adherence and differentiation

Objectives

Osseointegration of dental implants is a crucial prerequisite for long-term survival. Therefore, surface modifications are needed to interact with the extracellular environment and to trigger osteogenic cell responses such as cell proliferation, adherence, and differentiation. The purpose of this study was to investigate different surface modifications in vitro over 2 weeks.

Differential chemotactic effect of cementum attachment protein on periodontal cells

Selective re-population of the root surface by periodontal ligament cells is considered a key factor in Periodontal regeneration. A recently isolated cementum attachment protein (CAP) has been shown to enhance fibroblast attachment. In the present study the potential of CAP to selectively attract periodontal ligament cells (PLC) was studied in vitro in a micro-chemotaxis system. Human periodontal ligament cells and gingival fibroblasts (GF) were compared for their chemotactic response to either cementum attachment protein or to fibronectin. Murine dermal fibroblasts (MDF) served as control, irrelevant to the periodontium. The chemotactic response of PLC to fibronectin at 10^?8 M was of a similar magnitude as that of GF (16 ± 5 and 11 ± 3 cells/field, respectively), but both were significantly lower than the response of MDF (28 ± 3 cells/field). The chemotactic response of periodontal ligament cells to the cementum attachment protein at 10^?7 M was higher (36 ± 5 cells/field) than that of gingival fibroblasts or murine dermal fibroblasts (14 ± 2 and 16 ± 2 cells/field, respectively). These results suggest that cementum attachment protein can influence the selective re-population of root surfaces by periodontal ligament cells.     

Effects of an air-powder abrasive system on plasma-sprayed titanium implant surfaces: an in vitro evaluation

The purpose of this study was to conduct an in vitro evaluation of the effects of an air-powder abrasive system, commonly used in clinical dentistry for periodontal maintenance, on the surfaces of plasma-sprayed titanium dental implants. Twenty-eight plasma-coated titanium implant specimens were divided into a sterile water-treated control group and an air-powder-abrasive-treated test group. All specimens were subjected to three different in vitro testing conditions and post-treatment evaluations by scanning electron microscopy (SEM): (1) Topographical features of implant surfaces were studied before and after direct exposure to the abrasive; (2) biocompatibility of treated implant surfaces was evaluated and compared with those of control specimens via in vitro fibroblast attachment studies; and (3) the attachment of a common oral microbe to the implant surface and its subsequent removal by exposure to the air-powder abrasive were also evaluated. Results indicate that exposure of implant specimens to the air-powder abrasive for various periods resulted in only slight changes in surface topography, i.e., rounding of angles and edges of the plasma-spray coating and occasional surface pitting. Examination by SEM and a statistical comparison of the difference between the mean numbers of attached fibroblasts between control and test groups revealed no statistical significance. In both specimen groups, fibroblasts exhibited uniform attachment over the entire implant surface. A comparison of test and control groups demonstrated 100% removal of bacteria from the surfaces of test specimens exposed to the air-powder abrasive and approximately a 75% removal from control specimens exposed to sterile water.     

The Teeth of Alligator Mississippiensis Daud: VI. Periodontium

Abstract

Objective. Although dental implants are commonly used for tooth restoration, there is a lack of studies of treatment regimens for preventing extra-oral infection and decreasing osseointegration failures by establishing early peri-implant soft tissue seals on titanium dental implant abutments. In this study, air atmospheric-pressure plasma-jet (AAPPJ) treatment was applied to titanium disks to assay the potential for early peri-implant soft tissue seals on titanium dental implant abutment. Materials and methods. After titanium disks were treated with AAPPJ for 10 s at 250, 500, 1000 and 1500 sccm, surface analysis was performed; the control group received air only or no treatment. Human gingival fibroblasts (HGF) were seeded onto the specimens for evaluating cell attachment and proliferation and adherent-cell morphology was visualized via confocal microscopy. Results. In AAPPJ-treated specimens, the water contact angle decreased according to increased flow rate. Oxygen composition increased in XPS, but no topographical changes were detected. The effect of AAPPJ treatment at 1000 sccm was apparent 2 mm from the treated spot, with a 20% increase in early cell attachment and proliferation. Adherent HGF on AAPPJ-treated specimens displayed a stretched phenotype with more vinculin formation than the control group. Conclusions. Within the limitations of this study, the results indicate that AAPPJ treatment may enhance the early attachment and proliferation of HGF for establishing early peri-implant soft tissue seals on titanium dental implant abutments with possible favorable effects of osseointegration of dental implant.     

碱性成纤维细胞生长因子对牙周膜成纤维细胞整合素β1亚单位mRNA表达的影响

目的研究碱性成纤维细胞生长因子（bFGF）对人牙周膜成纤维细胞整合素β1亚单位mRNA表达的影响,探讨bFGF在牙周组织再生中的意义。方法体外培养人牙周膜成纤维细胞,分别用质量浓度0.1、1.0、10.0 ng·mL-1的bFGF刺激细胞,培养24、48、72 h,采用实时荧光定量聚合酶链反应法检测牙周膜成纤维细胞内整合素β1亚单位mRNA表达的变化。结果bFGF可促进人牙周膜成纤维细胞内整合素β1亚单位mRNA的合成,在培养24、48、72 h时,1.0 ng·mL-1组整合素β1亚单位表达均明显高于对照组;培养72 h时各实验组整合素β1亚单位表达均明显高于培养24、48 h时。结论bFGF通过提高整合素β1亚单位mRNA的表达,促进牙周膜成纤维细胞的黏附,在牙周组织修复再生中起作用。     

Periodontal ligament formation around titanium implants using cultured periodontal ligament cells: a pilot study

The aim of this study was to investigate whether a new periodontal ligament attachment will form on titanium implants when they are implanted with cultured periodontal ligament cells. Periodontal ligament cells obtained from the teeth of 3 dogs were cultured and attached to the surface of titanium implants. The implants with the cultured autologous periodontal ligament cells were placed in the mandibles of the dogs. After 3 months of healing, histologic examination revealed that, on some implant surfaces, a layer of cementum-like tissue with inserting collagen fibers had been achieved. These results demonstrated that cultured periodontal ligament cells can form tissue resembling a true periodontal ligament around implants.     

Ultrastructure of the Interface between Periodontal Tissues and Titanium Mini-Implants

Objective:To describe the ultrastructure of the interface between periodontal tissues and titanium mini-implants in rat mandibles.Materials and Methods:A titanium mini-implant was placed between the buccal roots of the mandibular first molar of 24 adult rats. After 21, 30, 45, 60, 90, and 120 days of implantation, the mandibular portion was removed and fixed in cacodylate-buffered 2% glutaraldehyde + 2.5% formaldehyde. The material was decalcified and processed for scanning and transmission electron microscopy.Results:Ultrastructural analysis revealed a thin cementum-like layer at longer times after implantation at the areas in which the periodontal ligament was in contact with the implant.Conclusions:The alveolar bone and the periodontal ligament reorganized their constituents around the implant, and a thin cementum-like layer was formed at longer times after implantation at the areas in which the periodontal ligament was in contact with the implant.     

Adhesion of human fibroblasts to root-end-filling materials

This study evaluated the attachment of cultured explants of human periodontal ligament fibroblasts and gingival fibroblasts to different root-end-filling materials. Although periodontal ligament and gingival fibroblasts initially attached avidly to Geristore, these same cells displayed no significant attachment to ProRoot, Tytin amalgam, or SuperEBA. With further incubation on Geristore, the attachment of both periodontal ligament and gingival fibroblasts improved and these cells proliferated. In contrast, no improvement in attachment or proliferation was observed for cells incubated for greater times with ProRoot, Tytin amalgam, or SuperEBA. Because the attachment characteristics of these two groups of fibroblasts were identical, we examined the potential role of the extracellular matrix family of receptors (integrins) on the attachment of gingival fibroblasts. Gingival fibroblast attachment to collagen type I was determined to be dependent on alpha1beta1 and alpha2beta1 integrins, whereas their attachment to the RGD-binding sequence of fibronectin and vitronectin was partially inhibited by antibodies to the beta1 and alphaV integrin subunits. However, attachment of gingival fibroblasts to Geristore was not reduced by the addition of any of the attachment-perturbing anti-integrin antibodies examined. Thus, gingival fibroblasts attach to Geristore, but this attachment was mediated by mechanisms other than integrins.     

New formation of periodontal tissues around titanium implants in a novel dentin chamber model

Direct bone-to-implant contact, defined as "osseointegration", is considered most optimal for long-term stability and survival of dental implants. However, the possibility of the formation of a tooth-like attachment apparatus around implants has also been demonstrated. The purpose of this study was to explore the formation of periodontal tissues around titanium implants using a novel and unique experimental model. After resection of the crowns of the maxillary canine teeth in nine mongrel dogs, the roots were hollowed to a depth of 5 mm leaving a thin dentinal wall. Slits were prepared in the cavity wall to create passages from the chamber to the periodontal ligament area. A custom-made, titanium implant was placed into the center of each chamber. Machined, titanium plasma sprayed (TPS) and sand blasted with large grit and acid attacked (SLA) surfaces were used. A collagen barrier was placed over the submerged chamber. Following 4 months of healing, jaw sections were processed for histology. Newly formed periodontal ligament, alveolar bone, and root cementum filled the space between the implant and the wall of the chamber. Ingrown bone was neither in contact with dentin nor with the implant. Thus, an interposed soft connective tissue layer was present. Healing by fibrous encapsulation was observed around most implants. However, cellular cementum was deposited on one TPS and one SLA implant and on the dentinal walls of the chamber. This study shows a remarkable capacity for new periodontal tissue formation at a site where no such tissues ever existed. Maintenance of original periodontal tissue domains most likely prevented osseointegration of the implants. The cementum layer deposited on two implants was likely formed through cementoconductivity rather than by differentiation of periodontal ligament cells upon contact with the implant surface.     

Biofunctionalization of titanium implants with a biomimetic active peptide (P‐15) promotes early osseointegration

Objectives: The early stages of peri‐implant bone formation play an essential role in the osseointegration and long‐term success of dental implants. By incorporating bioactive coatings, this biofunctionalization of implant surfaces may enhance the attachment of the implant to the surrounding bone and stimulate bone regeneration. Material and methods: To demonstrate faster osseointegration, the surfaces of dental implants were grit‐blasted and acid‐etched. They were then coated with hydroxyapatite (HA) and experimental implants were further coated with a biomimetic active peptide (P‐15) in one of two concentrations. These biofunctionalized samples and controls with no peptide were placed in the forehead region of 12 adult pigs. Six animals were evaluated for a period of 14 or 30 days. Results: Histomorphometric analysis demonstrated that the implants with the high concentration of P‐15 had significantly higher percentage of bone‐to‐implant contact (BIC) at 14 (P=0.018) and 30 (P=0.015) days compared with the other groups. Both concentrations of P‐15 showed increased peri‐implant bone density compared to the control group at 30 days. Conclusion: Biofunctionalization of the implant surface with a biomimetic active peptide leads to significantly increased BIC rates at 14 and 30 days and higher peri‐implant bone density at 30 days. To cite this article:
Lutz R, Srour S, Nonhoff J, Weisel T, Damien CJ, Schlegel KA. Biofunctionalization of titanium implants with a biomimetic active peptide (P‐15) promotes early osseointegration.
Clin. Oral Impl. Res. 21 , 2010; 726–734.
doi: 10.1111/j.1600‐0501.2009.01904.x     

Ultrastructural changes of smooth and rough titanium implant surfaces induced by metal and plastic periodontal probes

Objectives

To determine the ultrastructural changes of titanium surfaces of dental implants induced by the tip of periodontal probes.

Materials and methods

A total of 40 samples of smooth and rough surfaces of titanium implants were randomly assigned for the treatment with metal or plastic periodontal probes under application angles of 20° and 60°. Titanium surfaces have been evaluated with CLSM prior and following to experimental probing determining various standardized 2D and 3D roughness parameters.

Results

The average profile and surface roughness (Ra and Sa) showed no significant difference between treated and untreated samples on smooth and rough surface areas irrespective of the probe material. On smooth surfaces several amplitude roughness parameters were increased with metal probes but reached significance only for Rp (p = 0.007). Rough surface parts showed a slight but not significant reduction of roughness following to the contact with metal probes. The surface roughness remained almost unchanged on smooth and rough implant surfaces using plastic probes. The surface roughness on implant surfaces was not dependent on the application angle irrespective of the probe material.

Conclusion

Probing of titanium implants with metal probes and even less with plastic probes causes only minor changes of the surface roughness. The clinical significance of these changes remains to be elucidated.

Clinical relevance

Using plastic probes for the clinical evaluation of the peri-implant sulcus might avoid ultrastructural changes to titanium implant surfaces.

     

A Laboratory Investigation to Compare Enamel Preparation by Sandblasting or Acid Etching Prior to Bracket Bonding

Abstract

A laboratory investigation to compare the mean shear debonding force and mode of bond failure of metallic brackets bonded to sandblasted and acid-etched enamel is described. The buccal surfaces of 30 extracted human premolars were sandblasted for 5 seconds with 50 µ alumina and the buccal surfaces of a further 30 human premolars were etched with 37 per cent phosphoric acid for 15 seconds. Following storage for 24 hours at 37°C in distilled water, shear debonding forcewas measured using an Instron Universal Testing Machine with a cross-head speed of 10 mm/minute. Mean shear debonding force was significantly lower for brackets bonded to sandblasted enamel compared to acid etched enamel (P<0.001). Weibull analysis showed that at a given stress the probability of failure was significantly greater for brackets bonded to sandblasted enamel. Brackets bonded to etched enamel showed a mixed mode of bond failure whereas following sandblasting, failure was adhesive at the enamel/composite interface (P<0.01).     

Bioengineered periodontal tissue formed on titanium dental implants

The ability to use autologous dental progenitor cells (DPCs) to form organized periodontal tissues on titanium implants would be a significant improvement over current implant therapies. Based on prior experimental results, we hypothesized that rat periodontal ligament (PDL)-derived DPCs can be used to bioengineer PDL tissues on titanium implants in a novel, in vivo rat maxillary molar implant model. Analyses of recovered implants revealed organized PDL tissues surrounding titanium implant surfaces in PDL-cell-seeded, and not in unseeded control, implants. Rat PDL DPCs also exhibited differentiative potential characteristic of stem cells. These proof-of-principle findings suggest that PDL DPCs can organize periodontal tissues in the jaw, at the site of previously lost teeth, indicating that this method holds potential as an alternative approach to osseointegrated dental implants. Further refinement of this approach will facilitate the development of clinically relevant methods for autologous PDL regeneration on titanium implants in humans.     

Effect of various implant coatings on biological responses in MG63 using cDNA microarray

During the process of bone formation, titanium (Ti) surface is an important factor in the modulation of osteoblastic function. This study was conducted in order to determine the effects of different Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on smooth (S), sandblasted large-grit and acid etching (SLA), hydroxyapatite (HA), hydroxyfluoride (HF), titanium nitrate (TIN), and diamond-like carbon (DLC) Ti. The morphology of these cells were assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1152 elements). The appearances of the surfaces observed by SEM were different on each of the six dental substrate types. The SLA and HA surfaces were determined to be rougher than the others. MG63 cells cultured on SLA and HA exhibited cell-matrix interactions. In the expression of genes involved in osseointegration, several genes, including bone morphogenetic protein, cadherin, integrin, and insulin-like growth factors, were upregulated on the different surfaces. Several genes, including fibroblast growth factor receptor 4, Bcl 2-related protein, and collagen, were downregulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface roughness of the dental materials used.     

Titanium endosseous implant-soft tissue interface: a literature review

Background information about normal periodontal anatomy and titanium used in endosseous implant fabrication is provided. Literature is reviewed concerning epithelial and connective tissue attachment to titanium. Information about the adequacy of cell attachment to implants, possible mechanisms of cell attachment formation, and the effect of implant surface properties on attachment is presented. A chemical attachment between titanium implant surface oxide layer and epithelium has been demonstrated in vitro and in vivo. This attachment is mediated by a glycoprotein similar to that seen between epithelium and natural tooth surfaces. While only minimal histological evidence exists, connective tissue fibers adjacent to titanium implanted surfaces may bring the tissue in tight apposition to the implant without an absolute biologic attachment between the implant and connective tissue. Alteration of the titanium surface morphology may selectively enhance the attachment of either epithelial cells or fibroblasts, theoretically enhancing the formation of a biologic seal between the implanted titanium surface and its adjacent tissue. A greater understanding of the mechanisms of attachment and of the factors which enhance the integrity of the biologic seal between implant and soft tissues should permit an improved prognosis for functioning titanium implants.     

Physico-chemical characterization of the surface of 9 dental implants with 3 different surface treatments

There are many surface treatments applied to dental implants. The aim of the present investigation is to compare the physicochemical characteristics of titanium dental implant surfaces with different surface treatments. 9 dental implants from the same batch were divided in 3 groups and received 3 different surface treatments: machined, acid etched and a new chemical surface treatment called Avantblast. Scanning electron microscopy and confocal microscopy were used to image the treated surfaces, and energy-dispersive spectrometry and X-ray photoelectron spectrometry to provide a chemical characterization of the surfaces. RESULTS: The acid etched and chemical etched surfaces had an increased roughness over the machined one. Surface chemical composition had differences between processes, as the surface with the new treatment presented a reduced level of impurities and increased thickness of the titanium oxide layer. CONCLUSIONS: Surface roughness of titanium dental implants and thickness of the titanium oxide layer can be increased with a suitable surface treatment.