Effects of type I collagen coating on titanium osseointegration: histomorphometric, cellular and molecular analyses

The investigation of titanium (Ti) surface modifications aiming to increase implant osseointegration is one of the most active research areas in dental implantology. This study was carried out to evaluate the benefits of coating Ti with type I collagen on the osseointegration of dental implants. Acid etched Ti implants (AETi), either untreated or coated with type I collagen (ColTi), were placed in dog mandibles for three and eight weeks for histomorphometric, cellular and molecular evaluations of bone tissue response. While the histological aspects were essentially the same with both implants being surrounded by lamellar bone trabeculae, histomorphometric analysis showed more abundant bone formation in ColTi, mainly at three weeks. Cellular evaluation showed that cells harvested from bone fragments in close contact with ColTi display lower proliferative capacity and higher alkaline phosphatase activity, phenotypic features associated with more differentiated osteoblasts. Confirming these findings, molecular analyses showed that ColTi implants up-regulates the expression of a panel of genes well known as osteoblast markers. Our results present a set of evidences that coating AETi with collagen fastens the osseointegration by stimulating bone formation at the cellular and molecular levels, making this combination of morphological and biochemical modification a promising approach to treat Ti surfaces.

Implant Osseointegration in Circumferential Bone Defects Treated with Latex-Derived Proteins or Autogenous Bone in Dog's Mandible

Background: In sites with diminished bone volume, the osseointegration of dental implants can be compromised. Innovative biomaterials have been developed to aid successful osseointegration outcomes. Purpose: The aim of this study was to evaluate the osteogenic potential of angiogenic latex proteins for improved bone formation and osseointegration of dental implants. Materials and Methods: Ten dogs were submitted to bilateral circumferential defects (5.0 x 6.3 mm) in the mandible. Dental implant (3.3 x 10.0 mm, TiUnite MK3 (TM), Nobel Biocare AB, Goteborg, Sweden) was installed in the center of the defects. The gap was filled either with coagulum (Cg), autogenous bone graft (BG), or latex angiogenic proteins pool (LPP). Five animals were sacrificed after 4 weeks and 12 weeks, respectively. Implant stability was evaluated using resonance frequency analysis (Osstell Mentor T, Osstell AB, Goteborg, Sweden), and bone formation was analyzed by histological and histometric analysis. Results: LPP showed bone regeneration similar to BG and Cg at 4 weeks and 12 weeks, respectively (p >= 3.05). Bone formation, osseointegration, and implant stability improved significantly from 4 to 12 weeks (p <= 2.05). Conclusion: Based on methodological limitations of this study, Cg alone delivers higher bone formation in the defect as compared with BG at 12 weeks; compared with Cg and BG, the treatment with LPP exhibits no advantage in terms of osteogenic potential in this experimental model, although overall osseointegration was not affected by the treatments employed in this study.

Short implants (6 mm) installed immediately into extraction sockets: An experimental study in dogs

Aim: To evaluate the effect of implant length (6 mm vs. 11 mm) on osseointegration (bone-toimplant contact) of implants installed into sockets immediately after tooth extraction. Material and methods: In six Labrador dogs, the pulp tissue of the mesial roots of P-3(3) was removed and the root canals were filled. Flaps were elevated bilaterally, the premolars hemisectioned and the distal roots removed. Recipient sites were prepared in the distal alveolus and a 6 mm or an 11 mm long implant was installed at the test and control sites, respectively. Non-submerged healing was allowed. After 4 months of healing, block sections of the implant sites were obtained for histological processing and peri-implant tissue assessment. Results: No statistically significant differences were found between test and control sites both for hard and soft tissue parameters. The bone-to-implant contact evaluated at the apical region of the implants was similar as well. Although not statistically significant, the location of the top of the bony crest at the buccal aspect was more apical in relation to the implant shoulder at the test compared with the control sites (2.0 +/- 1.4 and 1.2 +/- 1.1 mm, respectively). Conclusions: Shorter implants (6 mm) present with equal osseointegration than do longer implants (11 mm).

Role of teeth adjacent to implants installed immediately into extraction sockets: an experimental study in the dog

Aim: To evaluate the influence of the presence of both adjacent teeth on the level of alveolar bony crest at sites where implants were installed into the socket immediately after tooth extraction. Material and methods: Six Labrador dogs were used. Extractions of all teeth from the second premolar to the first molar were performed in the right side of the mandible, after full-thickness flap elevation. In the left side of the mandible, an endodontic treatment of the mesial root of the third and fourth premolars was performed. Full-thickness flaps were elevated, the teeth hemisected, and the distal roots removed. Immediately after, implants were bilaterally installed with the margin flush to the buccal bony crest. The implants were placed in the center of the alveolus at the third premolars and toward the lingual bony plate of the alveolus at the fourth premolars. After 3 months of healing, the animals were euthanized. Results: All implants were integrated in mature bone. More bone resorption was observed at the test compared to the control sites. At the buccal aspect, a resorption of 2.8 +/- 0.5 and 1.6 +/- 0.4 mm at the third premolars and of 2.4 +/- 0.6 and 0.8 +/- 0.7 mm at the fourth premolars were found, at the test and control sites, respectively. At the lingual aspect, the bony crest was apically located in relation to the implant shoulder 1.5 +/- 0.3 and 0.5 +/- 0.5 mm at the third premolars and 1.6 +/- 0.6 and 0.3 +/- 1.1 mm at the fourth premolars, at the test and control sites, respectively. A lower buccal bone resorption was found at the control implants placed lingually. Conclusion: Multiple extractions of teeth adjacent to a socket into which implants were installed immediately after, tooth extraction induced more alveolar bone recession compared to sites where the adjacent teeth were preserved. Moreover, an implant placed more lingually yielded less recession of the buccal aspect of the implant.

Buccal Bone Plate Remodeling After Immediate Implant Placement With and Without Synthetic Bone Grafting and Flapless Surgery: Radiographic Study in Dogs

Recent studies in animals have shown pronounced resorption of the buccal bone plate after immediate implantation. The use of flapless surgical procedures prior to the installation of immediate implants, as well as the use of synthetic bone graft in the gaps represent viable alternatives to minimize buccal bone resorption and to favor osseointegration. The aim of this study was to evaluate the healing of the buccal bone plate following immediate implantation using the flapless approach, and to compare this process with sites in which a synthetic bone graft was or was not inserted into the gap between the implant and the buccal bone plate. Lower bicuspids from 8 dogs were bilaterally extracted without the use of flaps, and 4 implants were installed in the alveoli in each side of the mandible and were positioned 2.0 mm from the buccal bone plate (gap). Four groups were devised: 2.0-mm subcrestal implants (3.3 x 8 mm) using bone grafts (SCTG), 2.0-mm subcrestal implants without bone grafts (SCCG), equicrestal implants (3.3 x 10 mm) with bone grafts (EGG), and equicrestal implants without bone grafts (ECCG). One week following the surgical procedures, metallic prostheses were installed, and within 12 weeks the dogs were sacrificed. The blocks containing the individual implants were turned sideways, and radiographic imaging was obtained to analyze the remodeling of the buccal bone plate. In the analysis of the resulting distance between the implant shoulder and the bone crest, statistically significant differences were found in the SCTG when compared to the ECTG (P = .02) and ECCG (P = .03). For mean value comparison of the resulting linear distance between the implant surface and the buccal plate, no statistically significant difference was found among all groups (P > .05). The same result was observed in the parameter for presence or absence of tissue formation between the implant surface and buccal plate. Equicrestally placed implants, in this methodology, presented little or no loss of the buccal bone. The subcrestally positioned implants presented loss of buccal bone, even though synthetic bone graft was used. The buccal bone, however, was always coronal to the implant shoulder.

Bone healing pattern in surgically created circumferential defects around submerged implants: an experimental study in dog

Objective: To describe the healing of marginal defects below or above 1 mm of dimension around submerged implants in a dog model. Material and methods: In 12 Labrador dogs, all mandibular premolars and first molars were extracted bilaterally. After 3 months of healing, full-thickness flaps were elevated in the edentulous region of the right side of the mandible. Two recipient sites were prepared and the marginal 5mm were widened to such an extent to obtain, after implant installation, a marginal gap of 0.5mm at the mesial site (small defect) and of 1.25mm at the distal site (large defect). Titanium healing caps were affixed to the implants and the flaps were sutured allowing a fully submerged healing. The experimental procedures were subsequently performed in the left side of the mandible. The timing of the experiments and sacrifices were planned in such a way to obtain biopsies representing the healing after 5, 10, 20 and 30 days. Ground sections were prepared and histomorphometrically analyzed. Results: The filling of the defect with newly formed bone was incomplete after 1 month of healing in all specimens. Bone formation occurred from the base and the lateral walls of the defects. A larger volume of new bone was formed in the large compared with the small defects. Most of the new bone at the large defect was formed between the 10- and the 20-day period of healing. After 1 month of healing, the outline of the newly formed bone was, however, located at a similar distance from the implant surface (about 0.4mm) at both defect types. Only minor newly formed bone in contact with the implant, starting from the base of the defects, was seen at the large defects (about 0.8mm) while a larger amount was detected at the small defects (about 2.2 mm). Conclusion: Marginal defects around titanium implants appeared to regenerate in 20-30 days by means of a distance osteogenesis. The bone fill of the defects was, however, incomplete after 1 month.

Use of a titanium device in lateral sinus floor elevation: an experimental study in monkeys

Aim: To evaluate the effect of a space-maintaining device fixed to the lateral wall of the maxillary sinus after the elevation of the sinus mucosa on bone filling of the sinus cavity. Material and methods: Immediately after the elevation of the maxillary sinus Schneiderian membrane accomplished through lateral antrostomy in four monkeys, a titanium device was affixed to the lateral sinus wall protruding into the sinus cavity to maintain the mucosa elevated without the use of grafting material. The healing of the tissue around the implants was evaluated after 3 and 6 months. Ground sections were prepared and analyzed histologically. Results: The void under the elevated sinus membrane, originally filled with the blood clot, was reduced after 3 as well as after 6 months of healing of about 56% and 40.5%, respectively. In seven out of eight cases, the devices had perforated the sinus mucosa. The formation of mineralized bone and bone marrow amounted to about 42% and 69% after 3 and 6 months, respectively. The connective tissue represented about 53% and 23% of the newly formed tissue after 3 and 6 months, respectively. Conclusions: New bone formation was found below the devices. However, shrinkage of the newly formed tissue was observed both after 3 and 6 months of healing. Hence, the space-maintaining function of the devices used in the present study has to be questioned.

Early peri-implant endosseous healing of two implant surfaces placed in surgically created circumferential defects. A histomorphometric and fluorescence study in dogs

Objective Several implant surfaces are being developed, some in the nanoscale level. In this study, two different surfaces had their early healing properties compared in context of circumferential defects of various widths. Material and methods Six dogs had the mandibular premolars extracted. After 8weeks, four implants were placed equicrestally in each side. One acted as control, while the others were inserted into sites with circumferential defects of 1.0, 1.5 and 2.0mm wide and 5mm deep. A nano-modified surface was used on one side and a micro-rough on the other. Bone markers were administered on the third day after implant placement and then after 1, 2, 4weeks to investigate the bone formation dynamic through fluorescence analysis. Ground sections were prepared from 8-week healing biopsies and histomorphometry was performed. Results The fluorescence evaluation of the early healing showed numerically better results for the nano-modified group; however this trend was not followed by the histomorphometric evaluation. A non-significant numerical superiority of the micro-rough group was observed in terms of vertical bone apposition, defect bone fill, bone-to-implant contact and bone density. In the intra-group analysis, the wider defects showed the worse results while the control sites showed the best results for the different parameters, but without statistical relevance. Conclusion Both surfaces may lead to complete fill of circumferential defects, but the gap width has to be considered as a challenge. The nano-scale modification was beneficial in the early stages of bone healing, but the micro-rough surface showed numerical better outcomes at the 8-week final period.

The effect of implant design on insertion torque and immediate micromotion

Objectives: To evaluate the effect of insertion torque on micromotion to a lateral force in three different implant designs. Material and methods: Thirty-six implants with identical thread design, but different cutting groove design were divided in three groups: (1) non-fluted (no cutting groove, solid screw-form); (2) fluted (901 cut at the apex, tap design); and (3) Blossomt (Patent pending) (non-fluted with engineered trimmed thread design). The implants were screwed into polyurethane foam blocks and the insertion torque was recorded after each turn of 901 by a digital torque gauge. Controlled lateral loads of 10N followed by increments of 5 up to 100N were sequentially applied by a digital force gauge on a titanium abutment. Statistical comparison was performed with two-way mixed model ANOVA that evaluated implant design group, linear effects of turns and displacement loads, and their interaction. Results: While insertion torque increased as a function of number of turns for each design, the slope and final values increased (Po0.001) progressively from the Blossomt to the fluted to the non-fluted design (M +/- standard deviation [SD] = 64.1 +/- 26.8, 139.4 +/- 17.2, and 205.23 +/- 24.3 Ncm, respectively). While a linear relationship between horizontal displacement and lateral force was observed for each design, the slope and maximal displacement increased (Po0.001) progressively from the Blossomt to the fluted to the non-fluted design (M +/- SD 530 +/- 57.7, 585.9 +/- 82.4, and 782.33 +/- 269.4 mm, respectively). There was negligible to moderate levels of association between insertion torque and lateral displacement in the Blossomt, fluted and non-fluted design groups, respectively. Conclusion: Insertion torque was reduced in implant macrodesigns that incorporated cutting edges, and lesser insertion torque was generally associated with decreased micromovement. However, insertion torque and micromotion were unrelated within implant designs, particularly for those designs showing the least insertion torque.

Biomechanical and histologic evaluation of non-washed resorbable blasting media and alumina-blasted/acid-etched surfaces

Objectives: To compare the biomechanical fixation and histomorphometric parameters between two implant surfaces: non-washed resorbable blasting media (NWRBM) and alumina-blasted/acid-etched (AB/AE), in a dog model. Material and methods: The surface topography was assessed by scanning electron microscopy, optical interferometry and chemistry by X-ray photoelectron spectroscopy (XPS). Six beagle dogs of similar to 1.5 years of age were utilized and each animal received one implant of each surface per limb (distal radii sites). After a healing period of 3 weeks, the animals were euthanized and half of the implants were biomechanically tested (removal torque) and the other half was referred to nondecalcified histology processing. Histomorphometric analysis considered bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO). Following data normality check with the Kolmogorov-Smirnov test, statistical analysis was performed by paired t-tests at 95% level of significance. Results: Surface roughness parameters Sa (average surface roughness) and Sq (mean root square of the surface) were significantly lower for the NWRBM compared with AB/ AE. The XPS spectra revealed the presence of Ca and P in the NWRBM. While no significant differences were observed for both BIC and BAFO parameters (P>0.35 and P>0.11, respectively), a significantly higher level of torque was observed for the NWRBM group (P = 0.01). Bone morphology was similar between groups, which presented newly formed woven bone in proximity with the implant surfaces. Conclusion: A significant increase in early biomechanical fixation was observed for implants presenting the NWRBM surface.

Retrospective Clinical Study of 988 Dual Acid-Etched Implants Placed in Grafted and Native Bone for Single-Tooth Replacement

Purpose: To evaluate the influence of sex, implant characteristics, and bone grafting on the survival rate of dual acid-etched (DAE) implants. Materials and Methods: Patients treated with internal-hex DAE implants for single-tooth replacement in a military dental clinic between January 2005 and December 2010 were included in this study. Clinical data related to implant characteristics, implant location, presence of grafted bone, and implant failures were collected. The primary outcome was implant loss. The survival rate was analyzed using the Kaplan-Meier method. Cox regression modeling was used to determine which factors would predict implant failure. Results: DAE implants were evaluated in a total of 988 patients (80.3% men). Twenty-four (2.4%) implants failed, most were cylindric (54.2%) with regular platforms (70.8%) and were 10 mm long (58.3%). The failure rate was 2.4% for the anterior maxilla, 3.3% for the posterior maxilla, 1.6% for the anterior mandible, and 2.0% for posterior mandible. The cumulative survival rate was 97.6%. The failure rate was 8.8% in implants placed after sinus augmentation, 7.3% in bone block-grafted areas, and 1.6% in native bone. Based on multivariable analysis (Cox regression), sinus augmentation and bone block grafting had a statistically significant effect on implant failure; the hazard ratios were 5.5 and 4.6, respectively. Conclusion: The results revealed that DAE implants had high survival rates, and no influence of sex, location, shape, diameter, or length on failure rates could be observed. However, a significant association was observed between failure and presence of bone graft in the implant area. Int J Oral Maxillofac Implants 2012;27:1243-1248

In vitro analysis of the influence of surface treatment of dental implants on primary stability

Surface treatment interferes with the primary stability of dental implants because it promotes a chemical and micromorphological change on the surface and thus stimulates osseointegration. This study aimed to evaluate the effects of different surface treatments on primary stability by analyzing insertion torque (IT) and pullout force (PF). Eight samples of implants with different surface treatments (TS - external hexagon with acid surface treatment; and MS - external hexagon, machined surface), all 3.75 mm in diameter x 11.5 mm in length, were inserted into segments of artificial bones. The IT of each sample was measured by an electronic torquemeter, and then the pullout test was done with a universal testing machine. The results were subjected to ANOVA (p < 0.05), followed by Tukey's test (p < 0.05). The IT results showed no statistically significant difference, since the sizes of the implants used were very similar, and the bone used was not highly resistant. The PF values (N) were, respectively, TS = 403.75 +/- 189.80 and MS = 276.38 +/- 110.05. The implants were shown to be different in terms of the variables of maximum force (F = 4.401, p = 0.0120), elasticity in maximum flexion (F = 3.672, p = 0.024), and relative stiffness (F = 4.60, p = 0.01). In this study, external hexagonal implants with acid surface treatment showed the highest values of pullout strength and better stability, which provide greater indication for their use.

Long-term rabbits bone response to titanium implants in the presence of inorganic bovine-derived graft

This study evaluated bone responses to titanium implants in the presence of an inorganic graft material. The bilateral mandible incisors of 24 rabbits were surgically extracted and one of the exposed sockets, chosen at random, was filled with an inorganic xenogenic bone graft (Gen-ox (R)), whereas the remaining socket was left to heal naturally and served as a control. After 60 days, titanium implants were inserted in the specific areas, and on days 0, 30, 60, and 180 after the implant insertions, six animals of each group were killed. Digital periapical radiography of implant region was obtained and vertical bone height (VBH) and bone density (BD) were evaluated by digital analysis system. In the undecalcified tissue cuts, bone-to-implant contact (BIC) and bone area (BA) within the limits of the implant threads were evaluated and compared statistically by means of two-way ANOVA and Tukey's test (rho < 0.05). No significant differences were detected in VBH and BA, either between groups or between different experimental intervals. The BD was significantly higher in the experimental group than in the control group in all the intervals tested, but there were no significant differences by interval. The BIC was statistically lower in the control group on day 0; however, a significant increase was observed on days 60 and 180 (rho < 0.05). The use of an inorganic xenograft prior to insertion of a titanium implant did not interfere with the course of osseointegration.

Bone tissue, cellular, and molecular responses to titanium implants treated by anodic spark deposition

A myriad of titanium (Ti) surface modifications has been proposed to hasten the osseointegration. In this context, the aim of this study was to perform histomorphometric, cellular, and molecular analyses of the bone tissue grown in close contact with Ti implants treated by anodic spark deposition (ASD-AK). Acid-etched (AE) Ti implants either untreated or submitted to ASD-AK were placed into dog mandibles and retrieved at 3 and 8 weeks. It was noticed that both implants, AE and ASD-AK, were osseointegrated at 3 and 8 weeks. Histomorphometric analysis showed differences between treatments only for bone-to-implant contact, being higher on AE implants. Although not backed by histomorphometric results, gene expression of key bone markers was higher for bone grown in close contact with ASD-AK and for cells harvested from these fragments and cultured until subconfluence. Cell proliferation at days 7 and 10 and alkaline phosphatase activity at day 10 was higher on AE surfaces. No statistical significant difference was noticed for extracellular matrix mineralization at 17 days. Our results have shown that the Ti fixtures treated by ASD-AK allowed in vivo osseointegration and induced higher expression of key markers of osteoblast phenotype, suggesting that this surface treatment could be considered to produce implants for clinical applications. (c) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A:30923098, 2012.

The impact of healing time before loading on orthodontic mini-implant stability: a histomophometric study in minipigs

Objective: To evaluate healing time before loading, areas compression and tension and location of insertion on mini-implant stability. Design: Six minipigs were used. Each animal received 3 mini-implants in each quadrant: 1 mini-implant was used as an unloaded control (G1, n = 24); the other 2 were loaded with 150 g-force at three time intervals (G2: immediate loading, G3: after 15 days and G4: after 30 days), with 16 mini-implant in each experimental group. After 120 days, tissue blocks of the areas of interest were harvested. Clinical analysis (exact Fisher test) determined the survival rate. Histological analysis (Kontron KS 300TM, Zeiss) quantified the fractional bone-toimplant contact (%BIC) and bone area (%BA) at each healing time point, areas of interest, and insertion site (ANOVA and t tests for dependent and independent samples). Results: The mini-implant survival rates were G1: 71%, G2: 50%, G3: 75% and G4: 63%, with no statistical differences between them. The groups presented similar %BIC and %BA. There were no differences between the compression and tension sides or maxillary and mandibular insertion sites. Conclusions: These results suggest that low-intensity immediate or early orthodontic loading does not affect mini-implant stability, because similar histomorphometric results were observed for all the groups, with partial osseointegration of the mini-implants present.