In vitro analysis with human bone marrow stem cells on Ti-15Mo alloy for dental and orthopedic implants application

Aim: Nowadays, research on orthopedic and dental implants is focused on titanium alloys for their mechanical properties and corrosion resistance in the human body environment. Another important aspect to be investigated is their surface topography, which is very important to osseointegration. With laser beam irradiation for roughening the implants surface an easier control of the microtopography is achieved, and surface contamination is avoided. The aim of this study was to assess human bone marrow stem cells response to a newly developed titanium alloy, Ti-15Mo, with surface topography modified by laser beam irradiation. Materials and methods: A total of 10 Ti machined disks (control), 10 Ti-15Mo machined disks and 10 Ti-15Mo disks treated by laser beam-irradiation were prepared. To study how Ti-15Mo surface topografy can induce osteoblast differentiation in mesenchymal stem cells, the expression levels of bone related genes and mesenchymal stem cells marker were analyzed, using real time Reverse Transcription-Polymerase Chain Reaction. Results: In Test 1 (comparison between Ti-15Mo machined disks and Ti-machined disks) quantitative real-time RT-PCR showed a significant induction of ALPL, FOSL1 and SPP1, which increase 20% or more. In Test 2 (comparison between Ti-15Mo laser treated disks and Ti-machined disks) all investigated genes were up-regulated. By comparing Test 1 and Test 2 it was detected that COL1A1, COL3A1, FOSL1 and ENG sensibly increased their expression whereas RUNX2, ALPL and SPP1 expression remained substantially unchanged. Conclusion: The present study demonstrated that laser treated Ti-15Mo alloys are promising materials for implants application.

Cyclosporine-a and bone density around titanium implants: A histometric study in rabbits

Aim: Cyclosporine A (CsA) is an immunosuppressive agent commonly used to prevent organ transplantation rejection. It has been demonstrated that CsA may negatively affect osseointegration around dental implants. Therefore, the aim of this study was to evaluate the effect of CsA administration on bone density around titanium dental implants. Materials and Methods: Fourteen New Zealand rabbits were randomly divided into 2 groups with seven animals each. The test group (CsA) received daily subcutaneous injection of CsA (10mg/kg body weight) and the control group (CTL) received saline solution by the same route of administration. Three days after the beginning of immunosuppressive therapy, one machined dental implant (7.00 mm in lenght and 3.75 mm in diameter) was inserted bilaterally at the region of the tibial methaphysis. After 4 and 8 weeks the animals were sacrificed and the histometrical procedures were performed to analyse the bone density around the first four threads of the coronal part of the implant. Results: A significant increase in the bone density was observed from the 4- to the 8 week-period in the control group (37.41% + 14.85 versus 58.23% + 16.38 - p <0.01). In contrast, bone density consistently decreased in the test group overtime (46.31% + 17.38 versus 16.28 + 5.08 - p <0.05). In the 8-week period, there was a significant difference in bone density between the control and the test groups (58.23 + 16.38 eand16.28 + 5.08 - p= 0.001). Conclusion: Within the limits of this study, long-term CsA administration may reduce bone density around titanium dental implants during the osseointegration process.

A CAD/CAM flapless surgical technique and immediate prosthesis: A clinical report

Background: This article describes a clinical report with a new system for guided surgical treatment and immediate load prosthesis in the flapless surgical technique. Case report: Based on a computed tomography (CT) of a 64 - year-old edentulous patient, the cross sections were reformatted and used to construct a virtual planning of the implants and a guide template in Dental Slice. Six dental implants were placed in the maxilla and mandible using a Slice Guide System. Following a 30-month in maxilla and 24-month in mandible healing period, the clinical and radiographic evaluation and computed tomography (CT) showed good clinical stability. The Slice Guide System proved satisfactory for the Flapless Surgical Technique in dental implants.

The effect of laminin-1-doped nanoroughened implant surfaces: Gene expression and morphological evaluation

Aim. This study aimed to observe the morphological and molecular effect of laminin-1 doping to nanostructured implant surfaces in a rabbit model. Materials and Methods. Nanostructured implants were coated with laminin-1 (test; dilution, 100 g/mL) and inserted into the rabbit tibiae. Noncoated implants were used as controls. After 2 weeks of healing, the implants were removed and subjected to morphological analysis using scanning electron microscopy (SEM) and gene expression analysis using the real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Results. SEM revealed bony tissue attachment for both control and test implants. Real-time RT-PCR analysis showed that the expression of osteoblast markers RUNX-2, osteocalcin, alkaline phosphatase, and collagen I was higher (1.62-fold, 1.53-fold, 1.97-fold, and 1.04-fold, resp.) for the implants modified by laminin-1 relative to the control. All osteoclast markers investigated in the study presented higher expression on the test implants than controls as follows: tartrate-resistant acid phosphatase (1.67-fold), calcitonin receptor (1.35-fold), and ATPase (1.25-fold). The test implants demonstrated higher expression of inflammatory markers interleukin-10 (1.53-fold) and tumour necrosis factor-α (1.61-fold) relative to controls. Conclusion. The protein-doped surface showed higher gene expression of typical genes involved in the osseointegration cascade than the control surface. © 2012 Humberto Osvaldo Schwartz-Filho et al.

Comparative in vivo study of commercially pure Ti implants with surfaces modified by laser with and without silicate deposition: Biomechanical and scanning electron microscopy analysis

The purpose of this study was to evaluate commercially pure titanium implant surfaces modified by laser beam (LS) and LS associated with sodium silicate (SS) deposition, and compare them with machined surface (MS) and dual acid-etching surfaces (AS) modified. Topographic characterization was performed by scanning electron microscopy-X-ray energy dispersive spectroscopy (SEM-EDX), and by mean roughness measurement before surgery. Thirty rabbits received 60 implants in their right and left tibias. One implant of each surface in each tibia. The implants were removed by reverse torque for vivo biomechanical analysis at 30, 60, and 90 days postoperative. In addition, the surface of the implants removed at 30 days postoperative was analyzed by SEM-EDX. The topographic characterization showed differences between the analyzed surfaces, and the mean roughness values of LS and SS were statistically higher than AS and MS. At 30 days, values removal torque LS and SS groups showed a statistically significant difference (p < 0.05) when compared with MS and AS. At 60 days, groups LS and SS showed statistically significant difference (p < 0.05) when compared with MS. At 90 days, only group SS presented statistically higher (p < 0.05) in comparison with MS. The authors can conclude that physical chemistry properties and topographical of LS and SS implants increases bone-implant interaction and provides higher degree of osseointegration when compared with MS and AS. © 2012 Wiley Periodicals, Inc.

Biomechanical effect of one session of low-level laser on the bone-titanium implant interface

Low-level laser (LLL) has been used on peri-implant tissues for accelerating bone formation. However, the effect of one session of LLL in the strength of bone-implant interface during early healing process remains unclear. The present study aims to evaluate the removal torque of titanium implants irradiated with LLL during surgical preparation of implant bed, in comparison to non-irradiation. Sixty-four Wistar rats were used. Half of the animals were included in LLL group, while the other half remained as control. All animals had the tibia prepared with a 2 mm drill, and a titanium implant (2.2 × 4 mm) was inserted. Animals from LLL group were irradiated with laser (gallium aluminum arsenide), with a wavelength of 808 nm, a measured power output of 50 mW, to emit radiation in collimated beams (0.4 cm2), for 1 min and 23 s, and an energy density of 11 J/cm2. Two applications (22 J/cm 2) were performed immediately after bed preparation for implant installation. Flaps were sutured, and animals from both groups were sacrificed 7, 15, 30, and 45 days after implant installation, when load necessary for removing implant from bone was evaluated by using a torquimeter. In both groups, torque values tended to increase overtime; and at 30 and 45 days periods, values were statistically higher for LLL group in comparison to control (ANOVA test, p < 0.0001). Thus, it could be suggested that a single session of irradiation with LLL was beneficial to improve bone-implant interface strength, contributing to the osseointegration process. © 2012 Springer-Verlag London Ltd.

Alveolar bony crest preservation at implants installed immediately after tooth extraction: An experimental study in the dog

Aim: To evaluate the influence of deproteinized bovine bone mineral in conjunction with a collagen membrane, at implants installed into sockets in a lingual position immediately after tooth extraction, and presenting initial horizontal residual buccal defects <2 mm. Material and methods: The pulp tissue of the mesial roots of 4P4 was removed in six Labrador dogs, and the root canals were filled with gutta-percha and cement. Flaps were elevated, and the buccal and lingual alveolar bony plates were exposed. The premolars were hemi-sectioned, and the distal roots were removed. Implants were installed in a lingual position and with the margin flush with the buccal bony crest. After installation, defects resulted at about 1.7 mm in width at the buccal aspects, both at the test and control sites. Only in the left site (test), deproteinized bovine bone mineral (DBBM) particles were placed into the defect concomitantly with the placement of a collagen membrane. A non-submerged healing was allowed. Results: After 3 months of healing, one implant was found not integrated and was excluded from the analysis together with the contralateral control implant. All remaining implants were integrated into mature bone. The bony crest was located at the same level of the implant shoulder, both at the test and control sites. At the buccal aspect, the most coronal bone-to-implant contact was located at a similar distance from the implant margin at the test (1.7 ± 1.0 mm) and control (1.6 ± 0.8 mm) sites, respectively. Only small residual DBBM particles were found at the test sites. Conclusion: The placement of an implant in a lingual position into a socket immediately after tooth extraction may favor a low exposure of the buccal implant surface. The use of DBBM particles, concomitantly with a collagen membrane, did not additionally improve the outcome obtained at the control sites. © 2011 John Wiley & Sons A/S.

Healing outcomes at implants installed in sites augmented with particulate autologous bone and xenografts. An experimental study in dogs

Aim: To evaluate the integration of implants installed at the interface of pristine and grafted tissue augmented with particulate autologous bone or deproteinized bovine bone mineral (DBBM), concomitantly with a collagen membrane. Material and methods: In 6 Labrador dogs, the distal root of 3P3 and 4P4 was endodontically treated and hemi-sected, and the mesial roots extracted concomitantly with the extraction of 2P2. The buccal bony walls were removed, and two box-shaped defects, one larger and one smaller, were created. After 3 months, flaps were elevated, and the defects were filled with particulate autologous bone or DBBM in the right and left side of the mandible, respectively. Collagen membranes were used to cover the grafted areas. Three months later, flaps were elevated, and a customized device was used as surgical guide to prepare the recipient sites at the interface between grafts and pristine bone. One implant was installed in each of the four defects. After 3 months, biopsies were harvested and ground sections prepared for histological evaluation. Results: The augmentation technique was effective at all sites and all the foreseen implants were installed. In the histological analysis, all implants were integrated in mature bone, at both the buccal and lingual aspects. The most coronal bone-to-implant contact and the top of the buccal bony crest were located at a similar distance between test and control implants. However, these distances were higher at the larger compared with the smaller defects. Especially in the large defect, residual particles of DBBM were found embedded into connective tissue and located outside the bony crest. Conclusions: Particulate autologous bone as well as DBBM particles used to augment horizontally the alveolar bony process allowed for the osseointegration of implants installed after 3 months of healing. © 2012 John Wiley & Sons A/S.

Bone-healing pattern at the surface of titanium implants: An experimental study in the dog

Objective: To study the early sequential stages of osseointegration at implants installed in alveolar bony. Materials 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. Implants were installed, and the flaps were sutured to allow a fully submerged healing. The timing of the installations in the left side of the mandible and of sacrifices were performed with a schedule that various observation periods to sacrifice from 5, 10, 20, and 30 days were available so that n = 6 was obtained per each healing period. Ground sections were prepared and analyzed. Results: Newly formed bone in contact with the implant surface was found after 10 days of healing and the percentage increased up to 50% after 1 month of healing. A higher percentage was found in the trabecular compared with the cortical bony compartment. Old bone decreased by about 50% during healing, being still present after 1 month (16%). The proportions of bone debris and bone particles were at 27% after 5 days and decreased during healing to 6% after 1 month. Conclusion: Osseointegration (new bone-to-implant contact) developed at various rates for cortical and trabecular compartments, respectively. In the trabecular region, mesenchymal cells were identified, subsequently developing into new bone in contact with the implant surface. In the cortical compartment, however, resorptive processes were observed throughout all periods of healing. The proportion of newly formed bone percentage was lower compared with that of the trabecular area. Old bone was still present after 1 month of healing in both compartments. Bone debris and small bone particles appeared to be involved in initial bone formation. © 2013 John Wiley & Sons A/S.

Comparisons between Bio-Oss® and Straumann® Bone Ceramic in immediate and staged implant placement in dogs mandible bone defects

Objective: To compare immediate and staged approach implant placement in circumferential defects treated with deproteinized bovine bone mineral (DBBM); hidroxyapatite/tricalcium phosphate (HA/TP); autogenous bone (Ab); and coagulum (Cg); upon implant stability, osseointegration and alveolar crest maintenance. Materials and methods: Six dogs underwent extractions of lower premolars, bilaterally. Twelve weeks later four bone defects (6 mm wide/4 mm long) were drilled at one side and randomly filled with DBBM; HA/TP; Ab; and Cg, respectively, and left to heal (staged approach). Eight weeks later one implant (Osseospeed™, AstraTech) was placed in experimental sites. At the same session four defects were drilled on contra-lateral side and implants were inserted immediately after biomaterials grafting (immediate approach). Animals were euthanized 8 weeks later. Implant stability was measured by resonance frequency analysis (RFA) at installation and after sacrifice. Ground sections were prepared for bone contact (BIC); bone area (BA); distance implant shoulder-bone crest (IS-C); distance implant shoulder first bone contact (IS-B); and areas occupied by soft tissue. Results: The BA and BIC were superior in the staged approach. The Cg exhibited higher BIC and BA as compared with other materials at the total implant body (P = 0.004 and 0.012, respectively). The DBBM, HA/TP and Ab groups rendered similar BA and BIC. The immediate approach resulted in less crest resorption compared to staged approach. The biomaterials did not affect the IS-C and IS-B measurements. Particles area tended to be higher in DBBM group than HA/TP (P = 0.15), while soft tissue infiltrate was higher in DBBM group when used in the immediate approach (P = 0.04). The RFA indicated gain in stability in the staged approach (P = 0.002). The correlation test between RFA vs. BIC and BA demonstrated inferior stability for DBBM group in immediate approach (P = 0.01). Conclusions: Implants placed in healed defects resulted in better stability as a consequence of higher BIC and BA. The Cg alone rendered increased BIC compared to other materials in both approaches. Immediate approach should be preferable to staged approach in terms of alveolar crest maintenance. The BIC and BA values did not vary between micro and macro-threads in this experimental model. Implants installed in sites filled with DBBM in immediate approach were less stable. © 2011 John Wiley & Sons A/S.

Influence of presence or absence of teeth adjacent to implants installed immediately into extraction sockets on peri-implant hard tissue levels: An experimental study in the dog

Aim: To evaluate the influence of the presence or absence of adjacent teeth on the level of the mesial and distal alveolar bony crest following healing at sites where implants were installed immediately into extraction sockets. Material and methods: Six Labrador dogs were used. In the right side of the mandible, full-thickness flaps were elevated, and the second, third, and fourth premolars and first molars were extracted. In the left side of the mandible, endodontic treatments of the mesial roots of the third and fourth premolars as well as of the first molars were performed. Full-thickness flaps were elevated, the teeth were hemi-sected, and the distal roots were removed. The second premolars were extracted as well. Subsequently, implants were bilaterally installed with the implant shoulder flush with the buccal bony crest. Implants were placed in the center of the alveoli, but at the fourth premolars, they were placed toward the lingual bony plate of the alveoli. After 3 months of healing, the animals were euthanized and histological sections of the sites prepared. Results: Larger bony crest resorption was observed at the test compared with the control sites, both at the bucco-lingual and mesio-distal aspects. The differences between test and controls for the coronal level of osseointegration were smaller than those for resorption. When data from all mesial and distal sites facing an adjacent tooth were collapsed and compared with those opposing an edentulous zone, lower bony crest resorption and deeper residual marginal defects were found at the sites with neighboring teeth. Conclusion: The extraction of teeth adjacent to a socket into which implants were installed immediately after tooth extraction caused more alveolar bone resorption both for the bucco-lingual and at the mesio-distal aspects compared with sites adjacent to a maintained tooth. © 2012 John Wiley & Sons A/S.

Healing of buccal dehiscence defects at implants installed immediately into extraction sockets - an experimental study in dogs

Aim: To evaluate the influence of implant positioning into extraction sockets on bone formation at buccal alveolar dehiscence defects. Material and Methods: In six Labrador dogs the pulp tissue of the mesial roots of 4P4 was removed and the root canals were filled. Flaps were elevated bilaterally, the premolars hemi-sectioned and the distal roots removed. The implants were placed in contact with either the buccal (test site) or with the lingual (control site) bony wall of the extraction sockets. Healing abutments were affixed and triangular buccal bony dehiscence defects, about 2.7 mm deep and 3.5 mm wide, were then prepared. No regenerative procedures were done and a 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: After 4 months of healing, the bony crest and the coronal border of osseointegration at the test sites were located 1.71 ± 1.20 and 2.50 ± 1.21 mm apically to the implant shoulder, respectively. At the control sites, the corresponding values were 0.68 ± 0.63 and 1.69 ± 0.99 mm, respectively. The differences between test and control reached statistical significance (P < 0.05). Residual marginal bone defects were found both at the test and control sites. A statistically significant difference between test and control sites was only found at the lingual aspects (depth 2.09 ± 1.01 and 1.01 ± 0.48 mm, respectively). Similar heights of the buccal biological width were observed at both sites (about 5.1 mm). Conclusions: The placement of implants in a lingual position of the extraction sockets allowed a higher degree of bone formation at buccal alveolar dehiscence defects compared with a buccal positioning. © 2012 John Wiley & Sons A/S.

Ridge preservation at implants installed immediately after molar extraction. An experimental study in the dog

Aim: To evaluate the influence of deproteinized bovine bone mineral (DBBM), in conjunction with a collagen membrane, on bone resorption at implants installed in a lingual position immediately into extraction sockets with horizontal residual buccal defects >2.0 mm. Material & methods: The pulp tissue of the mesial roots of 1M1 was removed in six Labrador dogs, and the root canals were filled with gutta-percha and cement. Flaps were elevated. The molars were hemi-sectioned and the distal roots removed. Implants were installed in a lingual position and with the shoulder flush with the buccal bony crest. After installation, defects of about 2.5 and 2.7 mm in width resulted at the buccal aspects of the test and control sites, respectively. Only in the left site (test), deproteinized bovine bone mineral (DBBM) particles were placed into the defect concomitantly with the placement of a collagen membrane. On the control sites, no biomaterials were applied. A non-submerged healing was allowed. Results: After 3 months of healing, one control implant was not integrated and was excluded from the analysis, together with the contralateral test implant. All remaining implants were integrated into mature bone. The buccal alveolar bony crest was resorbed more at the test compared with the control sites, 2.2 ± 0.9 mm and 1.5 ± 1.3 mm, respectively. The vertical resorption of the lingual plate was 1.6 ± 1.5 mm and 1.5 ± 1.1 mm at the test and control sites, respectively. Only small residual DBBM particles were found at the test sites (1.4%). Conclusion: The use of DBBM particles to fill buccal defects of ≥2.5 mm at implants installed immediately into alveolar extraction sockets did not preserve the buccal bony wall. © 2012 John Wiley & Sons A/S.

The use of polylactic and polyglycolic copolymer biomaterial in a pre-clinical model of compromised primary stability

Aim Primary implant stability can be compromised by overdrilling of the implant bed. Filling the gap between the implant and the bone with a highly viscous copolymer of polylactic and polyglycolic acid (PLA/PGA) might stabilize the implant and thus supply osseointegration. The aim of this study was to evaluate implants installed in overdrilled beds associated with PLA/PGA in rats tibia model by means of removal torque test and fluorochrome analysis. Materials and methods For this experiment two groups were selected: in the test group 0.4 mm overdrilled defects (2.0 in diameter and 3 mm long ) were produced in the right tibia of seven rats and implants were placed covered with PLA/PGA biomaterial to fill the gap; the control group was not overdrilled and the implants were placed without the biomaterial. Implants of 1.6 mm in diameter and 3 mm long where placed into all defects. Calcein, alizarin and oxytetracyclin were injected at 7, 15 and 21 postoperative days, respectively, and the animals were sacrificed at 35 postoperative day. Results The results showed that all the implants achieved osseointegration. There were no statistical significance differences in torque-reverse and fluorocrome analysis (P>0.05). Conclusion We can conclude that overdrilled defects filled with PLA/PGA did not disturb osseointegration in this experimental model. © ARIESDUE.

Bone apposition to laminin-1 coated implants: Histologic and 3D evaluation

Laminin-1 has been reported as one of the factors responsible for the nucleation of calcium phosphates and, in vitro, has been reported to selectively recruit osteoprogenitors. This article focused on its in vivo effects, and evaluated the effect of laminin-1 local application on osseointegration. Polished cylindrical hydroxyapatite implants were coated with laminin-1 (test) and the bone responses in the rabbit tibiae after 2 and 4 weeks were evaluated and compared to the non-coated implants (control). Before the samples were processed for histological sectioning, they were three-dimensionally analysed with micro computed tomography (μCT). Both evaluation methods were analysed with regards to bone area around the implant and bone to implant contact. From the histologic observation, new bone formation around the laminin-1 coated implant at 2 weeks seemed to have increased the amount of supporting bone around the implant, however, at 4 weeks, the two groups presented no notable differences. The two-dimensional and three-dimensional morphometric evaluation revealed that both histologic and three-dimensional analysis showed some tendency in favour of the test group implants, however there was no statistical significance between the test and control group results. © 2012 International Association of Oral and Maxillofacial Surgeons.