Alveolar ridge augmentation using implants coated with recombinant human bone morphogenetic protein-2: histologic observations

Background: Studies using ectopic rodent, orthotopic canine, and non‐human primate models show that bone morphogenetic proteins (BMPs) coated onto titanium surfaces induce local bone formation. The objective of this study was to examine the ability of recombinant human BMP‐2 (rhBMP‐2) coated onto a titanium porous oxide implant surface to stimulate local bone formation including osseointegration and vertical augmentation of the alveolar ridge. Material and Methods: Bilateral, critical‐size, 5 mm, supra‐alveolar, peri‐implant defects were created in 12 young adult Hound Labrador mongrel dogs. Six animals received implants coated with rhBMP‐2 at 0.75 or 1.5 mg/ml, and six animals received implants coated with rhBMP‐2 at 3.0 mg/ml or uncoated control. Treatments were randomized between jaw quadrants. The mucoperiosteal flaps were advanced, adapted and sutured to submerge the implants for primary intention healing. The animals received fluorescent bone markers at weeks 3, 4, 7 and 8 post‐surgery when they were euthanized for histologic evaluation. Results: Jaw quadrants receiving implants coated with rhBMP‐2 exhibited gradually regressing swelling that became hard to palpate disguising the contours of the implants. The histologic evaluation showed robust bone formation reaching or exceeding the implant platform. The newly formed bone exhibited characteristics of the adjoining resident Type II bone including cortex formation for sites receiving implants coated with rhBMP‐2 at 0.75 or 1.5 mg/ml. Sites receiving implants coated with rhBMP‐2 at 3.0 mg/ml exhibited more immature trabecular bone formation, seroma formation and peri‐implant bone remodelling resulting in undesirable implant displacement. Control implants exhibited minimal, if any, bone formation. Thus, implants coated with rhBMP‐2 at 0.75, 1.5 and 3.0 mg/ml exhibited significant bone formation (height and area) compared with the sham‐surgery control averaging (±SD) 4.4±0.4, 4.2±0.7 and 4.2±1.2 versus 0.8±0.3 mm; and 5.0±2.2, 5.6±2.2 and 7.4±3.5 versus 0.7±0.3 mm², respectively (p<0.01). All the treatment groups exhibited clinically relevant osseointegration. Conclusions: rhBMP‐2 coated onto titanium porous oxide implant surfaces induced clinically relevant local bone formation including vertical augmentation of the alveolar ridge and osseointegration. Higher concentrations/doses were associated with untoward effects.     

Bone formation at recombinant human bone morphogenetic protein-2-coated titanium implants in the posterior maxilla (Type IV bone) in non-human primates

Background: Studies using ectopic rodent and orthotopic canine models (Type II bone) have shown that titanium porous oxide (TPO) surface implants adsorbed with recombinant human bone morphogenetic protein‐2 (rhBMP‐2) induce local bone formation including osseointegration. The objective of this study was to evaluate local bone formation and osseointegration at such implants placed into Type IV bone. Material and Methods: rhBMP‐2‐coated implants were installed into the edentulated posterior maxilla in eight young adult Cynomolgus monkeys: four animals each received three TPO implants adsorbed with rhBMP‐2 (2.0 mg/ml) and four animals each received three TPO implants adsorbed with rhBMP‐2 (0.2 mg/ml). Contra‐lateral jaw quadrants received three TPO implants without rhBMP‐2 (control). Treatments were alternated between left and right jaw quadrants. Mucosal flaps were advanced and sutured to submerge the implants. The animals received fluorescent bone markers at weeks 2, 3, 4, and at week 16 when they were euthanized for histologic analysis. Results: Clinical healing was uneventful. Extensive local bone formation was observed in animals receiving implants adsorbed with rhBMP‐2 (2.0 mg/ml). The newly formed bone exhibited a specific pinpoint bone–implant contact pattern regardless of rhBMP‐2 concentration resulting in significant osseointegration; rhBMP‐2 (2.0 mg/ml): 43% and rhBMP‐2 (0.2 mg/ml): 37%. Control implants exhibited a thin layer of bone covering a relatively larger portion of the implant threads. Thus, TPO control implants bone exhibited significantly greater bone–implant contact (～75%; p<0.05). There were no statistically significant differences between rhBMP‐2‐coated and control implants relative to any other parameter including peri‐implant and intra‐thread bone density. Conclusion: rhBMP‐2‐coated TPO implants enhanced/accelerated local bone formation in Type IV bone in a dose‐dependent fashion in non‐human primates resulting in significant osseointegration. rhBMP‐2‐induced de novo bone formation did not reach the level of osseointegration observed in native resident bone within the 16‐week interval.     

Alveolar ridge augmentation using implants coated with recombinant human bone morphogenetic protein-2: radiographic observations

Objectives: Effective carrier technologies and dosing appear critical for the successful use of bone morphogenetic proteins (BMPs). This study evaluated radiographically the potential of a purpose‐designed titanium porous‐oxide implant surface combined with recombinant human BMP‐2 (rhBMP‐2) to stimulate alveolar ridge augmentation. Material and methods: Twelve young‐adult Labrador dogs were used. Three 10‐mm titanium implants per jaw quadrant were placed 5 mm into the alveolar ridge following extraction of the premolar teeth and reduction of alveolar ridge. Six animals received implants coated with rhBMP‐2 at 0.75 or 1.5 mg/ml randomized to contralateral jaw quadrants. Another six animals received implants coated with rhBMP‐2 at 3 mg/ml or uncoated control using the same split‐mouth design. The mucoperiosteal flaps were advanced, adapted, and sutured to submerge the implants. Radiographic registrations were made immediately postsurgery (baseline), and at weeks 4 and 8 (end of study). Results: rhBMP‐2‐coated implants exhibited robust radiographic bone formation extending to and above the implant platform from week 4 (P<0.01). Some rhBMP‐2‐coated implants showed voids within the newly formed bone that gradually resolved and/or implant displacement, being severe in two animals receiving implants coated with rhBMP‐2 at 3 mg/ml. Controls showed limited, if any, new bone formation at weeks 4 and 8 postsurgery. There were no significant differences among the rhBMP‐2 groups in bone gain. Conclusions: The titanium porous‐oxide surface serves as an effective carrier for rhBMP‐2, showing a clinically significant potential to stimulate local bone formation. With the carrier technology used, therapeutic dosage appears to be in the range of 0.75–1.5 mg/ml.     

Alveolar ridge augmentation using implants coated with recombinant human bone morphogenetic protein-7 (rhBMP-7/rhOP-1): radiographic observations

Aim: The objective of this study was to radiographically evaluate the potential of a purpose‐designed titanium porous‐oxide implant surface coated with recombinant human bone morphogenetic protein‐7 (rhBMP‐7), also known as recombinant human osteogenic protein‐1 (rhOP‐1), to stimulate alveolar ridge augmentation. Material and Methods: Six young‐adult Hound Labrador mongrel dogs were used. Three 10 mm titanium oral implants per jaw quadrant were placed 5 mm into the alveolar ridge in the posterior mandible following surgical extraction of the pre‐molar teeth and reduction of the alveolar ridge leaving 5 mm of the implants in a supra‐alveolar position. The implants had been coated with rhBMP‐7 at 1.5 or 3.0 mg/ml and were randomized to contralateral jaw quadrants using a split‐mouth design. The mucoperiosteal flaps were advanced, adapted, and sutured to submerge the implants. Radiographic registrations were made immediately post‐surgery (baseline), and at weeks 4 and 8 (end of study). Results: rhBMP‐7‐coated implants exhibited robust radiographic bone formation. At 8 weeks, bone formation averaged 4.4 and 4.2 mm for implants coated with rhBMP‐7 at 1.5 and 3.0 mg/ml, respectively. There were no significant differences between the rhBMP‐7 concentrations at any observation interval. A majority of the implant sites showed voids within the newly formed bone at week 4 that generally resolved by week 8. The newly formed bone assumed characteristics of the resident bone. Conclusions: The titanium porous‐oxide implant surface serves as an effective carrier for rhBMP‐7 showing a clinically significant potential to stimulate local bone formation.     

Space-providing expanded polytetrafluoroethylene devices define alveolar augmentation at dental implants induced by recombinant human bone morphogenetic protein 2 in an absorbable collagen sponge carrier

Background: Surgical implantation of recombinant human bone morphogenetic protein 2 (rhBMP‐2) in an absorbable collagen sponge carrier (ACS) significantly enhances bone regeneration in horizontal alveolar defects; however, sufficient quantities of bone for implant dentistry are not routinely obtained. Purpose: The objective of this proof‐of‐principle study was to evaluate the potential of a space‐providing macroporous expanded polytetrafluoroethylene (ePTFE) device to control volume and geometry of rhBMP‐2/ACS‐induced alveolar bone augmentation. Materials and Methods: Bilateral critical‐size supra‐alveolar periimplant defects were created in four Hound‐Labrador mongrel dogs. Two turned and one surface‐etched 10 mm titanium dental implants were placed 5 mm into the surgically reduced alveolar ridge creating 5 mm supra‐alveolar defects. rhBMP‐2/ACS (0.4 mg rhBMP‐2) was placed around the exposed dental implants. Additionally, one jaw quadrant in each animal was randomly assigned to receive the domeshaped macroporous ePTFE device. Mucoperiosteal flaps were advanced for primary wound closure. The animals were euthanized at 8 weeks post surgery for histometric analysis. Results: The space‐providing macroporous ePTFE device defined the volume and geometry of rhBMP‐2/ACS‐induced bone formation, whereas bone formation at sites receiving rhBMP‐2/ACS alone varied considerably. Vertical bone gain at turned dental implants averaged (SD) 4.7 ± 0.2 mm at sites receiving rhBMP‐2/ACS and the ePTFE device compared with 3.5 ± 0.9 mm at sites receiving rhBMP‐2/ACS only. The corresponding values for rhBMP‐2/ACS‐induced bone area were 9.6 ± 0.7 mm ² and 7.5 ± 6.2 mm ². There was a highly significant correlation between induced bone area and the space provided by the ePTFE device (p .001). There was no difference in induced bone density or bone‐implant contact between the two technologies. These observations were consistent with those observed at surface‐etched dental implants. Conclusions: The data from this study suggest that a space‐providing macroporous ePTFE device defines rhBMP‐2/ACS‐induced alveolar augmentation to provide adequate bone quantities for implant dentistry. The dental implant surface technology does not appear to substantially influence bone formation.     

RhBMP-2/alphaBSM induces significant vertical alveolar ridge augmentation and dental implant osseointegration

Background: Recombinant human bone morphogenetic protein 2 (rhBMP‐2) in a carrier has been shown to induce significant bone formation. Several candidate carriers, however, lack structural integrity to offset compressive forces that may compromise rhBMP‐2 bone induction, in particular, for challenging onlay indications such as alveolar ridge augmentation. Purpose: The objective of this study was to evaluate rhBMP‐2 in a calcium‐phosphate cement carrier, αBSM, for vertical alveolar ridge augmentation and immediate dental implant Osseointegration. Materials and Methods: Six adult Hound Labrador mongrels with 5 mm critical size supra‐alveolar peri‐implant defects were used. Three animals received rhBMP‐2/αBSM (rhBMP‐2 at 0.40 and 0.75 mg/mL) in contralateral jaw quadrants (total implant volume/defect ～ 1.5 mL). Three animals received αBSM without rhBMP‐2 (control group). The animals were euthanized at 16 weeks post surgery, and block biopsies were processed for histologie and histometric analysis. Results: rhBMP‐2/αBSM induced substantial augmentation of the alveolar ridge. Control sites exhibited limited new bone formation. Vertical bone augmentation averaged (SD) 4.9 ± 1.0 mm (rhBMP‐2 at 0.40 mg/mL), 5.3 ± 0.3 mm (rhBMP‐2 at 0.75 mg/mL), and 0.4 ± 0.4 mm (control); new bone area 8.5 ± 4.2 mm ², 9.0 ± 1.9 mm ², and 0.5 ± 0.4 mm ²; new bone density 55.1 ± 6.4%, 61.1 ± 6.0%, and 67.7 ± 9.5%; and new bone‐implant contact 26.9 ± 17.5%, 28.5 ± 1.4%, and 24.6 ± 16.1%, respectively. Residual αBSM comprised 1% of the new bone. Bone density for the contiguous resident bone ranged from 65 to 71%, and bone‐implant contact ranged from 49 to 64%. Conclusions: Surgical implantation of rhBMP‐2/αBSM appears an effective protocol for vertical alveolar ridge augmentation procedures and immediate dental implant Osseointegration and for onlay indications of lesser complexity.     

Comparative Study of Chinese Hamster Ovary Cell Versus Escherichia coli‐Derived Bone Morphogenetic Protein‐2 Using the Critical‐Size Supraalveolar Peri‐Implant Defect Model

Background: Chinese hamster ovary (CHO) cell‐derived recombinant human bone morphogenetic protein‐2 (rhBMP‐2) has been introduced for spine, long bone, and craniofacial indications. Escherichia coli‐ (E. coli) derived rhBMP‐2 displays comparable efficacy to CHO cell‐derived rhBMP‐2 in vitro and in small‐animal models. The objective of this study is to evaluate the efficacy of E. coli‐derived rhBMP‐2 compared to the benchmark CHO cell‐derived rhBMP‐2 using an established large‐animal model. Methods: Contralateral, critical‐size supraalveolar peri‐implant defects in six adult male Hound Labrador mongrel dogs received CHO cell‐ or E. coli‐derived rhBMP‐2 (0.2 mg/mL) in an absorbable collagen sponge (ACS) carrier. In each quadrant, three dental implants were placed. A titanium mesh device was used to support space provision. The animals received fluorescent bone markers for qualitative evaluations. Animals were euthanized at 8 weeks for histopathologic and histometric evaluation. Results: Clinical healing included significant swelling, but none of the animals experienced wound dehiscences. CHO cell‐ and E. coli‐derived rhBMP‐2 supported comparable bone formation (new bone area, 35.8 ± 3.6 versus 30.1 ± 2.2 mm²; bone density, 31.8% ± 1.6% versus 35.6% ± 2.5%; and osseointegration, 32.9% ± 7.4% versus 33.7% ± 8.1%) without statistically significant differences between treatments. Newly formed immature delicate trabecular bone in fibrovascular marrow filled the space underneath the titanium mesh and extended coronally above the mesh. Seroma formation was frequently observed. There were no discernable qualitative histologic differences between treatments. Conclusion: CHO cell‐ and E. coli‐derived rhBMP‐2 in an ACS carrier appear equally effective at inducing local bone formation in support of dental implant osseointegration.     

Flapless Implant Surgery Using a Mini‐Incision

Background: Traditional flapless implant surgery using a soft tissue punch device requires a circumferential excision of keratinized tissue at the implant site. A new flapless implant technique that can submerge implant fixtures is needed. Purpose: This article describes a flapless implant surgery method using a mini‐incision and compares the effects of soft tissue punch and mini‐incision surgery on both the amount of osseointegration and the bone height around the implants using a canine mandible model. Materials and Methods: Bilateral, edentulated, flat alveolar ridges were created in the mandibles of six mongrel dogs. After a 3‐month healing period, two implants were placed on each side of the mandible using either soft tissue punch or mini‐incision procedures. After an additional 3‐month healing period, a second stage surgery and transmucosal abutment attachment was performed for mini‐incision implant cases. Following a 2‐month healing period, the dogs were sacrificed to evaluate the osseointegration and bone height around the implants. Results: Average bone height was 9.6 ± 0.4 mm in the soft tissue punch group and 9.8 ± 0.3 mm in the mini‐incision group (p > .05). Average osseointegration was 70.4 ± 6.3% in the soft tissue punch group and 71.2 ± 7.1% in the mini‐incision group (p > .05). No significant differences were noted between the two groups in vertical alveolar ridge height or bone/implant contact. Conclusions: Our findings support the clinical use of mini‐incision implant surgery at sites where implants need to be protected below the soft tissue during the early phase of healing, particularly for patients with poor bone quality and/or low primary implant stability.     

Influence of homeopathic treatment with comfrey on bone density around titanium implants: a digital subtraction radiography study in rats

Objective: The objective of this study was to evaluate the influence of homeopathic treatment with comfrey (Shymphytum officinalis 6CH) on radiographic bone density and area around titanium implants. Material and methods: Forty‐eight rats were divided into two groups of 24 animals each: a control group (C) and a test group (SO). Each animal received one titanium micro‐implant placed in the tibia. The animals in Group SO were subjected to 10 drops of comfrey 6CH per day mixed into their drinking water until the day of sacrifice. Eight animals of each group were sacrificed at 7, 14 and 28 days post‐surgery, respectively. Standardized digital radiographs were obtained on the day of implant installation (baseline images) and on the day of sacrifice (final images). Digital subtraction of the two corresponding images was performed to evaluate changes in bone density and the area related to change around the implant between baseline and final images. Results: Subtraction images demonstrated that a significant difference existed in mean shade of gray at 14 days post‐surgery between Group SO (mean 175.3±14.4) and Group C (mean 146.2±5.2). Regarding the area in pixels corresponding to the bone gain in Group SO, the differences observed between the sacrifice periods and groups were only significant at 7 days sacrifice between Group SO (mean 171.2±21.9) and Group C (mean 64.5±60.4). Conclusion: Within the limits of this study, comfrey administration promotes an increase in radiographic bone density around titanium implants in the initial period of bone healing.     

Bone formation at titanium porous oxide (TiUnite) oral implants in type IV bone

BACKGROUND: Several oral implant design advances have been suggested to overcome poor bone quality, an impediment for successful implant treatment. A novel titanium porous oxide (TPO) surface has been shown to offer favorable results in several settings. The objective of this study was to evaluate the local bone formation and osseointegration at TPO-modified implants in type IV bone. METHOD: Three TPO surface-modified implants (TiUnite) were installed into the edentulated posterior maxilla in each of 8 Cynomolgus monkeys. The animals were injected with fluorescent bone labels at 2, 3, 4 and 16 weeks post-surgery and were euthanized at week 16 when block biopsies were collected for histologic analysis. RESULTS: The predominant observation of the TPO implant surface was a thin layer of new bone covering most of the implant threads. Mean (+/-SE) bone-implant contact for the whole study group was 74.1 +/- 4.8%. There was a significant variability in bone-implant contact between animals (P = 0.0003) and between sites of the same animal (P < 0.0001). The variance in bone-implant contact was 30% larger among sites of the same animal than between different animals (187.5 vs. 144.8, respectively). There was a small but significant difference in bone density immediately outside, compared to within the threaded area of the implants (37.1 +/- 3.2% vs. 32.1 +/- 3.2%, P < 0.0001). Bone density outside the implant threads was significantly correlated (beta = 0.682, P < 0.0001) with the bone density within the threaded area. Bone density within the threaded area was significantly correlated (beta = 0.493, P = 0.0002) with bone-implant contact, whereas bone density outside the implant threads did not have a significant effect (beta = 0.232, P = 0.1). CONCLUSIONS: The results suggest that the TPO surface possesses a considerable osteoconductive potential promoting a high level of implant osseointegration in type IV bone in the posterior maxilla.     

Implant-guided vertical bone growth in the mini-pig

Objective: To attain and describe guided vertical bone regeneration around titanium (Ti) and titanium zirconium (Ti–Zr) dental implants utilizing non‐glycosylated recombinant human bone morphogenetic protein‐2 (ng/rhBMP‐2), biomaterial scaffolds and a scaffold retainer. Materials and methods: Thirty‐two modified Straumann TE implants were partially embedded in the mandibles of eight adult mini‐pigs. Pre‐shaped resorbable scaffolds were placed around the implant and shielded and stabilized with a newly developed Ti custom scaffold retainer (umbrella) or wide‐neck (WN) healing caps to stabilize the scaffold. Ng/rhBMP‐2 (50 μg) was applied to the supracrestal portion of the implant or incorporated within the scaffold. At 9 weeks, soft tissue healing was assessed. Vertical bone regeneration outcomes including bone height, bone‐to‐implant contact (BIC) and bone volume were assessed by micro‐computed tomography and histology. Results: Soft tissue healing at the test sites (+ng/rhBMP‐2/+scaffold) appeared to be substantially better than the control sites (?ng/rhBMP‐2/?scaffold). Bone height, BIC percentage and bone volume were all similar regardless of whether WN healing caps or umbrella scaffold stabilization was used for all biomaterial scaffolds tested. WN healing cap test sites showed greater new bone height and BIC as compared with aggregate data from the control sites (P=0.05). Comparison of aggregate data from the umbrella test sites showed greater BIC and new bone volume as compared with aggregate data from the control sites(P=0.05). Conclusion: Vertical bone regeneration was successfully attained utilizing ng/rhBMP‐2, biomaterial scaffolds and a scaffold retainer.     

Mesenchymal stem cells and platelet-rich plasma enhance bone formation in sinus grafting: a histomorphometric study in minipigs

Objectives: Autologous, allogenic, and alloplastic materials for sinus augmentation have specific drawbacks, which has stimulated an ongoing search for new materials and tissue‐engineering constructs. We investigated whether mesenchymal stem cells (MSCs) and platelet‐rich plasma (PRP) seeded on a fluorohydroxyapatite (FH) scaffold can improve bone formation and bone‐to‐implant contact (BIC) in maxillary sinus grafting. Material and Methods: Bilateral sinus augmentation procedures were performed in eight minipigs. MSCs, PRP, and FH scaffold (test site) or FH alone (control site) were grafted in each maxillary sinus. Distal to the osteotomy, one dental implant per sinus was placed in the grafting material through the facial sinus wall. The animals were killed 3 months after grafting, and block sections of the implant sites were harvested and prepared for histomorphometric analysis. Results: After 12 weeks, a significant increase in bone formation occurred in the test sites compared with the control sites (42.51%versus 18.98%; p=0.001). In addition, BIC was significantly greater in the test sites compared with the control sites in the regenerated area (23.71%versus 6.63%; p=0.028). Conclusions: These findings show that sinus augmentation with MSCs–PRP, combined with FH may enhance bone formation and osseointegration of dental implants compared with FH alone in minipigs.     

Evaluation of implant osseointegration with different regeneration techniques in the treatment of bone defects around implants: an experimental study in a rabbit model

Aim: The aim of this study was to evaluate the osseointegration of implants placed in areas with artificially created bone defects, using three bone regeneration techniques. Material and methods: The experimental model was the rabbit femur (16), where bone defects were created and implants were placed. The peri‐implant bone defects were filled with a deproteinized bovine bone mineral, NuOss^? (N), NuOss^? combined with plasma rich in growth factors (PRGF) (N+PRGF), NuOss^? covered by an RCM⁶ membrane (N+M), or remained unfilled (control group [C]). After 4 and 8 weeks, the animals were euthanized and bone tissue blocks with the implants and the surrounding bone tissue were removed and processed according to a histological protocol for hard tissues on non‐decalcified ground sections. The samples were studied by light and electron scanning microscopy, histometric analysis was performed to assess the percentage of bone in direct contact with the implant surface and a statistical analysis of the results was performed. Results: In the samples analyzed 4 weeks after implantation, the percentage of bone tissue in direct contact with the implant surface for the four groups were 57.66±24.39% (N), 58.62±20.37% (N+PRGF), 70.82±20.34 % (N+M) and 33.07±5.49% (C). In the samples with 8 weeks of implantation time, the percentage of bone in direct contact was 63.35±27.69% (N), 58.42±24.77% (N+PRGF), 78.02±15.13% (N+M) and 40.28±27.32% (C). In terms of the percentage of bone contact, groups N and N+M presented statistically significant differences from group C in the 4‐week trial test (P<0.05; ANOVA). For the 8‐week results, only group N+M showed statistically significant differences when compared with group C (P<0.05; ANOVA). Conclusion: In conclusion, the NuOss^? granules/RCM⁶ membrane combination presented a percentage of bone contact with the implant surface statistically greater than in the other groups. To cite this article:
Guerra I, Branco FM, Vasconcelos M, Afonso A, Figueiral H, Zita R. Evaluation of implant osseointegration with different regeneration techniques in the treatment of bone defects around implants: an experimental study in a rabbit model.
Clin. Oral Impl. Res. 22 , 2011; 314–322.
doi: 10.1111/j.1600‐0501.2010.02002.x     

Histological and biomechanical evaluation of phosphorylcholine-coated titanium implants

Objective: Compounds considered for drug delivery from oral implant surfaces in support of local bone formation might themselves influence osseointegration. Phosphorylcholine (PC) polymers have been shown to enhance the biocompatibility of medical devices and to serve as drug delivery systems. The objective of this study was to evaluate local bone formation and osseointegration at PC and positively charged PC (PC+)‐coated endosseous implants in an established rabbit model. Material and Methods: Sixteen adult female New Zealand White rabbits were used. Eight animals received PC‐coated and control titanium porous oxide surface implants placed in the left and right distal femural condyle (trabecular bone) and proximal tibial metaphysis (cortical bone) using aseptic routines. The remaining eight animals similarly received PC+ and control implants. One implant was placed in each femural condyle and two implants in each tibial metaphysis. Experimental and control implants were alternated between the left and right hind legs. Fascia and skin were closed in layers. The animals were euthanized following a 6‐week healing interval for biomechanical (removal torque) and histometric analyses. Results: Peri‐implant bone density was considerably greater at tibial compared with femoral sites within as well as immediately outside the implant threads. However, there were no significant differences in bone density among PC, PC+, and control implants. Nevertheless, bone–implant contact was significantly lower at PC compared with PC+ and control implants in cortical bone (p<0.05). Numerical differences in trabecular bone did not reach statistical significance. The removal torque evaluation revealed significantly lower values for PC compared with PC+ and control sites (p<0.05). Conclusion: The histometric and biomechanical analyses suggest that PC coating may influence biological processes and ultimately osseointegration of endosseous implants. Apparently, incorporation of cationic charges may reverse or compensate for this scenario. Nevertheless, both PC coatings exhibited clinically acceptable osseointegration. In perspective, PC technology appears to be a viable candidate delivery system for agents in support of local bone formation at endosseous implant surfaces.     

Peri-implant bone reactions to immediate implants placed at different levels in relation to crestal bone. Part I: a pilot study in dogs

Purpose: The aim of the present study was to evaluate bone remodeling and bone‐to‐implant contact (BIC) after immediate placement at different levels in relation to the crestal bone of Beagle dogs. Materials and methods: The mandibular bilateral second, third and fourth premolars of six Beagle dogs were extracted and six implants were immediately placed in the hemi‐arches of each dog. Randomly, three cylindrical and three tapered implants were inserted crestally (control group) and 2 mm subcrestally (experimental group). Both groups were treated with a minimal mucoperiosteal flap elevation approach. A gap from the buccal cortical wall to the implant was always left. Three dogs were allowed a 4‐week submerged healing period and the other three an 8‐week submerged healing period. The animals were sacrificed and biopsies were obtained. Biopsies were processed for ground sectioning. Histomorphometric analysis was carried out in order to compare buccal and lingual bone height loss, and BIC between the two groups. Results: All implants osseointegrated clinically and histologically. Healing patterns examined microscopically at 4 and 8 weeks for both groups (crestal and subcrestal) yielded similar qualitative bone findings. The distance from the top of the implant collar to the first BIC in the lingual crest (A–Lc) showed a significant difference (P=0.0313): 1.91 ± 0.2 mm in the control group and 1.08 ± 0.2 mm in the experimental group. There was less bone resorption in subcrestal implants than crestal implants. The mean percentage of newly formed BIC was greater with the cylindrical implant design (46.06 ± 4.09%) than with the tapered design (32.64 ± 3.72%). Conclusion: These findings suggest that apical positioning of the top of the implant does not jeopardize bone crest and peri‐implant tissue remodeling. However, less resorption of the Lc may be expected when implants are placed 2 mm subcrestally. To cite this article:
Negri B, Calvo‐Guirado JL, Pardo‐Zamora G, Ramírez‐Fernández MP, Delgado‐Ruíz RA, Muñoz‐Guzón F. Peri‐implant bone reactions to immediate implants placed at different levels in relation to crestal bone. Part I: a pilot study in dogs.
Clin. Oral Impl. Res. 23 , 2012; 228–235.
doi: 10.1111/j.1600‐0501.2011.02158.x     

Effects of Simvastatin on bone healing around titanium implants in osteoporotic rats

Objectives: Osteoporosis is known to impair the process of implant osseointegration. The recent discovery that statins (HMG‐CoA reductase inhibitors) act as bone anabolic agents suggests that statins can be used as potential agents in the treatment of osteoporosis. Therefore, we hypothesized that statins will promote osteogenesis around titanium implants in subjects with osteoporosis. Material and methods: Fifty‐four female Sprague Dawley rats, aged 3 months old, were randomly divided into three groups: Sham‐operated group (SHAM; n=18), ovariectomized group (OVX; n=18), and ovariectomized with Simvastatin treatment group (OVX+SIM; n=18). Fifty‐six days after being ovariectomized (OVX), screw‐shaped titanium implants were inserted into the tibiae. Simvastatin was administered orally at 5 mg/kg each day after the placement of the implant in the OVX+SIM group. The animals were sacrificed at either 28 or 84 days after implantation and the undecalcified tissue sections were obtained. Bone‐to‐implant contact (BIC) and bone area (BA) within the limits of implant threads were measured around the cortical (zone A) and cancellous (zone B) bone regions. Furthermore, bone density (BD) of zone B in a 500 μm wide zone lateral to the implants was also measured. Results: There were no significant differences in BIC and BA measurements in zone A in any of the three groups at either 28 or 84 days after implantation (P>0.05). By contrast, in zone B, significant differences in the measurement of BIC, BA, and BD were observed at 28 and 84 days between all three groups. Bone healing decreased with lower BIC, BA, and BD around implant in OVX group compared with other two groups, and Simvastatin reversed the negative effect of OVX on bone healing around implants with the improvement of BIC, BA, and BD in zone B. Conclusion: Osteoporosis can significantly influence bone healing in the cancellous bone around titanium implants and Simvastatin was shown to significantly improve the osseointegration of pure titanium implants in osteoporotic rats.     

Influence of two barrier membranes on staged guided bone regeneration and osseointegration of titanium implants in dogs. Part 2: augmentation using bone graft substitutes

Objectives: To assess the influence of two barrier membranes and two bone graft substitutes on staged guided bone regeneration and osseointegration of titanium implants in dogs. Materials and methods: Saddle‐type defects were prepared in the lower jaws of 6 fox hounds and randomly filled with a natural bone mineral (NBM) and a biphasic calcium phosphate (SBC) and allocated to either an in situ gelling polyethylene glycol (PEG) or a collagen membrane (CM). At 8 weeks, modSLA titanium implants were inserted and left to heal in a submerged position. At 8+2 weeks, respectively, dissected blocks were processed for histomorphometrical analysis (e.g., mineralized tissue [MT], bone‐to‐implant contact [BIC]). Results: The mean MT values (mm²) and BIC values (%) tended to be higher in the PEG groups (MT: NBM [3.4±1.7]; SBC [4.2±2]/BIC: NBM [67.7±16.9]; SBC [66.9±17.8]) when compared with the corresponding CM groups (MT: NBM [2.5±0.8]; SBC [2.3±1.6]/BIC: NBM [54.1±22.6]; SBC [61±8.7]). These differences, however, did not reach statistical significance. Conclusion: It was concluded that all augmentation procedures investigated supported bone regeneration and staged osseointegration of modSLA titanium implants. To cite this article :
Mihatovic I, Becker J, Golubovic V, Hegewald A, Schwarz F. Influence of two barrier membranes on staged guided bone regeneration and osseointegration of titanium implants in dogs. Part 2: augmentation using bone graft substitutes.
Clin Oral Impl Res. 23 , 2012; 308–315.
doi: 10.1111/j.1600‐0501.2011.02238.x     

Peri-implant bone organization under immediate loading conditions: collagen fiber orientation and mineral density analyses in the minipig model

Background: Mechanical properties of bones are greatly influenced by percentages of organic and mineral constituents. Nevertheless, information about mineralization level on a microscopic scale and collagen fiber organization in peri‐implant bone after immediate loading is scarce. Purpose: The aim of this work was to analyze and compare the degree of mineralization and collagen fiber orientation in alveolar bone (AB) and peri‐implant bone of immediately loaded (IL) and unloaded (NL) implants. Materials and Methods: A total of 25 dental implants of 3.8 mm in diameter and 11 mm in length were used in the present study. In five minipigs, three premolars and the first molar were removed from the left side of the mandible. Three months later, five implants for each animal were inserted. Four implants were loaded immediately with a fixed restoration, while one implant was left unloaded. After a 4‐month healing period, all implants were retrieved. Circularly polarized light and scanning electron microscope with backscattered electron imaging were used to analyze both peri‐implant and AB retrieved 5 mm from the implant. Results: The bone/implant contact ratio (BIC %) was 77.8 ± 5.9% for the IL implants and 78.0 ± 5.8% for the NL implants; the difference was not statistically significant (p = 0.554). In the peri‐implant bone, the area related to transverse collagen fibers was 112,453 ± 4,605 pixels for IL implants and 87,256 ± 2,428 pixels for NL implants. In the AB, the area related to transverse collagen fibers was 172,340 ± 3,892 pixels. The difference between groups was statistically significant (p < .001). The degree of mineralization of peri‐implant bone was 137 ± 19 gray level for IL implants and 115 ± 24 gray level for NL implants, while in the AB, the degree of mineralization was 125 ± 26 gray level. This difference was statistically significant (p < .001). Conclusion: In this study, it was found that IL and NL implants showed the same degree of osseointegration. The bone matrix around IL implants had a higher quantity of transverse collagen fibers and presented a higher level of mineralization.     

Influence of two barrier membranes on staged guided bone regeneration and osseointegration of titanium implants in dogs: part 1. Augmentation using bone graft substitutes and autogenous bone

Objectives: To assess the influence of two barrier membranes and two bone graft substitutes mixed with autogenous bone (AB) on staged guided bone regeneration and osseointegration of titanium implants in dogs. Materials and methods: Four saddle‐type defects each were prepared in the upper jaw of six fox hounds and randomly filled with a natural bone mineral (NBM)+AB and a biphasic calcium phosphate (SBC)+AB and allocated to either an in situ gelling polyethylene glycol (PEG) or a collagen membrane (CM). At 8 weeks, modSLA titanium implants were inserted and left to heal in a submerged position. At 8+2 weeks, dissected blocks were processed for histomorphometrical analysis (e.g., treated area [TA], bone‐to‐implant contact [BIC]). Results: The mean TA values (mm²) and BIC values (%) tended to be higher in the PEG groups(TA: NBM+AB [10.4 ± 2.5]; SBC+AB [10.4 ± 5.8]/BIC: NBM+AB [86.4 ± 20.1]; SBC+AB [80.1 ± 21.5]) when compared with the corresponding CM groups (TA: NBM+AB [9.7 ± 4.8]; SBC+AB [7.8 ± 4.3]/BIC: NBM+AB [71.3 ± 20.8]; SBC+AB [72.4 ± 20.3]). A significant difference was observed for the mean TA values in the SBC+AB groups. Conclusion: It was concluded that all augmentation procedures investigated supported bone regeneration and staged osseointegration of modSLA titanium implants. However, the application of PEG may be associated with increased TA values. To cite this article:
Schwarz F, Mihatovic I, Golubovic V, Hegewald A, Becker J. Influence of two barrier membranes on staged guided bone regeneration and osseointegration of titanium implants in dogs: part 1. Augmentation using bone graft substitutes and autogenous bone.
Clin. Oral Impl. Res. 23 , 2012; 83–89.
doi: 10.1111/j.1600‐0501.2011.02187.x     

Interface reaction at dental implants inserted in condensed bone

Purpose: The influence of the osteotome technique on the interface reaction of cylinder implants (SLA, ITI^®) was compared with the interface reaction of conventional implant insertion in an animal model. Material and methods: A total of 64 implants were placed in the cranial and caudal tibia of 8 Göttinger minipigs. The implant site was prepared either by a conventional technique with drills (control group A) or by the osteotome technique (experimental group B). Bone tissue responses were evaluated by histomorphometry, fluorescence microscopy and scanning electron microscopy after 7 and 28 days of osseointegration. Results: The average initial (7 days) bone‐to‐implant contact ratio was not statistically significantly different for the osteotome technique (35.88±2.94%) than for the control group (43.78±3.39%, P<0.095). After 28 days, the bone‐to‐implant contact ratio became statistically significantly higher when implants were inserted by conventional preparation (44.81±3.07% (group B), 63.47±4.87% (group A), P=0.003). Whereas fluorescence and immunhistologic examination revealed new bone formation with osteocalcin deposition directly at the implant surface in both groups, the extent of direct bone/implant contact was enhanced in conventionally prepared implant sites. SEM analysis confirmed an intimate bone to implant bond without fibrous tissue formation in places of direct contact at an ultrastructured level. Conclusion: Implant placement in conventionally prepared implantation sites is accompanied by an improved interface formation at an early stage of implantation.