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.     

Bone formation at recombinant human bone morphogenetic protein-2-coated titanium implants in the posterior mandible (Type II bone) in dogs

Background: Conventional oral/maxillofacial implants reach osseointegration over several months during which the titanium fixtures interact with alveolar bone. The objective of this study was to determine if adsorbing recombinant human bone morphogenetic protein‐2 (rhBMP‐2) onto a titanium porous oxide (TPO) implant surface might enhance or accelerate local bone formation and support osseointegration in a large animal oral/maxillofacial orthotopic model. Material and Methods: Endosseous implants with a TPO surface were installed into the edentulated posterior mandible in eight adult Hound Labrador mongrel dogs. The implant surface had been adsorbed with rhBMP‐2 at 0.2 or 4.0 mg/ml. TPO implants without rhBMP‐2 served as control. Treatments were randomized between jaw quadrants. Mucosal flaps were advanced and sutured leaving the implants submerged. Clinical and radiographic evaluations were made immediately post‐surgery, at day 10 (suture removal), and week 4 and 8 post‐surgery. The animals received fluorescent bone markers at week 3, 4, and at week 8 post‐surgery, when they were euthanized for histologic analysis. Results: TPO implants coated with rhBMP‐2 exhibited dose‐dependent bone remodelling including immediate resorption and formation of implant adjacent bone, and early establishment of clinically relevant osseointegration. The resulting bone–implant contact, although clinically respectable, appeared significantly lower for rhBMP‐2‐coated implants compared with the control [rhBMP‐2 (0.2 mg/ml) 43.3±10.8%versus 71.7±7.8%, p<0.02; rhBMP‐2 (4.0 mg/ml) 35.4±10.6%versus 68.2±11.0%, p<0.03]. Conclusions: rhBMP‐2 adsorbed onto TPO implant surfaces initiates dose‐dependent peri‐implant bone re‐modelling resulting in the formation of normal, physiologic bone and clinically relevant osseointegration within 8 weeks.     

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.     

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.     

Tissue engineering with recombinant human bone morphogenetic protein-2 for alveolar augmentation and oral implant osseointegration: experimental observations and clinical perspectives

Surgical placement of oral implants is governed by the prosthetic design and by the morphology and quality of the alveolar bone. Nevertheless implant placement often appears difficult, if at all possible, due to aberrations of the alveolar ridge. Hence prosthetically dictated implant positioning often entails augmentation of the alveolar ridge and adjoining structures. In this review we discuss recent observations of the biologic potential, clinical relevance, and perspectives of application of recombinant human bone morphogenetic protein-2 (rhBMP-2) technologies for alveolar bone augmentation and oral implant osseointegration. Using discriminating critical-size supraalveolar defects and clinical modeling in dogs, we show that rhBMP-2 has a substantial potential for augmenting alveolar bone and supporting osseointegration of titanium oral implants. Moreover, using clinical modeling, we demonstrate re-osseointegration in advanced periimplantitis defects and long-term functional loading of titanium oral implants placed into rhBMP-2-induced bone. Our studies suggest that inclusion of rhBMP-2 for alveolar bone augmentation and oral implant fixation will not only enhance the predictability of existing clinical protocol but also allow new approaches to these procedures.     

Peri-implant bone regeneration using recombinant human bone morphogenetic protein-2 in a canine model: a dose-response study

The objective of this study was to evaluate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) dose on alveolar ridge augmentation and dental implant osseointegration. Bilateral, 5 mm supraalveolar, peri-implant defects were surgically created in 6 beagle dogs. rhBMP-2 (0.05, 0.1 or 0.2 mg/ml) in an absorbable collagen sponge (ACS) carrier was molded around the fixtures and wounds were closed. Treatment variations were alternated between animals (incomplete block design). Animals were sacrificed at week 8 postsurgery. Nine of twelve jaw quadrants healed uneventfully. Two jaw quadrants exhibited wound failure by week 4 and one by week 8 postsurgery. Radiographic bone regeneration was observed in defects without wound failure from week 4 postsurgery. Radiolucent voids of variable size and shape were observed and regressed over time. In weeks 6 through 8, there was an apparent increase in bone density and trabecular structure, while bone height and volume decreased. Histometric analysis revealed limited differences in bone regeneration between experimental conditions. Bone regeneration area averaged (+/- SD) 1.0 +/- 0.5, 3.5 +/- 1.4 and 2.3 +/- 0.4 mm2 for the 0.05, 0.1 and 0.2 mg/ml dose, respectively. There were no significant differences in osseointegration. Osseointegration in newly formed bone averaged 19 +/- 4%, 18 +/- 10% and 21 +/- 6% for the 0.05, 0.1 and 0.2 mg/ml rhBMP-2 sites, respectively. Collectively, the data suggest that there are no dramatic differences in bone induction and osseointegration within the selected dose and observation interval.     

Effect of recombinant human bone morphogenetic protein-2 on bone regeneration and osseointegration of dental implants

Recombinant human bone morphogenetic protein-2 (rhBMP-2) induced bone regeneration and osseointegration was evaluated in bony defects created within the hollow chamber of endosseous dental implants in 14 foxhound dogs. Bilateral extractions of mandibular premolars were performed and surgical implantation of 104 hollow cylinder implants followed after 8 weeks of healing. Experimental implants had their hollow chamber filled with 20 microg of rhBMP-2 delivered with a bovine collagen carrier, whereas the control implants had their apical chamber left empty. Dogs were followed for 2, 4, 8 and 12 weeks. Histomorphometric evaluation and immunohistochemical analysis were performed. Minimal bone was regenerated at 2 weeks for both groups. At 4 weeks, bone fill averaged 23.48% for the rhBMP-2 and 5.98% for the control group (P<0.05). At 8 weeks, mean bone fill was 20.94% and 7.75% for the rhBMP-2 and the controls, respectively (P<0.05). At 12 weeks, mean bone fill was 31.39% and 24.31% for the rhBMP-2 and control implants, respectively (P>0.05). Bone-implant contact (BIC) increased for both groups over time and at 8 weeks the rhBMP-2 BIC value was 18.65% and for the control 7.22% (P<0.05). At 12 weeks, the BIC was 43.78% and 21.05% for the rhBMP-2 and the control group, respectively (P<0.05). Immunohistochemical staining for type II collagen was positive only for parts of the collagen carrier and formation of cartilaginous intermediate was not observed in any of the specimens. The results suggest that, in confined defects adjacent to dental implants, rhBMP-2 can induce bone regeneration in close apposition to the implant surface.     

Sinus floor augmentation with simultaneous placement of dental implants in the presence of platelet-rich plasma or recombinant human bone morphogenetic protein-7

The aim of the present study was to evaluate the possible benefit of platelet-rich plasma (PRP) in sinus grafting as compared with recombinant human bone morphogenetic protein-7 (rhBMP-7). For this purpose, we performed a bilateral sinus augmentation with anorganic bovine bone and simultaneous insertion of a titanium screw implant in five miniature pigs. Six hundred microliters of PRP and 15%-vol. autologous bone, which was collected with a trap during preparation of the implant recipient site, were added to the right sinus and 420 microl rhBMP-7 to the left sinus. A polychrome sequential labeling was performed. The animals were sacrificed 6 weeks after surgery. Undecalcified ground sections were evaluated by microradiography, digitized histomorphometry and under fluorescent light. The mean bone-implant contact using rhBMP-7 was 45.8% and 5.7% under PRP (P=0.002). The mean height of newly mineralized bone in the augmented area using rhBMP-7 amounted to 8.3 mm as opposed to 3.6 mm under PRP (P=0.013). Using PRP, the mean area of the newly formed bone was enhanced (51.3%) as compared with rhBMP-7 (33.1%); however, this difference was not statistically significant (P=0.081). In conclusion, under the selected experimental conditions the use of rhBMP-7 led to superior outcomes with regard to the osseointegration of dental implants and the height of new bone as compared with the use of PRP.     

Long-term functional loading of dental implants in rhBMP-2 induced bone. A histologic study in the canine ridge augmentation model

Osseointegration [direct bone-implant contact (BIC)] is a primary goal following installation of endosseous dental implants. Such bone contact provides stability for the dental implant over time. The objective of this study was to evaluate bone formation and BIC at long-term, functionally loaded, endosseous dental implants placed into bone induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) in an absorbable collagen sponge (ACS) carrier. Mandibular, saddle-type, alveolar ridge defects (approximately 15 x 10 x 10 mm), two per jaw quadrant, were surgically induced in each of six young adult American fox hounds. The defects were immediately implanted with rhBMP-2/ACS. Two defects per animal additionally received a nonresorbable expanded polytetrafluoroethylene (ePTFE) membrane or a bioresorbable polyglycolide fiber membrane. Healing was allowed to progress for 3 months, when the ePTFE membrane was removed, and machined, threaded, titanium dental implants were installed into the rhBMP-2/ACS induced bone and into the adjacent resident bone. At 4 months of osseointegration, the implants were exposed to receive abutments and prosthetic treatment (two- or three-unit bridges). Some implants were removed for histologic analysis. The remainder of implants were exposed to functional loading for 12 months at which time the animals were killed for histometric analysis. One animal died prematurely due to kidney failure unrelated to the experimental protocol and was not included in the analysis. The 12-month block sections from a second animal were lost in the histological processing. Four sites receiving rhBMP-2/ACS and ePTFE or resorbable membranes experienced wound failure and membrane exposure, and subsequently exhibited limited bone formation. Defects without wound failure filled to contour with the adjacent alveolar bone. The newly formed bone exhibited features of the resident bone with a re-established cortex; however, it commonly included radiolucent areas that resolved over time. Dental implants block biopsied at 4 months exhibited limited, if any, crestal resorption, whereas those exposed to functional loading for 12 months exhibited some crestal resorption. Implants biopsied at 4 months exhibited a mean (+/- SD) BIC of 40.6 +/- 8.2% in rhBMP-2/ACS induced bone vs. 52.7 +/- 11.4% in resident bone. Dental implants exposed to 12 months of functional loading exhibited a mean BIC of 51.7 +/- 7.1% in rhBMP-2/ACS induced bone vs. 74.7 +/- 7.0% in resident bone. There were no significant differences between dental implants placed into rhBMP-2/ACS induced bone and resident bone for any parameter at any observation interval. In conclusion, rhBMP-2/ACS-induced bone allows installation, osseointegration, and long-term functional loading of machined, threaded, titanium dental implants in dogs.     

Sinus floor augmentation with simultaneous placement of dental implants using a combination of deproteinized bone xenografts and recombinant human osteogenic protein-1. A histometric study in miniature pigs

Maxillary sinus floor augmentation with autogenous bone has become a widely accepted procedure in implant dentistry. The use of osteoconductive bone substitutes in this indication is controversial, since their use can lead to a prolonged healing time, inhomogenous ossification, foreign body reaction, migration of particles and low bone-implant contact (BIC). The purpose of this study was to examine whether the combination of an osteoinductive protein (recombinant human osteogenic protein-1 (rhOP-1 = bone morphogenetic protein-7) with natural bovine bone mineral (BioOss) would improve ossification and the bone-implant contact (BIC) in a sinus floor augmentation with simultaneous placement of implants. In this study, the maxillary sinus floors in 5 miniature pigs were augmented with 3 ml BioOss containing 420 micrograms rhOP-1 on the test side and 3 ml BioOss alone on the control side. At the time of augmentation a titanium implant (ITI) was inserted from a laterocaudal direction. After 6 months of healing the sites of augmentation were removed and examined in non-decalcified sections by microradiography, fluorescence microscopy of sequentially labelled specimens and by histometry. On both sides, significant amounts of newly-formed bone were observed. However, on the test sites, the percentage of BIC in the augmented area was 80.0% versus 38.6% on control sites. It can be concluded that the application of bone morphogenetic proteins caused a more rapid and enhanced osseointegration of simultaneously placed implants when compared to the bone substitute alone. Therefore recombinant human osteogenic protein-1 delivered by natural bone mineral has the potential to become a clinical alternative for autogenous bone grafts in sinus floor augmentation.     

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.     

Enhanced vertical alveolar bone augmentation by recombinant human bone morphogenetic protein-2 with a carrier in rats

This study aimed to evaluate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on vertical bone regeneration of edentulous ridge. Bilateral upper first and second molars of 8-week-old Wistar rats were extracted and the ridges were allowed to heal for 3 weeks. Compressed poly(lactic-co-glycolic acid) copolymer/gelatin sponge (PGS) was used as a carrier of rhBMP-2. PGS alone (control group) or PGS with 5 mug rhBMP-2 (test group) was implanted at the top part of alveolar ridge. The sham group received no implantation. The rats were killed at 1, 2, 4, 8 and 12 weeks after implantation and examined histologically and histomorphometrically. In the test group, significant bone augmentation was evident on the alveolar ridge throughout the experimental period. Histomorphometric analysis revealed greater tissue volume and height of alveolar bone in the test group compared with the control and sham groups (P < 0.05) from 4 weeks onward and the augmented tissues (5 mm3 in tissue volume and 1.5 mm in bone height) were maintained until 12 weeks. Osteoblast surface increased at 2 and 4 weeks and osteoid thickness reached a peak (25 microm) at 2 weeks. Dynamic variables, which represented calcification, were higher in the test group than the control and sham groups at 4 and 8 weeks (P < 0.05). These results suggest that use of rhBMP-2/PGS may achieve vertical bone augmentation, and stabilizes denture prosthesis or makes up for inadequate bone mass for implant prosthesis.