The role of ultrasound in monitoring reconstruction of mandibular continuity defects using osteogenic protein-1 (rhOP-1)

Introducing bone bioengineering concepts in craniofacial surgery demands development of novel imaging strategies, which overcome the shortcomings of radiography such as exposure to ionizing radiation. This study is aimed to investigate the usefulness of ultrasonography (US) in monitoring reconstruction of continuity osteoperiosteal mandibular defects in sheep using rhOP-1. The study was conducted on six adult sheep in which a critical size defect was created at the body of the mandible and was reconstructed using rhOP-1 with type-I collagen as a carrier. Ultrasound images were used to assess onset of bone formation, contour, and surface topography. The results were then compared to corresponding plain radiographs and to post-mortem observations. US showed bone union in all the subjects that concurred with radiographic and post-mortem examinations. US was superior to plain radiography in monitoring early events of ossification. However, it was relatively less efficient in describing the contour of the newly formed bone. It was possible to describe the pattern of bone formation and the dynamic changes in contour and surface topography via US during the follow-up period. In experienced hands, ultrasonography can offer valuable information about bone healing comparable with those obtained by plain radiography. US may replace plain radiography in becoming a routinely used tool for monitoring bone healing in selected sites of the craniofacial skeleton.     

Acceleration of callus maturation using rhOP-1 in mandibular distraction osteogenesis in a rat model

The aim of this study was to assess if the application of rhOP-1 induces accelerated consolidation of the callus in mandibular distraction osteogenesis. In seven adult Wistar rats a bilateral osteotomy of the horizontal ramus of the mandible was performed in the molar region and a custom designed distractor was mounted to the mandible. With a rate of 0.7 mm per day the device was activated bilaterally after the seventh postoperative day. After seven days of distraction two times 50 microg rhOP-1 were injected on two subsequent days directly into the callus. The contralateral side received an injection of placebo solution. The animals were killed four weeks after the end of distraction. A three-point bending test revealed a significantly higher strength of the distracted mandible in the rhOP-1 side (66.3 N vs. 30.4 N, P=0.034, paired t-test). Undecalcified histological sections were examined using microradiography and fluorescence microscopy after sequential intravital polychromic labelling. A continuous bony bridging was seen in all rhOP-1 sites and in none of the control sites. The data indicate that rhOP-1 may be an option to accelerate callus maturation in mandibular distraction osteogenesis.     

Mechanical behavior of mandibular continuity defects reconstructed using combinations of hydroxylapatite and autogenous bone

The purpose of this investigation was to determine the mechanical behavior of primate (Macaca mulatta) mandibles with continuity defects reconstructed using varying ratios of an alloplastic hydroxylapatite (HA) implant material mixed with autogenous bone (AB). The defects were allowed to heal for 6 and 18 months before killing the animals and mechanical testing. Each animal had one side of the mandible restored with 100% AB as control. The fracture strength and section modulus of the opposite side of the mandible, containing the test ratio of HA-AB, were directly compared with the mechanical behavior of the control side. The fracture strength and section modulus were evaluated by a cantilever type of mechanical test that took into account the anisotropic, viscoelastic, and geometric nature of the mandible. The type of bone and tissue at the fracture site was examined by scanning electron microscopy and a direct correlation between the amount of bone and/or fibrous tissue growth around the HA implant material and the fracture strength and section modulus values was established. The average fracture strength of the AB graft sites at 18 months was 9,975 +/- 4,300 psi, with ratios of 25:75 and 50:50 HA-AB giving test results approaching those of the AB graft sites.     

Contour and volume assessment of repairing mandibular osteoperiosteal continuity defects in sheep using recombinant human osteogenic protein 1.

BACKGROUND: This study describes the contour and volume of reconstructed mandibles using recombinant human osteogenic protein 1. MATERIAL AND METHODS: The investigation was conducted on six adult sheep, where a unilateral 35 mm parasymphyseal osteoperiosteal continuity defect of the mandible was created. Recombinant human osteogenic protein 1 and type-I collagen (as carrier) were applied to the defects. Radiographic and ultrasonographic examinations were carried out at day 1 of the surgery and 2, 4, 8, and 12 weeks following the surgery. The animals were then sacrificed 3 months after the operation. Postmortem CT-scan was performed for volumetric, cross-sectional area, height and width measurements. RESULTS: Ultrasound was more efficient than radiographs in demonstrating early callus formation at 2 weeks, while radiographic evidence of bone formation was consistently detectable only after 4 weeks. Using the combination of recombinant human osteogenic protein type 1 and type-I collagen resulted in twice the volume, cross-sectional surface area, and height when compared with those of the corresponding region of the contra-lateral non-operated side of the mandible. CONCLUSION: Within 3 months, recombinant human osteogenic protein type 1 on type-I collagen carrier failed to restore the original contour and volume of mandibular osteoperiosteal continuity defects.     

Prefabrication of vascularized bone grafts using recombinant human osteogenic protein-1--part 3: dosage of rhOP-1, the use of external and internal scaffolds

In a previous study vascularized bone grafts were prefabricated with recombinant human osteogenic protein-1 (rhOP-1) using blocks of xenogenic bone mineral (BioOss) as scaffolds. The present study addressed the dosage of rhOP-1 and the combination of an external (mould) and internal scaffold (granular BioOss). In five G?ttingen minipigs six prefabrication sites in the latissimus dorsi muscles were randomly assigned to groups a-f. Moulds were prepared by shaping collagen/polylactide membranes in a cylindrical form which was filled with 1g BioOss granules and rhOP-1 (a: 0; b: 50; c, f, e: 250; d: 1000 microg of rhOP-1, a-e: cylinder open to muscle, e cylinder perforated, f: cylinder open to subcutaneous fat). After 6 weeks a dose dependency of bone density (a-d: 0%; 9.4%; 15.8%; 31.1%) and vessel density (a-d: 0.3; 2.4; 7.9; 25.4 counts/view) was observed histomorphometrically. Muscular surrounding was advantageous to subcutaneous tissue. Perforations of the membranes increased vessel density and did not impair bone formation. Bone density decreased in the proximity of the polylactide membranes. The membrane material was too soft and partly collapsed and therefore needs not to be reconsidered. The use of BioOss granules with 1000 microg rhOP-1 per gram proved to be a suitable concept for prefabrication of bone transplants.     

Histological assessment of bioengineered new bone in repairing osteoperiosteal mandibular defects in sheep using recombinant human bone morphogenetic protein-7

Numerous experimental studies have been published about osteoinductive bone morphogenetic proteins (BMPs). However, to our knowledge there has been no detailed histological study of a mandibular defect in a large mammal, reconstructed using BMPs. We describe here the histological features of rhBMP-7-induced bone in mandibular defects in sheep. METHODS: A 35 mm osteoperiosteal defect was created at the parasymphyseal region of the mandible in six adult sheep. The continuity of the mandible was maintained using a bony plate, and rhBMP-7 was applied on a type I collagen carrier. Bone labels were injected at selected time intervals during the follow-up period. The animals were killed after 3 months and bone samples were examined histologically, histomorphometrically, and by fluorescence microscopy. RESULTS AND CONCLUSIONS: We found a mixture of woven and lamellar bone that contained many cells with large nuclei. This had not reorganised to form cortical bone and the rhBMP-7-induced bone was more porous than the native bone. The newly-formed bone restored both endosteal and periosteal layers. rhBMP-7-induced bone was biocompatible and induced no ossification of soft tissue or abnormal growth of nearby vital structures. The mineral apposition rate was 1.98 microm/day (range 0.62-5.63 microm/day), a value close to that reported in humans. This suggests that BMPs have a limited effect in accelerating the rate of mineralisation, but promote the pre-mineralisation processes, and perhaps the formation of woven bone.     

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.     

A collagen membrane containing osteogenic protein-1 facilitates bone regeneration in a rat mandibular bone defect

ObjectivesOsteogenic protein-1 (OP-1) has shown osteoinductive activities and is useful for clinical treatments, including bone regeneration. Regenerative procedures using a bioabsorbable collagen membrane (BCM) are well established in periodontal and implant dentistry. We evaluated the subsequent effects of the BCM in combination with OP-1 on bone regeneration in a rat mandibular circular critical-sized bone defect in vivo.DesignWe used 8 rats that received surgery in both sides of the mandible, and created the total 16 defects which were divided into 4 groups: Group 1; no treatment, as a control, Group 2; BCM alone, Group 3; BCM containing low dose 0.5 μg of OP-1 (L-OP-1), and Group 4; BCM containing high dose 2.0 μg of OP-1 (H-OP-1). Newly formed bone was evaluated by micro computed tomography (micro-CT) and histological analyses at 8 weeks postoperatively. In quantitative and qualitative micro-CT analyses of the volume of new bone formation, bone density, and percentage of new bone area was evaluated.ResultsBCM with rhOP-1 significantly increased and accelerated bone volume, bone mineral density, and percentage of new bone area compared to control and BCM alone at 8 weeks after surgery; these enhancements in bone regeneration in the OP-1-treated groups were dose-dependent.ConclusionsOP-1 delivered with a BCM may have effective osteoinductive potency and be a good combination for bone regeneration. The use of such a combination device for osteogenesis may result in safer and more predictable bone regenerative outcomes in the future.     

Mechanical evaluation of a patient-specific additively manufactured subperiosteal jaw implant (AMSJI) using finite-element analysis

Edentulism with associated severe bone loss is a widespread condition that hinders the use of common dental implants. An additively manufactured subperiosteal jaw implant (AMSJI) was designed as an alternative solution for edentulous patients with Cawood and Howell class V–VIII bone atrophy. A biomechanical evaluation of this AMSJI for the maxilla in a Cawood and Howell class V patient was performed via finite-element analysis. Occlusal and bruxism forces were incorporated to assess the loading conditions in the mouth during daily activities. The results revealed a safe performance of the implant structure during the foreseen implantation period of 15 years when exerting average occlusion forces of 200 N. For the deteriorated state of class VIII bone atrophy, increased stresses on the AMSJI were evaluated, which predicted implant fatigue. In addition, excessive bruxism and maximal occlusion forces might induce implant failure due to fatigue. The models predicted bone ingrowth at the implant scaffolds, resulting in extra stability and secondary fixation. For all considered loading conditions, the maximal stresses were located at the AMSJI arms. This area is most sensitive to bending forces and, hence, allows for further design optimization. Finally, the implant is considered safe for normal daily occlusion activities.     

Mandibular reconstruction in miniature pigs with prefabricated vascularized bone grafts using recombinant human osteogenic protein-1: a preliminary study

A new technique of prefabrication of vascularized bone grafts in the latissimus dorsi muscle of miniature pigs, using recombinant human osteogenic protein-1 (rhOP-1) and xenogenic bone mineral as a carrier, is presented. The grafts were used to reconstruct a mandible using microsurgical anastomosis of the thoracodorsal vessels of the flap with a branch of the external carotid artery and the external jugular vein. The technique provided enough bone to reconstruct a mandible. The bone marrow stayed viable after transplantation, demonstrated by fluorescence microscopy and scintigraphy. Compared with identical defects of the contralateral side, which were treated with directly applied rhOP-1/bone mineral, a significantly better result was obtained with the prefabrication technique. The prefabrication technique may provide a means to treat irradiated patients with bone grafts produced by osteoinductive proteins and has a potential to become a clinical alternative to conventional vascularized bone grafts.     

Guided bone regeneration of large mandibular defects in a primate model

BACKGROUND: An earlier publication from our laboratory described the use of guided bone regeneration to fill large bone voids in the mandible created through en bloc resection in primates. The present report is an embellishment of this paper and describes bone regeneration experiments in 18 adult male Macaca mulatta monkeys to determine how long membranes must be in position to promote guided bone regeneration. METHODS: Thirty-six lesions were created in the mandibles of 18 monkeys in a standardized mandibular defect of 8 x 19 mm. Reinforced ePTFE membranes were placed in the animals and held in place with mini screws and sutures for anywhere from 1 to 12 months. No material was added to the defect. In addition to clinical studies, digital subtraction radiology and fluorescent labeling with tetracycline and histomorphometry are described. RESULTS: The results indicate that no bone gain was observed in membranes exposed for 1 month or less, but bone gain (approximately well over 90% of defects) was observed at 12 months when membranes were left in situ for 2 to 12 months (P <0.0001). No significant difference in the amount of bone gained at 12 months was observed for membranes left in place for intervals ranging from 2 to 12 months. A significant correlation between the amount of bone gain observed at 3 and 12 months was observed (P <0.0001). CONCLUSIONS: Data therefore suggest that membranes left in situ for 1 month or less result in minimal bone gain compared with membranes left in place from 2 to 12 months. In addition, labeling and stained sections clearly showed that the bone produced after 2 months of membrane placement is mature.     

原核表达重组人 OP-1促进拔牙创愈合的实验研究

目的：检测原核表达基因工程重组人OP-1(rhop—1)促进拔牙创愈合，减少牙槽嵴吸收的的效能方法：采用大白兔建立拔牙创的动物模型，以明胶海绵作为裁体，将大肠杆菌原核表达的重组人OP—1与之复合后植入即刻拔除牙齿的牙槽窝内进行干预治疗，通过观察其组织学改变、钙含量及碱性磷酸酶活性测定的变化，探讨原核表达的基因工程重组产品rhOP—1是否促进拔牙创的愈合修复。结果：大体观察结果显示：两组牙槽嵴高度的吸收差异有显著性；组织学HE切片显示：实验组的骨创愈合比对照组大约提前4～6周：碱性磷酸酶(AIP)活性及钙含量测定均显示实验组均明显高于对照组，差异有统计学意义：结论：原核表达基因工程重组人骨形态发生蛋白具有良好的促进骨创愈合、减少或防止牙槽骨吸收的作用     

对改良MGA功能矫治器线性矫治数据的评价

目的：评价改良MGA功能性矫治器对青少年安氏Ⅱ^1错[牙合]下颌功能性后缩、低面角硬组织矢状方向的矫治疗效。方法：以Pancherz分析方法，对12例患者矫治前后头颅定位侧位片重叠后硬组织变化的线形数据进行统计学分析对比。结果：就矢状基准平面OLp而言，pg／OLp，mi／OLp，ii／OLp，co／OLp＋pg／OLp，is／OLp—ii／OLp，ms／OLp—mi／OLp，is／OLp—ss／OLp（P〈0．01）；is／OLp，ii／OLp—pg／OLp（P〈0．05），其变化具有统计学意义。ss／OLp，co／OLp，ms／OLp，ms／OLp—ss／OLp，mi／OLp—pg／OLp的变化无统计学意义。结论：采用IMGA矫治器对青少年安氏Ⅱ^1错[牙合]下颌后缩低面角患者进行矢状矫治可产生明显的硬组织变化。     

Bone regeneration by recombinant human bone morphogenetic protein-2 in rat mandibular defects. An experimental model of defect filling.

BACKGROUND: Bone defects and irregularities are major problems for dental implant and periodontal therapies. METHODS: We investigated whether the application of recombinant human bone morphogenetic protein-2 (rhBMP-2) induces bone formation in through-and-through bone defects in the rat mandible. A round through-and-through bone defect (5 mm in diameter) was created in the angle of the mandible on both sides of the jaw using a steel round bur in each of 8 Long-Evans rats. In the experimental group, polylactic acid-polyglycolic acid copolymer/gelatin sponge (PGS) containing rhBMP-2 (6 microg/60 microl) was inserted in the bone defect. In the control group, the same carrier without rhBMP-2 was applied in the bone defect on the opposite side. Four weeks after application, the rats were sacrificed. Step serial sections stained with hematoxylin and eosin at intervals of 200 microm were prepared in a bucco-lingual direction. The size of the bone defects and new bone formation were evaluated histometrically. RESULTS: In all cases in the experimental group, a large quantity of newly formed bone was observed. The bone defects were completely filled with new bone in 4 of 8 rats in the experimental group. In the control group, small amounts of new bone formation were observed along the border of the original mandibular bone. Histometrical analysis revealed that the amount of new bone was significantly larger in the rhBMP-2 treated sites than in the control sites (P <0.0001; paired t-test). CONCLUSIONS: These results indicate that the rhBMP-2/PGS system induced effective bone regeneration on mandibular defects in rats. This procedure may be suitable as an experimental model for bone regeneration using various growth factors and effective for alveolar ridge augmentation followed by dental implant surgery.     

The in vitro evaluation of a local pedicled osteomyocutaneous mandibular flap for the reconstruction of composite mandibular defects.

PURPOSE: The purpose of this study was to develop and assess the potential feasibility of reconstructing composite defects of the mandible with a local pedicled osteomyocutaneous mandibular flap. MATERIALS AND METHODS: The flap design was established based on anatomic principles. A prospective evaluation of the flap was then performed in a fresh cadaver model, and, subsequently, its vascular integrity was documented with angiography. RESULTS: The pedicled osteomyocutaneous mandibular flap was technically simple to raise and had an exceptionally long arc of rotation, which should enable it to fill most compound segmental defects of the mandible. Angiographic studies of the harvested flaps done under fluoroscopic guidance confirmed that excellent vascularity of all components of the flap was present. CONCLUSIONS: The pedicled osteomyocutaneous mandibular flap appears to have a sound anatomic basis. Clinical evaluation is needed to fully elucidate its potential role in head and neck reconstruction.     

In situ osteogenesis: regeneration of 10-cm mandibular defect in porcine model using recombinant human bone morphogenetic protein-2 (rhBMP-2) and Helistat absorbable collagen sponge

Traditional bone grafting relies upon the incorporation of a bone-cell bearing structure into a recipient site. The graft serves as a scaffold that is eventually replaced and remodeled. This process is known as osteoconduction. Recombinant human bone morphogenetic protein-2 (rhBMP-2) is commercially available as an acellular implant in which the protein is bound to an absorbable collagen sponge (ACS). The rhBMP-2/ACS implant converts undifferentiated mesenchymal stem cells into osteoblasts and promotes an intense local neovascular response. This process, known as osteoinduction, produces bone via membranous, chondroid, or endochondral ossification. The type of bone synthesis depends upon the mesenchymal substrate and the local cellular environment. Using this simple technique, bone defects can be resynthesized with good outcomes and a significant reduction in donor site morbidity. Repair of a critical-sized mandibular resection defect with ISO is described. Basic science concepts of rhBMP-2, relevant histopathologic findings, and clinical application are described.     

Bone regeneration using recombinant human bone morphogenetic protein-2 (rhBMP-2) in alveolar defects of primate mandibles

The efficacy of bone morphogenetic protein (BMP) for bone reconstruction has been widely studied in numerous animal experiments, but insufficient information exists about its ability to regenerate bone in primates. The purpose of this study was to evaluate the effects of recombinant human BMP-2 (rhBMP-2) on bone formation in alveolar bone defects in the mandibles of young primates. Marginal bone defects were created in the mandibles of nine 5-year-old rhesus monkeys and rhBMP-2 permeated in a polylactic-co-glycolic acid-coated gelatin sponge (PGS) was implanted into the bone defects. The resected bone treated with rhBMP-2 regenerated completely at 12 weeks postoperatively, and remodelling and consolidation of new bone were seen histologically. This study provides evidence of considerable bone regeneration in alveolar defects after surgical implantation of rhBMP-2 in non-human primates. This technique may be an effective alternative to autogenous bone grafts for reconstructive surgery in clinical practice.     

Periodontal tissue regeneration by combined applications of recombinant human osteogenic protein-1 and bone morphogenetic protein-2. A pilot study in Chacma baboons (Papio ursinus)

Native and recombinant human bone morphogenetic/osteogenic proteins (BMPs/ OPs) singly initiate bone induction in vivo. The finding of synchronous but spatially different BMPs/OPs expression during periodontal tissue morphogenesis suggests novel therapeutic approaches using morphogen combinations based on recapitulation of embryonic development. Twelve furcation defects prepared in the first and second mandibular molars of three adult baboons (Papio ursinus) were used to assess whether qualitative histological aspects of periodontal tissue regeneration could be enhanced and tissue morphogenesis modified by combined or single applications of recombinant hOP-1 and hBMP-2. Doses of BMPs/OPs were 100 microg of each protein per 1 g of insoluble collagenous bone matrix as carrier. Approximately 200 mg of carrier matrix was used per furcation defect. Undecalcified sections cut for histological analysis 60 d after healing of hOP-1-treated specimens showed substantial cementogenesis with scattered remnants of the collagenous carrier. hBMP-2 applied alone induced greater amounts of mineralized bone and osteoid when compared to hOP-1 alone or to combined morphogen applications. Combined applications of hOP-1 and hBMP-2 did not enhance alveolar bone regeneration or new attachment formation over and above the single applications of the morphogens. The results of this study, which is the first to attempt to address the structure-activity relationship amongst BMP/OP family members, indicate that tissue morphogenesis induced by hOP-1 and hBMP-2 is qualitatively different when the morphogens are applied singly, with hOP-1 inducing substantial cementogenesis. hBMP-2 treated defects, on the other hand, showed limited cementum formation but a temporal enhancement of alveolar bone regeneration and remodelling. The demonstration of therapeutic mosaicism in periodontal regeneration will require extensive testing of ratios and doses of recombinant morphogen combinations for optimal tissue engineering in clinical contexts.