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Ortho- and heterotopic bone grafts in bifocal transport osteogenesis for craniofacial reconstruction--an experimental study in sheep.

Bifocal transport osteogenesis (BTO) is a promising technique for the reconstruction of extended osseous craniofacial defects. Aim of this study was to determine the potential of this technique related to various donor sites of the transport segment. In 10 adult sheep critical size defects of the calvaria were treated by gradual movement of a transport segment consisting either of autogenous regional free calvarial bone grafts (n=5) or autogenous illiac free bone grafts (n=5). Latency period was 5 days; the rate of distraction was 1mm per day and extended approximately 40 days. The consolidation period was 28 days. After harvesting, specimens were investigated by conventional radiography, CT-scans, histologically and by fluorescence. In both groups transport osteogenesis resulted in a complete closure of the critical size defect. Membranous bone formation and remodeling occurred during the entire period of distraction and consolidation. The volumes and thickness of newly formed bone at the defect site were increased significantly when calvarial bone grafts were used (P<0.05). Iliac bone grafts became progrediently smaller during distraction, while the volume of calvarial grafts remained relatively constant (P<0.05). In conclusion, transport segments consisting of calvarial and iliac bone resulted in a reliable closure of craniofacial critical size defects in adult organisms; the application of calvarial bone grafts resulted in an increased extend of bone formation.

Animals↗

Possible problems of moulding the regenerate in mandibular distraction osteogenesis -- experimental aspects in a canine model.

INTRODUCTION: Moulding of the regenerate created by distraction osteogenesis has been shown clinically to be efficient and good enough so that for complex three-dimensional deformities, final adjustments by moulding the regenerate may be part of the treatment plan. This study assessed possible drawbacks of moulding a regenerate, taking into consideration compressive and tensile forces acting simultaneously on the fresh callus. METHOD: Distraction osteogenesis in 15 Beagle mandibles was performed using custom made devices which allowed for lengthening as well as for angulation. After linear distraction of 10 mm, a defined 20 degrees angulation was performed in one step. The position of the fulcrum of the device allowed simultaneously compression and stretching of the regenerated bone. Effects on bone healing were assessed after 6 and 13 weeks of consolidation respectively and compared with a control group where only linear distraction was performed. RESULTS: Radiological and histological investigations demonstrated that no significant differences between the biological behaviour of the compressed and the stretched zones of the regenerate could be found. However, there were signs showing the more critical character of the stretched area. After 6 weeks of consolidation some specimens revealed delayed ossification of the stretched zone. Under stable conditions, this delay was compensated for after 13 weeks of consolidation and complete osseous healing occurred. CONCLUSION: Under stable conditions, a fresh regenerate can be moulded to a considerable extent without permanently endangering osseous healing. Nevertheless tensile forces acting on the regenerate should be minimized to prevent damage to the new bone. This can be achieved by overdistraction prior to callus moulding or by gradually changing the vector of distraction during the lengthening process.

Animals↗

The efficacy of the bone markers osteocalcin and the carboxyterminal cross-linked telopeptide of type-I collagen in evaluating osteogenesis in a canine crural lengthening model.

The aim of the present study was to determine the efficacy of the bone markers osteocalcin (OC) and carboxyterminal cross-linked telopeptide of type-I collagen (ICTP) in evaluating new bone formation in the dog, using commercially available immunoassay kits. Dogs were randomly divided into three groups and a circular external skeletal fixation system (CESF) was mounted on the tibia. In the first group a distraction osteogenesis procedure of the crus was performed. The second group received an osteotomy without crural lengthening, whereas the third group served as a sham-operated control. Bone formation was assessed using densitometric image analysis of crural radiographs. Despite significant differences in the amount of newly formed bone, this finding was not reflected in the plasma levels of OC and ICTP. In conclusion, OC and ICTP were not efficacious as markers of bone formation and resorption during osteogenesis in this canine model.

Absorptiometry, Photon↗

Physiologic weight-bearing increases new vessel formation during distraction osteogenesis: a micro-tomographic imaging study.

During distraction osteogenesis, large volumes of new bone are formed through the slow distraction of fracture callus. The newly formed bone is closely linked to angiogenesis and positively influenced by physiologic loading. In this study, a rat model was used to explore the correlation between these two observations. Unilateral femoral lengthenings were performed in 18 male Sprague-Dawley rats (400-500 g, age<6 months). Half of the animals were allowed to bear weight freely (WB) while the remaining animals were made non-weight-bearing via a through-knee amputation (NWB). After a seven-day latency period, femurs were lengthened 7 mm over 21 days. Animals were sacrificed at 7, 21, 35, and 49 days (0, 4.7, 7, and 7 mm of distraction) at which time lower extremity vessels were perfused with a 60% (w/v) barium sulfate suspension. High-resolution three-dimensional images of the vascular architecture were generated using a fan-beam micro-tomography machine by digitally separating the contrast-filled vessels from surrounding tissue. For two subvolumes, VOI(1), which included vessels in the medullary canal, cortex, and periosteum, and VOI(2), which included only vessels in the canal and cortex, the total tissue volume (TV), the volume of vessels (VV), and vessel diameter (VD) were determined. For the larger subvolume (VOI(1)), VV and vessel density (VV/TV) increased as a function of time (p<0.001) in WB animals. In NWB animals, VV increased significantly with time (p=0.029), while VV/TV did not (p=0.36). Increases in VV and VV/TV were significantly greater in WB animals than in NWB animals (p<0.01 and 0.05, respectively). VD was similar in both groups and did not change with time. Our data suggest that weight bearing stimulates new vessel formation during distraction osteogenesis.

Animals↗

Transplantability and therapeutic effects of bone marrow-derived mesenchymal cells in children with osteogenesis imperfecta.

In principle, transplantation of mesenchymal progenitor cells would attenuate or possibly correct genetic disorders of bone, cartilage and muscle, but clinical support for this concept is lacking. Here we describe the initial results of allogeneic bone marrow transplantation in three children with osteogenesis imperfecta, a genetic disorder in which osteoblasts produce defective type I collagen, leading to osteopenia, multiple fractures, severe bony deformities and considerably shortened stature. Three months after osteoblast engraftment (1.5-2.0% donor cells), representative specimens of trabecular bone showed histologic changes indicative of new dense bone formation. All patients had increases in total body bone mineral content ranging from 21 to 29 grams (median, 28), compared with predicted values of 0 to 4 grams (median, 0) for healthy children with similar changes in weight. These improvements were associated with increases in growth velocity and reduced frequencies of bone fracture. Thus, allogeneic bone marrow transplantation can lead to engraftment of functional mesenchymal progenitor cells, indicating the feasibility of this strategy in the treatment of osteogenesis imperfecta and perhaps other mesenchymal stem cell disorders as well.

Bone Density↗

Induced osteogenesis by periosteal distraction.

PURPOSE: The purpose of this project was to evaluate a novel technique for inducing osteogenesis through periosteal distraction in a rabbit model. MATERIALS AND METHODS: A periosteal distraction device was rigidly fixed to the lateral surface of the mandible in 10 adult rabbits. Periosteal distraction was started 7 days after placement of the periosteal distraction device. The periosteum was distracted 7 mm over 15 days. The unoperated, contralateral side of the mandible served as the control. The animals were killed at postoperative days 28, 35, 42, and 56. The specimens were then fixed, decalcified, and stained with hematoxylin and eosin. Histologic examination and histomorphometric analysis were performed on all specimens. RESULTS: Nine of 10 periosteal distraction devices remained rigidly fixed to the lateral surface of the mandible. On postoperative day 28, the histologic specimen from the experimental side showed periosteal proliferation and an increase in the number of osteoblasts. On postoperative days 35, 42, and 56, the experimental side showed an increase in the number of osteocytes per unit area, collagen fibers parallel to the vector of distraction, islands of osteoblasts surrounded by newly formed bone, and maturation of bone. An average of 2.86 +/- 0.56 mm of new bone height was formed. CONCLUSION: We report on a novel technique for generating bone by periosteal distraction. Our histologic analysis showed proliferation of the periosteum, an increase in the number of osteoblasts and osteogenesis.

Animals↗

Alveolar ridge augmentation by distraction osteogenesis.

Distraction osteogenesis is an alternative method for reconstructing atrophic alveolar bone. Fourteen patients underwent vertical alveolar distraction by the LEAD SYSTEM-Endosseous Alveolar Distraction System (Stryker Leibinger, Kalamazoo, MI). An alveolar segmental osteotomy was carried out and the vertical distraction device was mounted. In patients with an extensive alveolar defect, two distraction devices were placed in order to better control the vector of elongation in both bone edges. The distraction was started on the fourth postoperative day at a rate of 0.8 mm/day for 10-16 days, followed by a consolidation period of 60 days. Vertical distraction osteogenesis (VDO) was completed successfully in all patients with segment lengths in the range of 8 to 13 mm and with an average of 10.3 mm. Subsequently, the devices were removed and 23 threaded titanium dental implants were placed for osteointegration. Earlier mineralization in the vertically distracted area was seen radiographically during the consolidation period. In a follow up of 6-20 months after the distraction, 22 implants were successfully osteointegrated while one implant failed due to improper distracted segment stability. As a result of alveolar distraction, a segment of mature bone was transported vertically in order to lengthen the crest for better implant anchorage, either for aesthetic purposes or for functional prosthetic requirements. The main advantages of VDO are: (1) augmentation of alveolar bone height with new bone formation and simultaneous expansion of the soft tissues; (2) no bone harvesting is necessary; (3) the technique has a lower morbidity rate compared with conventional techniques; (4) it makes the insertion of longer dental implants feasible.

Adolescent↗

Mechanical tension in distraction osteogenesis regulates chondrocytic differentiation.

Differentiation of chondrocytes to cells of osteoblastic phenotype occurs during an interim period of bone development, fracture repair and distraction osteogenesis. To study the relationship between tension-stress and chondrogenesis, uniaxial strains (0 microstrains, 2000 microstrains, 20000 microstrains, 200000 microstrains, 300000 microstrains) were applied in a rabbit model of mandibular distraction osteogenesis. The results demonstrated that cell differentiation, apoptosis and tissue development in the newly formed gap tissue showed a correlation to the applied strain magnitudes. Only strains of 20000 microstrains resulted in a statistically significant (P<0.05) formation of cartilage struts with embedded chondrocyte-like cells. However, chondrocyte-like cells were rarely detected in samples distracted at lower or higher strain magnitudes. Osteoblasts appeared to replace cartilaginous matrix by mineralized bone matrix. The phenotypic change from chondrocytes to osteoblasts was accompanied by a decreased proteoglycan synthesis. a change in the expression from type II collagen towards type I and involved asymmetric cell divisions and apoptotic cell death. Therefore, we suggest that mechanical strain is an external stimulus responsible for phenotypic cell alterations.

Analysis of Variance↗

Effects of latency period in a rabbit mandibular distraction osteogenesis.

Recently, distraction osteogenesis has been widely used in the craniomaxillofacial skeleton. There are varying views, however, about the need for a latency period for mandibular distraction, which has been reported from 0 to 14 days. We used a rabbit model to determine the ideal latency period for mandibular distraction osteogenesis. We performed a subperiosteal corticotomy of the bilateral mandible and then applied an external device. We used 0-, 2-, 5-, and 10-day latency periods. Distraction was done at a rate of 0.5 mm per day (0.25 mm every 12 h) to elongate by 5 mm after each latency period. Hematoxylin-eosin stain was used for histologic observation, and bone labelling was observed with a fluorescence microscope. In the 0-day latency, the lengthened region was filled with fibrous tissue. Cartilage formed like isles along the lengthened edge. In the 10-day latency, the lengthened region was almost completely filled with woven bone. In the 2- and 5-day latency, there were mineralization zones at the periphery of this fibrous interzone. The primary osteon remodelling zones were located at both ends of the zones of mineralization zones. In the 5-day latency, new bone formation was observed around the periosteum.

Animals↗

Preliminary investigation into the effects of electrical stimulation on mandibular distraction osteogenesis in goats.

Craniofacial distraction osteogenesis (DO) was found to be a procedure with low preoperative and postoperative morbidity. Direct current electrical stimulation is used clinically to treat different orthopaedic problems. It provided a significant increase in new-bone formation, and a higher mechanical strength of healing. The purpose of this study was to test the effect of electric current on distraction osteogenesis and to establish the best period to apply an electric current during the different distraction phases. Twelve healthy adult goats were subjected to a vertical osteotomy in the symphyseal area. A distraction device was fixed to the bone. Animals were divided into two groups, Group I (three goats) and group II (nine goats). In group I, the device was activated 1mm per day for 10 days; while in group II, the device was activated 1mm per day for 10 days with the application of direct electric current stimulation of 10 microA either during the first 3 days of latency in a continuous mode (ESL group), or during the first 3 days of the activation period in a continuous mode (ESA group); or during the first 3 days of the consolidation period in a continuous mode (ESC group). After the activation period was completed, the distraction device left in place for additional 15 days for bone consolidation in all animals. Animals were then sacrificed, the mandible was harvested, and the distracted areas were removed and processed for mechanical and histological studies. The results of this study suggested that direct current electrical stimulation display synergism on mandibular distraction when this stimulation applied to the distraction zone during activation or consolidation periods.

Analysis of Variance↗

Unique rodent model of distraction osteogenesis of the mandible.

Despite the increasing use of distraction osteogenesis (DO) of the mandible, the molecular mechanisms regulating new bone formation during DO remain poorly understood. The purposes of this study were (1) to establish a unique rodent model of DO capable of outlining parameters for new bone formation at the distraction site and (2) to determine a critical-size defect to differentiate osteogenesis resulting from distraction from conventional fracture healing at the osteotomy site. Adult Sprague-Dawley rats were fitted successfully with this newly developed distraction device. Analyses demonstrated that the device could distract the rat mandible reliably to 5.1 mm with complete union. Acute intersegmental gaps of 2 mm resulted in complete bony union in a manner consistent with fracture healing, whereas 3-mm acute gaps resulted in varying degrees of bony union. Acute intersegmental gaps of 5.1 mm invariably resulted in fibrous nonunion. In summary, the authors have developed a rodent model of DO of the mandible. Their distraction protocols resulted successfully in advancement to 5.1 mm with bony consolidation. Notable fracture healing occurred at immediate intersegmental spaces as large as 3 mm. A gap of 5.1 mm was sufficient to act as a critical-size defect, resulting consistently in fibrous nonunion. These findings validate the effectiveness of this distraction device and establish the critical-size defect of a rat mandible at more than 3 mm. This novel model of DO provides an effective method of examining fundamental mechanisms responsible for new bone formation in the craniofacial skeleton.

Animals↗

The role of hyaluronic acid, chitosan, and calcium sulfate and their combined effect on early bony consolidation in distraction osteogenesis of a canine model.

The purpose of this project was to study the effect of hyaluronic acid, calcium sulfate, and chitosan on early bony consolidation in distraction osteogenesis of a canine model. Sixteen dogs were used for this study. The lateral surface of the mandibular body was exposed in the subperiosteal plane, and the vertical osteotomy on the mandibular body was extended downward. An external distraction device was applied to the mandibular body, and the mandibular distraction was started 5 days after the operation at a rate of 1 mm/d up to a 10-mm distraction. The experimental group was then divided into a control group, chitosan group, hyaluronic acid group, calcium sulfate combined with hyaluronic acid group, and calcium sulfate combined with chitosan group, depending on the type of implantation material in the distracted area. After completing the distraction, implantation material was injected into the distracted area, although no material was implanted into the distracted area of the control group. After implanting the materials, the distraction device was left in place for 6 weeks to allow for bony consolidation. Four dogs were allocated to each group. Two dogs in each group (total of 8 dogs) were killed 3 weeks after implantation of the material, and the other 8 dogs were killed after 6 weeks. New bone was generated in the distracted zone of all groups. In the calcium sulfate combined with chitosan group and calcium sulfate combined with hyaluronic acid group, the formation of active woven bone was observed throughout the distracted zone. Moreover, the new bone seemed to be nearly normal cortical bone at 6 weeks after implantation. In the chitosan group and hyaluronic acid group, the development of new bone was observed in the distracted zone at 6 weeks. The amount was less than that in the calcium sulfate combined with hyaluronic acid group and calcium sulfate combined with chitosan group. These findings suggest that calcium sulfate and its combined materials seem to be quite effective in early bony consolidation in distraction osteogenesis.

Animals↗

The bone regenerative effect of growth hormone on consolidation in mandibular distraction osteogenesis of a dog model.

The purpose of this project was to study the effect of growth hormone on early bony consolidation in distraction osteogenesis of a dog model. Sixteen dogs were used for this study. The vertical osteotomy on the mandibular body was extended downward. An external distraction device was applied to the mandibular body and the mandibular distraction was started 5 days after the operation at a rate of 1 mm/d up to a 10-mm distraction. The experimental group was divided into a control group and growth hormone group. Dogs in the growth hormone group received a daily subcutaneous injection of 100 microg (1 IU) of recombinant human growth hormone per kilogram of body weight. The daily administration of growth hormone was performed from the day of the osteotomy through the whole distraction period to the sacrifice. Normal saline was injected in the control group. Eight dogs were allocated to each group. Two dogs in each group, a total of four dogs, were killed at 2 weeks after completion of distraction, four dogs were killed at 4 weeks, and the other eight dogs were killed at 6 weeks. The level of serum IGF-I in the growth hormone group was elevated and peaked between 8 days and 12 days after systemic administration of growth hormone. Bone mineral density was higher in the growth hormone group and lower in the control group for the whole period. Bone mechanical strength was 300% higher in the growth hormone group than in the control group. However, results were more suggestive than conclusive. On histological examination, the formation of a substantial amount of active woven bone was observed throughout the distracted zone at six weeks in the growth hormone group. In the control group, new bone was generated from the edge to the center of the distracted zone. In addition, most of the central area of the distracted zone was filled with fibrous tissue at six weeks. In conclusion, these findings suggest that growth hormone appears to be effective in early bony consolidation in distraction osteogenesis.

Animals↗

Bone and cartilage formation in an experimental model of distraction osteogenesis.

OBJECTIVES: (a) To develop a reliable and reproducible system for distraction osteogenesis in the rat to establish a model for future investigations of bone repair and regeneration. (b) To describe and characterize the histological events in distraction osteogenesis in the rat and to determine whether cartilage development is a normal component of the process. STUDY DESIGN: Species-specific, longitudinal time study. METHODS: Twenty rats underwent production of a middiaphyseal femoral osteotomy and application of a monolateral external fixator specifically designed for distraction. Animals were divided into five groups based on the time and extent of lengthening. RESULTS: During distraction, gap tissue showed collagen bundles and fibroblasts that were oriented longitudinally to the direction of the distraction force. Woven bone appeared to be laid down on these collagen scaffolds, and the newly formed vascular sinuses appeared to be the sites from which bone formation was initiated within the distraction gap. All groups undergoing active distraction showed intramembranous ossification in the distraction gap and endochondral ossification peripherally. However, when distraction was discontinued, endochondral ossification was observed in the gap. CONCLUSION: Distraction produces an environment in the distraction gap that suppresses the formation of cartilage. The formation of cartilage by injured periosteum, however, is obligatory and does not appear to be influenced by distraction. Bone formation within the distraction gap occurs where angiogenesis develops.

Animals↗

Role of BMP, betaig-h3, and chitosan in early bony consolidation in distraction osteogenesis in a dog model.

The purpose of this investigation was to study the effect of bone morphogenetic protein (BMP), transforming growth factor beta-induced gene h3 (betaig-h3), and chitosan on early bony consolidation in distraction osteogenesis in a dog model. Sixteen dogs were used for this study. The lateral surface of the mandibular body was exposed in the subperiosteal plane and the vertical osteotomy on the mandibular body was extended downward. An external distraction device was applied to the mandibular body, and the mandibular distraction was started 5 days after the operation at a rate of 2 mm/day up to a 10-mm distraction after 5 days. The experimental group was then divided into a control group, a BMP group, a betaig-h3 group, and a chitosan group, depending on the type of implantation material used in the distracted area. On the same day after completing the distraction, BMP, betaig-h3, or chitosan was implanted into the distracted area. No material was implanted into the distracted area in the control group. After implanting the materials, the distraction device was left in place for 7 weeks to allow for bony consolidation. Four dogs were allocated to each group. Two dogs in each group, a total of eight dogs, were killed 4 weeks after completing the distraction and the other eight dogs were killed after 7 weeks. Serial radiographs were obtained every week after completing the distraction. New bone was generated in the distracted zone in all groups. In the BMP group, the formation of active woven bone was observed throughout the distracted zone, and the new bone appeared to be nearly normal cortical bone 7 weeks after implantation. In the betaig-h3 and chitosan groups, the development of new bone was observed in the distracted zone after 7 weeks; however, the amount was less than that in the BMP group. In the control group, the new bone was observed at the edges of the distracted zone. These findings suggest that BMP seems to be very effective in early bony consolidation in distraction osteogenesis.

Animals↗

Osteogenesis and angiogenesis in regenerating bone during transverse distraction: quantitative evaluation using a canine model.

For some patients with a bone defect, curved deformity, or small diameter bone, transverse distraction might be better indicated than bone transport or lengthening. However, detailed quantitative evaluation of osteogenesis or angiogenesis during transverse distraction by creating a well-controlled animal model has not been done. We established a transverse distraction model of the canine tibia. A rectangular cortex was separated for a distraction fragment. Seven days after the operation, 0.5-mm distraction was applied twice daily for 14 days. After a 28-day consolidation period, the animals were sacrificed. Quantitative evaluations of the rates of mineralization and the increases of bone mineral density indicated faster mineralization and earlier corticalization of the regenerating bone in the initial stage of the consolidation period. The average blood vessel volume ratio in the distraction area was more than three times greater than in the intact contralateral tibiae. We hypothesized that adequate preservation of the marrow arteries and stability of the distraction site throughout an experimental period could induce this faster osteogenesis. Our results indicated that the transverse distraction technique was feasible. The transverse distraction technique could be indicated for patients with small-diameter bones or with massive bone defects.

Animals↗

Unexpected absence of effect of rhBMP-7 on distraction osteogenesis.

We asked whether locally applied rhBMP-7 with a Type I collagen carrier could enhance the consolidation phase in distraction osteogenesis. We performed unilateral transverse osteotomy of the tibia in 28 immature male rabbits. In half the animals (14 rabbits), 70 microg rhBMP-7 (28.5 mg rhBMP-7/CC, OP-1) was applied locally on the day of the osteotomy; the control group (14 rabbits) had no local application. After 7 days, 3 weeks of distraction was begun at a rate of 0.5 mm every 12 hours. Starting with Week 2 of distraction, we assessed radiographic, ultrasonographic, and densitometric parameters once per week. Animals were sacrificed after 3 weeks of distraction, and after 1, 2, and 3 weeks of additional consolidation. Radiographic evaluation revealed a tendency for increased bony union and bone differentiation in the control group compared with the rhBMP-7/CC group. The bone mineral content was higher in the control group at each time. Ultrasonographic evaluation revealed the development of a cyst in the regenerate in 92% of the animals in the rhBMP-7/CC group versus 0% in the control group. Ultrasonography and histomorphometry showed delayed regenerate development in the rhBMP-7/CC group. We suggest the poor results related to the relatively large bulk of the solid carrier of the OP-1 that might have acted as a mechanical obstacle to osteogenesis.

Administration, Topical↗

Vertical distraction osteogenesis in the human mandible: a prospective morphometric study.

Vertical distraction osteogenesis has received considerable interest as a way to augment bone prior to implant placement. However, very little is known regarding the appropriate distraction protocols in the human mandible. In this study, we evaluate the effect of the distraction rate and the duration of neutrofixation on bone formation and closure of the gap in the human mandible. Vertical distraction was performed in the atrophic mandible of 16 edentulous patients, aged 62+/-6 years. The bone was distracted for approximately 10 mm at a rate of either 0.5 or 1 mm/day. Bone biopsies were taken after 7-20 weeks of neutrofixation. Histological analysis demonstrated newly formed bone in the distraction gap in all biopsies. The bone was predominantly of the woven type. After 10 weeks of neutrofixation, the gap was bridged by new bone in two out of three intact samples in the 0.5 mm/day group, but not in two intact samples of the 1 mm/day group. Histomorphometry revealed longer bone trabeculae (P=0.02) and a somewhat increased bone volume in the area where new bone formation started (P=0.07) in the group of patients having the 0.5 mm/day of distraction rate than in the 1 mm/day group. We conclude that in elderly patients, a distraction rate of 0.5 mm/day results in faster osteogenesis in the distraction gap than a rate of 1 mm/day. A minimum of 10 weeks of neutrofixation seems to be needed to close a 10 mm gap after cessation of distraction.

Aged↗