Growth potential in suture bone inlay grafts: a comparison of vascularized and free calvarial bone grafts

The present study investigates transsutural growth in vascularized and free calvarial bone grafts and notes the effects of such growth on craniofacial development. The temporalis myoosseous flap served as a model of vascularized graft. In ten 8-week-old dogs, a standardized skeletal defect, including a segment of the zygomatico-maxillary suture, was created. The defect was reconstructed with a vascularized graft in half the animals and a corresponding free graft in the remaining animals. Growth was assessed by means of serial cephalometric radiography and direct osteometry. Vascularized bone grafts demonstrated persistent transsutural growth following transplantation. Growth at the recipient site was preserved, resulting in less restriction of vertical maxillary development.     

Long-term remodeling of vascularized and nonvascularized onlay bone grafts: a macroscopic and microscopic analysis

The present study was performed to compare vascularized and nonvascularized onlay bone grafts to investigate the potential effect of graft-to-recipient bed orientation on long-term bone remodeling and changes in thickness and microarchitectural patterns of remodeling within the bone grafts. In two groups of 10 rabbits each, bone grafts were raised bilaterally from the supraorbital processes and placed subperiosteally on the zygomatic arch. The bone grafts were oriented parallel to the zygomatic arch on one side and perpendicular to the arch on the contralateral side. In the first group, vascularized bone grafts were transferred based on the auricularis anterior muscle, and in the second group nonvascularized bone grafts were transferred. Fluorochrome markers were injected during the last 3 months of animal survival, and animals were killed either 6 or 12 months postoperatively. The nonvascularized augmented zygoma showed no significant change in thickness 6 months after bone graft placement and a significant decrease in thickness 1 year after graft placement (p < 0.01). The vascularized augmented zygoma showed a slight but statistically significant decrease in thickness 6 months after graft placement (p < 0.003), with no significant difference relative to its initial thickness 1 year after graft placement. In animals killed 6 months after bone graft placement, both the rate of remodeling and the bone deposition rate measured during the last 3 months of survival were significantly higher in the vascularized bone grafts compared with their nonvascularized counterparts (p < 0.02). By 1 year postoperatively, there were no significant differences in thickness, mineral apposition rate, or osteon density between bone grafts oriented perpendicular and parallel to the zygomatic arch. These findings indicate that the vascularity of a bone graft has a significant effect on long-term thickness and histomorphometric parameters of bone remodeling, whereas the direction of placement of a subperiosteal graft relative to the recipient bed has minimal effect on these parameters. In vascularized bone grafts, both bone remodeling and deposition are accelerated during the initial period following graft placement. Continued bone deposition renders vascularized grafts better suited for the long-term maintenance of thickness and contour relative to nonvascularized grafts.     

Cleft lip nose correction with onlay calvarial bone graft and suture suspension in Oriental patients

To correct the secondary cleft lip nose deformity in Oriental patients, many alar cartilage mobilization and suspension techniques have been developed. However, these techniques have critical limitations. One of the limitations is the suspension vector, and another is suspension power. The suspension vector is from inferior to superior and from the deformed alar cartilage to the normal alar cartilage. Thus, the vector is not suitable for normal nasal tip projection. The suspension power is not satisfactory because Oriental people have underdeveloped, thin alar cartilages and thick skin. So, the suspended, deformed alar cartilage may relapse and pull the normal alar cartilage to the deformed side. To overcome these limitations, the authors use the cantilever calvarial bone graft for tip projection; it also serves as a strong, rigid framework for cartilage and soft-tissue suspension. Using these techniques, the authors can create normal nasal tip projection and a normal looking nasal aperture.     

A torsional strength comparison of vascularized and nonvascularized bone grafts

Comparisons of torsional strength are made on the ulnae from the forelegs of short haired hounds where a nonvascularized graft was performed on one leg and a vascularized graft performed on the other. By using the forelegs of a dog as the experimental model and microsurgical techniques, a vascularized bone segment was used to graft a five centimeter nonunion in one leg and at the same time a conventional bone graft was performed on a similar nonunion in the other leg. Similar segments of rib bone were used for each graft. Torsional strength data are shown for nine experimental animals. A successful method for mounting the bones for testing of torsional strength in a torsion machine is given. In each case for which the bones healed properly, the vascularized bone graft proved to be significantly stronger in torsion.     

A biomechanical comparison of vascularized and conventional autogenous bone grafts

Vascularized and conventional autogenous rib grafts were used to reconstruct 6-cm ulnar defects in the forelegs of the nine dogs. Each dog served as its own control. Biomechanical torsional testing of the grafted ulnas showed that vascularized grafts were 234 percent stronger than the conventional grafts. Bone toughness (energy absorbed) was 483 percent greater in the vascularized grafts, and elastic modulus and proportional limits were 263 and 246 percent greater, respectively. We conclude that vascularized bone grafts are significantly stronger than conventional autogenous bone grafts after 3 months of healing in the dog ulna model.     

Electrical stimulation of onlay bone grafts

An animal model was developed to determine the ability of capacitively coupled electrical fields to enhance onlay bone graft survival in the craniofacial skeleton. Fifteen male New Zealand white rabbits were divided into control and stimulated groups. Blocks of iliac bone were transplanted as onlay grafts to the mandibular rami. In all animals a capacitor apparatus was attached externally over the right mandibular ramus; however, a 5-V peak-to-peak sinusoidal signal was applied only in the stimulated group. The experimental period was 6 weeks, with a total of 30 days of constant stimulation. Graft resorption in the stimulated animals was decreased a total of 24.8 percent (p less than 0.001). There appeared to be a trend toward decreased graft resorption caused by the apparatus alone, although this was not statistically significant. The electric field alone decreased resorption by 11.5 percent (p less than 0.05). No distinguishing features were demonstrated by fluorescent vital stains or routine histology.     

Bone grafts: a radiologic, histologic, and biomechanical model comparing autografts, allografts, and free vascularized bone grafts

We developed an experimental model to compare the efficacy of free vascularized bone grafts, conventional segmental autografts, matchstick autografts, and fresh segmental allografts in terms of their ability to reconstruct a 7-cm segmental diaphyseal defect created in the canine femur. Forty-five adult mongrel dogs were studied and followed for 6 to 12 months prior to sacrifice. Evaluation included radiologic assessment of graft incorporation and hypertrophy, histology, and biomechanical testing. These studies indicated that microsurgically revascularized autografts were superior to all other groups in terms of early incorporation, hypertrophy, and the highest mechanical strength to failure. Union of the bone graft to the recipient femur was achieved by 6 months in 25 of 26 autografts, and no difference in union rate was seen within the autograft group. However, only two of five allografts achieved bony union during this time interval. Arteriography, microangiography, fluorochrome, and histologic studies all supported the concept that microsurgically revascularized grafts, when successful, maintain their viability. However, the premise that all osteocytes survive in a successfully revascularized bone graft is open to question. While decalcified sections showed that all microsurgically revascularized grafts maintained normal viability in the central marrow and cancellous portions compared with the other three groups, the viability of cortical bone in the vascularized autografts was less clear. Undecalcified fluorochrome sections suggested that circulation was not preserved in all portions of the cortex. Revascularization of the nonvascularized autografts was complete at 3 months, while, in the avascular allografts, the process was not complete at 6 months.     

Small free vascularized iliac crest bone grafts in reconstruction of the scaphoid bone: a retrospective study in 60 cases.

Carpal instability may result in progressive degenerative arthritis of the wrist. The surgical goal of the reconstruction of scaphoid nonunion is to achieve bone union and to restore the scaphoid. Many procedures are described to treat scaphoid nonunion for different indications. This retrospective study reports on the anatomical fundamentals, the operative procedure, and the results of 60 patients (21 with recalcitrant scaphoid nonunion that lasted longer than 4 years, 26 with an avascular pole fragment, and 13 with scaphoid nonunion after previous surgery) who were treated by a small free vascularized iliac crest bone graft. All 60 patients have routinely been followed up clinically and with magnetic resonance imaging. Union was achieved in 91.7 percent by improvement of stability and the compromised vascularity of the scaphoid. The bone flap loss rate and persisting nonunion was 8.3 percent, leading to progressive arthritis and carpal collapse. Complaints concerning discomforts caused by the scar were heard from 40.1 percent of the patients, and 31.7 percent complained of discomforts caused by the bony deformity. Bone deformations on the donor site were detected radiologically in 63.3 percent of the patients. In 31.7 percent, an impairment of the lateral femoral cutaneous nerve was noted. Reconstruction of the scaphoid by means of implantation of a vascularized iliac bone graft proved efficient to treat avascular recalcitrant scaphoid nonunion and pseudarthrosis with avascular proximal pole fragments.     

Cell fate specification during calvarial bone and suture development

In this study we have addressed the fundamental question of what cellular mechanisms control the growth of the calvarial bones and conversely, what is the fate of the sutural mesenchymal cells when calvarial bones approximate to form a suture. There is evidence that the size of the osteoprogenitor cell population determines the rate of calvarial bone growth. In calvarial cultures we reduced osteoprogenitor cell proliferation; however, we did not observe a reduction in the growth of parietal bone to the same degree. This discrepancy prompted us to study whether suture mesenchymal cells participate in the growth of the parietal bones. We found that mesenchymal cells adjacent to the osteogenic fronts of the parietal bones could differentiate towards the osteoblastic lineage and could become incorporated into the growing bone. Conversely, mid-suture mesenchymal cells did not become incorporated into the bone and remained undifferentiated. Thus mesenchymal cells have different fate depending on their position within the suture. In this study we show that continued proliferation of osteoprogenitors in the osteogenic fronts is the main mechanism for calvarial bone growth, but importantly, we show that suture mesenchyme cells can contribute to calvarial bone growth. These findings help us understand the mechanisms of intramembranous ossification in general, which occurs not only during cranial and facial bone development but also in the surface periosteum of most bones during modeling and remodeling.     

Integration of FGF and TWIST in calvarial bone and suture development

Mutations in the FGFR1-FGFR3 and TWIST genes are known to cause craniosynostosis, the former by constitutive activation and the latter by haploinsufficiency. Although clinically achieving the same end result, the premature fusion of the calvarial bones, it is not known whether these genes lie in the same or independent pathways during calvarial bone development and later in suture closure. We have previously shown that Fgfr2c is expressed at the osteogenic fronts of the developing calvarial bones and that, when FGF is applied via beads to the osteogenic fronts, suture closure is accelerated (Kim, H.-J., Rice, D. P. C., Kettunen, P. J. and Thesleff, I. (1998) Development 125, 1241-1251). In order to investigate further the role of FGF signalling during mouse calvarial bone and suture development, we have performed detailed expression analysis of the splicing variants of Fgfr1-Fgfr3 and Fgfr4, as well as their potential ligand Fgf2. The IIIc splice variants of Fgfr1-Fgfr3 as well as the IIIb variant of Fgfr2 being expressed by differentiating osteoblasts at the osteogenic fronts (E15). In comparison to Fgf9, Fgf2 showed a more restricted expression pattern being primarily expressed in the sutural mesenchyme between the osteogenic fronts. We also carried out a detailed expression analysis of the helix-loop-helix factors (HLH) Twist and Id1 during calvaria and suture development (E10-P6). Twist and Id1 were expressed by early preosteoblasts, in patterns that overlapped those of the FGF ligands, but as these cells differentiated their expression dramatically decreased. Signalling pathways were further studied in vitro, in E15 mouse calvarial explants. Beads soaked in FGF2 induced Twist and inhibited Bsp, a marker of functioning osteoblasts. Meanwhile, BMP2 upregulated Id1. Id1 is a dominant negative HLH thought to inhibit basic HLH such as Twist. In Drosophila, the FGF receptor FR1 is known to be downstream of Twist. We demonstrated that in Twist(+/)(-) mice, FGFR2 protein expression was altered. We propose a model of osteoblast differentiation integrating Twist and FGF in the same pathway, in which FGF acts both at early and late stages. Disruption of this pathway may lead to craniosynostosis.     

Comparison of residual osseous mass between vascularized and nonvascularized onlay bone transfers

Composite flaps containing vascularized frontal bone were transferred on muscle pedicles in immature rabbits. Vascular continuity was maintained on one side and interrupted on the other. Bone weights at 16 weeks following transfer were compared with those of unoperated controls. The conventional bone graft demonstrated significant reduction in osseous mass. The vascularized bone maintained its mass compared with unoperated controls. Vascularized bone transfer appears to be the preferred surgical technique whenever possible.     

Orbital floor and infraorbital rim reconstruction after total maxillectomy using a vascularized calvarial bone flap.

A number of techniques have been introduced to support the orbital floor after maxillectomy without orbital exenteration. These methods include skin graft or muscular sling, but they have resulted in severe complications, such as enophthalmos, global ptosis, diplopia, and facial deformity. Currently, advanced microvascular reconstruction using bone and soft tissue is performed by many surgeons. This usually results in the filling of the postmaxillectomy defect, but the lack of support for the orbital rim and floor by the bone flap may still cause the complications mentioned above. Vascularized calvarial bone flap was chosen in this study for reconstruction of the orbital floor and infraorbital rim to function as a buttress, to reconstruct recipient sites of poor vascular bed after radiation therapy, and to withstand further postoperative radiation. By providing a solid floor and rim, these complications can be prevented with satisfactory function and aesthetically acceptable results. From September of 1995 to July of 1998, we performed vascularized bone flap for the reconstruction of the orbital floor and infraorbital rim in four cases after total maxillectomy involving the orbital floor. With a follow-up period from 19 to 35 months (mean, 27 months), we obtained significant improvement of functional and aesthetically acceptable results without global ptosis, enophthalmos, diplopia, or severe facial contour deformity.