Sarah E. Henderson

Magnesium alloys as a biomaterial for degradable craniofacial screws.

Recently, magnesium (Mg) alloys have received significant attention as potential biomaterials for degradable implants, and this study was directed at evaluating the suitability of Mg for craniofacial bone screws. The objective was to implant screws fabricated from commercially available pure Mg and alloy AZ31 in vivo in a rabbit mandible. First, Mg and AZ31 screws were compared to stainless steel screws in an in vitro pull-out test and determined to have a similar holding strength (∼40N). A finite-element model of the screw was created using the pull-out test data, and this model can be used for future Mg alloy screw design. Then, Mg and AZ31 screws were implanted for 4, 8 and 12weeks, with two controls of an osteotomy site (hole) with no implant and a stainless steel screw implanted for 12weeks. Microcomputed tomography was used to assess bone remodeling and Mg/AZ31 degradation, both visually and qualitatively through volume fraction measurements for all time points. Histological analysis was also completed for the Mg and AZ31 at 12weeks. The results showed that craniofacial bone remodeling occurred around both Mg and AZ31 screws. Pure Mg had a different degradation profile than AZ31; however, bone growth occurred around both screw types. The degradation rate of both Mg and AZ31 screws in the bone marrow space and the muscle were faster than in the cortical bone space at 12weeks. Furthermore, it was shown that by alloying Mg, the degradation profile could be changed. These results indicate the promise of using Mg alloys for craniofacial applications.

Animal Models of Temporomandibular Joint Disorders: Implications for Tissue Engineering Approaches

Animal models for temporomandibular joint disorder (TMD) or degradation are necessary for assessing the value of current and future tissue engineering therapies. After reviewing the literature, it is quite apparent that most TMD animal studies can be categorized into chemical approaches or surgical/mechanical approaches. Overall, it was found that the top five cited manuscripts for all chemical models were cited by almost 40% more manuscripts than the top five manuscripts for surgical/mechanical models. It is clear that the chemical approaches have focused on the inflammatory aspect of TMDs and its relationship to pain. However, chemical irritants must be tested in larger animal models, and the effect of short-term inflammation on the mechanical properties of the fibrocartilage must be examined. Nevertheless, therapeutic approaches aimed at reducing or controlling inflammation could use the established chemical methods. Surgical/mechanical methods can be used as negative controls for first generation TMJ tissue engineering approaches when the therapy is applied immediately after injury. Next generation tissue engineering approaches will require testing on tissues degenerated for a few months after the surgical/mechanical methods, with enhanced functional assessment techniques.

Na+/H+ exchanger regulatory factor 1 (NHERF1) directly regulates osteogenesis.

Background: The bone phenotype of NHERF1-null mice was ascribed to indirect actions. Results: With dietary supplementation to maintain normal serum phosphate, NHERF1-deficient mice showed aberrant bone mineralization and decreased bone quality. Osteoblast differentiation from mesenchymal stem cells was impaired. Conclusion: NHERF1 is expressed in mineralizing osteoblasts and directly regulates bone formation. Significance: We provide an experimentally validated mechanistic model of NHERF1 regulating bone formation. Bone formation requires synthesis, secretion, and mineralization of matrix. Deficiencies in these processes produce bone defects. The absence of the PDZ domain protein Na+/H+ exchange regulatory factor 1 (NHERF1) in mice, or its mutation in humans, causes osteomalacia believed to reflect renal phosphate wasting. We show that NHERF1 is expressed by mineralizing osteoblasts and organizes Na+/H+ exchangers (NHEs) and the PTH receptor. NHERF1-null mice display reduced bone formation and wide mineralizing fronts despite elimination of phosphate wasting by dietary supplementation. Bone mass was normal, reflecting coordinated reduction of bone resorption and formation. NHERF1-null bone had decreased strength, consistent with compromised matrix quality. Mesenchymal stem cells from NHERF1-null mice showed limited osteoblast differentiation but enhanced adipocyte differentiation. PTH signaling and Na+/H+ exchange were dysregulated in these cells. Osteoclast differentiation from monocytes was unaffected. Thus, NHERF1 is required for normal osteoblast differentiation and matrix synthesis. In its absence, compensatory mechanisms maintain bone mass, but bone strength is reduced.

BMP-2-regenerated calvarial bone: a biomechanical appraisal in a large animal model.

AbstractRecombinant human bone morphogenetic protein-2 (rhBMP-2) is gaining popularity in craniofacial applications. Calvarial defects are, under normal circumstances, subjected to only minimal levels of the biomechanical stresses known to play an important role in osteogenesis, yet regenerated calvarial bone must be capable of withstanding traumatic forces such that the underlying neurocapsule is protected. The aim of this study is to, for the first time, assess the biomechanical properties of calvarial bone regenerated with derivations of a commercially available rhBMP-2–based system. Standardized calvarial defects were created in 23 adult male canines. These defects were treated with rhBMP-2 on one of several carriers. After 24 weeks, the biomechanical properties of the rhBMP-2–generated bone were compared to those of controls with a modified punch-out test (Bluehill 2; Instron, Norwood, Mass) and compared using a paired nonparametric analyses (SPSS, 17.0, Chicago, Ill). In a previously published report, defects across all the rhBMP-2 therapy groups were observed to have a mean rate of 99.5% radio-opacity at 24 weeks indicating nearly full bony coverage of the calvarial defect (compared to 32.7% in surgical controls). For ultimate load, ultimate energy, and first peak energy, there were significant differences (P < 0.05) with the control native bone having more robust biomechanical properties than the rhBMP-2–generated bone. We conclude from these findings that rhBMP-2–generated calvarial bone is significantly less protective against trauma than native bone at 6 months. Further investigation is required to assess the efficacy of rhBMP-2 in healing calvarial defects in the longer term.

Temporomandibular joint fibrocartilage degeneration from unilateral dental splints

OBJECTIVE The objective of this study was to determine the extent to which altered loading in the temporomandibular joint (TMJ), as might be associated with a malocclusion, drives degeneration of articulating surfaces in the TMJ. We therefore sought to quantify the effects of altered joint loading on the mechanical properties and biochemical content and distribution of TMJ fibrocartilage in the rabbit. DESIGN Altered TMJ loading was induced with a 1mm splint placed unilaterally over the maxillary and mandibular molars for 6 weeks. At that time, TMJ fibrocartilage was assessed by compression testing, biochemical content (collagen, glycosaminoglycan (GAG), DNA) and distribution (histology), for both the TMJ disc and the condylar fibrocartilage. RESULTS There were no changes in the TMJ disc for any of the parameters tested. The condylar fibrocartilage from the splinted animals was significantly stiffer and the DNA content was significantly lower than that in control animals. There was significant remodeling in the condylar fibrocartilage layers as manifested by a change in GAG and collagen II distribution and a loss of defined cell layers. CONCLUSIONS A connection between the compressive properties of TMJ condylar fibrocartilage after 6 weeks of splinting and the changes in histology was observed. These results suggest a change in joint loading leads to condylar damage, which may contribute to pain associated with at least some forms of TMJ disease.

Functional analysis of the rabbit temporomandibular joint using dynamic biplane imaging

The dynamic function of the rabbit temporomandibular joint (TMJ) was analyzed through non-invasive, three-dimensional skeletal kinematics, providing essential knowledge for understanding normal joint motion. The objective of this study was to evaluate and determine repeatable measurements of rabbit TMJ kinematics. Maximal distances, as well as paths were traced and analyzed for the incisors and for the condyle-fossa relationship. From one rabbit to another, the rotations and translations of both the incisors and the condyle relative to the fossa contained multiple clear, repeatable patterns. The slope of the superior/inferior incisor distance with respect to the rotation about the transverse axis was repeatable to 0.14 mm/deg and the right/left incisor distance with respect to the rotation about the vertical axis was repeatable to 0.03 mm/deg. The slope of the superior/inferior condylar translation with respect to the rotational movement about the transverse axis showed a consistent relationship to within 0.05 mm/deg. The maximal translations of the incisors and condyles were also consistent within and between rabbits. With an understanding of the normal mechanics of the TMJ, kinematics can be used to compare and understand TMJ injury and degeneration models.

Analysis of pain in the rabbit temporomandibular joint after unilateral splint placement.

AIMS To determine whether behavioral, anatomical, and physiologic endpoints widely used to infer the presence of pain in rodent models of temporomandibular disorders (TMD) were applicable to the rabbit model of TMD associated with altered joint loading. METHODS Unilateral molar dental splints were used to alter temporomandibular joint (TMJ) loading. Changes in nociceptive threshold were assessed with a mechanical probing of the TMJ region on nine splinted and three control rabbits. Fos-like immunoreacitivty in the trigeminal subnucleus caudalis was assessed with standard immunohistochemical techniques from three splinted and six control animals. Retrogradely labeled TMJ afferents were studied with patch-clamp electrophysiologic techniques from three splinted and three control animals. Remodeling of TMJ condyles was assessed by histologic investigations of three splinted and three control animals. A Student t test or a Mann-Whitney U test was used with significance set at P < .05 to compare splinted to control samples. RESULTS While variable, there was an increase in mechanical sensitivity in splinted rabbits relative to controls. The increase in Fos+ cells in splinted rabbits was also significant relative to naïve controls (86 ± 8 vs 64 ± 15 cells/section, P < .05). The rheobase (364 ± 80 pA) and action potential threshold (-31.2 ± 2.0 mV) were higher in TMJ afferents from splinted rabbits compared to controls (99 ± 22 pA and -38.0 ± 2.0 mV, P < .05). There was significant remodeling in the condylar fibrocartilage layers as manifested by a change in glycosaminoglycan distribution and a loss of defined cell layers. CONCLUSION Behavioral and anatomical results were consistent with an increase in nociceptive signaling in concert with condylar remodeling driven by altered TMJ loading. Changes in excitability and action potential waveform were consistent with possible compensatory changes of TMJ afferents for an overall increase in afferent drive associated with joint degeneration. These compensatory changes may reflect pain-adaption processes that many patients with TMJ disorders experience.

Biomechanical Integrity in Craniofacial Surgery: Calvarial Reconstruction in Favorable and Infected Defects with Bone Morphogenetic Protein 2

Background: The limitations of autologous and alloplastic reconstruction for craniofacial bone defects have created a clinical need for viable tissue-engineering strategies. Recombinant human bone morphogenetic protein-2 (rhBMP-2) has shown promise in this setting. The aim of this study was to determine the long-term biomechanical properties of rhBMP-2–mediated calvarial reconstruction. Methods: Twelve-week-old New Zealand White rabbits underwent subtotal calvarectomy. Defects were repaired in one of several groups: immediate reconstruction with autologous graft, immediate reconstruction with cryopreserved bone graft, immediate reconstruction with rhBMP-2 (favorable), and delayed reconstruction with rhBMP-2 following infection and subsequent débridement (unfavorable). Cryopreserved reconstructions were measured at 6 weeks; autologous reconstructions were measured at 6 weeks and 6 months; and both favorable and unfavorable rhBMP-2 reconstructions were assessed at 6 weeks, 6 months, and 1 year after reconstruction. Healing was assessed with computed tomography. An unconfined compression test was performed for biomechanical analysis. Stress at 20 percent strain, percentage relaxation, tangent modulus, and final strain at 1800 N were compared between groups. Results: Nearly complete radiographic coverage was achieved by 6 months for autologous reconstruction and by 6 weeks for rhBMP-2 reconstruction. Favorable rhBMP-2 reconstruction demonstrated a larger final strain at 1800 N through 1 year compared with native bone. Bone in unfavorable rhBMP-2 reconstruction was more compressible than native bone, with a larger final strain at 1800 N at 1 year. There were no significant differences between favorable and unfavorable groups. Conclusions: Despite providing radiographic coverage, the biomechanical properties of rhBMP-2 bone differ from those of native bone. Further studies are warranted to determine how these properties affect overall strength and structural integrity.

Temporomandibular Joint Kinematics of the Rabbit Model With Mechanically Disrupted Occlusion

Degeneration of the articulating surfaces and pain associated with temporomandibular joint (TMJ) dysfunction are the primary symptoms of TMJ disorders (TMDs), where normal life activities such as eating, talking, and even sleeping may be drastically impaired [1–3]. To accelerate the discovery of effective therapeutic interventions for the treatment of TMD pain, we have been establishing a novel non-invasive approach for objectively assessing the presence of joint hypersensitivity. Our approach to identify chronic joint pain is based on evidence that all of the etiological factors associated with TMD pain implicate remodeling and degeneration of the joint in response to alterations in motion and loading. The injury model used for this study was a reversible, mechanical model through splint placement on the molars. It is hypothesized that arthrokinematic analysis will identify a specific pattern of functional changes that constitute a signature for the presence of irreversible damage.Copyright