GDF-5 deficiency in mice leads to disruption of tail tendon form and function

Although the biological factors which regulate tendon homeostasis are poorly understood, recent evidence suggests that Growth and Differentiation Factor-5 (GDF-5) may play a role in this important process. The purpose of this study was to investigate the effect of GDF-5 deficiency on mouse tail tendon using the brachypodism mouse model. We hypothesized that GDF-5 deficient tail tendon would exhibit altered composition, ultrastructure, and biomechanical behavior when compared to heterozygous control littermates. Mutant tail tendons did not display any compositional differences in sulfated glycosaminoglycans (GAG/DNA), collagen (hydroxyproline/DNA), or levels of fibromodulin, decorin, or lumican. However, GDF-5 deficiency did result in a 17% increase in the proportion of medium diameter (100-225 nm) collagen fibrils in tail tendon (at the expense of larger fibrils) when compared to controls (p < 0.05). Also, mutants exhibited a trend toward an increase in irregularly-shaped polymorphic fibrils (33% more, p > 0.05). While GDF-5 deficient tendon fascicles did not demonstrate any significant differences in quasistatic biomechanical properties, mutant fascicles relaxed 11% more slowly than control tendons during time-dependent stress-relaxation tests (p < 0.05). We hypothesize that this subtle alteration in time-dependent mechanical behavior is most-likely due to the increased prevalence of irregularly shaped type I collagen fibrils in the mutant tail tendons. These findings provide additional evidence to support the conclusion that GDF-5 may play a role in tendon homeostasis in mice.     

Effect of Y2 Receptor Deletion on Whole Bone Structural Behavior in Mice

Recent evidence has shown that mice deficient in the NPY Y2 receptor have an increase in trabecular bone volume as well as cortical bone mass due to increased osteoblast activity. However, the mechanical phenotype of Y2 ?/? bone has not yet been assessed. Thus, the aim of the present study was to examine the effect of Y2 deletion on murine cortical bone structural behavior, as well as to assess the material and geometric contributions to that behavior. The results of this study indicate that Y2 ?/? mice on a 129 SV × Balb/c background strain are smaller in body mass and have smaller bones than wild‐type controls. As expected based on smaller bone cross‐sectional properties, cortical bone structural strength was lower in ?/? animals. Surprisingly, the structural stiffness of ?/? bones was comparable to that of +/+ bones despite their smaller cross‐sectional geometry. Comparable structural stiffness appeared to be achieved by means of an elevated effective shear modulus, which was associated with a small, but statistically significant, higher ash content in Y2 ?/? bones. These data represent the first comprehensive characterization of the effect of Y2 deletion on cortical bone structural and material behavior to date. Anat Rec, 291:14–18, 2007. © 2007 Wiley‐Liss, Inc.     

Ultrastructural Determinants of Murine Achilles Tendon Strength During Healing

The mechanisms by which tendon strength is established during growth and development and restored following injury are not completely understood and are likely to be complex, multifactorial processes. Several studies examining the relationship between mechanical behavior and ultrastructural characteristics of tendons and ligaments during growth and maturation suggest that collagen fibril diameter is strongly correlated with tendon strength. Because of the similarities between development and repair processes of musculoskeletal tissues, increases in tendon strength during healing may be related to increases in fibril ultrastructural parameters such as fibril size, numerical density, and area fraction. In this study, we compared murine Achilles tendons at various time points after tenotomy with sham-operated controls in tensile tests to failure and examined tendons using electron microscopy to assess collagen fibril ultrastructure. We found that in the 6-week period following Achilles tenotomy, fibril mean diameter remained significantly smaller than sham-side diameter by a factor of 2–3. Despite the persistently small fibril size, increasing numerical density resulted in a gradual increase in fibril area fraction. Biomechanical strength did not reach that of intact tendons until some time between 5 and 7 weeks, approximately the same time period when fibril area fraction began to approach sham values. These data suggest that parameters other than collagen fibril size are most responsible for increased tendon strength during healing.     

Sex matters in the establishment of murine tendon composition and material properties during growth

The existence of sex‐based differences in tendon and ligament injury rates has led investigators to test the hypothesis that sex plays a significant role in modulating tendon and ligament composition and material properties. To date, no studies have attempted to characterize how such differences develop during the course of normal tissue maturation and growth. Thus, the primary aim of the present study was to use a murine model to test the hypothesis that sex‐based differences in the normal age‐related development of tendon composition and material properties exist by assessing these parameters in the Achilles and tail tendons from 4‐, 6‐, 9‐, 12‐, and 15‐week‐old male and female C57Bl/6J mice. Despite significantly lower levels of total collagen content in females subsequent to sexual maturity (p < 0.0001), as well as a significant effect of sex on glycosaminoglycan content (p < 0.0001), Achilles tendon elastic modulus was not compromised in females. Female Achilles tendons did exhibit a significantly higher failure strain (p = 0.0201) and strain energy density (p = 0.0004) than did males, as well as a trend toward higher ultimate strength (p = 0.0556). In contrast to the high load‐bearing environment of the Achilles tendon site, sex did not have a statistically significant effect on any compositional or material property in the low load‐bearing tendon fascicles of the tail. These data support recent studies by others, which suggest that male and female tendons have a differential adaptational response to their local mechanical loading environment. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:631–638, 2010     

Evaluation of undifferentiated carcinoma of nasopharyngeal type with thallium-201 and technetium-99m MIBI SPECT.

OBJECTIVE: To determine the value of 201-thallium (201-Tl) and technetium-99m sestamibi (Tc-99m MIBI) single photon emission computed tomography (SPECT) for detecting primary undifferentiated carcinoma of nasopharyngeal type (UCNT), residual/recurrent tissue, and lymph node involvement. STUDY DESIGN AND SETTING: SPECT of head and neck was prospectively performed in 46 patients with a history of UCNT (201-Tl in 24 patients, Tc-99m MIBI in 22). CT/MRI findings, clinical follow-up, and pathohistological verification served as a gold standard for calculating sensitivity, specificity, and accuracy of each scintigraphic technique. Tumor-to-background index (T/Bg) was derived when SPECT findings were positive. RESULTS: Sensitivity of 201-Tl SPECT was 87 percent, with 78 percent specificity, 83 percent accuracy, and T/Bg of 4.05 +/- 1.50. Tc-99m MIBI SPECT had 85 percent sensitivity, 78 percent specificity, 82 percent accuracy, and T/Bg of 4.45 +/- 1.27. CONCLUSION AND SIGNIFICANCE: 201-Tl SPECT and Tc-99m MIBI SPECT are useful for detecting primary UCNT, residual/recurrent disease, and lymph node involvement. This use is particularly valuable after chemoradiotherapy when CT/MRI may be ambiguous.     

GDF-5 deficiency in mice alters the ultrastructure, mechanical properties and composition of the Achilles tendon.

Acromesomelic dysplasia of the Hunter-Thompson and Grebe types are rare human disorders based on growth/differentiation factor (GDF)-5/CDMP-1 genetic mutations. Numerous skeletal abnormalities are present in these individuals, including shortened limb bones and severe dislocations of the knee. In the GDF-5 deficient brachypodism mouse, similar, although less severe, phenotypes are observed. It is unknown whether the joint dislocations observed in these disorders are due to a defect in the original formation of joints such as the knee, or to abnormalities in the tendons and ligaments themselves. We hypothesized that tendons from GDF-5 deficient mice would exhibit altered composition, mechanical properties, and ultrastructure when compared with heterozygous control littermates. GDF-5 deficient Achilles tendons were structurally weaker than controls, and structural strength differences appeared to be caused by compromised material properties: after normalizing by collagen per unit length, mutant tendons were still 50% weaker (P < 0.0001) and 50% more compliant (P < 0.001) than controls. Despite comparable levels of skeletal maturity in the two cohorts, the majority of mutant tendon failures occurred in the mid-substance of the tendon (64% of all failures), whereas the majority of control failures occurred via avulsion (92% of all failures). Mutant Achilles tendons contained 40% less collagen per microgram of DNA when compared to controls (P = 0.004). No significant difference in glycosaminoglycan (GAG)/DNA was detected. Ultrastructural analyses indicated a slight trend toward increased frequency of small diameter (30-100 nm) collagen fibrils in the mutant Achilles. Our findings suggest that increased tendon and ligament laxity may be the cause of the joint dislocations seen in patients with Hunter-Thompson and Grebe type dysplasia, rather than developmental abnormalities in the joints themselves.     

Altered hypertrophic chondrocyte kinetics in GDF-5 deficient murine tibial growth plates.

The growth/differentiation factors (GDFs) are a subgroup of the bone morphogenetic proteins best known for their role in joint formation and chondrogenesis. Mice deficient in one of these signaling proteins, GDF-5, exhibit numerous skeletal abnormalities, including shortened limb bones. The primary aim of this study was determine whether GDF-5 deficiency would alter the growth rate in growth plates from the long bones in mice and, if so, how this is achieved. Stereologic and cell kinetic parameters in proximal tibial growth plates from 5-week-old female GDF-5 -/- mice and control littermates were examined. GDF-5 deficiency resulted in a statistically significant reduction in growth rate (-14%, p=0.03). The effect of genotype on growth rate was associated with an altered hypertrophic phase duration, with hypertrophic cells from GDF-5 deficient mice exhibiting a significantly longer phase duration compared to control littermates (+25%, p=0.006). These data suggest that one way in which GDF-5 might modulate the rate of endochondral bone growth could be by affecting the duration of the hypertrophic phase in growth plate chondrocytes.     

The Outback Catheter: A New Device for True Lumen Re-entry After Dissection During Recanalization of Arterial Occlusions

To report the initial experience with a new catheter system (The Outback catheter) designed to allow fluoroscopically controlled re-entry of the true arterial lumen after subintimal guidewire passage during recanalization procedures of arterial occlusions. The catheter was used in 10 patients with intermittent claudication caused by chronic segmental occlusions of the superficial femoral or popliteal arteries. In all patients, conventional guidewire recanalization had failed. In 8 patients, successful true lumen re-entry was achieved with the Outback catheter. Percutaneous transluminal angioplasty was successfully performed in these patients without complications. Two technical failures occurred in heavily calcified arteries. The Outback catheter was safe and effective when used in complicated recanalization procedures in the superficial femoral and popliteal artery and the tibial trunk.     

Giant pseudoaneurysm from Vieussens' arterial ring

A giant coronary pseudoaneurysm of uncertain cause, arising from Vieussens' arterial ring, was preoperatively diagnosed in an oligosymptomatic female patient. Successful off-pump surgical excision without additional bypass grafting was performed. Difficulties in diagnostic algorithm, as well as possible cause and extremely rare localization were discussed.