Kimerly A. Powell

Spine Fusion Using Cell Matrix Composites Enriched in Bone Marrow-Derived Cells

Bone marrow-derived cells including osteoblastic progenitors can be concentrated rapidly from bone marrow aspirates using the surface of selected implantable matrices for selective cell attachment. Concentration of cells in this way to produce an enriched cellular composite graft improves graft efficacy. The current study was designed to test the hypothesis that the biologic milieu of a bone marrow clot will significantly improve the efficacy of such a graft. An established posterior spinal fusion model and cancellous bone matrix was used to compare an enriched cellular composite bone graft alone, bone matrix plus bone marrow clot, and an enriched bone matrix composite graft plus bone marrow clot. Union score, quantitative computed tomography, and mechanical testing were used to define outcome. The union score for the enriched bone matrix plus bone marrow clot composite was superior to the enriched bone matrix alone and the bone matrix plus bone marrow clot. The enriched bone matrix plus bone marrow clot composite also was superior to the enriched bone matrix alone in fusion volume and in fusion area. These data confirm that the addition of a bone marrow clot to an enriched cell-matrix composite graft results in significant improvement in graft performance. Enriched composite grafts prepared using this strategy provide a rapid, simple, safe, and inexpensive method for intraoperative concentration and delivery of bone marrow-derived cells and connective tissue progenitors that may improve the outcome of bone grafting.

Initial experience with the Cosgrove-Edwards annuloplasty system

BACKGROUND The mitral valve has a nonplanar shape and a sphincter action. Pathologic dilatation occurs along the posterior annulus. To preserve the physiologic function and correct annular dilatation, we developed an annuloplasty system that is universally flexible and produces a measured plication of the posterior annulus (Cosgrove-Edwards Annuloplasty System). METHODS The results of 150 consecutive mitral valve repairs using this system were analyzed. Mean age was 58 +/- 13 years; 59% were men. The cause of the valve disease was degenerative in 75% of the patients, rheumatic in 13%, ischemic in 8%, and infectious in 4%. Associated procedures were performed in 61 patients (41%). RESULTS Echocardiographic mitral regurgitation decreased from 3.7 +/- 0.6 before repair to 0.2 +/- 0.4 after repair (p < 0.0001). There were no hospital deaths and no cases of hemodynamically significant systolic anterior motion or other annuloplasty-related complications. Follow-up was 100% complete at a mean of 3.1 +/- 3.6 months. There were three late deaths, three transient ischemic attacks, and one episode of endocarditis. Five patients (3.3%) have undergone reoperation for recurrent mitral insufficiency; no reoperations were related to the annuloplasty system. At a mean of 9 months, three-dimensional reconstruction of the mitral annulus from multiple echocardiographic images confirmed the nonplanar shape and sphincter mechanism of the annulus. Annular orifice area decreased 19% during the cardiac cycle from a mean of 10.3 cm2 in diastole to 8.6 cm2 in systole. CONCLUSIONS This annuloplasty system is effective for repair of insufficiency secondary to all causes, preserves physiologic annulus function, and is associated with a low incidence of valve-related complication.

Quantitative classification of breast tumors in digitized mammograms.

The goal of this study was to develop a technique to distinguish benign and malignant breast lesions in secondarily digitized mammograms. A set of 51 mammograms (two views/patient) containing lesions of known pathology were evaluated using six different morphological descriptors: circularity, mu R/sigma R (where mu R = mean radial distance of tumor boundary, sigma R = standard deviation); compactness, P2/A (where P = perimeter length of tumor boundary and A = area of the tumor); normalized moment classifier; fractal dimension; and a tumor boundary roughness (TBR) measurement (the number of angles in the tumor boundary with more than one boundary point divided by the total number of angles in the boundary). The lesion was segmented from the surrounding background using an adaptive region growing technique. Ninety-seven percent of the lesions were segmented using this approach. An ROC analysis was performed for each parameter and the results of this analysis were compared to each other and to those obtained from a subjective review by two board-certified radiologists who specialize in mammography. The results of the analysis indicate that all six parameters are diagnostic for malignancy with areas under their ROC curves ranging from 0.759 to 0.928. We observed a trend towards increased specificity at low false-negative rates (0.01 and 0.001) with the TBR measurement. Additionally, the diagnostic accuracy of a classification model based on this parameter was similar to that of the subjective reviewers.

Selective retention of bone marrow-derived cells to enhance spinal fusion.

Connective tissue progenitors can be concentrated rapidly from fresh bone marrow aspirates using some porous matrices as a surface for cell attachment and selective retention, and for creating a cellular graft that is enriched with respect to the number of progenitor cells. We evaluated the potential value of this method using demineralized cortical bone powder as the matrix. Matrix alone, matrix plus marrow, and matrix enriched with marrow cells were compared in an established canine spinal fusion model. Fusions were compared based on union score, fusion mass, fusion volume, and by mechanical testing. Enriched matrix grafts delivered a mean of 2.3 times more cells and approximately 5.6 times more progenitors than matrix mixed with bone marrow. The union score with enriched matrix was superior to matrix alone and matrix plus marrow. Fusion volume and fusion area also were greater with the enriched matrix. These data suggest that the strategy of selective retention provides a rapid, simple, and effective method for concentration and delivery of marrow-derived cells and connective tissue progenitors that may improve the outcome of bone grafting procedures in various clinical settings.

Estrogen Mediated-Activation of miR-191/425 Cluster Modulates Tumorigenicity of Breast Cancer Cells Depending on Estrogen Receptor Status

MicroRNAs (miRNAs), single-stranded non-coding RNAs, influence myriad biological processes that can contribute to cancer. Although tumor-suppressive and oncogenic functions have been characterized for some miRNAs, the majority of microRNAs have not been investigated for their ability to promote and modulate tumorigenesis. Here, we established that the miR-191/425 cluster is transcriptionally dependent on the host gene, DALRD3, and that the hormone 17β-estradiol (estrogen or E2) controls expression of both miR-191/425 and DALRD3. MiR-191/425 locus characterization revealed that the recruitment of estrogen receptor α (ERα) to the regulatory region of the miR-191/425-DALRD3 unit resulted in the accumulation of miR-191 and miR-425 and subsequent decrease in DALRD3 expression levels. We demonstrated that miR-191 protects ERα positive breast cancer cells from hormone starvation-induced apoptosis through the suppression of tumor-suppressor EGR1. Furthermore, enforced expression of the miR-191/425 cluster in aggressive breast cancer cells altered global gene expression profiles and enabled us to identify important tumor promoting genes, including SATB1, CCND2, and FSCN1, as targets of miR-191 and miR-425. Finally, in vitro and in vivo experiments demonstrated that miR-191 and miR-425 reduced proliferation, impaired tumorigenesis and metastasis, and increased expression of epithelial markers in aggressive breast cancer cells. Our data provide compelling evidence for the transcriptional regulation of the miR-191/425 cluster and for its context-specific biological determinants in breast cancers. Importantly, we demonstrated that the miR-191/425 cluster, by reducing the expression of an extensive network of genes, has a fundamental impact on cancer initiation and progression of breast cancer cells.

Changes in rotator cuff muscle volume, fat content, and passive mechanics after chronic detachment in a canine model

BACKGROUND Long-standing tears of the rotator cuff can lead to substantial and perhaps irreversible changes in the affected rotator cuff muscles. We developed a chronic rotator cuff tear in a canine model to investigate and quantify the time-related changes in passive mechanics, volume, and fat of the infraspinatus muscle. We hypothesized that infraspinatus muscle stiffness would increase, volume would decrease, and fat content would increase at twelve weeks following tendon detachment. METHODS The right infraspinatus tendon of eight adult mongrel dogs were surgically detached from the proximal part of the humerus. The uninvolved left shoulder served as a control. Muscle volume changes were quantified with use of magnetic resonance imaging. At twelve weeks, the passive mechanical properties of the chronically detached and control muscles were determined intraoperatively with use of a custom-designed device. Intramuscular fat was evaluated histologically at the time that the animals were killed. RESULTS After twelve weeks of detachment, the stiffness was significantly increased in the detached infraspinatus muscles relative to that in the controls (p < 0.0001). Magnetic resonance image analysis demonstrated that the detached muscle volumes decreased by an average of 32% in the first six weeks and remained constant thereafter. Intramuscular fat increased significantly in the detached muscles and to a greater extent in the lateral regions (p < 0.05). CONCLUSIONS The chronically detached muscle is not merely a smaller version of the original muscle but, rather, a different muscle. The detached muscle becomes stiffer, and the passive loads required to repair it can become excessive. A significant reduction in muscle volume occurs within days to weeks following tendon detachment (p < 0.0001). The nonuniformity of changes in muscle fat suggests that fat content should be used cautiously as an indicator of muscle quality.

Regional heterogeneity of function in nonischemic dilated cardiomyopathy

OBJECTIVE To quantify regional three-dimensional (3D) motion and myocardial strain using magnetic resonance (MR) tissue tagging in patients with non-ischemic dilated cardiomyopathy (DCM). METHODS MR grid tagged images were obtained in multiple short- and long-axis planes in thirteen DCM patients. Regional 3D displacements and strains were calculated with the aid of a finite element model. Five of the patients were also imaged after LV volume reduction by partial left ventriculectomy (PLV), combined with mitral and tricuspid valve repair. RESULTS DCM patients showed consistent, marked regional heterogeneity. Systolic lengthening occurred in the septum in both circumferential (%S(C) -5+/-7%) and longitudinal (%S(L) -2+/-5%) shortening components (negative values indicating lengthening). In contrast, the lateral wall showed relatively normal systolic shortening (%S(C) 12+/-6% and %S(L) 6+/-5%, P<0.001 lateral vs. septal walls). A geometric estimate of regional stress was correlated with shortening on a regional basis, but could not account for the differences in shortening between regions. In the five patients imaged post-PLV, septal function recovered (%S(C) 9+/-5%,%S(L) 6+/-5%, P<0.02 pre vs. post) with normalization of wall stress, whereas lateral wall shortening was reduced (%S(C) 7+/-6%,%S(L) 3+/-3%, P<0.02 pre vs. post) around the site of surgical resection. CONCLUSIONS A consistent pattern of regional heterogeneity of myocardial strain was seen in all patients. Reduced function may be related to increased wall stress, since recovery of septal function is possible after PLV. However, simple geometric stress determinants are not sufficient to explain the functional heterogeneity observed.

Pressure Recovery in Bileaflet Heart Valve Prostheses: Localized High Velocities and Gradients in Central and Side Orifices With Implications for Doppler-Catheter Gradient Relation in Aortic and Mitral Position

BACKGROUND We investigate pressure recovery in central and side orifices of St Jude valves and the effect of mitral versus aortic position on the relation between Doppler- and catheter-derived pressure gradients. METHODS AND RESULTS Maximum, transvalvular, and net pressure gradients are calculated and compared with Doppler-derived gradients in an in vitro model. Pressure recovery and pressure loss coefficients are calculated. Simultaneous Doppler and catheter gradients are obtained intraoperatively in five patients undergoing mitral valve replacement. Centerline Doppler gradients correspond closely with maximum catheter gradients but are higher than transvalvular and net pressure gradients. Thirty-six percent of the initial pressure drop is recovered between the valve leaflets and is independent of valve size or configuration. A variable amount of postvalvular pressure recovery is observed depending on aortic or mitral configuration. Side orifice velocities are 85 +/- 4% of the centerline velocities. Incorporation of the pressure loss coefficient in the simplified Bernoulli equation shows close agreement between centerline Doppler gradients and transvalvular gradients (r = .99, y = 1.11x-0.19). CONCLUSIONS Gradients across the St Jude valve measured by Doppler ultrasound are higher than transvalvular or net catheter gradients due to downstream pressure recovery. This is more marked for Doppler gradients based on centerline velocities than side orifice velocities and is more pronounced for valves in an aortic than a mitral configuration. Therefore, to be comparable with invasive transvalvular catheter gradients, either Doppler gradients should be calculated based on side orifice velocity measurements or the Doppler gradient calculation should include the pressure loss coefficient when based on central orifice velocities.

Normal glenoid vault anatomy and validation of a novel glenoid implant shape

Current glenoid implants are designed to be secured to the articular surface. When the articular surface is compromised, a glenoid component could be implanted if it obtained fixation from the endosteal surface of the glenoid vault. The first step for designing such a glenoid implant is to define the normal three-dimensional anatomy of the glenoid vault. The purpose of this study was to define the variations in glenoid vault shape in a large group of cadaver scapula. Computed tomographic (CT) scans of 61 normal scapulae (mean, 25-34 years) from the Haman-Todd Osteological Collection, with a wide range of sizes, were examined to define the normal glenoid vault anatomy. A custom software program was used to manipulate and measure the scans to determine the morphologic variations among the different glenoid vaults. From these data, we defined a unique glenoid vault shape and empirically developed 5 sizes to represent the study population of the 61 scapulae. A second group of 11 cadaver scapulae were used to validate the shape defined using the other 61. Prototype implants were placed into the real 11 scapulae using standard surgical techniques and then CT-scanned to analyze the shape of the glenoid vault. In the 61 scapulae, 85% of the points defining the endosteal surfaces vary among scapulae by less than 2 mm. For each of the 11 cadaver scapulae, the implant size used in the virtual computer implantation was the same size used for the plastic components placed into the cadaver scapulae. Fifty percent of the measured distances between the outer dimensions of the plastic models was within 2.4 mm of the glenoid endosteal surface. Eighty percent of the surface area of the plastic models was within 3.1 mm of the glenoid endosteal surface. Five percent of the dimensions were less than 1 mm and were considered to be areas of point contact. Before designing implants that can be used in pathologic glenoids, the shape of the normal glenoid vault must first be defined. This study defined a normal glenoid vault shape that can accommodate different sized scapula with 5 sizes. This glenoid shape may be used as a template to design a glenoid implant that obtains fixation within the glenoid vault.

Cooperation of two ADAMTS metalloproteases in closure of the mouse palate identifies a requirement for versican proteolysis in regulating palatal mesenchyme proliferation.

We have identified a role for two evolutionarily related, secreted metalloproteases of the ADAMTS family, ADAMTS20 and ADAMTS9, in palatogenesis. Adamts20 mutations cause the mouse white-spotting mutant belted (bt), whereas Adamts9 is essential for survival beyond 7.5 days gestation (E7.5). Functional overlap of Adamts9 with Adamts20 was identified using Adamts9+/–;bt/bt mice, which have a fully penetrant cleft palate. Palate closure was delayed, although eventually completed, in both Adamts9+/–;bt/+ and bt/bt mice, demonstrating cooperation of these genes. Adamts20 is expressed in palatal mesenchyme, whereas Adamts9 is expressed exclusively in palate microvascular endothelium. Palatal shelves isolated from Adamts9+/–;bt/bt mice fused in culture, suggesting an intact epithelial TGFβ3 signaling pathway. Cleft palate resulted from a temporally specific delay in palatal shelf elevation and growth towards the midline. Mesenchyme of Adamts9+/–;bt/bt palatal shelves had reduced cell proliferation, a lower cell density and decreased processing of versican (VCAN), an extracellular matrix (ECM) proteoglycan and ADAMTS9/20 substrate, from E13.5 to E14.5. Vcan haploinsufficiency led to greater penetrance of cleft palate in bt mice, with a similar defect in palatal shelf extension as Adamts9+/–;bt/bt mice. Cell density was normal in bt/bt;Vcanhdf/+ mice, consistent with reduced total intact versican in ECM, but impaired proliferation persisted in palate mesenchyme, suggesting that ADAMTS-cleaved versican is required for cell proliferation. These findings support a model in which cooperative versican proteolysis by ADAMTS9 in vascular endothelium and by ADAMTS20 in palate mesenchyme drives palatal shelf sculpting and extension.