Brian D. Snyder

Bone tissue engineering using human mesenchymal stem cells: effects of scaffold material and medium flow.

We report studies of bone tissue engineering using human mesenchymal stem cells (MSCs), a protein substrate (film or scaffold; fast degrading unmodified collagen, or slowly degrading cross-linked collagen and silk), and a bioreactor (static culture, spinner flask, or perfused cartridge). MSCs were isolated from human bone marrow, characterized for the expression of cell surface markers and the ability to undergo chondrogenesis and osteogenesis in vitro, and cultured for 5 weeks. MSCs were positive for CD105/endoglin, and had a potential for chondrogenic and osteogenic differentiation. In static culture, calcium deposition was similar for MSC grown on collagen scaffolds and films. Under medium flow, MSC on collagen scaffolds deposited more calcium and had a higher alcaline phosphatase (AP) activity than MSC on collagen films. The amounts of DNA were markedly higher in constructs based on slowly degrading (modified collagen and silk) scaffolds than on fast degrading (unmodified collagen) scaffolds. In spinner flasks, medium flow around constructs resulted in the formation of bone rods within the peripheral region, that were interconnected and perpendicular to the construct surface, whereas in perfused constructs, individual bone rods oriented in the direction of fluid flow formed throughout the construct volume. These results suggest that osteogenesis in cultured MSC can be modulated by scaffold properties and flow environment.

Clinical trial of a farnesyltransferase inhibitor in children with Hutchinson–Gilford progeria syndrome

Hutchinson–Gilford progeria syndrome (HGPS) is an extremely rare, fatal, segmental premature aging syndrome caused by a mutation in LMNA that produces the farnesylated aberrant lamin A protein, progerin. This multisystem disorder causes failure to thrive and accelerated atherosclerosis leading to early death. Farnesyltransferase inhibitors have ameliorated disease phenotypes in preclinical studies. Twenty-five patients with HGPS received the farnesyltransferase inhibitor lonafarnib for a minimum of 2 y. Primary outcome success was predefined as a 50% increase over pretherapy in estimated annual rate of weight gain, or change from pretherapy weight loss to statistically significant on-study weight gain. Nine patients experienced a ≥50% increase, six experienced a ≥50% decrease, and 10 remained stable with respect to rate of weight gain. Secondary outcomes included decreases in arterial pulse wave velocity and carotid artery echodensity and increases in skeletal rigidity and sensorineural hearing within patient subgroups. All patients improved in one or more of these outcomes. Results from this clinical treatment trial for children with HGPS provide preliminary evidence that lonafarnib may improve vascular stiffness, bone structure, and audiological status.

Role of trabecular morphology in the etiology of age-related vertebral fractures

SummaryOsteoporotic compression fractures of the spine differ from most other age-related fractures in that they usually are associated with minimal trauma and with loads no greater than those encountered during normal activities of daily living. With aging and osteoporosis, there is progressive resorption of bone, resulting in reductions in bone density, thinning of trabeculae, and loss of trabecular contiguity. These changes in trabecular bone structure are associated with losses in bone strength which are disproportionate to the reductions in bone mass alone. To explain this disproportionate loss of bone strength, the prevailing opinion is that density reductions in the vertebral centrum are accompanied by a reduction in the number of trabeculae, by preferential resorption of horizontal trabeculae, and by hypertrophy of the remaining vertical trabeculae. To evaluate this view of vertebral morphology, we performed three-dimensional stereological analysis of trabecular bone extracted from midsagittal sections of first lumbar vertebral bodies from 12 donors spanning an age of 27–81 years. We found that both the number (R2 = 0.63,P < 0.01) and thickness (R2 = 0.91,P < 0.01) of trabeculae decreased linearly with density (as expressed by bone volume fraction) whereas the spacing between the trabeculae (R2 = 0.61,P < 0.01) increased reciprocally. There were more vertical trabeculae with transverse trabeculae at all densities, and the number of vertical trabeculae changed with density at twice the rate of the number of transverse trabeculae (P < 0.001). These data do not support the prevailing view that there is preferential resorption of horizontal trabeculae or hypertrophy of the remaining vertical trabeculae. Bone density was also a strong (R2 = 0.90,P < 0.01) power law function of the ratio of trabecular thickness to mean intertrabecular spacing. From buckling theory, the critical buckling load of a trabecula is related to this ratio of trabecular thickness to effective length. The changes in trabecular morphology observed with decreasing bone density thus pose a “triple threat” to the strength and stability of vertebral trabecular bone, as not only are there fewer trabeculae, but the remaining trabeculae are both thinner and longer.

Lateral entry compared with medial and lateral entry pin fixation for completely displaced supracondylar humeral fractures in children. A randomized clinical trial.

BACKGROUND Closed reduction and percutaneous pin fixation is the treatment of choice for completely displaced (type-III) extension supracondylar fractures of the humerus in children, although controversy persists regarding the optimal pin-fixation technique. The purpose of this study was to compare the efficacy of lateral entry pin fixation with that of medial and lateral entry pin fixation for the operative treatment of completely displaced extension supracondylar fractures of the humerus in children. METHODS This prospective, randomized clinical trial had sufficient power to detect a 10% difference in the rate of loss of reduction between the two groups. The techniques of lateral entry and medial and lateral entry pin fixation were standardized in terms of the pin location, the pin size, the incision and position of the elbow used for medial pin placement, and the postoperative course. The primary study end points were a major loss of reduction and iatrogenic ulnar nerve injury. Secondary study end points included radiographic measurements, clinical alignment, Flynn grade, elbow range of motion, function, and complications. RESULTS The lateral entry group (twenty-eight patients) and the medial and lateral entry group (twenty-four patients) were similar in terms of mean age, sex distribution, and preoperative displacement, comminution, and associated neurovascular status. No patient in either group had a major loss of reduction. There was no significant difference between the rates of mild loss of reduction, which occurred in six of the twenty-eight patients treated with lateral entry and one of the twenty-four treated with medial and lateral entry (p = 0.107). There were no cases of iatrogenic ulnar nerve injury in either group. There were also no significant differences (p > 0.05) between groups with respect to the Baumann angle, change in the Baumann angle, humerocapitellar angle, change in the humerocapitellar angle, Flynn grade, carrying angle, elbow flexion, elbow extension, total elbow range of motion, return to function, or complications. CONCLUSIONS With use of the specific techniques employed in this study, both lateral entry pin fixation and medial and lateral entry pin fixation are effective in the treatment of completely displaced (type-III) extension supracondylar fractures of the humerus in children. LEVEL OF EVIDENCE Therapeutic Level I. See Instructions to Authors for a complete description of levels of evidence.

Multicenter study of patients with angiotensin- converting enzyme inhibitor-induced angioedema who present to the emergency department

BACKGROUND Recent data are lacking about the number of patients with angiotensin-converting enzyme inhibitor (ACEI)-induced angioedema who present to the emergency department (ED). Current management of the condition and clinical outcomes also are not known. OBJECTIVE To describe the clinical epidemiology of ACEI-induced angioedema in patients who present to the ED. METHODS We performed a medical record review of ACEI-induced angioedema in patients who presented to 5 EDs in the Emergency Medicine Network. A structured data abstraction form was used to collect each patients demographic factors, medical history, and details about the angioedema that prompted the ED visit. The medical record review also focused on treatment provided in the ED and subsequent need for hospitalization. RESULTS We identified a total of 220 patients with ACEI-induced angioedema. The frequency of ACEI-induced angioedema among all patients with angioedema who presented to the ED was 30% (95% confidence interval, 26%-34%). The annual rate of visits for ACEI-induced angioedema was 0.7 per 10,000 ED visits. The most frequent presenting signs were shortness of breath, lip and tongue swelling, and laryngeal edema. Most patients (58%) were sent home directly from the ED, whereas 12% were regular inpatient admissions, 11% were admitted to the intensive care unit, and 18% were admitted under observation status (<24 hours). Pharyngeal swelling and respiratory distress were independent predictors of hospital admission and longer length of stay. CONCLUSION ACEI-induced angioedema accounted for almost one-third of angioedema treated in the ED, although it remains a rare ED presentation. A subgroup of these patients still needs inpatient hospitalization for management of upper airway angioedema.

NSAID Exposure and Risk of Nonunion: A Meta-Analysis of Case–Control and Cohort Studies

Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used for postoperative pain control. However, concerns regarding the potential deleterious effects of NSAIDs on bone healing have compelled many physicians to avoid NSAIDs in patients with healing fractures, osteotomies, and fusions. We systematically reviewed and analyzed the best clinical evidence regarding the effects of NSAID exposure on bone healing. Medline, Embase, and Cochrane electronic databases were searched for prospective and retrospective clinical studies of fracture, osteotomy, and fusion studies of patients with NSAID exposure and nonunion as an outcome. Study quality was assessed using the Newcastle–Ottawa Scale. Data on study design, patient characteristics, and risk estimates were extracted. Pooled effect estimates were calculated. Subanalyses were performed by bone type and by NSAID dose, duration, and route of administration. In the initial analysis of 11 cohort and case-control studies, the pooled odds ratio for nonunion with NSAID exposure was 3.0 (95% confidence interval 1.6–5.6). A significant association between lower-quality studies and higher reported odds ratios for nonunion was observed. When only higher-quality studies were considered, seven spine fusion studies were analyzed, and no statistically significant association between NSAID exposure and nonunion was identified (odds ratio = 2.2, 95% confidence interval 0.8–6.3). There was no increased risk of nonunion with NSAID exposure when only the highest-quality studies were assessed. Randomized controlled trials assessing NSAID exposure in fracture, fusion, and osteotomy populations are warranted to confirm or refute the findings of this meta-analysis of observational studies.

Directed assembly of PEGylated-peptide coatings for infection-resistant titanium metal.

Appropriate surface chemistry between a material and its surrounding biological environment is crucial to the eventual integration and performance of any implant, whether metal, plastic, or ceramic. A robust peptide-based coating technology capable of easily modifying the surface of titanium (Ti) metal through noncovalent binding is described. A short peptide possessing affinity for Ti was identified using a phage display screening process and subjected to an amino acid substitution exercise using solid-phase chemical synthesis. Through these studies, the HKH tripeptide motif was elucidated as an important contributor to Ti binding within the Ti-binding peptide. This peptide spontaneously and selectively adsorbs onto a Ti surface from dilute aqueous solution with submicromolar binding affinities as determined by ELISA and quartz crystal microbalance with dissipation monitoring (QCM-D), through a process largely dominated by electrostatic interactions. Atomic force microscopy (AFM) reveals a densely packed peptide adlayer with an average height of approximately 0.5 nm. Subsequently, a PEGylated analogue of the peptide was shown to rapidly coat Ti to afford a nonfouling surface that efficiently blocked the adsorption of fibronectin and significantly reduced the extent of Staphylococcus aureus attachment and biofilm formation in vitro. These PEGylated-peptide coatings show promise in terms of resolving two major hurdles common to implanted metals: (i) nonspecific protein adsorption and (ii) bacterial colonization. At the same time, the facile one-step modification process will facilitate the point-of-care application of these coatings in the surgical suite.

Quantitative micro-computed tomography: A non-invasive method to assess equivalent bone mineral density

One of the many applications of micro computed tomography (microCT) is to accurately visualize and quantify cancellous bone microstructure. However, microCT based assessment of bone mineral density has yet to be thoroughly investigated. Specifically, the effects of varying imaging parameters, such as tube voltage (kVp), current (microA), integration time (ms), object to X-ray source distance (mm), projection number, detector array size and imaging media (surrounding the specimen), on the relationship between equivalent tissue density (rhoEQ) and its linear attenuation coefficient (micro) have received little attention. In this study, in house manufactured, hydrogen dipotassium phosphate liquid calibration phantoms (K2HPO4) were employed in addition to a resin embedded hydroxyapatite solid calibration phantoms supplied by Scanco Medical AG Company. Variations in current, integration time and projection number had no effect on the conversion relationship between micro and rhoEQ for the K2HPO4 and Scanco calibration phantoms [p>0.05 for all cases]. However, as expected, variations in scanning tube voltage, object to X-ray source distance, detector array size and imaging media (referring to the solution that surrounds the specimen in the imaging vial) significantly affected the conversion relationship between mu and rhoEQ for K2HPO4 and Scanco calibration phantoms [p<0.05 for all cases]. A multivariate linear regression approach was used to estimate rhoEQ based on attenuation coefficient, tube voltage, object to X-ray source distance, detector array size and imaging media for K2HPO4 liquid calibration phantoms, explaining 90% of the variation in rhoEQ. Furthermore, equivalent density values of bovine cortical bone (converted from attenuation coefficient to equivalent density using the K2HPO4 liquid calibration phantoms) samples highly correlated [R2=0.92] with the ash densities of the samples. In conclusion, Scanco calibration phantoms can be used to assess equivalent bone mineral density; however, they cannot be scanned with a specimen or submerged in a different imaging media. The K2HPO4 liquid calibration phantoms provide a cost effective, easy to prepare and convenient means to perform quantitative microCT analysis using any microCT system, with the ability to choose different imaging media according to study needs. However, as with any liquid calibration phantom, they are susceptible to degradation over time.

Relationships among musculoskeletal impairments and functional health status in ambulatory cerebral palsy.

Orthopedic surgery for patients with cerebral palsy addresses motion impairments, assuming that this will improve motor function. This study evaluates the relationships among clinical impairment measures with standardized assessments of function and disability as an initial step in testing this assumption. A total of 129 ambulatory children and adolescents across six institutions participated in a prospective evaluation that consisted of passive motion and spasticity examination of the lower extremities, three-dimensional gait temporal-spatial and kinematic analysis, and administration of the Gross Motor Function Measure (GMFM) and the Pediatric Outcomes Data Collection Instrument (PODCI). The analysis found that isolated impairment measures of motion and spasticity were only weakly related to motor function in cerebral palsy and even when averaged across multiple joints yielded no more than a fair correlation with functional scores, nor did a combination of impairments emerge that could predict substantial variance in motor function. These findings suggest that caution should be exercised when anticipating functional change through the treatment of isolated impairment and that addressing multiple impairments may be needed to produce appreciable effects.

Predicting Fracture Through Benign Skeletal Lesions with Quantitative Computed Tomography

BACKGROUND There are no proven radiographic guidelines for predicting fracture risk in children and young adults with a benign skeletal lesion. An in vivo diagnostic study was conducted to determine whether a reduction in the load-carrying capacity of a bone measured with quantitative computed tomography was more accurate than current radiographic guidelines for predicting pathologic fracture in patients with a benign skeletal lesion. METHODS Eighteen patients who presented with a fracture through a benign skeletal lesion were compared with eighteen patients who had a benign skeletal lesion that had been thought to be at increased risk for fracture on the basis of currently used radiographic criteria but had not fractured over a two-year period. Structural analysis was performed to calculate the resistance of the affected bones to compressive, bending, and torsional loads with use of serial transaxial quantitative computed tomography data obtained along the length of the bone containing the lesion and from homologous cross sections through the contralateral, normal bone. At each cross section, the ratio of the structural rigidity of the affected bone divided by that of the normal, contralateral bone was determined. The cross section with the greatest reduction in compressive, bending, and torsional rigidity was identified as that most likely to fracture. RESULTS The mean age (and standard deviation) of the thirty-six patients was 12.5 +/- 3.6 years. Twenty lesions were located in the femur; eleven, in the tibia; three, in the humerus; one, in the ulna; and one, in the pelvis. A combination of the minimum bending and torsional rigidities calculated from the tomographic data provided optimal performance in differentiating between the fracture and non-fracture groups (100% sensitivity and 94% specificity). In contrast, plain radiographic criteria demonstrated 28% to 83% sensitivity and 6% to 78% specificity. CONCLUSIONS The combination of bending and torsional rigidity measured noninvasively with quantitative computed tomography was more accurate (97%) for predicting pathologic fracture through benign bone lesions in children than were standard radiographic criteria (42% to 61% accuracy). We believe that this method can provide accurate objective criteria for planning treatment of benign bone lesions and monitoring treatment response.