Faculty Bibliography

The bone organ is poorly represented in comparative research on mammalian mass-specific metabolic rates. As a first order attempt to remedy this, from the literature we collected mass-specific metabolic rates for all major organs except for the bone organ, and by subtraction infer the rate for the bone organ. The scaling relationships are given of each whole-organ mass-specific metabolic rate and of the relationship between whole-organ metabolic rate and body mass. Scaling of the lung, adipose depot and bone organ with body mass is higher than would be expected by ¾ power scaling. We interpret the similar scalings of bone and the adipose depot in light of their evolved regulation of whole-body metabolism. We also briefly examine the supra-¾ power scaling of the lung as well as the independence of the mass-specific metabolic rate of the heart from body mass. The bone organ exhibits relatively high energy expenditure with increasing body size. The bone marrow and its medullary adipocyte store may be responsible for engendering the greater share of the bone organ's energetic cost.

This study aimed to investigate the ultrastructural anatomy of the developing ACL tibial enthesis. We hypothesized that enthesis architecture would progressively mature and remodel, eventually resembling that of the adult by the early postnatal stage. Five fresh-frozen human pediatric cadaveric knees aged 1-36 months underwent anatomical dissection to harvest the ACL insertion and underlying tibial chondroepiphysis. The samples were prepared for scanning electron microscopy (SEM) to examine the ultrastructural anatomy of the enthesis and underwent histological staining for circular polarized light (CPL) and light microscopy imaging. SEM analysis of the 1- and 8-month-old samples revealed a shallow interdigitation between the dense fibrous (ligamentous) tissue and unmineralized chondrogenic tissues, with a minimal transition zone. By 11-month, a more complex transition zone was present. By age 19- and 36-month-old, a progressively more complex and defined fibrocartilage zone was observed. CPL analysis revealed distinct collagen fiber continuity, alignment, and organization changes over time. By 19 and 36 months, the samples exhibited complex fiber arrangements and a progression toward uniform fiber orientation. Similarly, histological analysis demonstrated progressive remodeling of the enthesis with increasing age. Our results suggest that the ACL enthesis of the developing knee begins to mimic that of an adult as early as 19 months of age, as a more complex transition between ligamentous and chondro-epiphyseal tissue can be appreciated. We hypothesize that the observed changes are likely due to mechanical loading of the enthesis with the onset of weightbearing. Future investigations of ACL reconstruction and repair will benefit from improved understanding of the chondro-epiphyseal/ACL regions.

OBJECTIVES/OBJECTIVE:Age at death estimation is a key element to many research questions in biological anthropology, archeology, and forensic science. Dental cementum is a tissue of choice for the estimation of age at death in adult individuals as it continues deposition for the entirety of an individual's life. Previous works have devised regression formulas correlating cementum thickness to age at death. However, interpopulation variances are unknown, and it is therefore not clear whether regressions based on a single population are applicable to individuals with different ancestries. MATERIALS AND METHODS/METHODS:Here, we use a sample (n = 52) of teeth from individuals with known age at tooth extraction/death of European, African, and East Asian ancestry to assess whether there are interpopulations differences in cementum growth rate. We measured growth rate in four different areas (2nd and 5th decile of both the lingual and buccal aspect of the root) of each tooth and used nonparametric tests to evaluate population differences in growth rate between homologous regions of the teeth. RESULTS:The results of the analyses show that, even after controlling for tooth size, individuals of European ancestry have significantly lower growth rates than those of both African and East Asian ancestry across all four tooth areas. DISCUSSION/CONCLUSIONS:These results call into question the applicability of the regression formulas derived from European ancestry individuals to individuals of other ancestries.

Dental caries are treated by the surgical removal of infected tissue where the biological mineral, hydroxyapatite, has been eroded. For early carious lesions, surgical methods have increasingly been replaced by minimally invasive dentistry protocols to arrest the lesion progression by controlling plaque formation and promoting dentin remineralization. Zinc phosphate mineral deposition in dentinal tubules was studied as a modality for the treatment of dental caries. Extracted permanent human molars, with and without carious lesions, were employed to study the coverage and depth of mineral deposition with in situ mixing of zinc and phosphate salt solutions. Milled hydroxyapatite was employed as a surrogate for dentin in the study of mineral formation in tubules. The mineral composition was identified by X-ray powder diffraction. Scanning electron microscopy and energy-dispersive X-ray spectroscopy revealed the deposition of zinc phosphate minerals that effectively occlude dentinal tubules by crystallization within dentinal tubules. Mineral deposition was similarly observed at the site of a carious lesion, which highlights the feasibility of zinc phosphate deposition for the treatment of dental caries.

This ex-vivo study investigated the effect of a light-emitting diode (LED) curing light on the depth of penetration of Silver Diamine Fluoride (SDF) into carious lesions. Twenty-four primary teeth with untreated caries lesions were allocated into groups and treated within 5 min after extraction: (1) n = 6 treated for 1 min with one drop of SDF followed by 10 sec rinse with tap water, (2) n = 6 treated for 10 sec with one drop of SDF and exposed to LED light for 20 sec (30 sec total SDF exposure) followed by 10 sec rinse with tap water, (3) n = 6 treated for 10 sec with one drop of SDF followed by a 10 sec rinse with tap water, (4) n = 3 untreated, and (5) n = 3 untreated but exposed to LED light for 20 sec. Samples were prepared, embedded, sectioned and silver penetration was measured using backscattered electron imaging in the scanning electron microscope and energy-dispersive X-ray spectroscopy analysis. Results were expressed as the average relative depth of penetration (%) = Ag depth/lesion depth × 100 from 5 sites in each lesion. Group means were compared using mixed model analysis. Mean ± standard deviation (SD) penetration was: 86.4 ± 20.7% in Group 1, 94.3 ± 13.7% in Group 2, and 26.7 ± 13.9% in Group 3. Groups 1 and 2 were statistically similar and different from Group 3 (p < 0.001). Groups 4 and 5 had no silver present. Use of LED light for 20 sec after 10 sec SDF application appears to facilitate silver penetration, similar to a 1 min SDF application. Clinical studies are needed to define the role of silver penetration in sustained caries arrest.

Sodium glucose cotransporter-2 inhibitors (SGLT2is) promote urinary glucose excretion and decrease plasma glucose levels independent of insulin. Canagliflozin (CANA) is an SGLT2i, which is widely prescribed, to reduce cardiovascular complications, and as a second-line therapy after metformin in the treatment of type 2 diabetes mellitus. Despite the robust metabolic benefits, reductions in bone mineral density (BMD) and cortical fractures were reported for CANA-treated subjects. In collaboration with the National Institute on Aging (NIA)-sponsored Interventions Testing Program (ITP), we tested skeletal integrity of UM-HET3 mice fed control (137 mice) or CANA-containing diet (180 ppm, 156 mice) from 7 to 22 months of age. Micro-computed tomography (micro-CT) revealed that CANA treatment caused significant thinning of the femur mid-diaphyseal cortex in both male and female mice, did not affect trabecular bone architecture in the distal femur or the lumbar vertebra-5 in male mice, but was associated with thinning of the trabeculae at the distal femur in CANA-treated female mice. In male mice, CANA treatment is associated with significant reductions in cortical bone volumetric BMD by micro-CT, and by quantitative backscattered scanning electron microscopy. Raman microspectroscopy, taken at the femur mid-diaphyseal posterior cortex, showed significant reductions in the mineral/matrix ratio and an increased carbonate/phosphate ratio in CANA-treated male mice. These data were supported by thermogravimetric assay (TGA) showing significantly decreased mineral and increased carbonate content in CANA-treated male mice. Finally, the sintered remains of TGA were subjected to X-ray diffraction and showed significantly higher fraction of whitlockite, a calcium orthophosphate mineral, which has higher resorbability than hydroxyapatite. Overall, long-term CANA treatment compromised bone morphology and mineral composition of bones, which likely contribute to increased fracture risk seen with this drug.

In the Iron Age IIA Philistine cemetery at Ashkelon (modern Israel), roughly 11% of individuals exhibit severe and highly variable dental wear, which we explore here at two timescales: wear that accumulates over days and weeks (microwear) and wear that accumulates over months and years (macrowear). Using teeth from both adult and nonadult individuals, we first established categories of dental macrowear patterns and sorted individuals within them. We then made replicas of the teeth from 27 individuals having both typical and atypical dental wear and performed metrology by noncontact profilometry on a reflected light microscope. We then calculated each tooth's surface roughness (Sa) and collected qualitative observations of teeth within each macrowear category. Our findings show no macrowear or microwear pattern exclusive to sex or age group. Likewise, there are no statistically significant differences (p < 0.05) in Sa between adult males and females, and sampled nonadults fall within two standard deviations of the pooled adult means. The microscopic surfaces of all teeth show a wide variety of textures on the occlusal surfaces, including wavy striations, deep parallel striations, globular pitting, and newly described rectangular pits. These results indicate that individuals used their teeth as a third hand while manipulating objects and that children also participated in these activities. Due to the similarities in dental wear between Ashkelon and other coastal populations, we conclude that the observed wear patterns arose from the performance of specialized tasks for a marine-based economy.

Overcoming the non-specificity of histological accentuated growth lines in hard tissues is an ongoing challenge. Identifying season at death and reproductive events has profound implications for evolutionary, ecological and conservation studies. Dental cementum is a mineralized tissue with yearly periodicity that continues deposition from tooth formation until death, maintaining a record spanning almost the entire life of an individual. Recent work has successfully employed elemental analysis of calcified incremental tissues to detect changes in extrinsic conditions such as diet and climate and to identify two important life-history milestones: weaning and sexual maturity. Here, we employ field-emission scanning electron microscopy and energy-dispersive X-ray analysis to measure the relative concentrations of calcium, phosphorous, oxygen, magnesium and sodium in the cementum of 34 teeth from seven male and female rhesus macaques with known medical and life-history information. We find that changes in relative magnesium concentrations correspond with reproductive events in females and breastfeeding in infants. Additionally, we observe seasonal calcium patterns in 77.3% of the samples.

BACKGROUND:While autografts to date remain the "gold standard" for bone void fillers, synthetic bone grafts have garnered attention due to their advantages such as ability to be tailored in terms of its physical and chemical properties. Bioactive glass (BG), an inorganic material, has the capacity to form a strong bond with bone by forming a bone-like apatite surface, enhancing osteogenesis. Coupled with three-dimensional printing it is possible to maximize bone regenerative properties of the BG. OBJECTIVE:The objective of this study was to synthesize and characterize 3D printed mesoporous bioactive glass (MBG), BG 45S5, and compare to β-Tricalcium phosphate (β-TCP) based scaffolds; test cell viability and osteogenic differentiation on human osteoprogenitor cells in vitro. METHODS:MBG, BG 45S5, and β-TCP were fabricated into colloidal gel suspensions, tested with a rheometer, and manufactured into scaffolds using a 3D direct-write micro-printer. The materials were characterized in terms of microstructure and composition with Thermogravimetric Analyzer/Differential Scanning Calorimeter (TGA/DSC), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Micro-Computed Tomography (μ-CT), Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Spectroscopy (EDS), and Mattauch-Herzog-Inductively Coupled Plasma-Mass Spectrometry (MH-ICP-MS). RESULTS:Scaffolds were tested for cell proliferation and osteogenic differentiation using human osteoprogenitor cells. Osteogenic media was used for differentiation, and immunocytochemistry for osteogenic markers Runx-2, Collagen-I, and Osteocalcin. The cell viability results after 7 days of culture yielded significantly higher (p < 0.05) results in β-TCP scaffolds compared to BG 45S5 and MBG groups. CONCLUSION/CONCLUSIONS:All materials expressed osteogenic markers after 21 days of culture in expansion and osteogenic media.