Faculty Bibliography

The calvaria (top part of the skull) is made of pieces of bone as well as multiple soft tissue joints called sutures. The latter is crucial to the growth and morphogenesis of the skull, and thus a loss of calvarial sutures can lead to severe congenital defects in humans. During embryogenesis, the calvaria develops from the cranial mesenchyme covering the brain, which contains cells originating from the neural crest and the mesoderm. While the mechanism that patterns the cranial mesenchyme into bone and sutures is not well understood, function of Lmx1b, a gene encoding a LIM-domain homeodomain transcription factor, plays a key role in this process. In the current study, we investigated a difference in the function of Lmx1b in different parts of the calvaria using neural crest-specific and mesoderm-specific Lmx1b mutants. We found that Lmx1b was obligatory for development of the interfrontal suture and the anterior fontanel along the dorsal midline of the skull, but not for the posterior fontanel over the midbrain. Also, Lmx1b mutation in the neural crest-derived mesenchyme, but not the mesoderm-derived mesenchyme, had a non-cell autonomous effect on coronal suture development. Furthermore, overexpression of Lmx1b in the neural crest lineage had different effects on the position of the coronal suture on the apical part and the basal part. Other unexpected phenotypes of Lmx1b mutants led to an additional finding that the coronal suture and the sagittal suture are of dual embryonic origin. Together, our data reveal a remarkable level of regional specificity in regulation of calvarial development.

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.

BACKGROUND:The bulk metallic glass (BMG), Pd79Ag3.5P6Si9.5Ge2, has a high fracture toughness and has been found to accommodate post-yield stress, unlike most other BMG. Moreover, due to its greater noble gas composition it has a intrinsic corrosion resistance, ideal for dental and orthopedic implants. OBJECTIVE:This present study aimed to evaluate the in vivo application of Pd79Ag3.5P6Si9.5Ge2 in a large translational sheep model to assess its efficacy to be utilized as an endosteal device. METHODS:Twelve implants in the form of cylindrical rods (3 mm in diameter) were produced through rapid quenching. Each sheep (n = 12) received one osteotomy in the mandibular region using rotary instrumentation, which was subsequently filled with Pd79Ag3.5P6Si9.5Ge2. After 6- and 24-weeks the animals were euthanized, and samples collected en bloc to conduct histomorphometric analysis. The level/degrees of osseointegration were assessed through bone-to-implant contact (BIC). RESULTS:Favorable BIC was observed with fibrous connective tissue layers at both 6- and 24-weeks. Bone along with interfacial remodeling was observed in proximity with the metallic glass surface at 6 weeks with higher degrees of bone organization being observed at the later healing time, 24 weeks. CONCLUSIONS:The introduced BMG revealed potential to serve as an alternative biomaterial to commonly used Ti alloys given its unique combination of toughness and strength.

The purpose of this study was to evaluate the influence of implant design features on osseointegration parameters. Two different implant macrogeometries and surface treatments were evaluated as follows: (1) progressive buttress threads possessing the SLActive surface (SLactive/BL), and (2) inner and outer trapezoidal threads possessing nano-hydroxyapatite coating over a dual acid-etched surface (Nano/U). Implants were placed in the right ilium of 12 sheep, and histologic/metric analyses were conducted after 12 weeks in vivo. The percentage of bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO) within the threads were quantified. Histologic observations showed more intimate BIC in the SLactive/BL group compared to the Nano/U group. In contrast, the Nano/U group depicted woven bone formation generated between the wall of the osteotomy and implant threads within the healing chambers, while bone remodeling was evident at the tip of the outer thread. The SLActive/BL group presented higher BIC than the Nano/U group. On the other hand, significantly higher BAFO was observed at 12 weeks in the Nano/U group compared to the SLactive/BL group (P < .042). Differences in implant design features influenced the osseointegration pathway, which supports the need for further investigations to describe the clinical performance and differences in a timely fashion.

The aim of this study was to clinically evaluate the guided bone regeneration (GBR) potential of allograft, xenograft, and alloplastic materials in combination with resorbable membranes in extraction sockets. The qualitative and quantitative assessments of this prospective study were accomplished through histologic and histomorphometric analysis. Three experimental groups and 1 control group for comparison (n = 8) received either an allograft (human cancellous bone, freeze dried, Deutsches Institut für Zell und Gewebeersatz, Berlin, Germany), xenograft (BioOss, Geistlich Pharma AG, Wolhusen, Switzerland), or alloplast (biphasic calcium sulphate, Bondbone, MIS Implants Technologies Ltd., Charlotte, NC). The negative control group received no regenerative material. Tissue samples were then qualitatively and quantitatively evaluated as a function of percentage of new vital bone, graft particles content, soft tissue, and bone marrow over time. All 3 study groups presented bone volume suitable for the successful placement of a dental implant. The xenograft group yielded significantly less amount of vital bone compared with the allograft and alloplast groups. When comparing the percentage of residual graft particles, there was significantly greater amounts associated with the xenograft group in contrast to the allograft and alloplast groups. Similarly, a significantly increased amount of soft tissue percentage was observed within the xenograft group relative to all other groups. No significant differences were observed in the percentage of residual graft particles between the allograft and alloplast groups. There were also no significant differences detected in vital bone percentage between the allograft, alloplast, and control groups. When evaluating the bone marrow percentage, the only significant difference detected was between the xenograft and alloplast materials. Overall, no complications (ie, fever, malaise, purulence or fistula) were observed during the entirety of clinical trial among all patients. The greatest GBR potential was associated with the allograft material because of the greater degree of vital bone and the lowest percentage of residual graft particles. All studied bone substitute materials resulted in bone apposition for efficient use in alveolar ridge preservation procedures.

The aim of this study was to evaluate the in vivo performance of two different deproteinized bovine bone (DBB) grafting materials: DBBB (Bio-Oss®) and DBBL (Laddec®), for the regeneration of critically sized (8 mm) defects in rabbit's calvaria. Three round-shaped defects were surgically created in the calvaria of 13 New Zealand White rabbits proximal to the coronal suture in the parietal bone. Two of the defects were filled with one of the grafting materials while a third was left empty to serve as a negative control. Bone regeneration properties were evaluated at 4- and 8-weeks after implantation by means of histological and histomorphometrical analyses. Statistical analyses were performed through a mixed model analysis with fixed factors of time and material. Histological evaluation of the control group evidenced a lack of bridging bone formation across the defect sites at both evaluation time points. For the experimental groups, new bone formation was observed around the defect periphery and to progress radially inwards to the center of the defect site, regardless of the grafting material. Histomorphometric analyses at 4 weeks demonstrated higher amount of bone formation through the defect for DBBB group. However, at 8 weeks, DBBL and DBBB demonstrated osteoconductivity and low resorption rates with evidence of statistically similar bone regeneration through the complete boney defect. Finally, DBBB presented lower soft tissue migration within the defect when compared to DBBL at both evaluation time points. DBBB and DBBL presented similar bone regeneration performance and slow resorption rates. Although both materials promoted bone regeneration through the complete defect, DBBB presented lower soft tissue migration within the defects at 4- and 8-weeks.

Leukocyte-platelet-rich fibrin (L-PRF) has been suggested for gap management for immediate implant placement when the distance is greater than 2 mm. However, there remains a paucity in hierarchically designed research to support this application. The present study aimed to evaluate the effect of L-PRF on the osseointegration parameters of dental implants placed after conventional osteotomy of surgically created bone defects that simulate post extraction sockets in a canine model after 3, 6, and 12 weeks in vivo. Eighty dental implants (Intra-Lock, Boca Raton, FL) were placed in the radius of 13 beagle dogs. The experiment consisted of 4 groups (n = 20 implants/group): 1) Regular osteotomy (Reg n/L-PRF); 2) Regular osteotomy and implant placement with L-PRF membrane (Reg L-PRF); 3) Wide osteotomy with no gap management performed, where an osteotomy/bony defect (6 mm of diameter and ~5 mm deep) was created to simulate immediate implant placement in post-extraction sockets, and the gap was left for spontaneous healing (Wide nL-PRF); and 4) Wide osteotomy with L-PRF gap management (Wide L-PRF). L-PRF membranes were obtained by blood drawn from each subject and centrifuged at 2700 rpm (408 RCF-clot) for 12 min. In the experimental groups where L-PRF was utilized, the membrane was inserted into the osteotomy site prior to implant placement. Six dogs had implants placed in the radius for 3 weeks; and 7 dogs had implants placed in the left radius for 6 weeks and in the right radius for 12 weeks. At the corresponding experimental time points, samples were harvested, and subjected to histological processing for qualitative and quantitative analyses, via bone-to-implant contact (BIC) and bone-area-fraction occupancy (BAFO). Qualitative analysis demonstrated increased amounts of bone formation around the implant and within the healing chambers over time for all groups. While comparable histological features were observed for both Reg groups (L-PRF and nL-PRF), the gap management performed in Wide L-PRF group resulted in effective gap filling with improved bone growth in close proximity to the implant surface. Quantitative analyses of BIC and BAFO yielded higher values for both variables at 3 weeks for Wide L-PRF (~38% and ~56% respectively) compared to Wide nL-PRF (~20% for BIC and BAFO) (p < .03). No statistical differences were detected between Wide groups at 6 and 12 weeks, neither between Reg groups, independent of the association with or without the L-PRF membrane at all healing times. L-PRF placed within wide osteotomies, prior to implant placement, resulted in increased early bone formation compared to unfilled wide osteotomies at the early healing time (3 weeks in vivo).

To evaluate the effect of fatigue and aging on the crystalline content and reliability of a zirconia-toughened-alumina (ZTA) composite compared to its individual counterpart materials (3Y-TZP and Al₂O₃). Thirty-six disc-shaped specimens per group were obtained to comply with ISO 6872:2015. Crystalline content, microstructure and reliability of experimental groups were evaluated in four stages: 1) immediate; 2) aged; 3) fatigued; 4) aged + fatigue. Aging was performed in autoclave and Step-Stress-Accelerated-Life-Testing (SSALT) was performed using three stress profiles. Weibull statistics were used to determine Weibull parameters and life-expectancy. A significant increase in monoclinic phase in 3Y-TZP was observed after aging (19.31%), fatigue (17.88%) and aging + fatigue (55.81%), while ZTA evidenced minimal variation among all conditions (<5.69%). 3Y-TZP presented higher reliability than ZTA at 300 and 500 MPa, and ZTA outperformed Al₂O₃ at the same stress missions. None of the ceramics yielded acceptable reliability at 800 MPa. A higher characteristic strength was observed for 3Y-TZP, followed by ZTA and Al₂O₃. While after aging ZTA and Al₂O₃ remained stable, 3Y-TZP exhibited a significant increase in the characteristic stress. Aging did not affect the reliability of ZTA and Al₂O₃. 3Y-TZP demonstrated an increase in monoclinic content and characteristic strength after aging.

PURPOSE/OBJECTIVE:This study evaluated the probability of survival and failure mode of endodontically treated incisors without ferrule restored with CAD/CAM FRC post-cores. METHODS:Root canals of bovine incisors were treated, leaving post preparations of ∼10 mm. Teeth were allocated into three groups: (i) cast metal post-core, (ii) FRC prefabricated post with a direct resin core build-up, and (iii) CAD/CAM FRC post-core. Posts and zirconia crowns were cemented using resin cement. Specimens were subjected to step-stress accelerated-life fatigue testing in water. Use level probability Weibull curves, probability of survival for a mission of 100,000 cycles at 25, 50, and 100 N, Weibull modulus, and characteristic strength were calculated and plotted. Failure mode was examined under a stereomicroscope. RESULTS:Restored incisors demonstrated high probability of survival (93-100%) for missions estimated at 25 and 50 N, irrespective of post-core foundation. At 100 N, incisors restored with metal posts presented significantly higher probability of survival (99%) relative to CAD/CAM posts (79%), whereas FRC groups demonstrated no significant difference. Weibull analysis indicated no significant difference on the Weibull modulus (m = 3.38-5.92). Incisors reconstructed with metal post-cores (431 N) presented significantly higher characteristic strength relative to prefabricated (200 N) and CAD/CAM (202 N) FRC post-cores. While post fracture was the chief failure mode for prefabricated and CAD/CAM FRC post-cores, post and/or root fracture were the main event for metal post-cores. CONCLUSION/CONCLUSIONS:Endodontically treated incisors without ferrule restored with CAD/CAM FRC post-cores presented promising probability of survival for loads compatible with anterior masticatory forces and favorable failure modes.

Fused Filament Fabrication (FFF)-based 3D printing is an efficient technique for developing medical implants, but it is not very useful in developing small yet mechanically robust design-specific fixtures such as dental implants (<15 mm). Specifically, it is challenging to 3D print robust Polyetheretherketone (PEEK) small implants due to PEEK's high melting temperature and melt viscosity. However, in this study, we efficiently utilize high-temperature FFF to develop the first-of-its-kind patient-specific robust PEEK dental implants with high print resolution. Specifically, we explore the effects of critical FFF processing conditions on the mechanical properties of the implants and subsequently determine an optimized set of processing conditions that are essential in developing durable dental implant systems. Our results indicate that the 3D printed dental implants exhibit good fatigue properties and suffice the clinical and industrial requirements for dental implants. Furthermore, we prove that the 3D printed implants exhibit adequate mechanical durability even after simulated (accelerated) aging of 30 years.