Faculty Bibliography

Skeletal conditions represent a considerable challenge to health systems globally. Barriers to effective therapeutic development include a lack of accurate preclinical tissue and disease models. Most recently, work was attempted to present a novel whole organ approach to modeling human bone and cartilage tissues. These self-assembling skeletal organoids mimic the cellular milieu and extracellular organization present in native tissues. Bone organoids demonstrated osteogenesis and micro vessel formation, and cartilage organoids showed evidence of cartilage development and maturation. Skeletal organoids derived from both bone and cartilage tissues yielded spontaneous polarization of their cartilaginous and bone components. Using these hybrid skeletal organoids, we successfully generated "mini joint" cultures, which we used to model inflammatory disease and test Adenosine (A_2A ) receptor agonists as a therapeutic agent. The work and respective results indicated that skeletal organoids can be an effective biological model for tissue development and disease as well as to test therapeutic agents.

HYPOTHESIS/OBJECTIVE:Injuries requiring resection of tissue followed by autogenous bone transfer may be prone to infection by Staphylococcus aureus, impeding recovery and increasing medical costs. For critical sized defects, the common approach to reconstruction is a tissue transfer procedure but is subject to limitations (e.g., donor site morbidity, cost, operating time). Utilizing beta tricalcium phosphate (β-TCP) as bone grafting material augmented with silver (Ag), a custom graft may be 3D printed to overcome limitations and minimize potential infections. EXPERIMENTS/METHODS:) groups followed by electron microscopy, thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC) to gather information of chemical and physical properties. Preliminary biocompatibility and bactericidal capacity of the scaffolds were tested using human osteoprogenitor (hOP) cells and methicillin-sensitive S. aureus strain, respectively. RESULTS:groups, whereas electron microscopy showed a decrease in Ca and an increase in Ag ions, decreasing Ca/P ratio with increasing surfactant concentrations. PrestoBlue assays yielded an increase in fluorescence cell counts among experimental groups with lower concentrations of Ag characterized by their characteristic trapezoidal shape whereas cytotoxicity was observed at higher concentrations. Similar observations were made with alkaline phosphatase assays. Antimicrobial evaluation showed reduced colony-forming units (CFU) among all experimental groups when compared to 100% β-TCP. β-TCP scaffolds augmented with Ag ions facilitate antibacterial effects while promoting osteoblast adhesion and proliferation.

Bone defects are often linked to congenital disorders, high impact traumas, tumors or oncological resections. Potential treatment options include autografts, allografts, or synthetic grafts, such as bioactive ceramic-based materials which have been successfully utilized in an effort to regenerate bone. β-tricalcium phosphate (β-TCP), is a commonly utilized bioactive ceramic for regenerative purposes with favorable osteoconductive properties. Alternatively, Synthetic Bone Mineral (SBM) has been previously utilized in in vivo experiments as a supplement for bone loss treatment. As a potential alternative to β-TCP, it is also a bioactive ceramic, which consists of a carbonate hydroxyapatite with ionic substitutions such as F^−, Zn²⁺ and Mg²⁺. The objective of this work was to characterize the physiochemical properties of the colloidal gel obtained from a formulation of SBM and compare the properties directly to β-TCP. Mechanical properties were evaluated for both materials in bulk, using Biaxial Flexural Strength tests. Scanning electron microscopy and micro-computed tomography were utilized to explore the structure of the bulk material and the three dimensionally (3D) printed scaffolds. Inductive coupled plasma (ICP), X-ray diffraction (XRD), and Fourier transform infrared spectrometry (FT-IR), were utilized to determine the calcium-phosphorous ratio (Ca:P), quantitative analysis of crystalline phases, and functional groups, respectively. Thermogravimetric analysis (TGA) was used to quantify the weight percent of water, organic components, carbonate and mineral in the SBM colloidal gel. Flexural strength of SBM discs sintered at 700°C were statistically analogous to β-TCP sintered at 900°C. The Ca:P ratio of the sintered SBM was found to be 1.47 ± 0.04, statistically different from β-TCP sintered at higher temperatures. The carbonate content of the SBM was determined to be ~2.8% ± 0.9. The novel SBM colloidal gel has hence been characterized chemically and physically for its potential use in 3D printing grafts to repair critical sized bone defects.

PURPOSE/OBJECTIVE:This study evaluated the effect of different tightening protocols on the probability of survival of screw-retained implant-supported anterior crowns. MATERIALS AND METHODS/METHODS:Seventy-two implants with internal conical connections (4.0 × 10mm, Ti-6Al-4V, Colosso, Emfils) were divided into four groups (n = 18 each): 1) Manufacturer's recommendations torque (25 N.cm for abutment's screw and 30 N.cm for crown's screw) (MaT); 2) Retightening after 10 min (ReT); 3) Torque 16% below recommended to simulate an uncalibrated wrench (AgT), and; 4) Temporary crown simulation (TeT), where crowns were torqued to 13 N.cm to simulate manual tightening, subjected to 11,200 cycles to simulate temporary crown treatment time (190 N), and then retightened to manufacturer torque (TeT). All specimens were subjected to cyclic fatigue in distilled water with a load of 190 N until 250,000 cycles or failure. The probability of survival (reliability) to complete a mission of 50,000 cycles was calculated and plotted using the Weibull 2-Parameter analysis. Weibull modulus and number of cycles at which 62.3% of the specimens would fail were also calculated and plotted. The failure mode was characterized in stereo and scanning electron microscopes (SEM). RESULTS:The probability of survival was 69.3% for MaT, 70% for ReT, 54.8% for AgT, and 40.3% for TeT, all with no statistically significant difference. Weibull modulus was approximately 1.0 for all groups. The characteristic number of cycles for failure was 105,000 cycles for MaT, 123,000 for ReT, 82,000 cycles for AgT, and 54,900 cycles for TeT, with no significant difference between groups. The chief failure mode for MaT, ReT, AgT groups was crown screw fracture, whereas abutment screw fracture was the chief failure mode for the TeT group. CONCLUSION/CONCLUSIONS:Tightening protocol did not influence the probability of survival of the screw-retained anterior crowns supported by internal conical implants (Ti-6Al-4V, Colosso, Emfils).

BACKGROUND:β-Tricalcium phosphate (β-TCP) is one of the most common synthetic bone grafting materials utilized in craniofacial reconstruction; however, it is limited by a slow degradation rate. The aim of this study was to leverage 3-dimensional (3D) printing in an effort to accelerate the degradation kinetics of β-TCP. METHODS:Twenty-two 1-month-old New Zealand white rabbits underwent creation of calvarial and alveolar defects, repaired with 3D-printed β-TCP scaffolds coated with 1000 μM of osteogenic agent dipyridamole. Rabbits were euthanized after 2, 6, and 18 months after surgical intervention. Bone regeneration, scaffold degradation, and bone mechanical properties were quantified. RESULTS:Histological analysis confirmed the generation of vascularized and organized bone. Microcomputed tomography analysis from 2 to 18 months demonstrated decreased scaffold volume within calvarial (23.6% ± 2.5%, 5.1% ± 2.2%; P < 0.001) and alveolar (21.5% ± 2.2%, 0.2% ± 1.9%; P < 0.001) defects, with degradation rates of 54.6%/year and 90.5%/year, respectively. Scaffold-inducted bone generation within the defect was volumetrically similar to native bone in the calvarium (55.7% ± 6.9% vs 46.7% ± 6.8%; P = 0.064) and alveolus (31.4% ± 7.1% vs 33.8% ± 3.7%; P = 0.337). Mechanical properties between regenerated and native bone were similar. CONCLUSIONS:Our study demonstrates an improved degradation profile and replacement of absorbed β-TCP with vascularized, organized bone through 3D printing and addition of an osteogenic agent. This novel additive manufacturing and tissue engineering protocol has implications to the future of craniofacial skeletal reconstruction as a safe and efficacious bone tissue engineering method.

PURPOSE/OBJECTIVE:The aim of this study was to qualitatively and quantitatively assess the effect of osteotomy preparation by conventional, subtractive, or osseodensification instrumentation on osteotomies, treated with or without endosteal implants, and healing capacity. MATERIALS AND METHODS/METHODS:Seven sheep were used, and 56 osteotomies were made in the left and right ilium of the sheep (n = 8/sheep [4 per side/time point (3 and 6 weeks)]). Two different instrumentation techniques were used: (1) conventional/regular drilling in a three-step series of a 2-mm pilot and 3.2-mm and 3.8-mm twist drills and (2) osseodensification drilling with a Densah Bur 2.0-mm pilot and 2.8-mm and 3.8-mm multi-fluted tapered burs. Drilling was performed at 1,100 rpm with saline irrigation. RESULTS:Qualitative histomorphometric evaluation of the osteotomies after 3 and 6 weeks did not indicate any healing impairment due to the instrumentation. In all samples, histologic examination suggested bone remodeling and growth (empty and treated with an implant), irrespective of preparation technique. Osteotomies prepared using the osseodensification instrumentation showed the existence of bone chips autografted into the trabecular spaces along the length of the osteotomy wall. CONCLUSION/CONCLUSIONS:The osseodensification group yielded higher osseointegration rates, as distinguished through qualitative assessment, bone-to-implant contact, and bone-area-fraction occupancy, indicating an increased osteogenic potential in osteotomies prepared using the osseodensification technique.

OBJECTIVE:To characterize the biomechanical performance of fiber-reinforced composite 5-unit implant-supported fixed dental prostheses (FDPs) receiving individually milled crowns by insilico and fatigue analyses. METHODS:Eighteen implant-supported five-unit fiber-reinforced composite frameworks with an individually prepared abutment design were fabricated, and ninety resin-matrix ceramic crowns were milled to fit each abutment. FDPs were subjected to step-stress accelerated-life testing with load delivered at the center of the pontic and at 2nd molar and 1st premolar until failure. The reliability of the prostheses combining all loaded data and of each loaded tooth was estimated for a mission of 50,000 cycles at 300, 600 and 900 N. Weibull parameters were calculated and plotted. Fractographic and finite element analysis were performed. RESULTS:Fatigue analysis demonstrated high probability of survival at 300 N, with no significant differences when the set load was increased to 600 and 900 N. 1st and 2nd molar dataset showed high reliability at 300 N, which remained high for the higher load missions; whereas 1st premolar dataset showed a significant decrease when the reliability at 300 N was compared to higher load missions. The characteristic-strength of the combined dataset was 1252 N, with 1st molar dataset presenting higher values relative to 2nd molar and 1st premolar, both significantly different. Failure modes comprised chiefly cohesive fracture within the crown material originated from cracks at the occlusal area, matching the maximum principal strain location. SIGNIFICANCE/CONCLUSIONS:Five-unit implant-supported FDP with crowns individually cemented in a fiber-reinforced composite framework presented a high survival probability. Crown fracture comprised the main failure mode.

BACKGROUND/PURPOSE/OBJECTIVE:The utilization of three-dimensionally (3D)-printed bioceramic scaffolds composed of beta-tricalcium phosphate in conjunction with dipyridamole have shown to be effective in the osteogenesis of critical bone defects in both skeletally immature and mature animals. Furthermore, previous studies have proven the dura and pericranium's osteogenic capacity in the presence of 3D-printed scaffolds; however, the effect galea aponeurotica on osteogenesis in the presence of 3D scaffolds remains unclear. METHOD/DESCRIPTION/UNASSIGNED:Critical-sized (11 mm) bilateral calvarial defects were created in 35-day old rabbits (n = 7). Two different 3D scaffolds were created, with one side of the calvaria being treated with a solid nonporous cap and the other with a fully porous cap. The solid cap feature was designed with the intention of preventing communication of the galea and the ossification site, while the porous cap permitted such communication. The rabbits were euthanized 8 weeks postoperatively. Calvaria were analyzed using microcomputed tomography, 3D reconstruction, and nondecalcified histologic sectioning in order assess differences in bone growth between the two types of scaffolding. RESULTS:Scaffolds with the solid (nonporous) cap yielded greater percent bone volume (P = 0.012) as well as a greater percent potential bone (P = 0.001) compared with the scaffolds with a porous cap. The scaffolds with porous caps also exhibited a greater percent volume of soft tissue (P < 0.001) presence. There were no statistically significant differences detected in scaffold volume. CONCLUSION/CONCLUSIONS:A physical barrier preventing the interaction of the galea aponeurotica with the scaffold leads to significantly increased calvarial bone regeneration in comparison with the scaffolds allowing for this interaction. The galea's interaction also leads to more soft tissue growth hindering the in growth of bone in the porous-cap scaffolds.

BACKGROUND:The goal of this study was to evaluate hard tissue response following guided bone regeneration using commercially available bovine bone grafts and collagen membranes; bilayer collagen membrane and porcine pericardium-based membrane, by means of a non-destructive three-dimensional (3D) computerized volumetric analysis following microtomography reconstruction. MATERIAL AND METHODS/METHODS:Bone regenerative properties of various bovine bone graft materials were evaluated in the Göttingen minipig model. Two standardized intraosseous defects (15mm x 8mm x 8mm) were created bilaterally of the mandible of eighteen animals (n=72 defects). Groups were nested within the same subject and randomly distributed among the sites: (i) negative control (no graft and membrane), (ii) bovine bone graft/bilayer collagen membrane (BOB) (iii) Bio-Oss® bone graft/porcine pericardium-based membrane (BOJ) and (iv) cerabone® bone graft/porcine pericardium-based membrane (CJ). Samples were harvested at 4, 8, and 12-week time points (n=6 animal/time point). Segments were scanned using computerized microtomography (μCT) and three dimensionally reconstructed utilizing volumetric reconstruction software. Statistical analyses were performed using IBM SPSS with a significance level of 5%. RESULTS:From a temporal perspective, tridimensional evaluation revealed gradual bone ingrowth with the presence of particulate bone grafts bridging the defect walls, and mandibular architecture preservation over time. Volumetric analysis demonstrated no significant difference between all groups at 4 weeks (p>0.127). At 8 and 12 weeks there was a higher percentage of new bone formation for control and CJ groups when compared to BOB and BOJ groups (p<0.039). The natural bovine bone graft group showed more potential for graft resorption over time relative to bovine bone graft, significantly different between 4 and 8 weeks (p<0.003). CONCLUSIONS:Volumetric analysis yielded a favorable mandible shape with respect to time through the beneficial balance between graft resorption/bone regenerative capacity for the natural bovine bone graft.