Faculty Bibliography

Bone defects are associated with trauma, congenital disorders, non-unions, or infections following surgical procedures. Defects which are unable to heal spontaneously are categorized as "critical sized" and are commonly treated using bone grafts in an effort to facilitate bone regeneration and stabilization. Grafting materials can be either natural or synthetic, each having their respective advantages and disadvantages. Synthetic bone grafts are favored due to their ability to be tailored to exhibit desired properties and geometric configurations. β-tricalcium phosphate (β-TCP) is a synthetic grafting material that has been widely utilized for regenerative purposes due to its favorable osteoconductive properties. In combination with 3D printing, grafting materials can be further customized with respect to their macro and micro features. One way to customize devices is by using 3D printing and varying the surface area, by varying the internal component measurements. The objective of this study was to compare the effect of porosity and surface area of 3D printed β-TCP scaffolds with different strut diameters and the effect on cell proliferation in vitro. ß-TCP scaffolds were printed using a custom-built 3D direct-write micro printer with syringes equipped with different extrusion tip diameters (fdiameter: 200 µm, 250 µm and 330 µm). After sintering and post processing, scaffolds were subjected to micro-computed tomography (µCT) and a Scanning Electron Microscope (SEM) to evaluate surface area and porosity, respectively. Compressive strength was assessed using a universal testing machine. Cell proliferation was assessed through cellular viability, using human osteoprogenitor cells. The surface area of the scaffolds was found to increase with smaller strut diameters. Statistically significant differences (p<0.05) were detected for cellular proliferation, between the smallest extrusion diameter, 200 μm, and the largest diameter, 330 μm, after 48-, 72-, and 168-hours. No statistical significances were detected (p>0.05) with regards to the mechanical properties between groups. This study demonstrated that a smaller diameter rod yielded a higher surface area resulting in increased levels of cellular proliferation. Therefore, tailoring rod dimensions has the capacity to enhance cellular adhesion and ultimately, proliferation.

BACKGROUND:Computerized surgical planning (CSP) in osseous reconstruction of head and neck cancer defects has become a mainstay of treatment. However, the consequences of CSP-designed titanium plating systems on planning adjuvant radiation remains unclear. METHODS:Two patients underwent head and neck cancer resection and maxillomandibular free fibula flap reconstruction with CSP-designed plates and immediate placement of osseointegrated dental implants. Surgical treatment was followed by adjuvant intensity modulated radiation therapy (IMRT). RESULTS:Both patients developed osteoradionecrosis (ORN), and one patient had local recurrence. The locations of disease occurred at the areas of highest titanium plate burden, possibly attributed to IMRT dosing inaccuracy caused by the CSP-designed plating system. CONCLUSION/CONCLUSIONS:Despite proven benefits of CSP-designed plates in osseous free flap reconstruction, there may be an underreported risk to adjuvant IMRT treatment planning leading to ORN and/or local recurrence. Future study should investigate alternative plating methods and materials to mitigate this debilitating outcome.

BACKGROUND:We aimed to histomorphometrically evaluate the effects of Leucocyte-Platelet-Rich Fibrin (L-PRF), with and without the combination of a bone grafting material, for alveolar ridge preservation using an in vivo canine model. MATERIAL AND METHODS/METHODS:Seven dogs (Female Beagles, ~18-month-old) were acquired for the study. L-PRF was prepared from each individual animal by drawing venous blood and spinning them through a centrifuge at 408 RCF-clot (IntrasSpin, Intra-Lock, Boca Raton, FL). L-PRF membranes were obtained from XPression fabrication kit (Biohorizons Implant Systems, Inc., AL, USA). A split mouth approach was adopted with the first molar mesial and distal socket defects treated in an interpolated fashion of the following study groups: 1) Empty socket (negative control); 2) OSS filled defect 3) L-PRF membrane; and 4) Mix of Bio-Oss® with L-PRF. After six weeks, samples were harvested, histologically processed, and evaluated for bone area fraction occupancy (BAFO), vertical/horizontal ridge dimensions (VRD and HRD, respectively), and area of coronal soft tissue infiltration. RESULTS:BAFO was statistically lower for the control group in comparison to all treatment groups. Defects treated with Bio-Oss® were not statistically different then defects treated solely with L-PRF. Collapsed across all groups, L-PRF exhibited higher degrees of BAFO than groups without L-PRF. Defects filled with Bio-Oss® and Bio-Oss® with L-PRF demonstrated greater maintenance of VRD relative to the control group. Collapsed across all groups, Bio-Oss® maintained the VRD and resulted in less area of coronal soft tissue infiltration compared to the empty defect. Soft tissue infiltration observed at the coronal area was not statistically different among defects filled with L-PRF, Bio-Oss®, and Bio-Oss® with L-PRF. CONCLUSIONS:Inclusion of L-PRF to particulate xenograft did not promote additional bone heading at 6 weeks in vivo. However, we noted that L-PRF alone promoted alveolar socket regeneration to levels comparable to particulate xenografts, suggesting its potential utilization for socket preservation.

PURPOSE/OBJECTIVE:To evaluate the effect of composition, fabrication mode, and thermal cycling on the mechanical properties of different polymeric systems used for temporary dental prostheses. MATERIALS AND METHODS/METHODS:Standard bar-shaped specimens (25 × 2 × 2 mm) were fabricated of six polymeric systems of varying compositions and fabrication modes (n = 10/group): conventional PMMA (Alike, GC) - group CGC; conventional PMMA (Dêncor, Clássico) - group CD; bis-acryl (Tempsmart, GC) - group BGC; bis-acryl (Yprov, Yller) - group BY; milled PMMA (TelioCAD, Ivoclar) - group MI; 3D printed bis-acryl - (Cosmos Temp, Yller) group PY. Half of the specimens were subjected to 5000 thermal cycles (5 °C to 55 °C). Three-point bending tests were performed using a universal testing machine with a crosshead speed set to 0.5 mm/min. Flexural strength and elastic modulus were calculated from the collected data. FTIR spectra were recorded pre and post curing and after thermal cycling to evaluate material composition and degree of conversion. Energy-dispersive spectroscopy (EDS) and scanning electron microscope (SEM) were utilized to examine the composition and micromorphology of the systems, respectively. Data were analyzed using two-analysis of variance and Tukey tests (α = 0.05). RESULTS:FTIR spectra indicated that BGC, BY and PY groups corresponded to urethane dimethacrylate systems (bis-acryl), while CGC, CD, and MI groups corresponded to monomethacrylate systems, polymethyl methacrylate (PMMA). Bis-acryl BGC system yeilded the highest flexural strength (80 MPa), followed by the milled PMMA MI system (71 MPa), both statistically significant different relative to other groups. Bis-acryl BY exhibited the lowest flexural strength (27 MPa). Thermocycling significantly increased the flexural strength of all polymeric systems (∼10-15 MPa), except for the 3D-printed PY group. Bis-acryl BGC (1.89 GPa) and conventional PMMA CGC (1.66 GPa) groups exhibited the highest elastic modulus, followed by milled PMMA MI group (1.51 GPa) and conventional PMMA CD (1.45 GPa) systems, with significant difference detected between BGC group and MI and CD groups. The 3D printed PY (0.78 GPa) and bis-acryl BY (0.47 GPa) systems presented the lowest elastic modulus. Thermocycling did not have a significant influence on the elastic modulus. FTIR spectra indicate water sorption and release of unreacted monomers as well as increased degree of conversion (∼5-12%) after thermal cycling. CONCLUSION/CONCLUSIONS:Composition and fabrication mode and thermal cycling significantly affected the mechanical properties of polymeric systems used for temporary dental prostheses.

The present study aimed to evaluate the effect of parathormone (PTH) administered directly to the implant's surface prior to insertion, using a large translational animal model. Sixty titanium implants were divided into four groups: (i) Collagen, control group, where implants were coated with Type-I Bovine-collagen, and three experimental groups, where implants received varying doses of PTH: (ii) 12.5, (iii) 25, and (iv) 50 μg, prior to placement. Fifteen female sheep (~2 years old, weighing ~65 kg) received four implants in an interpolated fashion in C3, C4 or C5 vertebral bodies. After 3-, 6- and 12-weeks, samples were harvested, histologically processed, qualitatively and quantitatively assessed for bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO). BIC yielded lower values at 6-weeks for 50 μg relative to the control group, with no significant differences, when compared to the 12.5- and 25-μg. No significant differences were detected at 6-weeks between collagen, 12.5- and 25-μg groups. At 3- and 12-weeks, no differences were detected for BIC among PTH groups. With respect to BAFO, no significant differences were observed between the control and experimental groups independent of PTH concentration and time in vivo. Qualitative observations at 3-weeks indicated the presence of a more mature bone near the implant's surface with the application of PTH, however, no significant differences in new bone formation or healing patterns were observed at 6- and 12-weeks. Single local application of different concentrations of PTH on titanium implant's surface did not influence the osseointegration at any time-point evaluation in low-density bone.

OBJECTIVES/OBJECTIVE:To characterize the mechanical and biological properties of three commercially available resins, which are currently used for provisional restorations and to compare them to an experimental resin intended for definitive fixed dental prostheses. MATERIALS AND METHODS/METHODS:Three commercially available resins: Crowntec (CT, Saremco), Temporary C&B (FL, Formlabs), C&B MFH (ND, Nextdent), and the experimental resin: Permanent Bridge (PB, Saremco) were printed and subjected to biaxial flexural strength test, finite element analysis, Weibull analysis, scanning electron microscopy, cell proliferation, immunohistochemistry and cytotoxicity assays. Samples from CT, PB, and ND were provided directly from the manufacturers ensuring ideal workflow. FL was printed using the workflow as recommended by the manufacturer, using a Formlabs 2 printer and their post-processing units Form Wash and Form Cure. RESULTS:From the tested resins, PB yielded the best overall results in terms of mechanical properties. Cell proliferation and cytotoxicity did not show any significant differences among materials. PB showed higher values for probability of survival predictions (35%) when subjected to 250 MPa loads, whereas the other materials did not reach 10%. SIGNIFICANCE/CONCLUSIONS:Despite mechanical differences between the evaluated materials, the outcomes suggest that 3D printed provisional resins may be used in clinical settings, following the manufacturers indications. New materials intended for long-term use, such as the PB resin, yielded higher mechanical properties compared to the other materials. Alternative printing and post-processing methods have not yet been evaluated and should be avoided until further literature is available. CLINICAL SIGNIFICANCE/CONCLUSIONS:3D printed resins for provisional restorations have become popular with the emergence of new technologies. In this study, we evaluated three different commercially available resins for provisional restorations and one new experimental resin. The results from this study indicate that commercially available resins could be used in clinical settings under certain conditions and limited periods of time. Following the manufacturers protocols is of paramount importance to not compromise these properties.

Rubber band ligation is a commonly used method for the removal of tissue abnormalities. Most often, rubber band ligation is performed to remove internal hemorrhoids unresponsive to first line treatments to avoid surgery. While the procedure is considered safe, patients experience mild to significant pain and discomfort until the tissue sloughs off. As patients often require multiple bandings and sessions, reducing these side effects can have a considerable effect on patient adherence and quality of life. To reduce pain and discomfort, we developed drug-eluting rubber bands for ligation procedures. We investigated the potential for a band to elute anesthetics and drug combinations to durably manage pain for a period of up to 5 days while exhibiting similar mechanical properties to conventional rubber bands. We show that the rubber bands retain their mechanical properties despite significant drug loading. Lidocaine, released from the bands, successfully altered the calcium dynamics of cardiomyocytes in vitro and modulated heart rate in zebrafish embryos, while the bands exhibited lower cytotoxicity than conventional bands. Ex vivo studies demonstrated substantial local drug release in enteric tissues. These latex-free bands exhibited sufficient mechanical and drug-eluting properties to serve both ligation and local analgesic functions, potentially enabling pain reduction for multiple indications.

The current standard of care for an alveolar cleft defect is an autogenous bone graft, typically from the iliac crest. Given the limitations of alveolar bone graft surgery, such as limited supply, donor site morbidity, graft failure, and need for secondary surgery, there has been growing interest in regenerative medicine strategies to supplement and replace traditional alveolar bone grafts. Though there have been preliminary clinical studies investigating bone tissue engineering methods in human subjects, lack of consistent results as well as limitations in study design make it difficult to determine the efficacy of these interventions. As the field of bone tissue engineering is rapidly advancing, reconstructive surgeons should be aware of the preclinical studies informing these regenerative strategies. We review preclinical studies investigating bone tissue engineering strategies in large animal maxillary or mandibular defects and provide an overview of scaffolds, stem cells, and osteogenic agents applicable to tissue engineering of the alveolar cleft. An electronic search conducted in the PubMed database up to December 2021 resulted in 35 studies for inclusion in our review. Most studies showed increased bone growth with a tissue engineering construct compared to negative control. However, heterogeneity in the length of follow up, method of bone growth analysis, and inconsistent use of positive control groups make comparisons across studies difficult. Future studies should incorporate a pediatric study model specific to alveolar cleft with long-term follow up to fully characterize volumetric defect filling, cellular ingrowth, bone strength, tooth movement, and implant support.

It is well known that Semaphorin 4D (Sema4D) inhibits IGF-1-mediated osteogenesis by binding with PlexinB1 expressed on osteoblasts. However, its elevated level in the gingival crevice fluid of periodontitis patients and the broader scope of its activities in the context of potential upregulation of osteoclast-mediated periodontal bone-resorption suggest the need for further investigation of this multifaceted molecule. In short, the pathophysiological role of Sema4D in periodontitis requires further study. Accordingly, attachment of the ligature to the maxillary molar of mice for 7 days induced alveolar bone-resorption accompanied by locally elevated, soluble Sema4D (sSema4D), TNF-α and RANKL. Removal of the ligature induced spontaneous bone regeneration during the following 14 days, which was significantly promoted by anti-Sema4D-mAb administration. Anti-Sema4D-mAb was also suppressed in vitro osteoclastogenesis and pit formation by RANKL-stimulated BMMCs. While anti-Sema4D-mAb downmodulated the bone-resorption induced in mouse periodontitis, it neither affected local production of TNF-α and RANKL nor systemic skeletal bone remodeling. RANKL-induced osteoclastogenesis and resorptive activity were also suppressed by blocking of CD72, but not Plexin B2, suggesting that sSema4D released by osteoclasts promotes osteoclastogenesis via ligation to CD72 receptor. Overall, our data indicated that ssSema4D released by osteoclasts may play a dual function by decreasing bone formation, while upregulating bone-resorption.

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.