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Item Alveolar Ridge Augmentation Around Exposed Mandibular Dental Implant With Histomorphometric Analysis(Wiley, 2021-03) Ibraheem, Ahmed Gamil; Blanchard, Steven B.; Periodontology, School of DentistryIntroduction Alveolar ridge augmentation either before or during implant placement is a predictable procedure under certain conditions. A major complication during the healing phase is incision line opening and membrane exposure, which may result in reduced bone gain and reduced implant survival. This case report describes alveolar bone regeneration around three dental implants despite membrane exposure that developed during healing post-surgically. Case Presentation A 72-year-old female presented requesting dental implants to replace tooth numbers 18, 19, and 20. A cone-beam computed tomography (CBCT) scan showed loss of horizontal and vertical ridge dimensions. All implants were placed with a variable degree of implant thread exposure on their buccal surfaces, ranging from 3 to 4.5 mm. Simultaneous bone grafting was performed using freeze dried bone allograft and deproteinized bovine bone mineral that was covered by a d-PTFE membrane that was secured with tacking screws. Primary closure was obtained, and flaps were sutured. Three weeks post-surgically, membrane exposure occurred. Exposure was monitored and patient was instructed to follow strict oral hygiene instructions around the exposed membrane. Membrane exposure gradually increased without infection and was removed at 16 weeks. Membrane removal revealed dense fibrous tissues covering all implant surfaces. At the second stage surgery, new bone was seen covering all the implants coronal to the cover screws. A trephine core biopsy specimen revealed significant new bone formation and connective tissue around any residual grafted bone. Conclusion d-PTFE membrane exposure does not necessarily lead to adverse healing outcomes for alveolar ridge augmentation if handled properly with close patient follow-up.Item Amnion–Chorion Allograft Barrier Used on Root Surface for Regenerative Procedures: Case Report(Wiley, 2020-12) Hamada, Yusuke; Yeh, Yu-Ting; Blanchard, Steven B.; Periodontology, School of DentistryIntroduction Guided tissue regeneration (GTR) has been well documented with combination of bone graft substitutes and biologic modifiers to improve the outcomes of periodontal regenerative procedures. Amnion-chorion allograft membrane (ACM) is a placenta-derived resorbable allograft membrane which contains growth factors found in the placenta. The primary purpose of the barrier membranes for GTR was to exclude the epithelial down-growth along with the root surface, however, the ACM can be used as an additional biologic modifier because of the release of growth factors from the ACM after placement. The aim of this case report is to evaluate the efficacy and the application of ACM on the previously diseased root surface to treat periodontal intrabony defect. Case Presentation A 60-year-old Caucasian male with deep and wide intrabony defect on mesial #19 was treated with a regenerative procedure with combination of application of ACM on the root surface and filling the intrabony defect with the corticocancellous freeze-dried bone allograft. The bone substitute was covered with another layer of ACM and primary closure was achieved. Wound healing process was uneventful, and the clinical and radiographic outcomes were favorable up to 18 months after the surgical procedure. Conclusion This case report demonstrated that the application of ACM on the root surface with a combination of bone substitute might enhance to the radiographic bone fill and the clinical attachment level gain and minimize the risk of post-operative gingival recession.Item The Application of a CO2 Laser in Implant Site Development: A Case Report(2019) Villanueva, Justin; Blanchard, Steven B.; Hamada, Yusuke; Periodontology, School of DentistryIntroduction: Ridge preservation procedures have been utilized to minimize the alveolar ridge dimensional changes following tooth extraction. This case report describes a novel approach to ridge preservation with an application of a carbon dioxide laser (CO2 laser) to stabilize blood clot formation and stimulate wound healing. Case presentation: A 76-year-old Caucasian male with history of type 2 diabetes was referred for extraction of #8 and placement of a dental implant to reconstruct the area. Extraction was performed in a minimal traumatic manner and a mineralized freeze-dried bone allograft was placed in the socket. A 10,600 nm wavelength CO2 laser was used in setting of 1.0 watts focused continuous wave energy to stabilize the blood clot over the extraction site. The laser was used until a char layer was formed. This char layer was a non-bleeding stable blood clot, and the end-point of laser application was set as “no blood flow from the clot are within 10 seconds.” No membranes or sutures were used. Eleven weeks after the extraction and ridge preservation, a 4.1x10mm dental implant was placed with >35N/cm of primary stability and Implant stability quotient (ISQ) value of 83. A provisional crown was delivered at the time of implant placement. Three months following implant placement, the final restoration was delivered. The soft and hard tissue healing were uneventful to achieve esthetic and functional outcomes. Conclusion: Within limits of this case report, the application of CO2 laser for “Laser-Assisted Blood Clot Formation” may enhance the soft and hard tissue healing following extraction for ridge preservations.Item Bone regeneration in novel porous titanium implants(2010) Khouja, Naseeba, 1981-; Chu, Tien-Min Gabriel; Brown, David T.; Platt, Jeffery A., 1958-; Blanchard, Steven B.; Levon, John A.The objective of this study was to evaluate the in vivo performance of the novel press-fit dental implant fabricated via electron beam melting (EBM, Southern Methodist Univ.) and compare it to a commercially-available porous-coated press-fit dental implant (Endopore, Innova Corp.). Twelve cylindrical shaped implants 3 mm in diameter x 5 mm long were made by EBM (Southern Methodist Univ.) using Ti6Al4V ELI alloy. Twelve commercial implants (Endopore, Innova Corp.) of the same geometry were used as controls. Samples were implanted in rabbit tibia and retrieved six weeks postoperatively. Six specimens from each implant type were embedded undecalcified, sectioned, and stained with toluidine blue (Sigma) for histomorphometry analysis. Bone-to-implant contact (BIC) was measured. On the six remaining samples from each implant type, the mechanical properties were evaluated by pushout test on a material testing machine. The samples were loaded at a loading rate of 1 mm/min. The pushout strength was measured and the apparent shear stiffness was calculated. The results were analyzed with a paired-t test. The histology shows osteointegration of surrounding bone with both implant types. Bone was found to grow into the porous space between the beads. Both the Endopore (Innova Corp.) and the EBM (Southern Methodist Univ.) showed similar BIC. The mean BIC for the Endopore (Innova Corp.) and EBM (Southern Methodist Univ.) implant were 35 ± 6% and 32 ± 9%, respectively. It failed to reach statistical significance (p > 0.05). The peak pushout force for Endopore (Innova Corp.) and EBM (Southern Methodist Univ.) implants were 198.80 ± 61.29 N and 243.21 ± 69.75 N, respectively. The apparent shear stiffness between bone and implant for the Endopore (Innova Corp.) and EBM (Southern Methodist Univ.) implants were 577.36 ± 129.99 N/mm; and 584.48 ± 146.63 N/mm, respectively. Neither the peak pushout force nor the apparent shear stiffness of the implants was statistically different between the two groups (p > 0.05). The results suggest that the implants manufactured by EBM (Southern Methodist Univ.) perform equally well as the commercial implant Endopore (Innova Corp.) in this current animal model.Item Comparative evaluation of mandibular canal visibility on crosssectional cone-beam CT images: a retrospective study(British Institute of Radiology, 2015) Miles, Mahogany S.; Parks, Edwin T.; Eckert, George J.; Blanchard, Steven B.; Department of Periodontics and Allied Dental Programs, IU School of DentistryOBJECTIVES: The purpose of this study was to determine the visibility of the mandibular canal (MC) in CBCT images and if the visibility of the MC is affected by gender, location and/or age. METHODS: CBCT images were evaluated for the visibility of the MC by a board-certified oral and maxillofacial radiologist, a board-certified periodontist and a periodontics resident. Representative slices were examined for the first premolar (PM1), second premolar (PM2), first molar (M1) and second molar (M2) sites by all examiners. The visibility of the MC was registered as either present or absent. RESULTS: 360 total CBCT cross-sectional images were examined, with the MC identified in 204 sites (56%). Age had a significant effect on MC visibility, but it differed by location: for PM1, age 47–56 had lower visibility than age 65+ (p = 0.0377). Gender also had a significant effect on canal visibility, where females had lower visibility than males overall (p = 0.0178) and had the most pronounced difference for PM1 (p = 0.0054). Location had a significant effect on visibility, but it differed by age and by gender: for age 65+, M2 had lower visibility than PM1 (p = 0.0411) and PM2 (p = 0.0180), while for females, PM1 had lower visibility than M1 (p = 0.0123) and M2 (p = 0.0419). CONCLUSIONS: The MC was visualized only in just over half of the CBCT images. Age, gender and location had significant effects on the visibility.Item Effect of HA-coating and HF etching on experemental zirconia implant evaluation using in vivo rabbit model(2010) Huang, Sung-En; Chu, Tien-Min Gabriel; John, Vanchit (Vanchit Kurien), 1965-; Kowolik, Michael J.; Zunt, Susan L., 1951-; Blanchard, Steven B.The objective of this study was to evaluate the in vivo performance of the hydroxyapatite (HA) coating and hydrofluoric acid (HF) etching zirconia (ZrO) implants and to compare the result with titanium (Ti) implants treated in a similar manner. A total of four different implant types were tested in this study. Threaded zirconia implants with HA coating (Test 1) and zirconia implants with HF-treated surfaces (Test 2) were used to compare to the same size of titanium implants treated in identical fashion (control 1 and control 2). All implants measured about 3.5 mm at the thread diameter and 7.0 mm in total length. Each rabbit received two zirconia and two titanium implants treated in the same manner (either HA-coated or HF-etched). The samples were implanted into the rabbit tibias and retrieved at 6 weeks. Upon retrieval, 24 specimens (6 samples for each group) were fixed and dehydrated. The samples were then embedded undecalcified in PMMA for histomorphometry to quantify the bone-to-implant contact (BIC). Another 24 samples were kept in 0.9% saline and were evaluated using removal torque (RT) analysis to assess the strength of the implant-to-bone interface. The histomorphometric examination demonstrated direct bone-to-implant contact for all four groups. HA particle separation from the implants surface was seen in a majority of the HA-coated samples. No signs of inflammation or foreign body reaction were found during examination. Due to the HA particle smear contamination in the ZrO-HA group, no data was collected in this group. The mean BIC at the first three threads of the Ti-HA, Ti-HF and ZrO-HF were 57.78±18.22%, 46.41±14.55% and 47.41±14.05%, respectively. No statistically significant difference was found pair-wise among these three groups. When comparing the BIC data with the machined-surface implants, a statistically significant difference was found between the Ti-HA versus Ti implant group and the Ti-HF versus Ti implant group. The mean bone area (BA) at the first three threads for Ti-HA, Ti-HF and ZrO-HF showed statistically significant difference (p<0.05) between the ZrO-HF and Ti-HA groups, favoring the ZrO-HF group. The value of the peak removal force could only be collected from the Ti-HA group during the removal torque test. The mean RT value for the Ti-HA group was 24.39±2.58 Ncm. When comparing the RT result with our pilot study using machined-surface implants, the Ti-HA group showed statistically significant (p<0.05) higher values than the machined-surface Ti implants. The result of this study proves the in vivo biocompatibility of all four implant types tested. In the three measurable implant groups, the histomorphologic analysis showed comparable osseointegration properties in this animal model.Item The Effects of Human Amniotic Fluid on Periodontal Ligament Fibroblast Cell Viability, Proliferation, and Cytokine/Growth Factor Expression(Wolters Kluwer, 2019) Ibraheem, Ahmed Gamil; Blanchard, Steven B.; Al-Hijji, Saleh Mohammed; Al-Nasr-Allah, Khaled; Windsor, L. Jack; Biomedical Sciences and Comprehensive Care, School of DentistryBackground: The importance of the amniotic fluid (AF) to the fetus is clear. However, very few studies have been published to examine the potential uses of this fluid in various areas such as tissue regeneration. AF contains epidermal growth factor, transforming growth factor-alpha, transforming growth factor beta-1, insulin-like growth factor-I, erythropoietin and granulocyte colony-stimulating factor, as well as hyaluronic acid and hyaluronic acid-stimulating factor. Previous studies suggest that AF can increase fibroblast proliferation and chemotaxis, and decrease apoptosis as well as promote wound healing. Furthermore, evidence showed that human AF inhibits hyaluronidase, elastase, and cathepsin. The current study examined the effects of human AF on periodontal ligament fibroblasts (PDLF) in terms of cell toxicity, cell proliferation, and cytokine/growth factor expression. Materials and Methods: Cytotoxicity of AF on PDLF was determined using lactate dehydrogenase assays. PDLF proliferation was determined using water-soluble tetrazolium-1 assays. Cytokine/growth factor expression was determined on AF-treated PDLF, AF alone, and PDLF alone utilizing protein arrays. Results: Human AF at 10% and below did not affect cell growth and was not toxic. AF-treated PDLF cells showed a decrease in cytokine/growth factor levels compared to the sum of cytokine/growth factor levels in AF only and cells only for 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased. Conclusion: Human AF at the examined concentrations was not toxic to PDLF cells and did not influence their proliferation. In addition, AF (10%) caused a decrease in the total protein levels of cytokines/growth factors expressed in 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased.Item The Effects of Nicotine on the Proteolytic Activity of Periodontal Pathogens(2011) Kaeley, Janice,1976-; Gregory, Richard L.; Blanchard, Steven B.; Kowolik, Michael J.; Windsor, L. Jack; Zunt, Susan L., 1951-Periodontal disease is the leading cause of tooth loss in adults. Bacterial biofilm on tooth surfaces is the primary initiator of periodontal disease. Various factors contribute to the severity of periodontal disease including the different virulence factors of the bacteria within the biofilm. In the progression of periodontal disease, the microflora evolves from a predominantly Gram positive microbial population to a mainly Gram negative population. Specific gram negative bacteria with pronounced virulence factors have been implicated in the etiology and pathogenesis of periodontal disease, namely Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola which form the red complex of bacteria. The orange complex bacteria become more dominant in the maturation process of dental plaque and act to bridge the early colonizers of plaque with the later more dominant red complex bacterial and consists of such bacteria as Campylobacter showae, Campylobacter rectus, Fusobacterium nucleatum and Prevotella intermedia. Perhaps the most investigated contributing factor is the relationship between smoking and periodontal disease. When examining the association between cigarette smoking and interproximal bone loss, greater bone loss is associated with higher cigarette consumption, longer duration (i.e., pack year history) and higher lifetime exposure. The presence of various virulence factors such as the production of a capsular material, as well as the proteolytic activity of the various periopathodontic bacteria has been associated with the pathogenesis of periodontitis. Even though many different enzymes are produced in large quantities by these periodontal bacteria, trypsin-like enzymes, chymotrypsin-like enzymes and elastase-like enzymes, as well as dipeptidyl peptidase-like enzymes, have been thought to increase the destructive potential of the bacterium and mediate destruction of the periodontal apparatus. More specifically, it is hypothesized that the proteolytic activity of other clinically important periodontal pathogens, such as Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas assacharolyticus, is increased in the presence of nicotine. The purpose of this study was to determine the effects of nicotine on F. nucleatum, P. intermedia and P. assacharolyticus proteolytic activity. Cultures were maintained on anaerobic blood agar plates containing 3% sheep blood. Bacterial cells were harvested from the plates and washed. Washed F. nucleatum, P. intermedia and P. assacharolyticus cells were incubated with 1 mg/ml of nicotine. Bacterial cells not incubated with nicotine were used as positive controls. Secreted enzymatic activity was measured using the synthetic chromogenic substrates glycyl-L-proline-p-nitroanilide (GPPNA), N-succinyl-L-alanyl-L-alanyl-L-alanyl-p-nitroanilide (SAAAPNA), N-succinyl-alanine-alanine-proline-phenylalanine-p-nitroanilide (SAAPPPNA) and N-α-benzoyl-L-arginine-p-nitroanilide (L-BAPNA) (Sigma-Aldrich Products, St. Louis, MO, USA). Appropriate means and standard deviations were determined for each of the enzymatic activities measured and analysis of variance (ANOVA) was used to compare the groups utilizing a 5% significance level for all comparisons. Results demonstrated that after 60 minutes of incubation of F. nucleatum, P. intermedia and P. assacharolyticus cells with 1 mg/ml of nicotine and the various synthetic substrates, had the following proteolytic activity for GPPNA: 0.83 ± 0.14, 0.72 ± 0.03 and 0.67 ± 0.10, respectively; SAAAPNA: 0.82 ± 0.06, 0.76 ± 0.05 and 0.68 ± 0.08, respectively; SAAPPPNA: 0.90 ± 0.13, 0.85 ± 0.17 and 0.72 ± 0.03, respectively; and BAPNA: 0.81 ± 0.15, 0.74 ± 0.13 and 0.74 ± 0.16, respectively. In conclusion, the results indicate that in the presence of 1 mg/ml of nicotine, the proteolytic activity of F. nucleatum and P. assacharolyticus was increased with all of the synthetic substrates (with statistical significance seen only in the increases with F. nucleatum and GPPNA, SAAAPNA and BAPNA). The proteolytic activity exhibited an increasing trend in activity for P. intermedia with SAAPPPNA and BAPNA but a decreasing trend in activity with GPPNA and SAAAPNA when incubated with 1 mg/ml of nicotine, once again demonstrating no statistical significance for any of the substrates. Therefore, it could be concluded that based on these results nicotine at a concentration of 1 mg/ml may increase the proteolytic activity of periodontal pathogens and thus may increase periodontal disease activity and subsequent periodontal breakdown. Further studies are needed to validate these results utilizing different concentrations of nicotine.Item EFFECTS OF ORTHODONTIC MINI-IMPLANT DIAMETER ON MICRODAMAGE.(Office of the Vice Chancellor for Research, 2012-04-13) Cruz, Enrique; Liu, Sean; Sun, Jun; Blanchard, Steven B.; Soto, Armando; Stewart, Kelton T.; Allen, Matthew R.; Liu, SeanMicrodamage reduces bone mechanical properties and thus could possi-bly contribute to implant failure. The objective of this study was to investi-gate whether the diameter of mini-implants (MI) affects microdamage gen-eration and whether this differs between the mandible and maxilla due to their contrasting cortical thicknesses. Methods: Maxillary and mandibular quadrants of 5 dogs were randomly assigned to receive, in situ, no interven-tion (control), pilot drilling only, or pilot drilling plus one of three diameters of MI: 1.4 (n=18), 1.6 (n=18), and 2.0 mm (n=18). Microdamage was as-sessed on basic fuchsin stained sections using epifluorescence microscopy. Results: No microdamage was found in the non-drilling controls. Pilot drill-ing produced only minimal microdamage in the maxilla but more microdamage in the mandible. There was significantly higher microdamage generated in the mandible, compared to the maxilla (p<0.05). In the maxil-la, although insertion of all implants produced higher microdamage than the control and pilot drilling, there were no differences between the 3 MI diame-ters. In the mandible, insertion of implants generated significantly higher microdamage than the control, but it did not produce higher microdamage than pilot drilling. Similarly, no differences in microdamage were found be-tween 3 MI diameters. Conclusion: Insertion of MIs in the mandible pro-duced higher microdamage than in the maxilla, which may explain that the higher MI failure rate in the mandible. Implant diameter did not affect over-all microdamage burden in either jaw. Microdamage was mostly generated by pilot drilling through the cortex in the mandible, while microdamage in the maxilla was mainly produced when manual inserting MIs after pilot drill-ing.Item Effects of platelet‐rich fibrin on human gingival and periodontal ligament fibroblast proliferation from chronic periodontitis versus periodontally healthy subjects(Wiley, 2021-08) Goel, Apoorv; Windsor, L. Jack; Gregory, Richard L.; Blanchard, Steven B.; Hamada, Yusuke; Periodontology, School of DentistryBackground: Platelet-rich fibrin (PRF), an autogenous blood concentrate, contains multiple growth factors and is used as an adjunct in the periodontal regeneration and implant site development procedures to stimulate wound healing. Patient-related factors such as chronic periodontitis may affect the quality of PRF. Objectives: This study aimed to investigate and compare PRF's effects from patients diagnosed with generalized moderate or severe chronic periodontitis to patients who presented with intact periodontium on human gingival fibroblast (HGF) and human periodontal ligament fibroblast (HPLF) proliferation. Materials and methods: A total of 33 ml of whole intravenous blood was collected from each subject and centrifuged at 2700 rpm for 12 min in three 10 ml tubes, and 3 ml of blood was used for Complete Blood Count analysis. Three PRF clots were compressed to produce the membranes and liquid exudate. PRF membrane and 10% liquid exudate were exposed to 20,000 HPLFs/well or 25,000 HGFs/well in triplets from each subject in a 48 cell well plate. After 72 h of incubation, the conditioned media were evaluated by Water Soluble Tetrazolium-1 assays to determine fibroblast proliferation. Controls included cells alone and media without cells. Complete blood counts were measured. Results: Subjects in both groups were age and gender-matched (intact 46.7 ± 11.4 years and periodontitis 54.8 ± 10.4 years, p-value = 0.1344). Body Mass Index and White Blood Corpuscles in the periodontitis group was significantly higher than the intact group (p = 0.0176 and p = 0.0038) whereas no differences were seen for Red Blood Corpuscles (p = 0.2020), Hemoglobin (p = 0.2290) and Platelets (p = 4,094). There were no significant differences in the HGF and HPLF proliferation with PRF exudates and membranes between intact periodontium and periodontitis groups (all p > 0.05). However, PRF exudates in both groups induced significant more cell proliferation when compared to PRF membranes. Conclusions: PRF exudates induced significant proliferation of fibroblasts and can play a vital role in wound healing. The current study concluded that PRF membranes, in combination with PRF exudates, can be utilized for their therapeutic and wound healing potential, not affected by the periodontal condition of the patient.