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Analysis of salivary flow rate, biochemical composition, and redox status in orchiectomized spontaneously hypertensive rats
(2023-08-01) de Oliveira, Ana Carolina Fujimori [UNESP]; Brito, Victor Gustavo Balera [UNESP]; Ramos, Guilherme Henrique Alves dos Santos [UNESP]; Werlang, Matheus Lima Cypriano [UNESP]; Fiais, Gabriela Alice [UNESP]; Dornelles, Rita Cássia Menegati [UNESP]; Antoniali, Cristina [UNESP]; Nakamune, Ana Cláudiade Melo Stevanato [UNESP]; Fakhouri, Walid D.; Chaves-Neto, Antonio Hernandes [UNESP]; Universidade Estadual Paulista (UNESP); School of Dentistry; University of Texas Health Science Center at Houston
Objective: This study aimed to analyze the salivary flow rate, biochemical composition, and redox status in orchiectomized spontaneously hypertensive rats (SHR) compared to normotensive Wistar rats. Design: Thirty-two young adult male SHR and Wistar (3-months-old) rats were randomly distributed into four groups; either castrated bilaterally (ORX) or underwent fictitious surgery (SHAM) as Wistar-SHAM, Wistar-ORX, SHR-SHAM, and SHR-ORX. Two months beyond castration, pilocarpine-induced salivary secretion was collected from 5-month-old rats to analyze salivary flow rate, pH, buffer capacity, total protein, amylase, calcium, phosphate, sodium, potassium, chloride, thiobarbituric acid reactive substances (TBARs), carbonyl protein, nitrite, and total antioxidant capacity. Results: The salivary flow rate was higher in the Wistar-ORX compared to the Wistar-SHAM group, while remaining similar between the SHR-SHAM and SHR-ORX groups. ORX did not affect pH and salivary buffer capacity in both strains. However, salivary total protein and amylase were significantly reduced in the Wistar-ORX and SHR-ORX compared to the respective SHAM groups. In both ORX groups, salivary total antioxidant capacity and carbonylated protein were increased, while lipid oxidative damage (TBARs) and nitrite concentration were higher only in the Wistar-ORX than in the Wistar-SHAM group. In the Wistar-ORX and SHR-ORX, the salivary calcium, phosphate, and chloride were increased while no change was detected in the SHAM groups. Only salivary buffering capacity, calcium, and chloride in the SHR-ORX adjusted to values similar to Wistar-SHAM group. Conclusion: Hypertensive phenotype mitigated the orchiectomy-induced salivary dysfunction, since the disturbances were restricted to alterations in the salivary biochemical composition and redox state.
Hand instrumentation provides improved tissue response over ultrasonic scaler and substantiates safe dental practice: An in vivo study in rats
(2023-05-01) de Almeida, Juliano Milanezi [UNESP]; de Araujo, Nathália Januario [UNESP]; Matheus, Henrique Rinaldi [UNESP]; de Abreu Furquim, Elisa Mara [UNESP]; Piovezan, Bianca Rafaeli [UNESP]; Fiorin, Luiz Guilherme [UNESP]; Ervolino, Edilson [UNESP]; Universidade Estadual Paulista (UNESP)
Objective The aim of this study was to evaluate the effectiveness of hand debridement (HD) versus ultrasonic dental scaler (UDS) for the treatment of experimental periodontitis (EP) in rats. Material and methods Thirty 3-month-old male rats were used. EP was induced around the mandibular first molars (right and left). Seven days after induction, the treatments with either HD (n = 30) or UDS (n = 30) were randomly performed in each molar. Euthanasia were performed at 7, 15, and 30 days after treatment. Histometric (percentage of bone in the furcation [PBF]), histopathological, and immunohistochemical (for detection of tartrate-resistant acid phosphatase [TRAP] and osteocalcin [OCN]). Parametric data (PBF and TRAP) was analyzed by One-way ANOVA followed by Tukey's post-test. OCN was analyzed by Kruskal-Wallis followed by Student-Newman-Keuls post-test. The level of significance was 5%. Results Group HD presented higher PBF and lower TRAP-immunolabeling at 30 days as compared with UDS in the same period (p≤0.05). Group HD presented higher OCN immunolabeling at 30 days as compared with 7 and 15 days (p≤0.05). Persistent and exacerbated inflammatory process was observed in some specimens from group UDS at 30 days, as well as the bone trabeculae presented irregular contour, surrounded by many active osteoclasts. Conclusion Nonsurgical periodontal therapy with HD resulted in higher PBF and lower expression of TRAP as compared with UDS. Also, HD increased the expression of OCN over time.
Role of junctional epithelium in maintaining dento-gingival adhesion and periodontal health
(2023-01-01) Theodoro, Letícia Helena [UNESP]; Garcia, Valdir Gouveia [UNESP]; Ervolino, Edilson [UNESP]; Holcroft, James; McCulloch, Christopher A.; Ganss, Bernhard; Universidade Estadual Paulista (UNESP); Latin American Institute of Dental Research and Education (ILAPEO); University of Toronto
The dento-gingival junction comprises multiple epithelia including the junctional epithelium (JE), which is the most coronally-located structural element of the dento-gingival junction that demarcates external from internal periodontal environments. After tooth eruption into the oral cavity, a specialized basal lamina is formed that provides firm attachment of the JE to the enamel. This attachment prevents microbial species and oral debris from entering subjacent periodontal tissues. Here we discuss the expression of certain JE adhesion molecules and enamel proteins that maintain the health of the dento-gingival junction but that are perturbed in the pathogenesis of periodontitis. We also consider how evolutionary processes have influenced the development of the JE as a specialized adhesion that is well-suited for protection of the dento-gingival junction. A detailed understanding of the biology of the JE will deepen current models of dento-gingival adhesion, potentially clarify inter-patient variability of susceptibility to periodontitis and help to identify new roles of enamel proteins in periodontal regeneration.
Higher expression of Th1/Th2-related cytokines in the intestine of Wistar rats with ligature-induced periodontitis
(2023-06-01) de Mello-Neto, João Martins; Elangovan, Gayathiri; Ervolino, Edilson [UNESP]; Johnson, Newell Walter; Gustafsson, Anders; da Silva Figueredo, Carlos Marcelo; Griffith University; James Cook University; Universidade Estadual Paulista (UNESP); Karolinska Institutet
Objective: This study aimed to investigate the impact of ligature-induced periodontitis (LIP) on histopathological and immunological outcomes in the colon of Wistar rats. Background: It has been repeatedly shown that inflammatory bowel disease (IBD) patients are at higher risk of developing periodontitis and presenting worse oral health than non-IBD patients. However, whether the chronic inflammatory process around teeth contributes to the pathophysiology of IBD needs to be further explored. Materials and Methods: Thirteen Wistar rats were allocated into LIP (n = 7) and controls (n = 6). Half of the colon was processed for histopathological analyses and immunohistochemical (CD45); the other half was homogenized for immunological analyses. Periodontal destruction was confirmed by measuring the distance from the cementum-enamel junction to the mandible's apical position of the mesial interproximal bone. The immunological analyses were performed with the Bio-Plex Th1/Th2 assay. Results: There was a significantly higher interproximal bone loss in LIP compared to controls. The LIP group showed a moderate infiltrate of inflammatory cells, predominantly mononucleated cells in the intestinal tissues. There was significantly higher expression of GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-12 (p70), IL-13, and TNF-α in the intestinal tissues of LIP group compared to controls. Conclusion: Ligature-induced periodontitis was associated with an overexpression of Th1/Th2-related cytokines in the colon of Wistar rats.
Polydioxanone Membrane Compared with Collagen Membrane for Bone Regeneration
(2023-02-01) Quirino, Lilian Caldas [UNESP]; de Azambuja Carvalho, Pedro Henrique [UNESP]; Neto, Renato Torres Augusto [UNESP]; Comachio, Cássio Amaro [UNESP]; Monteiro, Naara Gabriela [UNESP]; Ervolino-Silva, Ana Cláudia [UNESP]; Okamoto, Roberta [UNESP]; Pereira-Filho, Valfrido Antonio [UNESP]; Universidade Estadual Paulista (UNESP)
Guided bone regeneration (GBR) is an approach that induces osteopromotion through the regenerative membranes. These barriers exhibit bioactive behavior and mechanical function. Polydioxanone is a synthetic option, already used in medicine and dentistry, with good results in bone regeneration. This study aimed to evaluate bone repair in critical defects in rat calvaria using a polydioxanone membrane (Plenum® Guide) compared with a commercially available collagen-based membrane (Bio-Gide®). The bone defects were filled with Plenum® Osshp, a synthetic bone graft, hydroxyapatite:β-tricalcium phosphate, 70:30%, Group PG (Plenum® Guide + Plenum® Osshp), and Group BG (Geistlich Bio-Gide® + Plenum® Osshp). The specimens were submitted to immunohistochemical (RUNX2 and OPN), gene expression (RUNX2, IBSP, and VEGF), histometric, and microtomography analyses after 07, 15, 30, and 60 days postoperative. PG group showed greater immunolabeling area for RUNX2 and OPN, higher gene expression of VEGF (3.15 ± 0.85), and IBSP (24.9 ± 0.59). However, there was no statistical difference between groups in the histometric analysis regarding the percentage of connective tissue PG (0.83 ± 0.45), BG (0.70 ± 0.34), neoformed bone PG (0.60 ± 0.4), BG (0.65 ± 0.51), and remaining biomaterial PG (0.84 ± 0.31), BG (0.91 ± 0.33). In addition, there was no statistical difference between groups by micro-CT analysis. The absorbable-synthetic membrane, Plenum® Guide, is an effective membrane for guided bone regeneration.
Inflammatory profile of apical periodontitis exacerbated by cigarette smoke inhalation: Histological and immunohistochemical analysis in rats
(2023-04-01) Vasques, Ana Maria Veiga [UNESP]; da Silva, Ana Claudia Rodrigues [UNESP]; Bueno, Carlos Roberto Emerenciano [UNESP]; Cury, Marina Tolomei Sandoval [UNESP]; Ervolino, Edilson [UNESP]; Cintra, Luciano Tavares Angelo [UNESP]; Dezan Junior, Eloi [UNESP]; Universidade Estadual Paulista (UNESP)
Aim: The aim of this study was to evaluate the effects of cigarette smoke inhalation (CSI) on inflammation, pro-inflammatory mediators and haematological parameters in rats with induced apical periodontitis (AP). Methodology: Thirty-two 3-month-old male Wistar rats were divided into four experimental groups (n = 8): C—Control; S—rats with CSI; AP—rats with AP; and SAP—rats with CSI + AP. Animals in groups S and SAP inhaled cigarette smoke by remaining inside a smoking chamber for 8 min, three times daily, for 50 days. After 20 days of smoke inhalation, animals in AP and SAP groups had the pulps of the lower right first molar exposed to oral environment for 30 days to induce AP. In these subsequent 30 days, animals in group S and SAP continued with CSI. On Day 50, animals were euthanized and mandibles were histologically processed to assess inflammatory infiltrate, immunohistochemical interleukins (IL-1β, IL-6 and TNF-α), and blood samples collected for laboratory analysis. The Mann–Whitney test was performed for non-parametric data and the pairwise analyses of Student's t-test for parametric data, with a significance level of p <.050. Results: Inflammatory infiltrate was moderate in AP group and more severe in the SAP (p =.010). The interleukins IL-6, IL-1β and TNF-α were higher in SAP group (p <.001) when compared to the AP group. A greater number of red blood cells (p =.010), haemoglobin (p =.007) and neutrophils (p =.014) were observed in the SAP group in comparison with the AP group. Conclusion: Cigarette smoke inhalation induced a more severe inflammatory infiltrate, with increased levels of pro-inflammatory cytokines and changes in haematological parameters in rats with induced AP. Thus, CSI aggravated AP, exacerbating the inflammatory response.
Impact of Bifidobacterium animalis subsp. lactis HN019 probiotic in the prevention of periodontitis associated with immunosuppression
(2023-03-01) Maia, Luciana Prado; de Almeida Silva Levi, Yara Loyanne; Henrique Félix Silva, Pedro; Carla Wons, Luana; Pizzo Pitelli, Livia; Goulart de Castro, Jéssica; da Silva Dólens, Eder; Gregorio, Danielle; Gouveia Straioto, Fabiana; dos Santos Santinoni, Carolina; Casarin, Renato; Ervolino, Edilson [UNESP]; Aparecida Chaves Furlaneto, Flávia; Reis Messora, Michel; UNOESTE – University of Western São Paulo; UNOPAR – University Pitágoras UNOPAR; Universidade de São Paulo (USP); Universidade Estadual de Campinas (UNICAMP); Universidade Estadual Paulista (UNESP)
Background: This study evaluated the effects of the probiotic Bifidobacterium animalis subsp. lactis HN019 (HN019) in the development of experimental periodontitis (EP) in rats submitted to chemotherapy (5-fluorouracil [5FU]). Methods: Eighty male rats were divided into the following groups: control (C); treated with 5FU (60 mg/kg at day 30 and 40 mg/kg at day 32); treated with probiotic (HN019) (daily, for 44 days, starting at day 0); treatment with 5FU and probiotic (5FU-HN019); only EP (EP) (ligature placed on lower first molars at day 30, maintained for 14 days); EP and treatment with 5FU (EP-5FU); EP and treatment with probiotic (EP-HN019); and EP and treatment with 5FU and probiotic (EP-5FU-HN019). Euthanasia occurred at day 44. Morphometric, histomorphometric, microtomographic, immunohistochemical, immunoenzymatic, and gene expressions analyses were performed. The data obtained were statistically analyzed (p < 0.05). Results: The EP-5FU-HN019 group showed less bone and connective tissue loss when compared with EP-5FU group, while EP-HN019 and EP-5FU-HN019 groups had greater bone volume than EP and EP-5FU groups, respectively (p < 0.05). A decrease in immunostaining for tartrate-resistant acid phosphatase and RANKL, an increase for osteoprotegerin and lower interleukin-1β levels were observed in EP-5FU-HN019 group, when compared with EP-5FU group (p < 0.0001). Probiotic therapy led to an increase in the proportions of B. lactis in the feces (p = 0.0018), but not in the biofilm, and reduced the expression of Fusobacterium nucleatum and Prevotella intermedia in the biofilm (p < 0.0001). Conclusion: B. lactis HN019 reduced the severity of EP in rats submitted to chemotherapy, modulating immunoinflammatory parameters in periodontal tissues and reducing periodontopathogens expression on biofilm in rats submitted to chemotherapy.
Effectiveness of antimicrobial photodynamic therapy mediated by butyl toluidine blue in preventing medication-related osteonecrosis of the jaws in rats
(2022-12-01) Ervolino, Edilson [UNESP]; Olivo, Mariane Bocalon [UNESP]; Toro, Luan Felipe [UNESP]; Freire, Jéssica de Oliveira Alvarenga [UNESP]; Ganzaroli, Vinícius Franzão [UNESP]; Guiati, Isabella Zacarin [UNESP]; Nuernberg, Marta Aparecida Alberton [UNESP]; Franciscon, João Paulo Soares [UNESP]; Ângelo Cintra, Luciano Tavares [UNESP]; Garcia, Valdir Gouveia; Wainwright, Mark; Theodoro, Leticia Helena [UNESP]; Universidade Estadual Paulista (UNESP); Latin American Institute of Dental Research and Education (ILAPEO); Liverpool John Moores University
Background: Medication-related osteonecrosis of the jaws (MRONJ) is difficult to treat, therefore, prevention would be the ideal clinical approach. This study evaluated the effectiveness of antimicrobial photodynamic therapy (aPDT), mediated by butyl toluidine blue (BuTB) in the prevention of MRONJ-like lesions after tooth extraction in rats. Methods: Twenty-eight senescent female rats were distributed in groups: VEH and VEH-aPDT, treated with vehicle, ZOL and ZOL-aPDT, treated with 100 µg/Kg of zoledronate, both treatments every three days over seven weeks. After three weeks from the commencement of treatment, the mandibular first molar was extracted. For the VEH and ZOL groups, no local treatment was performed, while with the VEH-aPDT and ZOL-aPDT groups, photodynamic treatment was carried out at 0, 2, and 4 days after extraction. For aPDT, 500μl of BuTB solution was deposited on the dental extraction site (0.5 mg/ml; 60 s), followed by irradiation with low-level laser (InGaAIP; 660 nm; 35 mW; 74.2 J/cm²; 60 s). After 28 postoperative days, euthanasia was performed. The hemimandibles were processed to: (1) histological analysis of tissue repair; (2) histometric analysis of the percentage of newly formed bone tissue (PNFBT) and percentage of non-vital bone tissue (PNVBT); (3) immunohistochemical analysis for tartrate-resistant acid phosphatase (TRAP). Results: The ZOL and ZOL-aPDT groups showed less TRAP-positive cells when compared with VEH and VEH-aPDT. The ZOL group demonstrated great compromise in the tissue repair process, consistent with MRONJ-like lesions. VEH, VEH-aPDT and ZOL-aPDT presented a favorable tissue repair process. PNFBT in the ZOL group was lower than in the VEH, VEH-aPDT and ZOL-aPDT groups, whereas PNVBT in the ZOL group was higher than in the VEH, VEH-aPDT and ZOL-aPDT groups. Conclusion: aPDT mediated by BuTB prevented the occurrence of MRONJ-like lesions after tooth extraction in rats.
Colitis induced by dextran sulphate sodium causes histopathological and immunological changes in the periodontal tissues of Wistar rats
(2022-12-01) de Mello-Neto, João Martins; Elangovan, Gayathiri; Ervolino, Edilson [UNESP]; Johnson, Newell Walter; Gustafsson, Anders; da Figueredo, Carlos Marcelo; Griffith University; Universidade Estadual Paulista (UNESP); Karolinska Institutet
Objective: This study investigated the impact of colitis induced by dextran sulphate sodium (DSS)-induced colitis (DIC) on histopathological and immunological outcomes in the periodontal tissues of Wistar rats. Background: Inflammatory bowel diseases (IBD) and periodontitis have been reported to present a bidirectional relationship. However, the inflammatory pathway that connects both diseases needs further investigation. Material and Methods: Twenty-five male Wistar rats were allocated in four groups: unilateral ligature-induced periodontitis for 14 days: LIP (n = 7); dextran sulphate sodium-induced colitis only: DIC (n = 6); DIC + LIP (n = 6) and controls (n = 6). Digital images were obtained from the histological sections. In order to assess the attachment loss (AL), the linear distance between the cementoenamel junction (CEJ) and the alveolar bone crest was measured on the mesial root using histological photomicrography's ImageJ software. Immunological analyses of gingival tissues and plasma were performed by Bio-Plex Th1/Th2 Assay. Results: The DIC group showed inflammatory cells extending to the periodontal connective tissues, which contained significantly elevated expressions of IL-1α, IL-1β, IL-2, IL-6, IL-12, IL-13, GM-CSF, IFN-γ and TNF-α compared to controls. There was no significant difference in bone loss between controls and DIC. There were no significant histopathological differences between DIC + LIP and LIP. However, DIC + LIP presented a significantly lower IL-2 and IL-5 than the LIP group. There was no bone loss difference between LIP+DIC and LIP groups. DIC + LIP group presented significantly higher levels of GM-CSF in plasma. Conclusion: DSS-induced colitis was associated with an overexpression of Th1/Th2- related cytokines in the gingival tissue.
Beta tricalcium phosphate, either alone or in combination with antimicrobial photodynamic therapy or doxycycline, prevents medication-related osteonecrosis of the jaw
(2022-12-01) Hadad, Henrique [UNESP]; Kawamata de Jesus, Laís [UNESP]; Piquera Santos, Ana Flávia [UNESP]; Rinaldi Matheus, Henrique [UNESP]; de Souza Rodrigues, Letícia Gabriella [UNESP]; Paolo Poli, Pier; Marcantonio Junior, Elcio [UNESP]; Pozzi Semeghini Guastaldi, Fernando; Maiorana, Carlo; Milanezi de Almeida, Juliano [UNESP]; Okamoto, Roberta [UNESP]; Ávila Souza, Francisley [UNESP]; Universidade Estadual Paulista (UNESP); University of Milan; Harvard School of Dental Medicine
Surgical trauma in those under a prolonged use of bisphosphonates, can lead to mediation-related osteonecrosis of the jaw (MRONJ). This study aimed to evaluate the preventive therapies for MRONJ. Following four cycles of zoledronic acid administration, Wistar rats had their molar extracted, and were organized into nine treatment groups: negative control group (NCG), treated with saline solution and blood-clot in the alveolus; positive control group (PCG), with blood-clot in the alveolus; BG, β-tricalcium phosphate-based biomaterial; DG, 10% doxycycline gel; aG, antimicrobial photodynamic therapy; and DBG, aBG, aDG, and aDBG, using combination therapy. After 28 days, the lowest bone volume (BV/TV) was reported in PCG (42.17% ± 2.65), and the highest in aDBG (69.85% ± 6.25) (p < 0.05). The higher values of daily mineral apposition rate were recorded in aDBG (2.64 ± 0.48) and DBG (2.30 ± 0.37) (p < 0.001). Moreover, aDBG presented with the highest neoformed bone area (82.44% ± 2.69) (p < 0.05). Non-vital bone was reported only in the PCG (37.94 ± 18.70%). Owing to the key role of the biomaterial, the combination approach (aDBG) was the most effective in preventing MRONJ following tooth extraction.
Effect of maternal true combined endodontic-periodontal lesion on insulin and inflammatory pathway in adult offspring
(2023-04-01) Belardi, Bianca Elvira [UNESP]; Mattera, Maria Sara de Lima Coutinho [UNESP]; Tsosura, Thaís Verônica Saori [UNESP]; dos Santos, Rodrigo Martins [UNESP]; Cantiga-Silva, Cristiane [UNESP]; Tavares Ângelo Cintra, Luciano [UNESP]; Chiba, Fernando Yamamoto [UNESP]; Bravo, Lara Teschi [UNESP]; Carvalho, Nubia Ramos [UNESP]; Carnevali, Ana Carolina Nascimento [UNESP]; de Oliveira, Renan Akira Fujii [UNESP]; Ervolino, Edilson [UNESP]; Matsushita, Doris Hissako [UNESP]; Universidade Estadual Paulista (UNESP)
Background: Maternal periodontal disease (PED) and apical periodontitis (AP) are associated insulin resistance (IR), increased tumor necrosis factor-α (TNF-α) levels, and alterations in insulin signaling (IS) in the gastrocnemius muscle (GM) of adult offspring. TNF-α stimulates I kappa B kinase (IKK) and c-Jun N-terminal protein kinase (JNK), resulting in IS attenuation. However, studies that investigated the maternal true endodontic-periodontal lesion (EPL) in offspring are scarce, and in this case, the impact could be even higher. This study aimed to evaluate the effects of EPL on the IR, IS, and inflammatory pathways on the offspring GM. Methods: Female Wistar rats were distributed into control, AP, PED, and EPL groups. After 30 days of oral inflammation induction, rats from all groups were allowed to mate with healthy rats. The body weight of the offspring was assessed from birth to 75 days of age. After 75 days, the following measurements were performed: glycemia, insulinemia, IR, TNF-α content, and IKKα/β, JNK, pp185 (Tyr), and IRS-1 (Ser) phosphorylation status in the GM. Results: Maternal PED and EPL were associated with low birth weights. All maternal oral inflammations promoted IR and IS impairment in the GM and only maternal PED and EPL caused an increase in TNF-α content and IKKα/β phosphorylation status in the GM of offspring. The offspring of the rats with EPL group showed worsening of metabolic changes when compared with offspring of rats with AP or PED. Conclusion: Association of maternal AP and PED promoted a more pronounced worsening in the health of the adult offspring.
Assessment of redox state and biochemical parameters of salivary glands in streptozotocin-induced diabetic male rats treated with mate tea (Ilex paraguariensis)
(2022-11-01) Gomes, Marco Aurélio [UNESP]; Manzano, Carolina [UNESP]; Alves, Thayane Miranda [UNESP]; Fiais, Gabriela Alice [UNESP]; Freitas, Rayara Nogueira [UNESP]; Coutinho Mattera, Maria Sara de Lima [UNESP]; Dornelles, Rita Cássia Menegati [UNESP]; Matsushita, Doris Hissako [UNESP]; Stevanato Nakamune, Ana Cláudia de Melo [UNESP]; Chaves-Neto, Antonio Hernandes [UNESP]; Universidade Estadual Paulista (UNESP)
Objective: The study aimed to assess the effects of mate tea [Ilex paraguariensis] on the redox state and biochemical parameters of salivary glands in diabetic male rats. Design: Twenty-four male Wistar rats (3 months old) were randomly divided into groups (n = 8 per group): control rats that received water (C); diabetic rats that received water (D); diabetic rats treated with mate tea (DMT). The treated streptozotocin-induced diabetic rats were given mate tea powder by intragastric gavage at a dose of 20 mg/kg daily for 28 days. Content of total protein, amylase, oxidative lipid damage, measured as thiobarbituric acid reactive substances (TBARs), oxidative protein damage, measured as protein carbonyl, total antioxidant capacity, uric acid, reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were examined by the spectrophotometric method in the parotid and submandibular glands. Results: The D group showed lower total protein, amylase, TBARs, protein carbonyl, total antioxidant capacity, GSH, uric acid, and GPx than the C group in both salivary glands, as well as higher SOD and CAT activities. The DMT group showed higher total protein, amylase, total antioxidant capacity, GSH, uric acid, and GPx than the D group in both salivary glands. Moreover, mate tea increased SOD in the parotid gland and CAT in the submandibular gland of diabetic rats but did not influence TBARs and protein carbonyl in either salivary gland compared to D group. Conclusion: Mate tea increased tissue protein synthesis and improved antioxidant defenses in the salivary glands of streptozotocin-induced diabetic male rats.
Influence of age and gender on alveolar bone healing post tooth extraction in 129 Sv mice: a microtomographic, histological, and biochemical characterization
(2023-01-01) Biguetti, Claudia Cristina [UNESP]; Lakkasetter Chandrashekar, Bhuvana; Simionato, Gustavo Baroni [UNESP]; Momesso, Nataira Regina [UNESP]; Duarte, Marco Antonio Hungaro; Rodrigues, Danieli C.; Matsumoto, Mariza Akemi [UNESP]; University of Texas Rio Grande Valley; Universidade Estadual Paulista (UNESP); University of at Dallas; Universidade de São Paulo (USP)
Objectives: To analyze the effect of biological sex and aging on craniofacial bone features in 129 Sv mice and their influence on dental socket healing post tooth extraction. Materials and methods: A total of 52 129 Sv mice were used, of which 28 were young (3–4 months) and 24 were aged (17–18 months), equally distributed according to biological sex. After an upper right incisor extraction, mice specimens were collected at 7, 14, and 21-days post-surgery for microtomographic (microCT) and comprehensive histological analysis. Mandible, skull bones, and maxillae at 21 days were analyzed by microCT, while blood plasma samples were collected for the detection of key bone turnover markers (P1NP and CTX-1) by enzyme-linked immunosorbent (ELISA) assay. Results: Aged females depicted significantly decreased mineralized bone content in alveolar sockets in comparison to young females and aged males at day 7, and aged males at day 14. Mandible RCA and Ma.AR of aged females were also significantly decreased in comparison with young females. Histological evaluation revealed that all alveolar sockets healed at 21 days with inflammation resolution and deposition of new bone. Immunohistochemistry for TRAP revealed increased area density for osteoclasts in alveolar sockets of aged females when compared to young females at 21 days. While a significant increase in CTX-1 levels was detected in blood plasma of aged females when compared to young females, P1NP levels did not significantly change between young and older females. No significant changes were observed for males. Conclusions: Age and gender can significantly affect craniofacial bones of 129 Sv mice, especially maxilla and mandible in females. Considering the altered bone resorption parameters and delayed alveolar bone healing in older females, careful deliberation is necessary during development of pre-clinical models for craniofacial research. Clinical relevance: Aging can be a contributing factor to slower bone healing in craniofacial bones. However, there are no sufficient experimental studies that have addressed this phenomenon along with biological sex taken into consideration.
DNA hydrogels for bone regeneration
(2023-04-25) Athanasiadou, Dimitra; Meshry, Nadeen; Monteiro, Naara G. [UNESP]; Ervolino-Silva, Ana C. [UNESP]; Chan, Ryan Lee; McCulloch, Christopher A.; Okamoto, Roberta [UNESP]; Carneiro, Karina M.M.; University of Toronto; Universidade Estadual Paulista (UNESP)
DNA-based biomaterials have been proposed for tissue engineering approaches due to their predictable assembly into complex morphologies and ease of functionalization. For bone tissue regeneration, the ability to bind Ca2+ and promote hydroxyapatite (HAP) growth along the DNA backbone combined with their degradation and release of extracellular phosphate, a known promoter of osteogenic differentiation, make DNA-based biomaterials unlike other currently used materials. However, their use as biodegradable scaffolds for bone repair remains scarce. Here, we describe the design and synthesis of DNA hydrogels, gels composed of DNA that swell in water, their interactions in vitro with the osteogenic cell lines MC3T3-E1 and mouse calvarial osteoblast, and their promotion of new bone formation in rat calvarial wounds. We found that DNA hydrogels can be readily synthesized at room temperature, and they promote HAP growth in vitro, as characterized by Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, atomic force microscopy, and transmission electron microscopy. Osteogenic cells remain viable when seeded on DNA hydrogels in vitro, as characterized by fluorescence microscopy. In vivo, DNA hydrogels promote the formation of new bone in rat calvarial critical size defects, as characterized by micro-computed tomography and histology. This study uses DNA hydrogels as a potential therapeutic biomaterial for regenerating lost bone.
Resveratrol for preventing medication-related osteonecrosis of the jaws in rats
(2023-01-01) Vitale, Marcelo; Corrêa, Mônica Grazieli; Ervolino, Edilson [UNESP]; Cirano, Fabiano Ribeiro; Ribeiro, Fernanda Vieira; Monteiro, Mabelle Freitas; Casati, Marcio Zaffalon; Pimentel, Suzana Peres; Paulista University; Universidade Estadual Paulista (UNESP)
This study evaluated the effect of resveratrol (RES) on the prevention of medication-related osteonecrosis of the jaws (MRONJ) in ovariectomized (OVX) rats treated with zoledronate (ZOL). Fifty rats were distributed in five groups: SHAM (n = 10): non-ovariectomy + placebo; OVX (n = 10):ovariectomy + placebo; OVX + RES (n = 10):ovariectomy + resveratrol; OVX + ZOL (n = 10):ovariectomy + placebo + zoledronate; and OVX + RES + ZOL (n = 10):ovariectomy + resveratrol + zoledronate. The mandibles left sides were analyzed with micro-CT, histomorphometry, and immunohistochemistry. On the right side, bone markers gene expression was analyzed by qPCR. ZOL increased the percentage of necrotic bone and reduced the neo-formed bone compared to groups not receiving ZOL (p < 0.05). RES impacted the tissue healing pattern in OVX + ZOL + RES, reduced inflammatory cell infiltrate, and improved bone formation in the extraction site. Osteoblasts, alkaline phosphatase (ALP)-, and osteocalcin (OCN)-immunoreactive cells were lower in OVX-ZOL than in SHAM, OVX, and OVX-RES. The OXV-ZOL-RES had fewer osteoblasts and ALP- and OCN-cells than the SHAM and OVX-RES. The tartrate-resistant acid phosphatase (TRAP)-positive cells were reduced in the presence of ZOL (p < 0.05), while the TRAP mRNA levels increased with ZOL treatment, with or without resveratrol, compared with the other groups (p < 0.05). RES alone increased superoxide dismutase levels compared to OVX + ZOL and OVX + ZOL + RES (p < 0.05). In conclusion, resveratrol reduced the tissue impairment severity induced by ZOL; however, it could not prevent the occurrence of MRONJ.
Dose–response effect of Montelukast on post-extraction dental socket repair and skeletal phenotype of mice
(2023-01-01) Ribeiro, Kim Henderson Carmo [UNESP]; da Silva, Raquel Barroso Parra [UNESP]; Roseno, Ana Carolyna Becher [UNESP]; Barreto, Ana Julia Moreno [UNESP]; Bacelar, Ana Carolina Zucon [UNESP]; Ervolino, Edilson [UNESP]; Duarte, Marco Antônio Húngaro; Fakhouri, Walid D.; Chaves-Neto, Antonio Hernandes [UNESP]; Biguetti, Cláudia Cristina; Matsumoto, Mariza Akemi [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); The University of Texas at Health Science Center at Houston (UTH); The University of Texas at Rio Grande Valley (UTRGV)
Bone metabolism and repair are directly regulated by arachidonic acid metabolites. At present, we analyzed the dose–response effects of a selective cysteinyl leukotriene receptor type-1 antagonist during bone repair after tooth extraction and on non-injured skeleton. Sixty-three 129 Sv/Ev male mice composed the groups: C—Control (saline solution); MTK2—2 mg/Kg of Montelukast (MTK) and MTK4–4 mg/Kg of MTK, daily administered by mouth throughout all experimental periods set at 7, 14, and 21 days post-operative. Dental sockets were analyzed by computed microtomography (microCT), histopathology, and immunohistochemistry. Femurs, L5 vertebra and organs were also removed for observation. Blood was collected for plasma bone and liver markers. Histopathology and microCT analysis revealed early socket repair of MTK2 and MTK4 animals, with significant increased BV/TV at days 14 and 21 compared to C. Higher plasma calcium was detected at days 7 and 21 in MTK4 in comparison to C, while phosphate was significantly increased in MTK2 in the same periods in comparison to C and MTK4. No significant differences were found regarding plasma ALP and TRAP, neither for local TRAP and Runx2 immunolabeling at the healing sockets. Organs did not present histological abnormalities. Increased AST levels have been detected in distinct groups and periods. In general, femur phenotype was improved in MTK treated animals. Collectively, MTK promoted early bone formation after tooth extraction and increased bone quality of femurs and vertebra in a time-dose-dependent manner, and should be considered as an alternative therapy when improved post-extraction socket repair or skeleton preservation is required.
High-Intensity Interval Training Minimizes the Deleterious Effects of Arterial Hypertension on the Urinary Bladder of Spontaneously Hypertensive Rats
(2023-01-01) Batista, Victor Rogério Garcia [UNESP]; Correia, Rafael Ribeiro [UNESP]; Fernandes, Vítor S.; Veras, Allice Santos Cruz [UNESP]; Tavares, Maria Eduarda Almeida [UNESP]; Chaves-Neto, Antonio Hernandes [UNESP]; Fiais, Gabriela Alice [UNESP]; De Souza, Francilene Lima Agostinho [UNESP]; Pacagnelli, Francis Lopes; Suggett, Cara Beth; Teixeira, Giovana Rampazzo [UNESP]; Universidade Estadual Paulista (UNESP); Universidad Complutense de Madrid; University of Western São Paulo (UNOESTE); University of Guelph
Arterial hypertension promotes urological complications by modifying the functional capacity of the urinary bladder. On the other hand, physical exercise has been suggested as a nonpharmacological tool to improve blood pressure regulation. High-intensity interval training (HIIT) can effectively increase peak oxygen consumption, body composition, physical fitness, and health-related characteristics of adults; however, its action on the urinary bladder is little discussed. In the present study, we verified the effect of HIIT on the modulation of the redox state, morphology, and inflammatory and apoptotic processes of the urinary bladder of hypertensive rats. Spontaneously hypertensive rats (SHR) were divided into two groups: SHR sedentary and SHR submitted to HIIT. Arterial hypertension promoted an increase in the plasma redox state, modified the volume of the urinary bladder, and increased collagen deposition in detrusor muscle. It was also possible to identify, in the sedentary SHR group, an increase in inflammatory markers such as IL-6 and TNF-α in the urinary bladder, as well as a reduction in BAX expression. However, in the HIIT group, reduced blood pressure levels were observed, together with an improvement in morphology, such as a decrease in collagen deposition. HIIT also regulated the proinflammatory response, promoting increases in IL-10 and BAX expressions and in the number of plasma antioxidant enzymes. The present work highlights the intracellular pathways involved with the oxidative and inflammatory capacity of the urinary bladder and the potential effect of HIIT on the regulation of the urothelium and detrusor muscle of hypertensive rats.
Bactericidal potential of two species typical of Brazilian cerrado: Stryphnodendron obovatum and Lafoensia pacari
(2019-01-01) Dos Santos, Rodrigo Martins [UNESP]; Ferraz, Diego Emanuel; Neto, Maria José; Bernardes, Luiz Gustavo; Brito, Vinícius Martins; Gelle-Oliveira, Glória Maria; Universidade Federal de Mato Grosso do Sul (UFMS); Universidade Estadual Paulista (UNESP)
This study aimed to evaluate the bactericidal potential of plant extracts from medicinal plants of the Brazilian cerrado against microorganisms associated with gastrointestinal infections, through the disc-diffusion method. Two typical cerrado plants were selected: Stryphnodendron obovatum and Lafoensia pacari. The leaves were crushed and reduced to the powder state, and for the extraction of substances 100% methanol was used as the solvent followed by the proportion of 1 g and 2 g of vegetable powder 10 ml of solvent [extracts 1:10 and 2:10 (p/v)] and the microorganisms submitted to the test were Escherichia coli, Staphylococcus aureus and Klebsiella sp. The results showed that L. pacari presented a better percentage of bactericidal activity by the disc-diffusion method in all microorganisms tested with inhibition halos between 15 and 20 mm in both concentrations; S. obovatum showed only antimicrobial activity in S. aureus cultures at both concentrations with inhibition halos between 12 and 16 mm. Both species present great possibilities of being useful in the fight against multiresistant bacteria, allowing the investigation and the isolation guided by bioassays as well as the future identification of new bioactive compounds. It’s concluded that it is necessary to fill gaps related to the scientific knowledge of bioactive compounds produced by species of the Brazilian Cerrado biome.
Evaluation of Vitamin D isolated or Associated with Teriparatide in Peri-Implant Bone Repair in Tibia of Orchiectomized Rats
(2023-02-01) Gomes-Ferreira, Pedro Henrique Silva [UNESP]; Frigério, Paula Buzo [UNESP]; de Moura, Juliana [UNESP]; Duarte, Nathália Dantas [UNESP]; de Oliveira, Danila [UNESP]; Deering, Joseph; Grandfield, Kathryn; Okamoto, Roberta [UNESP]; Universidade Estadual Paulista (UNESP); McMaster University
(1) Background: The objective of this study was to evaluate the morphometry of peri-implant bone tissue in orchiectomized rats, treated with vitamin D isolated or associated with teriparatide. (2) Methods: 24 rats were divided into 4 groups: ORQ—orchiectomy, without drug treatment, ORQ+D—orchiectomy, treated with vitamin D, ORQTERI—orchiectomy, treated with teriparatide and ORQTERI+D—orchiectomy, treated with teriparatide + vitamin D. Each animal received an implant in the tibial metaphysis. Euthanasia occurred 60 days after implant surgery. Computed microtomography (micro-CT) was performed to evaluate the parameters of volume and percentage of bone volume (BV, BV/TV), trabecular thickness (Tb.Th), number and separation of trabeculae (Tb.N, Tb.Sp) and percentage of total porosity (Po-tot). Data were subjected to 1-way ANOVA and Tukey post-test, with a significance level of 5%. (3) Results: For the parameters BV, BV/TV, Tb.Th, the ORQTERI+D group showed the highest values in relation to the other groups and for Po-tot, the lowest values were for ORQTERI+D. For Tb.Sp and Tb.N, there was no statistically significant difference when comparing intragroup results (p > 0.05). (4) Conclusions: It was possible to conclude that treatment with vitamin D associated with teriparatide increases bone volume and improves bone quality.
Performance of Polydioxanone-Based Membrane in Association with 3D-Printed Bioceramic Scaffolds in Bone Regeneration
(2023-01-01) Pitol-Palin, Letícia [UNESP]; Frigério, Paula Buzo [UNESP]; Moura, Juliana [UNESP]; Pilatti, Livia; de Oliveira, Letícia Marques Jordão; Matsubara, Elaine Yoshiko; Tunchel, Samy; Shibli, Jamil Awad; Blay, Alberto; Saska, Sybele; Okamoto, Roberta [UNESP]; Universidade Estadual Paulista (UNESP); Odontológicos e Correlatos S.A; Guarulhos University
This study evaluated the bioactivity of 3D-printed β-tricalcium phosphate (β-TCP) scaffolds or hydroxyapatite (HA) scaffolds associated with polydioxanone (PDO) membrane (Plenum® Guide) for guided bone regeneration in rats. Fifty-four rats were divided into three groups (n = 18 animals): autogenous bone + PDO membrane (Auto/PG); 3D-printed β-TCP + PDO membrane (TCP/PG); and 3D-printed HA + PDO membrane (HA/PG). A surgical defect in the parietal bone was made and filled with the respective scaffolds and PDO membrane. The animals were euthanized 7, 30, and 60 days after the surgical procedure for micro-CT, histomorphometric, and immunolabeling analyses. Micro-CT showed an increase in trabecular thickness and a decrease in trabecular separation, even with similar bone volume percentages between TCP/PG and HA/PG vs. Auto/PG. Histometric analysis showed increased bone formation at 30 days in the groups compared to 7 days postoperatively. Immunolabeling analysis showed an increase in proteins related to bone formation at 30 days, and both groups showed a similar immunolabeling pattern. This study concludes that 3D-printed scaffolds associated with PDO membrane (Plenum® Guide) present similar results to autogenous bone for bone regeneration.

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