Publications by authors named "Paulo Sérgio Cerri"

49 Publications

Impaired macrophages and failure of steroidogenesis and spermatogenesis in rat testes with cytokines deficiency induced by diacerein.

Histochem Cell Biol 2021 Sep 13. Epub 2021 Sep 13.

Laboratory of Histology and Embryology, Department of Morphology, Genetics, Orthodontics and Pediatric Dentistry, Dental School - São Paulo State University (UNESP), Rua Humaitá, 1680, Araraquara, SP, CEP: 14801-903, Brazil.

The role of cytokines in testicular function under normal conditions has not been completely understood. Here, we evaluated testicular macrophages (TM), steroidogenesis by Leydig cells (LC) and seminiferous tubules integrity in cytokines-deficient rat testes induced by diacerein, an anti-inflammatory drug that inhibits interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α). Male rats received daily 100 mg/kg of diacerein (DIAG; n = 8) or saline (CG; n = 8) for 30 days. Serum testosterone (T) levels were measured and the seminiferous tubule (ST) area, epithelial area (EA), frequency of damaged ST and number of Sertoli cells (SC) were evaluated. TUNEL method and immunoreactions for detection of pro-IL-1β, TNF-α, steroidogenic acute regulatory protein (StAR), 17β-hydroxysteroid dehydrogenase (17β-HSD), androgen receptor (AR) and scavenger receptor for hemoglobin-haptoglobin complexes (CD163), a TM marker, were performed. Testicular AR, 17β-HSD and IL-1β levels were detected by Western blot. Data were submitted to Student t test (p ≤ 0.05). In DIAG, T and testicular AR, 17β-HSD and IL-1β levels decreased significantly (p < 0.05). The number of TUNEL-positive interstitial cells increased and LC showed weak StAR, 17β-HSD and AR immunoexpression in association with reduced IL-1β immunoexpression and number of CD163-positive TM in the interstitial tissue from diacerein-treated rats. Numerous damaged ST were found in DIAG, and reduction in the EA were associated with germ cells death. Moreover, the number of SC reduced and weak AR and TNF-α immunoexpression was observed in SC and germ cells, respectively. The cytokines deficiency induced by diacerein impairs TM, LC and spermatogenesis, and points to a role of IL-1β in steroidogenesis under normal conditions. In the ST, the weak AR and TNF-α immunoexpression in SC and germ cells, respectively, reinforces the idea that TNF-α plays a role in the SC androgenic control.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00418-021-02023-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8436873PMC
September 2021

Spatio-temporal immunolocalization of VEGF-A, Runx2, and osterix during the early steps of intramembranous ossification of the alveolar process in rat embryos.

Dev Biol 2021 Oct 7;478:133-143. Epub 2021 Jul 7.

São Paulo State University (UNESP), School of Dentistry, Araraquara - Department of Morphology, Genetics, Orthodontics and Pediatric Dentistry - Laboratory of Histology and Embryology, Araraquara, SP, Brazil. Electronic address:

Vascular endothelial growth factor A (VEGF-A) is expressed by several cell types and is a crucial factor for angiogenic-osteogenic coupling. However, the immunolocalization of VEGF-A during the early stages of the alveolar process formation remains underexplored. Thus, we analyzed the spatio-temporal immunolocalization of VEGF-A and its relationship with Runt-related transcription factor 2 (Runx2) and osterix (Osx) during the early steps of intramembranous ossification of the alveolar process in rat embryos. Embryo heads (E) of 16, 18 and 20-day-old rats were processed for paraffin embedding. Histomorphometry and immunohistochemistry to detect VEGF-A, Runx2, and Osx (osteoblast differentiation markers) were performed. The volume density of bone tissue including bone cells and blood vessels increased significantly in E18 and E20. Cells showing high VEGF-A immunoreactivity were initially observed within a perivascular niche in the ectomesenchyme; afterwards, these cells were diffusely located near bone formation sites. Runx2-and Osx-immunopositive cells were observed in corresponded regions of cells showing strong VEGF-A immunoreactivity. Although these immunostained cells were observed in all specimens, this immunolocalization pattern was more evident in E16 specimens and gradually decreased in E18 and E20 specimens. Double immunofluorescence labelling showed intracellular co-localization of Osx and VEGF-A in cells surrounding the developing alveolar process, indicating a crucial role of VEGF-A in osteoblast differentiation. Our results showed VEGF-A immunoexpression in osteoblasts and its precursors during the maxillary alveolar process formation of rat embryos. Moreover, the VEGF-A-positive cells located within a perivascular niche at the early stages of the alveolar process development suggest a crosstalk between endothelium and ectomesenchymal cells, reinforcing the angiogenic-osteogenic coupling in this process.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ydbio.2021.07.001DOI Listing
October 2021

Biocompatibility and bioactive potential of the NeoMTA Plus endodontic bioceramic-based sealer.

Restor Dent Endod 2021 Feb 17;46(1):e4. Epub 2020 Dec 17.

Department of Morphology, Genetics, Orthodontics and Pediatric Dentistry, Laboratory of Histology and Embryology, Dental School, São Paulo State University (UNESP), Araraquara, SP, Brazil.

Objectives: This study evaluated the biocompatibility and bioactive potential of NeoMTA Plus mixed as a root canal sealer in comparison with MTA Fillapex.

Materials And Methods: Polyethylene tubes filled with NeoMTA Plus ( = 20), MTA Fillapex ( = 20), or nothing (control group, CG; = 20) were inserted into the connective tissue in the dorsal subcutaneous layer of rats. After 7, 15, 30 and 60 days, the specimens were processed for paraffin embedding. The capsule thickness, collagen content, and number of inflammatory cells (ICs) and interleukin-6 (IL-6) immunolabeled cells were measured. von Kossa-positive structures were evaluated and unstained sections were analyzed under polarized light. Two-way analysis of variance was performed, followed by the Tukey test ( ≤ 0.05).

Results: At 7 days, the capsules around NeoMTA Plus and MTA Fillapex had more ICs and IL-6-immunostained cells than the CG. However, at 60 days, there was no significant difference in the IC number between NeoMTA Plus and the CG ( = 0.1137) or the MTA Fillapex group ( = 0.4062), although a greater number of IL-6-immunostained cells was observed in the MTA Fillapex group ( = 0.0353). From 7 to 60 days, the capsule thickness of the NeoMTA Plus and MTA Fillapex specimens significantly decreased, concomitantly with an increase in the collagen content. The capsules around root canal sealers showed positivity to the von Kossa stain and birefringent structures.

Conclusions: The NeoMTA Plus root canal sealer is biocompatible and exhibits bioactive potential.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5395/rde.2021.46.e4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7906839PMC
February 2021

Effects of obesity on periodontal tissue remodeling during orthodontic movement.

Am J Orthod Dentofacial Orthop 2021 Apr 6;159(4):480-490. Epub 2021 Feb 6.

Department of Diagnosis and Surgery, School of Dentistry at Araraquara, São Paulo State University, Araraquara, São Paulo, Brazil. Electronic address:

Introduction: Orthodontic movement triggers a sequence of cellular and molecular events that may be affected by different systemic conditions. This study evaluated the effect of obesity on rat periodontal tissue remodeling induced by mechanical orthodontic force.

Methods: Thirty-two Holtzman rats were distributed into 4 groups: control, obesity induction (O), orthodontic movement (M), and obesity induction and orthodontic movement (OM). Obesity was induced by a high-fat diet for 90 days. After 15 days of orthodontic movement, the animals were killed. Obesity induction was confirmed by animal body weight, adipose tissue weight, and serologic analysis. Periodontal tissue remodeling was evaluated using microcomputed tomography and histologic analysis. The gene expression of adipokines and cytokines in gingival tissues was evaluated.

Results: An increase in body and adipose tissue weight was observed in the obesity induction groups. The O group presented an increase in lipids and blood glucose. The OM group showed a decrease in bone volume fraction and bone mineral density compared with all other groups and a tendency for more rapid tooth movement than the M group. The OM group showed a higher quantity of inflammatory cells and higher Mmp1 expression than the O group. The O and OM groups showed higher Nampt expression than the control group and lower Nampt expression than the M group.

Conclusions: Obesity modulates periodontal tissue remodeling during orthodontic movement and results in more inflammation and bone loss than in nonobese animals.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ajodo.2019.12.025DOI Listing
April 2021

Immunoexpression pattern of autophagy mediators in alveolar bone osteoclasts following estrogen withdrawal in female rats.

J Mol Histol 2021 Apr 6;52(2):321-333. Epub 2021 Jan 6.

Araraquara - Laboratory of Histology and Embryology, School of Dentistry, São Paulo State University (UNESP), Araraquara, SP, Brasil.

It is known that estrogen deficiency increases osteoclast formation and activity. Autophagy, a cell survival pathway, has been shown to be crucial for osteoclast function. However, little is known about the effects of estrogen depletion on osteoclast autophagy. Here, we evaluated the effects of estrogen deficiency in the immunoexpression of autophagy mediators in alveolar bone osteoclasts of ovariectomized rats. Twelve adult female rats were ovariectomized (OVX-group) or SHAM-operated (SHAM-group). After three weeks, the rats were euthanized and maxillary fragments containing alveolar bone of the first molars were processed for light microscopy or transmission electron microscopy (TEM). Paraffin-sections were subjected to the TRAP method (osteoclast marker) or to the immunohistochemical detections of beclin-1, LC3α, and p62 (autophagy mediators); araldite-sections were processed for TEM. The number of TRAP-positive osteoclasts and the number of immunolabeled-multinucleated cells (MNCs) along the alveolar bone surface of the first molar were computed. The number of TRAP-positive osteoclasts and the number of beclin-1-, LC3α- and p62-immunolabelled osteoclasts were significantly higher in OVX-group than the SHAM-group. MNCs were frequently located juxtaposed to Howship lacunae along the alveolar bone surface, indicating that these cells are osteoclasts. TEM revealed osteoclasts exhibiting autophagosomes. Our data indicate that autophagy plays an important role during estrogen deficiency-induced osteoclastogenesis. Thus, our results contribute to a better understanding on the role of autophagy on osteoclasts under estrogenic deficiency, and reinforce the idea that modulation of autophagy may be a useful tool to inhibit excessive oral bone resorption in post-menopausal women.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10735-020-09953-xDOI Listing
April 2021

The effect of amoxicillin on dental enamel development in vivo.

Braz Oral Res 2020 Sep 4;34:e116. Epub 2020 Sep 4.

Universidade Estadual Paulista - Unesp, School of Dentistry, Department of Morphology, Araraquara, SP, Brazil.

The exposure to amoxicillin has been associated with molar incisor hypomineralization. This study aimed to determine if amoxicillin disturbs the enamel mineralization in in vivo experiments. Fifteen pregnant rats were randomly assigned into three groups to received daily phosphatase-buffered saline or amoxicillin as either 100 or 500 mg/kg. Mice received treatment from day 13 of pregnancy to day 40 postnatal. After birth, the offsprings from each litter continued to receive the same treatment according to their respective group. Calcium (Ca) and phosphorus (P) content in the dental hard tissues were analyzed from 60 upper first molars and 60 upper incisors by the complexometric titration method and colorimetric analysis using a spectrophotometer at 680 nm, respectively. Lower incisors were analyzed by X-ray microtomography, it was measured the electron density of lingual and buccal enamel, and the enamel and dentin thickness. Differences in Ca and P content and electron density among the groups were analyzed by one-way ANOVA. There was no significant difference on enamel electron density and thickness among the groups (p > 0.05). However, in incisors, the higher dose of amoxicillin decreased markedly the electron density in some rats. There were no statistically significant differences in Ca (p = 0.180) or P content (p = 0.054), although the higher dose of amoxicillin could affect the enamel in some animals. The amoxicillin did not significantly alter the enamel mineralization and thickness in rats.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1590/1807-3107bor-2020.vol34.0116DOI Listing
September 2020

Biocompatibility and Bioactive Potential of New Calcium Silicate-based Endodontic Sealers: Bio-C Sealer and Sealer Plus BC.

J Endod 2020 Oct 17;46(10):1470-1477. Epub 2020 Jul 17.

Department of Restorative Dentistry, School of Dentistry, São Paulo State University, Araraquara, São Paulo, Brazil.

Introduction: Bio-C Sealer (BC; Angelus, Londrina, PR, Brazil) and Sealer Plus BC (SPBC; MK Life, Porto Alegre, Brazil) are new ready-to-use bioceramic endodontic sealers. The aim of this study was to evaluate the biocompatibility and bioactive potential of BC and SPBC sealers in comparison with AH Plus (AHP; Dentsply DeTrey Gmbh, Konstanz, Germany) in subcutaneous tissue of rats.

Methods: Polyethylene tubes filled with materials and empty tubes, serving as the control group, were implanted in the subcutaneous tissues of rats. After 7, 15, 30, and 60 days, the tubes with connective tissue were removed, and inflammatory cells (ICs)/mm and immunolabeled cells for interleukin (IL)-6 were evaluated. Osteocalcin and von Kossa analysis were also performed. Data were submitted to analysis of variance and Tukey tests, with a significance level of 5%.

Results: At 7 days, SPBC showed lower ICs than BC (P < .05). AHP exhibited greater immunolabeled cells for IL-6 (P < .05). After 15 days, BC showed lower ICs and IL-6 compared with other materials. At 30 days, SPBC and AHP showed higher values for ICs (P < .05). After 60 days, calcium silicate sealers did not show statistical difference (P > .05) for ICs and IL-6, with values lower than AHP (P < .05). The materials showed positive structures to von Kossa staining. BC exhibited osteocalcin labeling in all periods. SPBC showed osteocalcin labeling from 15-60 days. AHP and the control group did not exhibit osteocalcin labeling.

Conclusions: BC and SPBC sealers are biocompatible and present bioactive potential.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.joen.2020.07.011DOI Listing
October 2020

Immunoinflammatory response and bioactive potential of GuttaFlow bioseal and MTA Fillapex in the rat subcutaneous tissue.

Sci Rep 2020 04 28;10(1):7173. Epub 2020 Apr 28.

Department of Morphology, Genetics, Orthodontics and Pediatric Dentistry - Laboratory of Histology and Embryology, Dental School - São Paulo State University (UNESP), Araraquara, SP, Brazil.

To evaluate the effect of GuttaFlow bioseal (GFB) and MTA Fillapex (MTAF) in comparison with Endofill (EF) in the subcutaneous tissue. Polyethylene tubes with GFB, MTAF, EF or empty tubes (control group; CG) were implanted into subcutaneous of rats. After 7, 15, 30 and 60 days, the capsule thickness, inflammatory reaction, interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), caspase-3, TUNEL-positive cells, von Kossa and ultrastructural features were evaluated. The data were statistically analyzed (p ≤ 0.05). At all periods, the number of IL-6- and VEGF-immunolabelled cells, and capsule thickness were lower in GFB than MTAF, which was lower than EF (p < 0.0001). At 60 days, the number of inflammatory cells was similar in GFB and MTAF (p = 0.58). Significant differences in the number of TUNEL- and caspase-3-positive cells were not observed among GFB, MTAF and CG whereas the highest values were found in EF specimens. The EF specimens exhibited several cells with condensed chromatin, typical of apoptosis. von Kossa-positive and birefringent structures were only observed in GFB and MTAF, suggesting the presence of calcite crystals. Taken together, these results show that cellular and structural damage induced by GFB and MTAF sealers were recovery over time. Moreover, these sealers express bioactive potential in subcutaneous tissue.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41598-020-64041-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7188821PMC
April 2020

Muscular atrophy, impaired epithelial autophagy and UCHL1 increase in androgen-deficient cauda epididymis.

Reproduction 2020 05;159(6):693-705

Laboratory of Histology and Embryology, Department of Morphology, Genetics, Orthodontics and Pediatric Dentistry, Dental School - São Paulo State University (UNESP/FOAr), Araraquara, São Paulo, Brazil.

In epididymis, cimetidine induces androgenic failure due to reduced sex hormone-binding globulin stromal levels and blockade of androgen receptor (AR) nuclear import. UCHL1, a hydrolase of ubiquitin-proteasome system (UPS), seems to play a role in autophagy and apoptotic pathway. However, the role of UPS and autophagy in epididymis has not been clarified. We evaluated UCHL1 and autophagy in epididymal cauda epithelium under androgenic deficiency induced by cimetidine, focusing on the interplay among these processes and apoptosis. The integrity of epididymal muscular layer was also evaluated. Male rats received cimetidine (CMTG) or saline (CG). Seminal vesicles were weighed, the expression of androgen-responsive genes Crisp1 and connexin 43 (Cx43) in cauda epididymis was evaluated, and cauda fragments were processed for light and transmission electron microscopy. The epithelium height and muscular thickness were measured. TUNEL, immunohistochemistry for caspase-3 and Cx43, and immunofluorescence for AR, Bcl-2, UCHL1, MAP LC3A, and p62/SQSTM1 (autophagic markers) were performed. Bcl-2, UCHL1, and Cx43 were detected by Western blot. In CMTG, the reduction in seminal vesicles weight accompanied by downregulation of Crisp1 and Cx43 confirmed epididymal androgenic failure. These results were associated with muscular atrophy, apoptosis and weak Cx43 and AR immunoexpression, supporting the androgenic dependence of muscular integrity. The high UCHL1 levels and reduction in Bcl-2 reinforce UCHL1 role in epithelial cells death. The intense immunoexpression of LC3A and p62/SQSTM1 indicates autophagic disturb, which in association with high UCHL1 levels, points to a role of UPS and autophagy in the regulation of epididymal epithelial cells viability under androgenic control.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1530/REP-19-0462DOI Listing
May 2020

Physical Properties, Antimicrobial Activity and In Vivo Tissue Response to Apexit Plus.

Materials (Basel) 2020 Mar 5;13(5). Epub 2020 Mar 5.

Laboratory of Histology and Embryology, Dental School - São Paulo State University (UNESP), Araraquara 14801-903, Brazil.

We investigated the physical properties, antimicrobial activity, and tissue reaction to Apexit Plus in comparison to Sealapex. Flow, radiopacity, setting time, and solubility were evaluated in each material. The antimicrobial activity against was performed. Polyethylene tubes containing Apexit Plus or Sealapex, and without material (control group) were implanted into the subcutaneous tissue of rats. At 7, 15, 30, and 60 days of implantation, the specimens were paraffin-embedded and the number of inflammatory cells (ICs) and the amount of birefringent collagen (BC) were quantified. The von Kossa reaction followed by immunohistochemistry for detection of alkaline phosphatase (ALP) was also performed. Statistical analysis was performed with ANOVA and Tukey test ( ≤ 0.05). The flow value of Apexit Plus was greater than Sealapex, whereas the radiopacity (3.44 mm Al) was lower than Sealapex (6.82 mm Al). Apexit Plus showed lower solubility and shorter initial and final setting ( 0.0001), whereas the antimicrobial activity was significantly greater than Sealapex. Although the number of ICs was higher in Apexit Plus ( = 0.0009) at 7 days, no significant difference was detected between Apexit Plus and Sealapex at 15, 30, and 60 days. All groups showed higher values for BC in the capsules over time. ALP-immunolabelled cells were observed, mainly around von Kossa-positive structures, either in the capsules of Apexit Plus or Sealapex. Therefore, our results revealed that Apexit Plus exhibited a greater effectiveness against and better physical properties than Sealapex, except for the radiopacity. In vivo findings indicate that Apexit Plus is biocompatible and presents potential bioactivity in the subcutaneous tissue.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/ma13051171DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7085033PMC
March 2020

Severe magnesium deficiency compromises systemic bone mineral density and aggravates inflammatory bone resorption.

J Nutr Biochem 2020 03 26;77:108301. Epub 2019 Nov 26.

Department of Diagnosis and Surgery, Sao Paulo State University, School of Dentistry at Araraquara-UNESP, Araraquara, 14801-930, Sao Paulo, Brazil.

We sought to evaluate the effects of magnesium (Mg) intake deficiency on bone metabolism in rats with induced periodontal disease (PD). Holtzman rats were randomly divided into two groups: Control - animals fed a standard diet and test - animals fed a diet with 90% Mg deficiency. After 60 days on the diets, all animals received ligature on the lower left first molars to induce PD. Animals were euthanized after 30 days following ligature placement. Blood and urine were collected for determination of serum concentrations of Mg, calcium, osteocalcin (OCN), alkaline phosphatase and parathyroid hormone (PTH) by enzyme-linked immunosorbent assay, and the urinary concentration of deoxypyridinoline (DPD). Systemic bone mineral density (BMD), bone volume and architectural bone parameters were evaluated by micro-CT in L4 lumbar vertebrae and mandible. Tartrate-resistant acid phosphatase staining and immunohistochemical (IHC) analysis of inducible nitric oxide synthase (iNOS), Runt-related transcription factor 2 (RUNX2), CD86, CD80, proliferating cell nuclear antigen, vascular endothelial growth factor, OCN and osteopontin were investigated. Reverse-transcription polymerase chain reaction was employed to assess mRNA expression of receptor-activator of nuclear factor-kB ligand, osteoprotegerin (OPG) and interleukin (IL)-6. Mg deficiency was associated with higher concentrations of PTH and DPD, and significant decrease on both systemic and mandibular BMD, as well as greater severity of alveolar and trabecular bone loss. Significant increase in osteoclasts was observed in the test group with PD. IHC analysis showed significant increase in the expression of iNOS and decreased expression of OCN and RUNX2. Increased IL-6 mRNA and decreased OPG mRNA expressions were evidenced in the test group with PD. Mg deficiency caused systemic effects indicative of altered bone metabolism in the vertebrae and affected both immune and stromal cells, aggravating inflammatory bone resorption in the ligature-induced model of periodontitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jnutbio.2019.108301DOI Listing
March 2020

Vitamin B Prevents Cimetidine-Induced Androgenic Failure and Damage to Sperm Quality in Rats.

Front Endocrinol (Lausanne) 2019 10;10:309. Epub 2019 Jul 10.

Laboratory of Histology and Embryology, Department of Morphology, Dental School - São Paulo State University (UNESP/FOAr), Araraquara, Brazil.

Cimetidine, used as an anti-ulcer and adjuvant treatment in cancer therapy, causes disorders in the male reproductive tract, including steroidogenesis. However, its effect on sperm quality and male fertility has been poorly addressed. Since vitamin B has demonstrated to recover spermatogonia number and sperm concentration in cimetidine-treated rats, we evaluated the impact of cimetidine on sperm quality and fertility potential and whether vitamin B is able to prevent the harmful effect of this drug on steroidogenesis and sperm parameters. Adult male rats were treated for 52 consecutive days as follows: cimetidine group (100 mg/kg of cimetidine), cimetidine/vitamin B group (100 mg/kg of cimetidine + 3 μg vitamin B), vitamin B group (3 μg vitamin B) and control group (saline). Serum testosterone levels and immunofluorescence associated to western blot for detection of 17β-HSD6 were performed. Sperm morphology and motility, mitochondrial activity, acrosome integrity, DNA integrity by Comet assay, lipid peroxidation as well as fertility potential were analyzed in all groups. Apoptotic spermatids were also evaluated by caspase-3 immunohistochemistry. In the cimetidine-treated animals, reduced serum testosterone levels, weak 17β-HSD6 levels and impaired spermiogenesis were observed. Low sperm motility and mitochondrial activity were associated with high percentage of sperm tail abnormalities, and the percentage of spermatozoa with damaged acrosome and DNA fragmentation increased. MDA levels were normal in all groups, indicating that the cimetidine-induced changes are associated to androgenic failure. In conclusion, despite the fertility potential of rats was unaffected by the treatment, the sperm quality was significantly impaired. Therefore, considering a possible sperm-mediated transgenerational inheritance, the long term offspring health needs to be investigated. The administration of vitamin B to male rats prevents the androgenic failure and counteracts the damage inflicted by cimetidine upon sperm quality, indicating that this vitamin may be used as a therapeutic agent to maintain the androgenic status and the sperm quality in patients exposed to androgen disrupters.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fendo.2019.00309DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6635670PMC
July 2019

Effects of an Er,Cr:YSGG Laser on Bone Regeneration in Critical-Sized Calvarial Defects of Rats Exposed to Inhalation of Cigarette Smoke.

Photobiomodul Photomed Laser Surg 2019 Aug 3;37(8):500-507. Epub 2019 Jul 3.

2Department of Diagnosis and Surgery, School of Dentistry at Araraquara, Sa∼o Paulo State University-Unesp, Araraquara, Brazil.

To evaluate the effect of the Er,Cr:YSGG laser on healing of critical-sized calvarial defects (CSDs) in rats submitted to inhalation of cigarette smoke. Smoking has been implicated with the delay in the bone healing after osteotomy procedures, then the use of the Er,Cr:YSGG laser for osteotomy in smokers could be an alternative to the conventional drills. One hundred animals were randomly allocated into four groups: trephine-the CSDs were made with a trephine drill in healthy rats; Er,Cr:YSGG-the CSDs were made with the Er,Cr:YSGG laser in healthy rats; Trephine-S-the CSDs were made with a trephine drill in rats exposed to cigarette smoke; and Er,Cr:YSGG-S-the CSDs were made with the Er,Cr:YSGG laser in rats exposed to cigarette smoke. The inhalation of cigarette smoke started 7 days before the surgical procedure until euthanasia (immediately, 7, 15, 30, or 60 days after the surgical procedure). A histometric analysis and a histological description were performed to evaluate (1) the residual linear lengths and bone formation in the CSDs; (2) the quality of bone healing. The use of Er,Cr:YSGG laser induces more bone formation compared with the trephine in smokers; however, the closure of the CSD was only superior in the Er,Cr:YSGG-S group compared to the Trephine-S group at the 60-day period. The use of the Er,Cr:YSGG laser stimulated the bone repair process after osteotomy procedures in animals submitted to exposure of inhalation of cigarette smoke.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/photob.2018.4546DOI Listing
August 2019

The influence of Cisplatin-based chemotherapy on the osseointegration of dental implants: An in vivo mechanical and histometrical study.

Clin Oral Implants Res 2019 Jul 11;30(7):603-616. Epub 2019 May 11.

Department of Diagnosis and Surgery, São Paulo State University (UNESP), School of Dentistry, Araraquara, Brazil.

Objectives: To evaluate the effect of Cisplatin on bone repair and mineralization around implants and on the mechanical properties of bone tissue.

Materials And Methods: Forty-three Wistar rats were randomly divided into two groups: Cisplatin (CIS, medication) and control (CTL, placebo solution), administered once a week for 4 weeks. After 4 weeks, implants were installed in both tibiae metaphysis. After 30 and 60 days, the animals were sacrificed and their femurs and tibiae were removed. Femurs were subjected to mechanical tests and tibiae for removal torque, arrangement and distribution of collagen fibers, morphometrical analyses (bone tissue in contact with the implant surface [BIC] and areal fraction between implant threads occupied by bone tissue [BAFO]) and scanning electron microscopy to calcium and calcium/phosphorus analysis. Data were analyzed by ANOVA or MANOVA, and Tukey or Games-Howell post hoc tests, respectively (α = 0.05).

Results: The CTL specimens had significantly higher values (0.0001 ≤ p≤0.036) of strength (N), removal torque (N/cm ), %BIC, and %BAFO than CIS specimens, being their best results at day 60. No significant differences were found among the groups regarding the values of deformation, percentage of calcium, and calcium/phosphorus ratio. In CIS groups, there was a reduction in the organization of collagen at the bone/implant interface, resulting in a trabecular bone with thin trabeculae and birefringent collagen and irregular arrangement.

Conclusions And Clinical Implications: Cisplatin interfered negatively in the repair and mineralization around dental implants, as well as on the quality of the bone tissue, mainly in the period of 30 days after the implant placement.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/clr.13445DOI Listing
July 2019

Systemic administration of curcumin or piperine enhances the periodontal repair: a preliminary study in rats.

Clin Oral Investig 2019 Aug 29;23(8):3297-3306. Epub 2018 Nov 29.

Department of Diagnosis and Surgery, School of Dentistry at Araraquara - Univ Estadual Paulista (UNESP), Rua Humaita, 1680 - Centro, Araraquara, SP, 14801-903, Brazil.

Objectives: Studies have documented the anti-inflammatory effects of spices, which may be related to treatment of chronic diseases. The purpose of this study was to evaluate the influence of curcumin and piperine and their association on experimental periodontal repair in rats.

Materials And Methods: Periodontitis was induced via the installation of a ligature around the first molar. After 15 days, the ligatures were removed, and the rats were separated into groups (12 animals per group): (i) curcumin, (ii) piperine, (iii) curcumin+piperine, (iv) corn oil vehicle, and (v) control group (animals had ligature-induced periodontitis but were not treated). The compounds were administered daily, for 15 days by oral gavage. Animals were euthanized at 5 and 15 days, and hemimaxillae and gingival tissues were harvested. Bone repair was assessed by μCT (microcomputer tomography). Histological sections were stained with hematoxylin/eosin (H/E) for the assessment of cellular infiltrate or picrosirius red for quantification of collagen content, and subjected to immunohistochemistry for detecting NF-ĸB. Gingival tissues were used to evaluate levels of TGF-β and IL-10 (ELISA).

Results: Curcumin and piperine increased the TGF-β level, significantly improved the collagen repair, and decreased the cellularity and activation of NF-ĸB in the periodontal tissues, but only curcumin caused a significant increase in early bone repair.

Conclusion: Curcumin and piperine promoted a substantive effect on tissue repair; however, there was not synergistic effect of compounds administered in combination.

Clinical Relevance: Curcumin and piperine stimulates the tissue repair and may be potential candidates for the treatment of periodontal disease.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00784-018-2755-9DOI Listing
August 2019

Matrix Metalloproteinase-1 and Acid Phosphatase in the Degradation of the Lamina Propria of Eruptive Pathway of Rat Molars.

Cells 2018 Nov 10;7(11). Epub 2018 Nov 10.

Laboratory of Histology and Embryology-Araraquara, School of Dentistry, São Paulo State University (UNESP), 1680 Centro, CEP 14801⁻903 Araraquara, SP, Brazil.

The comprehension of dental pathogenesis and disorders derived from eruption failure requires a deep understanding of the molecular mechanisms underlying normal tooth eruption. As intense remodelling is needed during tooth eruption, we hypothesize that matrix metalloproteinase-1 (MMP-1) and acid phosphatase (ACP) play a role in the eruptive pathway degradation. We evaluated MMP-1-immunoexpression and the collagen content in the lamina propria at different eruptive phases. Immunohistochemistry and ultrastructural cytochemistry for detection of ACP were also performed. In the maxillary sections containing first molars of 9-, 11-, 13-, and 16-day-old rats, the birefringent collagen of eruptive pathway was quantified. MMP-1 and ACP-2 immunohistochemical reactions were performed and the number of MMP-1-immunolabelled cells was computed. Data were analyzed by one-way ANOVA and Tukey post-test ( ≤ 0.05). ACP cytochemistry was evaluated in specimens incubated in sodium β-glycerophosphate. In the eruptive pathway of 13- and 16-day-old rats, the number of MMP-1-immunolabelled cells increased concomitantly to reduction of collagen in the lamina propria. Enhanced ACP-2-immunolabelling was observed in the lamina propria of 13- and 16-day-old rats. Fibroblasts and macrophages showed lysosomes and vacuoles containing fragmented material reactive to ACP. MMP-1 degrades extracellular matrix, including collagen fibers, being responsible for the reduction in the collagen content during tooth eruption. The enhanced ACP activity at the mucosal penetration stage indicates that this enzyme plays a role in the degradation of remnant material, which is engulfed by macrophages and fibroblasts of the eruptive pathway. Therefore, enzymatic failure in the eruptive pathway may disturbs tooth eruption.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/cells7110206DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6262441PMC
November 2018

Antimicrobial Photodynamic Therapy Mediated by Curcumin-Loaded Polymeric Nanoparticles in a Murine Model of Oral Candidiasis.

Molecules 2018 Aug 19;23(8). Epub 2018 Aug 19.

School of Dentistry, São Paulo State University (UNESP), Araraquara 14801-903, São Paulo, Brazil.

Antimicrobial photodynamic therapy (aPDT) has been proposed as an alternative method for oral candidiasis (OC), while nanocarriers have been used to improve the water solubility of curcumin (CUR). The aim of this study is to encapsulate CUR in polymeric nanoparticles (NPs) and to evaluate its photodynamic effects on a murine model of OC. Anionic and cationic CUR-NP is synthesized using poly-lactic acid and dextran sulfate and then characterized. Female mice are immunosuppressed and inoculated with (Ca) to induce OC. aPDT is performed by applying CUR-NP or free CUR on the dorsum of the tongue, followed by blue light irradiation for five consecutive days. Nystatin is used as positive control. Afterward, Ca are recovered and cultivated. Animals are euthanized for histological, immunohistochemical, and DNA damage evaluation. Encapsulation in NP improves the water solubility of CUR. Nystatin shows the highest reduction of Ca, followed by aPDT mediated by free CUR, which results in immunolabelling of cytokeratins closer to those observed for healthy animals. Anionic CUR-NP does not show antifungal effect, and cationic CUR-NP reduces Ca even in the absence of light. DNA damage is associated with Ca infection. Consecutive aPDT application is a safe treatment for OC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/molecules23082075DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6222858PMC
August 2018

Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions.

J Microbiol Methods 2018 09 11;152:52-60. Epub 2018 Jul 11.

Department of Dental Materials and Prosthodontics, Oral Rehabilitation Program, Araraquara School of Dentistry UNESP, Univ. Estadual Paulista, Centro, 14801903 Araraquara, SP, Brazil. Electronic address:

The aim of this study was to (i) design, develop and validate a practical and physiologically relevant reconstituted in vitro oral mucosa tissue model and (ii) to assess its applicability in in vitro host-pathogen interactions with C. albicans and S. aureus. Co-culture organotypic constructions were created by incorporating specific numbers of keratinocytes (NOK-si) onto cellularised, collagen gel scaffolds containing human gingival fibroblasts incubated in KGM media and cultured for 14 days. The detection of the appropriate oral mucosa/epithelial structure was evaluated by histology (hematoxylin and eosin (HE), periodic acid-Schiff (P.A.S.) and Picrosirius red), and immunocytochemistry (cytokeratin 13, cytokeratin 14, Ki-67 and collagen IV) compared to a normal human gingiva. The morphology of the reconstituted tissue was analyzed by Transmission Electron Microscopy. To further quantitate tissue damage, lactate dehydrogenase (LDH) was measured in the tissue supernatant. NOK-si grown upon a gingival scaffold provided an organotypic model in an in vitro setting and exhibited structural characteristics typically associated with normal oral mucosa. Immunocytochemistry revealed the detection of epithelial cytokeratins 13 and 14, Col IV and Ki-67 in the reconstituted oral mucosa model. Infection was detected after 8 h and 16 h. This study presents an in vitro cellularised, organotypic model of reconstituted oral mucosa, which enables close control and characterization of its structure and differentiation over a mid-length period of time in culture.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.mimet.2018.07.004DOI Listing
September 2018

Biodentine and MTA modulate immunoinflammatory response favoring bone formation in sealing of furcation perforations in rat molars.

Clin Oral Investig 2019 Mar 7;23(3):1237-1252. Epub 2018 Jul 7.

UNESP - Univ Estadual Paulista, Dental School, Department of Morphology, Laboratory of Histology and Embryology, Rua Humaitá, 1680, Centro, Araraquara, SP, 14801-903, Brazil.

Objectives: Evaluate the tissue reaction of periodontium subjacent to furcation perforations in rat molars sealed with Biodentine or mineral trioxide aggregate (MTA).

Materials And Methods: The pulp chamber floor of right upper first molars of 60 rats was perforated and filled with Biodentine, MTA, or cotton pellet (sham); the left first molars were used as control. After 7, 15, 30, and 60 days, maxillary fragments were processed for paraffin-embedding. The periodontal space (PS), volume density of inflammatory cells (VvIC) and fibroblasts (VvFb), number of osteoclasts, and collagen content were obtained. Interleukin-6 (IL-6) and osterix (osteoblast marker) were detected by immunohistochemistry. The data were submitted to ANOVA and Tukey's test (p ≤ 0.05).

Results: At 7 days, high values in VvIC, IL-6-immunolabeled cells, and osteoclasts were accompanied by reduced collagen content in enlarged PS of experimental groups. At all periods, VvIC, number of osteoclasts and IL-6, and PS were higher in sham than in Biodentine and MTA (p < 0.0001). From 7 to 60 days, significant reduction in VvIC, IL-6 immunoexpression, and osteoclasts was accompanied by significant increase in VvFb, osteoblasts, and collagen in Biodentine and MTA groups. At 60 days, significant differences in VvIC, PS, IL-6, osteoclasts, and osteoblasts were not found between Biodentine and MTA. Significant differences in the osteoclast number were not observed among Biodentine, MTA, and control groups while osteoblasts number was higher in Biodentine and MTA groups.

Conclusions: Despite the initial inflammatory reaction and bone resorption, the sealing of furcation perforations with Biodentine and MTA favors the repair of periodontal tissues.

Clinical Relevance: Biodentine and MTA exhibit potential as repair material in the treatment of furcation perforations.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00784-018-2550-7DOI Listing
March 2019

Differential effects of natural Curcumin and chemically modified curcumin on inflammation and bone resorption in model of experimental periodontitis.

Arch Oral Biol 2018 Jul 10;91:42-50. Epub 2018 Apr 10.

Department of Restorative Dentistry School of Dentistry of Riberão Preto University of São Paulo Riberão Preto SP Brazil. Electronic address:

Objective: The purpose of this study was to compare the effects of the oral administration of natural curcumin and a chemically modified curcumin (CMC2.24) on osteoclast-mediated bone resorption, apoptosis, and inflammation in a murine model of experimental periodontal disease.

Design: Fifty male rats were distributed among the following treatment groups: (i) 2% carboxymethylcellulose, (ii) CMC2.24 30 mg/kg body weight, (iii) Curcumin 100 mg/kg body weight and (iv) no treatment. Compounds were administered daily by oral intubation over a 15-day period of time. Periodontal disease was induced by injections of LPS (lipopolysaccharide) into the gingival tissues three times per week. Contralateral sides were injected with the same volume of PBS (phosphate buffered saline) vehicle. After 15 days, hemimaxillae and gingival tissues were harvested. Bone resorption was assessed by μCT (microcomputer tomography). Formalin-fixed, paraffin embedded histological sections were stained with haematoxylin/eosin (H/E) for the assessment of cellular infiltrate or subjected to immunohistochemistry for detecting TRAP (tartrate-resistant acid phosphatase)-positive cells and caspase-3. Apoptosis was assessed in the gingival tissues by DNA fragmentation.

Results: CMC2.24 and curcumin caused a significant reduction of the inflammatory cell infiltrate, however μCT analysis showed that only CMC2.24 reduced bone resorption and the number of TRAP-positive multinucleated cells (osteoclasts). Curcumin, but not CMC2.24, significantly reduced the number of apoptotic cells in the gingival tissues and of osteocytes in the alveolar bone crest.

Conclusions: The results suggest that CMC2.24 and curcumin inhibit inflammation by different mechanisms, but only CMC2.24 was capable of reducing alveolar bone resorption in the LPS-induced model of periodontitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.archoralbio.2018.04.007DOI Listing
July 2018

Reduced interleukin-6 immunoexpression and birefringent collagen formation indicate that MTA Plus and MTA Fillapex are biocompatible.

Biomed Mater 2018 02 20;13(3):035002. Epub 2018 Feb 20.

Department of Restorative Dentistry, São Paulo State University (UNESP), School of Dentistry, Araraquara, Brazil.

Considering that endodontic sealers release some components which may promote delay in the repair process, our purpose was to evaluate the tissue reaction promoted by MTA Plus and MTA Fillapex in comparison with AH Plus (standard control) and Endofill, which has a long clinical track record. One hundred rats were distributed into five groups: MTA Plus (Avalon Biom Inc., Bradenton, FL, USA), MTA Fillapex (Angelus, Londrina, PR, Brazil), AH Plus (Dentsply DeTrey GmbH, Konstanz, Germany), Endofill (Dentsply, Petrópolis, RJ, Brazil) and CG (control group, empty polyethylene tubes). The polyethylene tubes filled with sealers or empty (CG) were implanted into subcutaneous. After 7, 15, 30 and 60 days, the tubes surrounded by capsules were paraffin-embedded. In HE-stained sections, the volume density of inflammatory cells (VvIC) was estimated in the capsules. The number of interleukin-6-immunolabelled cells (IL-6), a pro-inflammatory cytokine, was also computed in the capsules. The birefringent collagen content was quantified in picrosirius-stained sections. Data were analysed by ANOVA and Tukey tests (p ≤ 0.05). At 7 days, the capsules showed moderate inflammatory reaction. In all groups, VvIC and IL-6-immunostained cells reduced significantly from 7 to 60 days. At 60 days, IL-6 immunoexpression was reduced significantly in MTA Plus and MTA Fillapex in comparison with AH Plus; no difference was found in the VvIC among MTA Plus, MTA Fillapex, AH Plus and CG whereas Endofill exhibited the highest VvIC. The reduction in VvIC was parallel to an increase in the collagen in all the groups, except Endofill. MTA Plus, MTA Fillapex and AH Plus induce a response that culminates in the regression of inflammation and formation of a fibrous capsule over time. The lower IL-6 immunoexpression in the capsules of MTA Plus and MTA Fillapex than AH Plus suggests that the immune response is suppressed more rapidly in the MTA-based sealers.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1088/1748-605X/aaa1f5DOI Listing
February 2018

Osteoporosis and autophagy: What is the relationship?

Rev Assoc Med Bras (1992) 2017 Feb;63(2):173-179

PhD, Adjunct Professor (Habilitation: BR. Livre-docente) of the Department of Morphology, Laboratory of Histology and Embryology, Faculty of Dentistry of Araraquara, Universidade Estadual Paulista (Unesp), Araraquara, SP, Brazil.

Autophagy is a survival pathway wherein non-functional proteins and organelles are degraded in lysosomes for recycling and energy production. Therefore, autophagy is fundamental for the maintenance of cell viability, acting as a quality control process that prevents the accumulation of unnecessary structures and oxidative stress. Increasing evidence has shown that autophagy dysfunction is related to several pathologies including neurodegenerative diseases and cancer. Moreover, recent studies have shown that autophagy plays an important role for the maintenance of bone homeostasis. For instance, in vitro and animal and human studies indicate that autophagy dysfunction in bone cells is associated with the onset of bone diseases such as osteoporosis. This review had the purpose of discussing the issue to confirm whether a relationship between autophagy dysfunction and osteoporosis exits.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1590/1806-9282.63.02.173DOI Listing
February 2017

Cimetidine Reduces Interleukin-6, Matrix Metalloproteinases-1 and -9 Immunoexpression in the Gingival Mucosa of Rat Molars With Induced Periodontal Disease.

J Periodontol 2017 01 2;88(1):100-111. Epub 2016 Sep 2.

Department of Morphology, Dental School, Laboratory of Histology and Embryology, University Estadual Paulista, Araraquara, São Paulo, Brazil.

Background: Histamine seems to act, via H receptor, on inflammatory processes by stimulating interleukin (IL)-6 and matrix metalloproteinase (MMP) release. As cimetidine is an H receptor antagonist, the authors hypothesize that this antiulcer drug reduces IL-6, MMP-1, and MMP-9 immunoexpression in gingiva with induced periodontal disease (PD). To confirm a possible modulatory role of IL-6 on MMPs, the relationship between IL-6/MMP-1 and IL-6/MMP-9 immunoexpression was evaluated.

Methods: Forty-six male rats were distributed into the cimetidine group (CimG: received daily intraperitoneal injections of 100 mg/kg of body weight of cimetidine) or the saline group (SG). PD was induced by cotton ligature around the maxillary left first molars (PDSG and PDCimG). The right molars were used as controls (SG and CimG). After 7, 15, 30, and 50 days, maxillary fragments were processed for paraffin embedding or for transmission electron microscopy. Tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in the alveolar process surface and number of IL-6, MMP-1, and MMP-9-immunolabeled cells in the gingival mucosa were quantified. Statistical analyses were performed (P ≤0.05).

Results: In PDSG and PDCimG, gingival mucosa exhibited few collagen fibers among numerous inflammatory cells. In PDCimG, the number of TRAP-positive osteoclasts and IL-6, MMP-1, and MMP-9-immunolabeled cells was significantly lower than in PDSG at all periods. A positive correlation between IL-6/MMP-1 and IL-6/MMP-9 was detected in PDSG and PDCimG.

Conclusion: Cimetidine decreases bone loss through reduction of osteoclast number and induces reduction of IL-6, MMP-1, and MMP-9 immunoexpression, reinforcing the idea that the beneficial effect of cimetidine in PD may be due to reduction of IL-6 immunolabeling in the inflamed gingival mucosa.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1902/jop.2016.160132DOI Listing
January 2017

Seminiferous epithelium damage after short period of busulphan treatment in adult rats and vitamin B efficacy in the recovery of spermatogonial germ cells.

Int J Exp Pathol 2016 08 31;97(4):317-328. Epub 2016 Aug 31.

Department of Morphology, Laboratory of Histology and Embryology, Dental School - São Paulo State University (UNESP), Araraquara, Brazil.

Several different strategies have been adopted in attempt to recover from chemotherapy-damaged spermatogenesis that is often seen in oncologic patients. In this study, we have evaluated the impact of short period of exposure to busulphan on the haemogram and seminiferous epithelium of adult rats, focusing on spermatogonial depletion and Sertoli cell (SC) integrity. We then examined whether vitamin B supplementation improves the haematological parameters and spermatogonia number. The animals received 10 mg/kg of busulphan (BuG) or busulfan+vitamin B (Bu/B G) on the first and fourth days of treatment. In H.E.-stained testicular sections, the areas of the seminiferous tubule (ST) and seminiferous epithelium were measured. The number of spermatogonia in H.E-stained and PCNA-immunolabelled testicular sections was quantified. The frequency of tubules with abnormal SC nuclei or TUNEL-positive SC was evaluated. Vimentin immunofluorescence in ST was also evaluated. In BuG and Bu/B G, the animals showed leukopenia and thrombocytopenia, but the body weight reduced only in BuG. The areas of ST and seminiferous epithelium decreased in Bu/B G and BuG. In BuG, the number of H.E.-stained and PCNA-immunolabelled spermatogonia reduced significantly. The frequency of tubules containing abnormal SC nuclei and TUNEL-positive SC increased and the vimentin immunoexpression pattern changed. In Bu/B G, the number of H.E.-stained or PCNA-immunolabelled spermatogonia increased fourfold in comparison with BuG. The structural changes in ST after 6 days of busulphan exposure may be associated with the potential effect of this anti-neoplastic agent on SC. The increased number of spermatogonia in the busulphan-treated animals receiving vitamin B indicates that this vitamin can be an adjuvant therapy to improve the fertility in male cancer patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/iep.12195DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5061760PMC
August 2016

Rheological Characterization and Safety Evaluation of Non-Ionic Lamellar Liquid Crystalline Systems Containing Retinyl Palmitate.

J Biomed Nanotechnol 2016 Feb;12(2):394-403

Retinyl palmitate (RP) is widely used as a special interest ingredient in dermatological formulations to improve the elasticity of the skin and to reduce wrinkles by stimulating collagen synthesis. Nanotechnology-based drug delivery systems, such as liquid crystalline systems (LCSs), can modulate drug permeation into skin and improve the drug action. The effects of such systems on the skin, however, are not completely known. Possible undesirable effects of these formulations on the skin can be detected and interpreted by histopathology and histomorphometry. The objective of this study was to perform a rheological characterization to evaluate the safety of RP used in a lamellar LCS in vitro and in vivo. LCSs containing polyether functional siloxane as a surfactant, silicon glycol copolymer as an oil phase and water at ratios of 60:10:30 and 40:30:30, with (F₁v and F₂v, respectively) and without (F₁ and F₂ respectively) RP, were investigated. The rheological characterization was performed using steady shear rate sweep tests and dynamic frequency sweep tests carried out for up to 30 days for various storage temperature conditions (25 ± 2 °C, 37 ± 2 °C and 5 ± 2 °C). Cytotoxic effects were evaluated using J-774 mouse macrophages as a cellular model system. The in vivo tests were conducted on rabbits that had areas of skin treated as follows for 15 days: C (Control); F₁; F₁v; F₂; and F₂v. Histomorphometric and histopathological techniques were used to estimate the thicknesses of the epidermis and stratum corneum and the numbers of fibroblasts and leukocytes in the papillary dermis. Mean values were compared by ANOVA, followed by the Tukey test (p < 0.05). The steady shear rate sweep and dynamic frequency sweep tests confirmed the high viscosity of the LCS and the typical pseudo-plastic characteristic of the lamellar system. The RP-unloaded LCS and the RP-loaded LCS did not produce cytotoxicity, nor did they provoke significant thickening of the epidermis and stratum corneum. The number of leukocytes in the treated areas did not change; however, the number of fibroblasts in the area treated with F₁v was higher than in the areas treated with the control and F₂. The histological analyses demonstrated that none of the formulations irritated the skin and that formulation F₁v significantly increased the number of fibroblasts in the dermis, which could result in an increase in the production of collagen.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1166/jbn.2016.2165DOI Listing
February 2016

Biology of Bone Tissue: Structure, Function, and Factors That Influence Bone Cells.

Biomed Res Int 2015 13;2015:421746. Epub 2015 Jul 13.

Department of Morphology, Laboratory of Histology and Embryology, Dental School, Universidade Estadual Paulista (UNESP), 14801-903 Araraquara, SP, Brazil.

Bone tissue is continuously remodeled through the concerted actions of bone cells, which include bone resorption by osteoclasts and bone formation by osteoblasts, whereas osteocytes act as mechanosensors and orchestrators of the bone remodeling process. This process is under the control of local (e.g., growth factors and cytokines) and systemic (e.g., calcitonin and estrogens) factors that all together contribute for bone homeostasis. An imbalance between bone resorption and formation can result in bone diseases including osteoporosis. Recently, it has been recognized that, during bone remodeling, there are an intricate communication among bone cells. For instance, the coupling from bone resorption to bone formation is achieved by interaction between osteoclasts and osteoblasts. Moreover, osteocytes produce factors that influence osteoblast and osteoclast activities, whereas osteocyte apoptosis is followed by osteoclastic bone resorption. The increasing knowledge about the structure and functions of bone cells contributed to a better understanding of bone biology. It has been suggested that there is a complex communication between bone cells and other organs, indicating the dynamic nature of bone tissue. In this review, we discuss the current data about the structure and functions of bone cells and the factors that influence bone remodeling.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1155/2015/421746DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4515490PMC
April 2016

Apoptosis and reduced microvascular density of the lamina propria during tooth eruption in rats.

J Anat 2015 Oct 30;227(4):487-96. Epub 2015 Jul 30.

Dental School - Laboratory of Histology and Embryology, UNESP - São Paulo State University, Araraquara, SP, Brazil.

During tooth eruption, structural and functional changes must occur in the lamina propria to establish the eruptive pathway. In this study, we evaluate the structural changes that occur during lamina propria degradation and focus these efforts on apoptosis and microvascular density. Fragments of maxilla containing the first molars from 9-, 11-, 13- and 16-day-old rats were fixed, decalcified and embedded in paraffin. The immunohistochemical detection of vascular endothelial growth factor (VEGF), caspase-3 and MAC387 (macrophage marker), and the TUNEL method were applied to the histological molar sections. The numerical density of TUNEL-positive cells and VEGF-positive blood vessel profiles were also obtained. Data were statistically evaluated using a one-way anova with the post-hoc Kruskal-Wallis or Tukey test and a significance level of P ≤ 0.05. Fragments of maxilla were embedded in Araldite for analysis under transmission electron microscopy (TEM). TUNEL-positive structures, fibroblasts with strongly basophilic nuclei and macrophages were observed in the lamina propria at all ages. Using TEM, we identified processes of fibroblasts or macrophages surrounding partially apoptotic cells. We found a high number of apoptotic cells in 11-, 13- and 16-day-old rats. We observed VEGF-positive blood vessel profiles at all ages, but a significant decrease in the numerical density was found in 13- and 16-day-old rats compared with 9-day-old rats. Therefore, the establishment of the eruptive pathway during the mucosal penetration stage depends on cell death by apoptosis, the phagocytic activity of fibroblasts and macrophages, and a decrease in the microvasculature due to vascular cell death. These data point to the importance of vascular rearrangement and vascular neoformation during tooth eruption and the development of oral mucosa.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/joa.12359DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4580106PMC
October 2015

Amoxicillin diminishes the thickness of the enamel matrix that is deposited during the secretory stage in rats.

Int J Paediatr Dent 2016 May 7;26(3):199-210. Epub 2015 Jul 7.

Department of Pediatric Dentistry, Dental School, UNESP - University Estadual Paulista, Araraquara, Brazil.

Background: The use of amoxicillin during early childhood has been associated with molar incisor hypomineralization.

Aim: The objective of this study was to determine whether the use of amoxicillin interferes with enamel development, during secretion and early mineralization stages.

Design: Fifteen pregnant rats were randomly assigned to three groups that received physiological solution (sham group), 100 mg/kg/day amoxicillin (A100G), and 500 mg/kg/day amoxicillin (A500G). After birth, the pups in each group received the same treatment until post-natal day 7 or 12. The upper first molars were analyzed histomorphometrical and immunostaining with amelogenin on day 7, and MMP-20 on day 12 was performed using a semiquantitative method (H-score).

Results: At 7 days, several vacuolar structures were observed in the ameloblasts in the A100G and A500G groups. A significant reduction of the enamel thickness (P < 0.001) was found in amoxicillin-treated rats compared with the sham group. Significant differences were not observed in enamel thickness (P > 0.05) between the groups of 12-day-old rats. Moreover, significant differences were not observed in the number of amelogenin- and MMP-20-immunolabeled ameloblasts (P > 0.05) between groups.

Conclusion: The present results suggest that amoxicillin interferes with the initial stages of amelogenesis by causing structural changes in the ameloblasts and a reduction of the enamel matrix.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/ipd.12184DOI Listing
May 2016

Treatment of periodontal disease with an Er,Cr:YSGG laser in rats exposed to cigarette smoke inhalation.

Lasers Med Sci 2015 Nov 27;30(8):2095-103. Epub 2015 Mar 27.

Araraquara Dental School-São Paulo University "Júlio de Mesquita Filho", Araraquara, São Paulo, Brazil.

The purpose of this study was to evaluate the erbium, chromium:yttrium-scandiumgallium-garnet (Er,Cr:YSGG) laser irradiation in the treatment of periodontitis in rats exposed to cigarette smoke inhalation (CSI). Ligatures were placed in the maxillary second molars. After a 15-day period, the ligatures were removed and 180 animals were randomly divided into six groups: (1) CSRP group--CSI and manual scaling and root planing (SRP) treatment; (2) CL group--CSI and Er,Cr:YSGG laser irradiation; (3) CSRP + L group-CSI, SRP, and Er,Cr:YSGG irradiation; (4) SRP group-manual SRP; (5) L group--Er,Cr:YSGG irradiation; (6) SRP + L group--SRP and Er,Cr:YSGG irradiation. At 7, 15, and 30 days after treatments, animals were euthanized and histologic, histometric, immunohistochemistry, and real-time PCR analyses were performed. Histometrically, no differences were observed in the SRP, L, and SRP + L groups exposed to CSI. The CSRP group showed more bone formation at 30 days than at 15 days (p < 0.01) but less bone at 30 days than the CL group at 30 days (p < 0.05). Immunohistochemical staining was positive for osteoblasts, fibroblasts, and osteoclasts. Real-time PCR showed more (vascular endothelial growth factor) VEGF expression in the L (p < 0.05) and SRP + L (p < 0.01) groups at 30 days than at 15 days and less VEGF expression in the CSRP group at 30 days than at 15 days (p < 0.05). There was no difference in fibroblast growth factor (FGF) expression. The Er,Cr:YSGG laser irradiation promotes favorable conditions for tissue repair even in animals exposed to CSI, with similar results as those achieved from manual scaling and root planing.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10103-015-1731-8DOI Listing
November 2015

Effect of Er,Cr:YSGG laser application in the treatment of experimental periodontitis.

Lasers Med Sci 2015 Apr 30;30(3):993-9. Epub 2014 Jan 30.

Araraquara Dental School-São Paulo State University "Júlio de Mesquita Filho", Araraquara, São Paulo, Brazil.

The purpose of this study was to evaluate the influence of an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser in the absence or presence of manual scaling and root planning (SRP) for the treatment of induced periodontitis in rats. Ligatures were placed in the subgingival region of the maxillary first molar. After a 7-day period, the ligatures were removed, and 40 rats were randomly divided into four groups (G), as follows: (GI) no treatment, (GII) scaling and root planning (SRP) with curettes, (GIII) Er,Cr:YSGG laser irradiation and (GIV) SRP with curettes followed by Er,Cr:YSGG laser irradiation. Seven and 30 days after the treatment, the animals were sacrificed and histologic, histometric and immunohistochemistry analyses were performed. All groups showed similar histopathological characteristics during the evaluation period. The histometric analysis was confirmed using Bonferroni and paired t tests. At 7 and 30 days, groups II, III and IV exhibited greater bone formation in the furcation area when compared to group I (p < 0.0001; p < 0.05). During the 7-day period, the groups irradiated with the laser (III and IV) showed a statistically larger new bone area than the group treated with SRP (II) (p < 0.01). Immunohistochemistry analysis revealed that the control group exhibited a higher expression of tartrate-resistant acid phosphatase (TRAP) and the receptor activator of nuclear factor κΒ ligand (RANKL) when compared to groups II, III and IV (p < 0.05). All treatments were able to reduce the inflammatory processes, consequently enabling the repair of periodontal tissues. The results achieved with the application of the Er,Cr:YSGG laser suggest that this laser can stimulate greater bone formation, especially over a shorter period of time.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10103-014-1526-3DOI Listing
April 2015
-->