AimTo evaluate whether the bioceramic materials Bio‐C Pulpo (Bio‐C, Angelus) and mineral trioxide... more AimTo evaluate whether the bioceramic materials Bio‐C Pulpo (Bio‐C, Angelus) and mineral trioxide aggregate (MTA) Repair HP (MTA‐HP, Angelus) induce fibroblast proliferation and release of interleukin‐10 (IL‐10), an anti‐inflammatory cytokine, stimulating connective tissue remodelling. The tissue response of Bio‐C and MTA‐HP was compared with the White MTA (WMTA; Angelus) since studies have demonstrated that WMTA induces tissue repair.MethodologyBio‐C, MTA‐HP and WMTA were inserted into polyethylene tubes and implanted in the subcutaneous tissue of Holtzman rats for 7, 15, 30 and 60 days. As a control group (CG), empty tubes were implanted subcutaneously. The number of fibroblasts (FB), Ki‐67‐, fibroblast growth factor‐1‐ (FGF‐1) and IL‐10‐immunolabelled cells and collagen content in the capsules was obtained. The data were subjected to two‐way anova followed by Tukey's test (p ≤ .05).ResultsAt 7 days, significant differences in the number of FB were not detected amongst Bio‐C, ...
Several synthetic and natural materials have been studied for the confection of temporary grafts ... more Several synthetic and natural materials have been studied for the confection of temporary grafts for application in regenerative medicine, however, the development of a material with adequate properties remains a challenge, mainly because its degradation kinetics in biological systems. Nature provides materials with noble properties that can be used as such for many applications, thus, taking advantage of the available morphology and assembled structures of plants, we propose to study the vegetable stems for use as temporary graft. Since the in vivo degradation is maybe one of the most important features of the temporary grafts, here we have implanted the plant stems from pumpkin, papaya, and castor into the subepithelial tissue of animals and followed their biodegradation process and the local inflammatory response. Mechanical tests, FTIR and contact angle with water were also analysed. The results indicated the mechanical properties and the contact angle were adequate for use in r...
Vascular endothelial growth factor A (VEGF-A) is expressed by several cell types and is a crucial... more 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.
AimTo evaluate the tissue response promoted by Bio‐C Pulpo (Bio), MTA Repair HP (MTA‐HP) and Whit... more AimTo evaluate the tissue response promoted by Bio‐C Pulpo (Bio), MTA Repair HP (MTA‐HP) and White MTA (WMTA) and whether these materials cause liver changes in a rat experimental model.MethodologyPolyethylene tubes filled with Bio, MTA‐HP and WMTA, and empty tubes (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days. Inflammatory reaction score (IRS), capsule thickness, number of inflammatory cells (IC), von Kossa reaction, interleukin‐6 (IL‐6) and alkaline phosphatase (ALP) immunohistochemistry reactions were performed. Combined methods, von Kossa followed by immunohistochemistry for detection of ALP, were performed. At 60 days, the serum glutamic‐oxaloacetic transaminase (GOT) and glutamic‐pyruvic transaminase (GPT) levels were measured and liver fragments were collected for histological analysis; the data were assessed by one‐way ANOVA analysis followed by Sidak's post‐test. The biocompatibility and bioactivity data were subjecte...
Venlafaxine, a norepinephrine and serotonin reuptake inhibitor, impairs rat sperm parameters, spe... more Venlafaxine, a norepinephrine and serotonin reuptake inhibitor, impairs rat sperm parameters, spermatogenesis and causes high intratesticular estrogen and testosterone levels, indicating that Leydig cells (LCs) may be a venlafaxine target. We evaluated the effect of venlafaxine treatment on LCs in vivo, focusing on adrenergic signaling, EGF immunoexpression and steroidogenesis. Germ cells mitotic/meiotic activity and UCHL1 levels were also evaluated in the seminiferous epithelium. Adult male rats received venlafaxine (30 mg/kg) or distilled water. In testicular sections, the seminiferous tubules, epithelium and the LCs nuclear areas were measured, and the immunoexpression of Ki-67, UCHL1, StAR, EGF, c-Kit and 17β-HSD was evaluated. UCHL1, StAR and EGF protein levels and Adra1a, Nur77 and Ndrg2 expression were analyzed. MDA and nitrite testicular levels, and serum estrogen and testosterone levels were measured. Venlafaxine induced LCs hypertrophy and Ndrg2 upregulation, in parallel to increased number of Ki-67, c-Kit- and 17β-HSD-positive interstitial cells, indicating that this antidepressant stimulates LCs lineage proliferation and differentiation. Upregulation of Adra1a and Nur77 could explain the high levels of StAR and testosterone levels, as well as aromatization. Enhanced EGF immunoexpresion in LCs suggests that this growth fact is involved in adrenergically-induced steroidogenesis, likely via upregulation of Nur77. Slight tubular atrophy and weak Ki-67 immunoexpression in germ cells, in association with high UCHL1 levels, indicate that spermatogenesis is likely impaired by this enzyme under supraphysiological estrogen levels. These data corroborate the unchanged MDA and nitrite levels. Therefore, venlafaxine stimulates LCs steroidogenesis via adrenergic signaling, and EGF may be involved in this process.
Venlafaxine (selective serotonin and norepinephrine reuptake inhibitor) use has increased worldwi... more Venlafaxine (selective serotonin and norepinephrine reuptake inhibitor) use has increased worldwide. However, the impact of venlafaxine on testes and sperm parameters has not been investigated.
To evaluate the inflammatory process induced by Biodentine and mineral trioxide aggregate (MTA) i... more To evaluate the inflammatory process induced by Biodentine and mineral trioxide aggregate (MTA) in rat subcutaneous tissues. A polyethylene tube filled with Biodentine (n = 20) or MTA (n = 20) was placed into the dorsal subcutaneous of forty male rats; in the control group (CG; n = 20), empty tubes were implanted. After 7, 15, 30 and 60 days, the polyethylene tubes surrounded by connective tissue were fixed and embedded in paraffin. The number of inflammatory cells was estimated in HE-stained sections; numerical density of interleukin-6 (IL-6)-immunolabelled cells was also performed. The differences amongst the groups were analysed statistically by Tukey's test (P ≤ 0.05). A high number of inflammatory cells and IL-6-positive cells were observed at 7 days, in all groups; however, in the Biodentine group, the number of inflammatory cells and IL-6-immunolabelled cells was significantly higher (P ≤ 0.05) in comparison with the other groups at 7 and 15 days. In the capsules of anima...
The cauda epididymidis is the major sperm storage region whose androgenic supply, essential for t... more The cauda epididymidis is the major sperm storage region whose androgenic supply, essential for the sperm viability, is provided by the vasculature and is dependent upon testosterone diffusion through the stromal tissue to reach the epithelial cells. We have focused our efforts on examining the regulation of this important epididymal region by evaluating the impact of the androgen disrupter cimetidine on the epithelial–stromal androgenic microenvironment. Male rats received 100 mg/kg cimetidine (CMTG) or saline (CG) for 50 days, serum testosterone levels were measured and the epididymal cauda region was processed for light and transmission electron microscopy. In the proximal cauda region, the duct diameter was measured and birefringent collagen in the stroma was quantified. TUNEL-labeled epithelial cells were quantified, and androgen receptor (AR), karyopherin alpha (KPNA) and sex hormone-binding globulin (SHBG) levels were analyzed by immunofluorescence and Western blot. CMTG show...
OBJECTIVES The present study aimed to compare two different models of orthodontic tooth movement ... more OBJECTIVES The present study aimed to compare two different models of orthodontic tooth movement (OTM) in rats by evaluating tooth movement efficiency and periodontal tissues remodelling. DESIGN Fifteen animals were randomly distributed into 3 groups: control group (untreated); ligature appliance (LA) as experimental OTM using a closed coil spring fixed around maxillary first molar by steel ligature; occlusal appliance (OA) as experimental OTM using a closed coil spring attached on the occlusal surface of the maxillary first molar. After 15 days, all animals were euthanized, and the maxilla of each animal was collected for qPCR, micro-computed tomography, and histological analyses. RESULTS Interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha gene expressions were significantly upregulated in the animals of the LA group as compared to the other groups. No significant difference was observed in tooth displacement between both methods. The LA group presented higher linear bone loss and lower values of bone volume fraction, bone mineral density, trabecular number and increased values of trabecular separation compared to the other groups. The birefringent collagen content in the tension side of the periodontal ligament contained significantly lower collagen content in the LA group than in the control group. Furthermore, on the pressure side, the collagen content was significantly lower in the LA and OA groups than in the control group. CONCLUSIONS The OA group presented little or no deleterious effect on periodontal tissues compared to the LA group, suggesting its use may be more reliable for OTM induction in rats for 15 days.
AimTo evaluate whether the bioceramic materials Bio‐C Pulpo (Bio‐C, Angelus) and mineral trioxide... more AimTo evaluate whether the bioceramic materials Bio‐C Pulpo (Bio‐C, Angelus) and mineral trioxide aggregate (MTA) Repair HP (MTA‐HP, Angelus) induce fibroblast proliferation and release of interleukin‐10 (IL‐10), an anti‐inflammatory cytokine, stimulating connective tissue remodelling. The tissue response of Bio‐C and MTA‐HP was compared with the White MTA (WMTA; Angelus) since studies have demonstrated that WMTA induces tissue repair.MethodologyBio‐C, MTA‐HP and WMTA were inserted into polyethylene tubes and implanted in the subcutaneous tissue of Holtzman rats for 7, 15, 30 and 60 days. As a control group (CG), empty tubes were implanted subcutaneously. The number of fibroblasts (FB), Ki‐67‐, fibroblast growth factor‐1‐ (FGF‐1) and IL‐10‐immunolabelled cells and collagen content in the capsules was obtained. The data were subjected to two‐way anova followed by Tukey's test (p ≤ .05).ResultsAt 7 days, significant differences in the number of FB were not detected amongst Bio‐C, ...
Several synthetic and natural materials have been studied for the confection of temporary grafts ... more Several synthetic and natural materials have been studied for the confection of temporary grafts for application in regenerative medicine, however, the development of a material with adequate properties remains a challenge, mainly because its degradation kinetics in biological systems. Nature provides materials with noble properties that can be used as such for many applications, thus, taking advantage of the available morphology and assembled structures of plants, we propose to study the vegetable stems for use as temporary graft. Since the in vivo degradation is maybe one of the most important features of the temporary grafts, here we have implanted the plant stems from pumpkin, papaya, and castor into the subepithelial tissue of animals and followed their biodegradation process and the local inflammatory response. Mechanical tests, FTIR and contact angle with water were also analysed. The results indicated the mechanical properties and the contact angle were adequate for use in r...
Vascular endothelial growth factor A (VEGF-A) is expressed by several cell types and is a crucial... more 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.
AimTo evaluate the tissue response promoted by Bio‐C Pulpo (Bio), MTA Repair HP (MTA‐HP) and Whit... more AimTo evaluate the tissue response promoted by Bio‐C Pulpo (Bio), MTA Repair HP (MTA‐HP) and White MTA (WMTA) and whether these materials cause liver changes in a rat experimental model.MethodologyPolyethylene tubes filled with Bio, MTA‐HP and WMTA, and empty tubes (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days. Inflammatory reaction score (IRS), capsule thickness, number of inflammatory cells (IC), von Kossa reaction, interleukin‐6 (IL‐6) and alkaline phosphatase (ALP) immunohistochemistry reactions were performed. Combined methods, von Kossa followed by immunohistochemistry for detection of ALP, were performed. At 60 days, the serum glutamic‐oxaloacetic transaminase (GOT) and glutamic‐pyruvic transaminase (GPT) levels were measured and liver fragments were collected for histological analysis; the data were assessed by one‐way ANOVA analysis followed by Sidak's post‐test. The biocompatibility and bioactivity data were subjecte...
Venlafaxine, a norepinephrine and serotonin reuptake inhibitor, impairs rat sperm parameters, spe... more Venlafaxine, a norepinephrine and serotonin reuptake inhibitor, impairs rat sperm parameters, spermatogenesis and causes high intratesticular estrogen and testosterone levels, indicating that Leydig cells (LCs) may be a venlafaxine target. We evaluated the effect of venlafaxine treatment on LCs in vivo, focusing on adrenergic signaling, EGF immunoexpression and steroidogenesis. Germ cells mitotic/meiotic activity and UCHL1 levels were also evaluated in the seminiferous epithelium. Adult male rats received venlafaxine (30 mg/kg) or distilled water. In testicular sections, the seminiferous tubules, epithelium and the LCs nuclear areas were measured, and the immunoexpression of Ki-67, UCHL1, StAR, EGF, c-Kit and 17β-HSD was evaluated. UCHL1, StAR and EGF protein levels and Adra1a, Nur77 and Ndrg2 expression were analyzed. MDA and nitrite testicular levels, and serum estrogen and testosterone levels were measured. Venlafaxine induced LCs hypertrophy and Ndrg2 upregulation, in parallel to increased number of Ki-67, c-Kit- and 17β-HSD-positive interstitial cells, indicating that this antidepressant stimulates LCs lineage proliferation and differentiation. Upregulation of Adra1a and Nur77 could explain the high levels of StAR and testosterone levels, as well as aromatization. Enhanced EGF immunoexpresion in LCs suggests that this growth fact is involved in adrenergically-induced steroidogenesis, likely via upregulation of Nur77. Slight tubular atrophy and weak Ki-67 immunoexpression in germ cells, in association with high UCHL1 levels, indicate that spermatogenesis is likely impaired by this enzyme under supraphysiological estrogen levels. These data corroborate the unchanged MDA and nitrite levels. Therefore, venlafaxine stimulates LCs steroidogenesis via adrenergic signaling, and EGF may be involved in this process.
Venlafaxine (selective serotonin and norepinephrine reuptake inhibitor) use has increased worldwi... more Venlafaxine (selective serotonin and norepinephrine reuptake inhibitor) use has increased worldwide. However, the impact of venlafaxine on testes and sperm parameters has not been investigated.
To evaluate the inflammatory process induced by Biodentine and mineral trioxide aggregate (MTA) i... more To evaluate the inflammatory process induced by Biodentine and mineral trioxide aggregate (MTA) in rat subcutaneous tissues. A polyethylene tube filled with Biodentine (n = 20) or MTA (n = 20) was placed into the dorsal subcutaneous of forty male rats; in the control group (CG; n = 20), empty tubes were implanted. After 7, 15, 30 and 60 days, the polyethylene tubes surrounded by connective tissue were fixed and embedded in paraffin. The number of inflammatory cells was estimated in HE-stained sections; numerical density of interleukin-6 (IL-6)-immunolabelled cells was also performed. The differences amongst the groups were analysed statistically by Tukey's test (P ≤ 0.05). A high number of inflammatory cells and IL-6-positive cells were observed at 7 days, in all groups; however, in the Biodentine group, the number of inflammatory cells and IL-6-immunolabelled cells was significantly higher (P ≤ 0.05) in comparison with the other groups at 7 and 15 days. In the capsules of anima...
The cauda epididymidis is the major sperm storage region whose androgenic supply, essential for t... more The cauda epididymidis is the major sperm storage region whose androgenic supply, essential for the sperm viability, is provided by the vasculature and is dependent upon testosterone diffusion through the stromal tissue to reach the epithelial cells. We have focused our efforts on examining the regulation of this important epididymal region by evaluating the impact of the androgen disrupter cimetidine on the epithelial–stromal androgenic microenvironment. Male rats received 100 mg/kg cimetidine (CMTG) or saline (CG) for 50 days, serum testosterone levels were measured and the epididymal cauda region was processed for light and transmission electron microscopy. In the proximal cauda region, the duct diameter was measured and birefringent collagen in the stroma was quantified. TUNEL-labeled epithelial cells were quantified, and androgen receptor (AR), karyopherin alpha (KPNA) and sex hormone-binding globulin (SHBG) levels were analyzed by immunofluorescence and Western blot. CMTG show...
OBJECTIVES The present study aimed to compare two different models of orthodontic tooth movement ... more OBJECTIVES The present study aimed to compare two different models of orthodontic tooth movement (OTM) in rats by evaluating tooth movement efficiency and periodontal tissues remodelling. DESIGN Fifteen animals were randomly distributed into 3 groups: control group (untreated); ligature appliance (LA) as experimental OTM using a closed coil spring fixed around maxillary first molar by steel ligature; occlusal appliance (OA) as experimental OTM using a closed coil spring attached on the occlusal surface of the maxillary first molar. After 15 days, all animals were euthanized, and the maxilla of each animal was collected for qPCR, micro-computed tomography, and histological analyses. RESULTS Interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha gene expressions were significantly upregulated in the animals of the LA group as compared to the other groups. No significant difference was observed in tooth displacement between both methods. The LA group presented higher linear bone loss and lower values of bone volume fraction, bone mineral density, trabecular number and increased values of trabecular separation compared to the other groups. The birefringent collagen content in the tension side of the periodontal ligament contained significantly lower collagen content in the LA group than in the control group. Furthermore, on the pressure side, the collagen content was significantly lower in the LA and OA groups than in the control group. CONCLUSIONS The OA group presented little or no deleterious effect on periodontal tissues compared to the LA group, suggesting its use may be more reliable for OTM induction in rats for 15 days.
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