Combined effects of bone morphogenetic protein-7 and mineral trioxide aggregate on the proliferation, migration, and differentiation of human dental pulp stem cells

Bioactive molecules present the potential to be used along with biomaterials in vital pulp therapy and regenerative endodontic treatment. Objective: The aim of this study was to assess the effects of the combined use of bone morphogenetic protein-7 (BMP-7) and mineral trioxide aggregate (MTA) on the proliferation, migration, and differentiation of human dental pulp stem cells (DPSCs). Methodology: For the proliferation analysis, DPSCs were incubated with a growth medium and treated with MTA and/or BMP-7 at different concentrations. For the following analyses, DPSCs were incubated with a differentiation medium and treated with MTA and/or BMP-7. Moreover, there were groups in which DPSCs were incubated with the growth medium (control), the differentiation medium, or DMEM/F12 containing fetal bovine serum, and not treated with MTA or BMP-7. Cell proliferation was analyzed using the WST-1 assay. The odontogenic/osteogenic differentiation was evaluated by immunocytochemistry, alkaline phosphatase (ALP) activity assay, alizarin red staining, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell migration was evaluated using a wound-healing assay. Data were analyzed using analysis of variance and Tukey test (p=0.05). Results: The use of BMP-7 with MTA presented no significant effect on cell proliferation in comparison with the treatment with MTA alone (p>0.05), but showed higher ALP activity, increased mineralization, and higher expression of DMP1 and DSPP when compared with other groups (p<0.05). Nestin expression was higher in the control group than in groups treated with MTA and/or BMP-7 (p<0.05). The cell migration rate increased after treatment with MTA when compared with other groups in all periods of time (p<0.05). At 72 hours, the wound area was smaller in groups treated with MTA and/or BMP-7 than in the control group (p<0.05). Conclusion: The use of BMP-7 with MTA increased odontogenic/osteogenic differentiation without adversely affecting proliferation and migration of DPSCs. The use of BMP-7 with MTA may improve treatment outcomes by increasing repair and regeneration capacity of DPSCs

Histopatološke i apoptotske promjene u bubrezima uzrokovane subakutnom izloženosti arokloru 1254 u štakora s nedostatnim i nadomjesnim unosom selenija

Aroclor 1254 (A1254), a mixture of polychlorinated biphenyls, exerts hepatic, renal, and reproductive toxicity in rodents. This study aimed to determine a protective role of selenium on histopathological changes, oxidative stress, and apoptosis caused by A1254 in rat kidney. It included a control group, which received regular diet containing 0.15 mg/kg Se (C), a Se-supplemented group (SeS) receiving 1 mg/kg Se, a Se-deficient group (SeD) receiving Se-deficient diet of ≤0.05 mg/ kg Se, an A1254-treated group (A) receiving 10 mg/kg of Aroclor 1254 and regular diet, an A1254-treated group receiving Se-supplementation (ASeS), and an A1254-treated group receiving Se-deficient diet (ASeD). Treatments lasted 15 days. After 24 h of the last dose of A1254, the animals were decapitated under anaesthesia and their renal antioxidant enzyme activities, lipid peroxidation (LP), glutathione, protein oxidation, and total antioxidant capacity levels measured. Histopathological changes were evaluated by light and electron microscopy. Apoptosis was detected with the TUNEL assay. Kidney weights, CAT activities, and GSH levels decreased significantly in all A1254-treated groups. Renal atrophic changes and higher apoptotic cell counts were observed in the A and ASeD groups. Both groups also showed a significant drop in GPx1 activities (A – 34.92 % and ASeD – 86.46 %) and rise in LP (A – 30.45 % and ASeD – 20.44 %) vs control. In contrast, LP levels and apoptotic cell counts were significantly lower in the ASeS group vs the A group. Histopathological changes and renal apoptosis were particularly visible in the ASeD group. Our findings suggest that selenium supplementation provides partial protection against renal toxicity of Aroclor 1254.Mješavina polikloriranih bifenila poznata pod nazivom aroklor 1254 (A1254) dokazano je toksična za jetru, bubrege i reprodukcijski sustav u glodavaca. Cilj ovoga istraživanja bio je na temelju histoloških promjena te parametara oksidacijskoga stresa i apoptoze utvrditi u kojoj mjeri selenij (Se) štiti bubrege od njegove toksičnosti. Istraživanje je obuhvatilo kontrolnu skupinu na normalnoj prehrani, koja je sadržavala 0,15 mg/kg Se, zatim skupinu čija je prehrana bila obogaćena selenijem (SeS) u dozi od 1 mg/kg, skupinu čija je prehrana bila osiromašena selenijem (SeD) (≤0.05 mg/ kg), skupinu koja je uz uobičajenu prehranu bila izložena arokloru 1254 (A) u dozi od 10 mg/kg tjelesne težine, skupinu koja je uz prehranu obogaćenu selenijem bila izložena arokloru 1254 (ASeS) i konačno skupinu koja je uz prehranu osiromašenu selenijem bila izložena arokloru 1254 (ASeD). Izloženost arokloru 1254 trajala je 15 dana. Životinje su 24 sata nakon primitka posljednje doze aroklora 1254 bile dekapitirane pod općom anestezijom te su im izmjerene vrijednosti bubrežnih enzimskih aktivnosti, lipidne peroksidacije (LP), glutationa (GSH), proteinske oksidacije i ukupnoga antioksidacijskoga kapaciteta. Histopatološke promjene utvrđene su pomoću svjetlosne i elektronske mikroskopije. Broj apoptotskih stanica utvrđen je TUNEL metodom. U svih skupina izloženih arokloru 1254 uočen je pad težine bubrega te aktivnosti katalaze i glutationa. U skupinama A i ASeD također je uočen povišen broj apoptotskih stanica i atrofija bubrežnoga tkiva, značajan pad aktivnosti GPX1 (u skupini A za 34,92 %, a u skupini ASeD za 86,46 %) te porast lipidnih peroksida (u skupini A za 30,45 %, a u skupini ASeD za 20,44 %) u odnosu na kontrolnu skupinu. Nasuprot skupini A, razine lipidnih peroksida i broj apoptotskih stanica bili su značajno niži u skupini ASeS, koja je primala hranu obogaćenu selenijem. Histopatološke promjene i apoptoza bubrežnih stanica osobito su se isticale u skupini ASeD na hrani osiromašenoj selenijem. Naši rezultati upućuju na to da nadomjesna primjena selenija pruža barem djelomičnu zaštitu od toksičnoga djelovanja aroklora 1254 na bubrege

Assessment of electromechanically stimulated bone marrow stem cells seeded acellular cardiac patch in a rat myocardial infarct model

In this study, we evaluated cardiomyogenic differentiation of electromechanically stimulated rat bone marrow-derived stem cells (rt-BMSCs) on an acellular bovine pericardium (aBP) and we looked at the functioning of this engineered patch in a rat myocardial infarct (MI) model. aBP was prepared using a detergent-based decellularization procedure followed by rt-BMSCs seeding, and electrical, mechanical, or electromechanical stimulations (3 millisecond pulses of 5 V cm-1at 1 Hz, 5% stretching) to enhance cardiomyogenic differentiation. Furthermore, the electromechanically stimulated patch was applied to the MI region over 3 weeks. After this period, the retrieved patch and infarct region were evaluated for the presence of calcification, inflammatory reaction (CD68), patch to host tissue cell migration, and structural sarcomere protein expressions. In conjunction with any sign of calcification, a higher number of BrdU-labelled cells, and a low level of CD68 positive cells were observed in the infarct region under electromechanically stimulated conditions compared with static conditions. More importantly, MHC, SAC, Troponin T, and N-cad positive cells were observed in both infarct region, and retrieved engineered patch after 3 weeks. In a clear alignment with other results, our developed acellular patch promoted the expression of cardiomyogenic differentiation factors under electromechanical stimulation. Our engineered patch showed a successful integration with the host tissue followed by the cell migration to the infarct region

Evaluation Of The Cytotoxic And Autophagic Effects Of Atorvastatin On Mcf-7 Breast Cancer Cells

PubMedWoSScopu

In Vitro Cytotoxicity Of Calcium Silicate-Based Endodontic Cement As Root-End Filling Materials

The aim of this study was to evaluate the cytotoxicity of three types of calcium silicate-based endodontic cement after different incubation periods with human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were cultured from extracted third molars and seeded in 96-well plates. MTA, calcium enriched mixture (CEM) cement, and Biodentine were prepared and added to culture insert plates which were immediately placed into 96-well plates containing cultured cells. After incubation periods of 24, 48, and 72 hours, cell viability was determined with WST-1 assay. Data were analysed statistically by ANOVA with repeated measures and Bonferroni tests. There was no significant difference in cell viability amongst the test materials after each incubation period (P > 0.05). MTA and CEM presented more than 90% cell viability after 24 and 48 hours of incubation and showed statistically significant decrease in cell viability after 72 hours of incubation (P < 0.05). Biodentine showed significantly less cell viability (73%) after 24 hours of incubation, whereas more than 90% cell viability was seen after 48 and 72 hours of incubation (P < 0.05). Despite the significant changes in cell viability over time, materials presented similar cytotoxicity profile. Biodentine and CEM can be considered as alternative materials for root-end surgery procedures.PubMedScopu

Cytotoxic Effect Of Endodontic Irrigants In Vitro

Background Cytotoxicity of root canal irrigants is important due to their close contact with host tissues. The aim of this study was to assess the cytotoxic effect of NaOCl 3%, Chx 2%, and MTAD on rat periodontal ligament fibroblasts, at 0.1 and 100 μl/mL, using WST-1 colorimetric method. Material/Method Rat ligamental fibroblasts were exposed to the irrigants and their viability was assessed after 1, 24, 48, and 72 h. The measurements were determined using WST-1 assay, using a micro ELISA reader. Results At 100 ml/L all 3 irrigants were strongly cytotoxic, although CHX was less so than NaOCl and MTAD. At the 0.1 ml/L concentration, NaOCl and MTAD were only moderately cytotoxic, whereas Chx was highly deleterious to cell viability at all time points. There was a significant influence of the dilution rate of the substance, because the odds ratio for cell viability being over 50% was increased 51 times between the 100 ml/L and 0.1 ml/L dilutions. Conclusions It seems that irrigating solutions should be used at lower concentrations to enhance cell viability.PubMe

The Early Histological Effects of Intravesical Instillation of Platelet-Rich Plasma in Cystitis Models

Purpose To evaluate the early histological effects of the intravesical instillation of platelet-rich plasma (PRP) in rabbit models of interstitial and hemorrhagic cystitis. Methods Thirty-six rabbits were classified into 6 groups: saline (S), S+PRP, hydrochloric acid (HCl), HCl+PRP, cyclophosphamide (CyP), and CyP+PRP. At 48 hours after induction, PRP was prepared and intravesically administered to the S+PRP, HCl+PRP, and CyP+PRP groups. Bladder sections were stained with toluidine blue for mast cell counting and with hematoxylin and eosin for histopathology and mitotic index determination. The proliferation index was determined by proliferating cell nuclear antigen (PCNA) immunolabeling. The nonparametric Mann-Whitney U-test was used for statistical analysis. Results No abnormalities were observed in the S group, whereas increased interstitial edema and increased average mitotic and proliferation indices were observed in the S+PRP group (P=0.023, P=0.004, and P=0.009, respectively). Intense epithelial loss, hemorrhage, and leukocyte infiltration were detected in the HCl and HCl+PRP groups, whereas a significantly increased average mitotic index was observed in the HCl+PRP group (P=0.002). When compared with its CyP counterpart, a significant reduction in hemorrhage and an increase in leukocyte infiltration and mitotic index were observed in the CyP+PRP group (P=0.006, P=0.038, and P=0.002, respectively). In addition, PCNA staining revealed a significantly increased proliferation index in the HCl+PRP and CyP+PRP groups (P=0.032 and P=0.015, respectively). Conclusions The intravesical instillation of PRP increased the mitotic index in the saline and cyclophosphamide groups while decreasing macroscopic bleeding.PubMedWoSScopu

The Early Histological Effects of Intravesical Instillation of Platelet-Rich Plasma in Cystitis Models

Purpose: To evaluate the early histological effects of the intravesical instillation of platelet-rich plasma (PRP) in rabbit models of interstitial and hemorrhagic cystitis

Effects of prenatal and lactational bisphenol a and/or di(2-ethylhexyl) phthalate exposure on male reproductive system

Bisphenol A (BPA) and phthalates are abundantly used endocrine disrupting chemicals (EDCs). The aim of this study was to evaluate the effects of single and combined exposures to BPA and/or di(2-ethylhexyl) phthalate (DEHP) in prenatal and lactational period on rat male reproductive system in later stages of life. Pregnant Sprague-Dawley rats were divided randomly to four groups (n = 3/group): Control (corn oil); DEHP (30 mg/kg/day); BPA (50 mg/kg/day); and BPA+ DEHP (30 mg/kg/day DEHP and 50 mg/kg/day BPA). Groups exposed to EDCs through 6–21 gestational days and lactation period by intragastric lavage. Male offspring (n = 6/group) from each mother were fed till adulthood and were then euthanized. Later, reproductive hormones, sperm parameters, and oxidative stress parameters were determined. In conclusion, we can suggest that prenatal and lactational exposure to BPA and DEHP may cause adverse effects in male reproductive system in later stages of life especially after combined exposure. © 2020 Informa UK Limited, trading as Taylor & Francis Group