In vitro evaluation of the action of irrigating solutions associated with intracanal medications on Escherichia coli and its endotoxin in root canals

Objective: The purpose of this study was to evaluate the efficacy of auxiliary chemical substances and intracanal medications on Escherichia coli and its endotoxin in root canals. Material and Methods: Teeth were contaminated with a suspension of E. coli for 14 days and divided into 3 groups according to the auxiliary chemical substance used: G1) 2.5% sodium hypochlorite (NaOCl); G2) 2% chlorhexidine gel (CLX); G3) pyrogen-free solution. After, these groups were subdivided according to the intracanal medication (ICM): A) Calcium hydroxide paste (Calen (R)), B) polymyxin B, and C) Calcium hydroxide paste+2% CLX gel. For the control group (G4), pyrogen-free saline solution was used without application of intracanal medication. Samples of the root canal content were collected immediately after biomechanical preparation (BMP), at 7 days after BMP, after 14 days of intracanal medication activity, and 7 days after removal of intracanal medication. The following aspects were evaluated for all collections: a) antimicrobial activity; b) quantification of endotoxin by the limulus Amebocyte lysate test (LAL). Results were analyzed by the kruskal-wallis and Dunn's tests at 5% significance level. Results: The 2.5% NaOCl and CLX were able to eliminate E. coli from root canal lumen and reduced the amount of endotoxin compared to saline. Conclusions: It was concluded that 2.5% NaOCl and CLX were effective in eliminating E. coli. Only the studied intracanal medications were to reduce the amount of endotoxin present in the root canals, regardless of the irrigant used.

Effects of acetylsalicylic acid and acetic acid solutions on VX2 liver carcinoma in rabbits: in vivo analysis

OBJETIVO: Analisar os efeitos das soluções de aspirina e de ácido acético, in vivo, em fígado de coelhos portadores de tumor hepático VX2, verificando o efeito histolítico e anatomo-patológico das soluções e eventuais alterações bioquímicas hepáticas. MÉTODOS: Utilizou-se 48 coelhos, divididos em 2 protocolos experimentais(3 e 4), subdivididos em 3 grupos cada. Após 4 dias da implantação do tumor no fígado, procedeu-se a laparotomia mediana, com injeção de 0,4 ml da solução de aspirina (5,0%), de ácido acético (5,0%) e solução salina; o sacrifício ocorreu apos 24 horas (protocolo 3) e 11 dias (protocolo 4); avaliou-se o peso, evolução clinica, dosagens bioquímicas, cavidade abdominal e torácica e microscopia do fígado. RESULTADOS: Não foram observadas alterações na evolução clinica, peso e nas dosagens bioquímicas, apenas elevação da fosfatase alcalina no grupo controle do protocolo 4. Observamos desaparecimento do tumor em ambos os protocolos. CONCLUSÃO: As soluções de ácido acético e ácido acetilsalicílico acarretam destruição do tumor hepático experimental.

Histological study of the effect of some irrigating solutions on bacterial endotoxin in dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of different irrigation solutions and calcium hydroxide on bacterial LPS

Aim To evaluate the effect of biomechanical preparation with different irrigating solutions and calcium hydroxide dressing in dog root canals containing bacterial endotoxin (lipopolysaccharides; LPS).Methodology One hundred and forty premolar roots from seven dogs were filled with Escherichia coli LPS for 10 days (three roots were lost during histological processing). The following irrigating solutions were used for biomechanical preparation: 1% (group I, n = 20), 2.5% (group II, n = 19) and 5% sodium hypochlorite (group III, n = 19), 2% chlorhexidine digluconate (group IV, n = 20) and physiological saline solution (group V, n = 19). In group VI (n = 20), the LPS solution was maintained in the root canal during the entire experiment and in group VII (n = 20), after biomechanical preparation with saline solution, the root canals were filled with a calcium hydroxide dressing (Calen; control). After 60 days, the animals were sacrificed and the following parameters of periapical disease were evaluated: (a) inflammatory infiltrate, (b) periodontal ligament thickness, (c) cementum resorption and (d) bone resorption. Scores were given and data were analysed statistically with the Kruskal-Wallis and Dunn tests (P < 0.05).Results Histopathological evaluation showed that groups I-VI had more inflammatory infiltrate, greater periodontal ligament thickening and greater cementum and bone resorption (P < 0.05) compared to group VII, which received the calcium hydroxide intracanal dressing.Conclusions Biomechanical preparation with the irrigating solutions did not inactivate the effects of the endotoxin but the calcium hydroxide intracanal dressing did appear to inactivate the effects induced by the endotoxin in vivo.

Inflammatory response to different endodontic irrigating solutions

Aim the aim of this study was to evaluate the inflammatory response to irrigating solutions injected into the peritoneal cavity of mice.Methodology Sixty mice received intra-peritoneal injections of 0.3 mL of 0.5% sodium hypochlorite, 2.0% chlorhexidine digluconate or phosphate buffered saline (PBS, control). Five animals of each group were sacrificed at 4, 24, 48 h and 7 days after the injection. Liquid from the peritoneal cavity of each animal was collected for the total and differential counting of inflammatory cells and protein leakage.Results the 0.5% sodium hypochlorite solution group had greater migration of neutrophils and mononuclear cells to the peritoneal cavity from 48 to 168 h (P < 0.05). There was a significant increase in protein leakage to the peritoneal cavity after 4 up to 48 h in the 0.5% sodium hypochlorite group compared to the control group. Protein leakage was similar in all groups at 168 h. The 2.0% chlorhexidine group had similar results to the control group at all time periods.Conclusions the 0.5% sodium hypochlorite solution induced an inflammatory response, however, the 2.0% chlorhexidine digluconate solution did not induce a significant inflammatory response.

Cytotoxic effects of cleansing solutions recommended for chemical lavage of pulp exposures

Purpose: To evaluate the in vitro cytotoxic effects of three cleansing solutions used for chemical lavage of pulp exposures. Materials and Methods: the immortalized odontoblast cell line (MDPC-23) was plated (30,000 cells/cm(2)) and incubated for 72 hrs in 24-well dishes. After counting the cell number under inverted light microscopy, 20 mul of the experimental and control solutions were added to 980 mul of fresh culture medium. Then, hydrogen peroxide (3%, H2O2), sodium hypochlorite (6%, NaOCl) or calcium hydroxide-saline solution (5g of Ca(OH)(2) in 10 mi of sterile distilled water) were added to wells for experimental Groups 1, 2 and 3, respectively. The positive and negative control groups received Syntac Sprint bonding agent (SS) and phosphate buffered saline (PBS), respectively. Following incubation for 120 min the cell number was counted again, the cell morphology was evaluated by scanning electron microscopy (SEM) and the cell metabolism was determined by the methyltetrazolium (MTT) assay. The scores obtained from cell counting and MTT assay were analyzed with an ANOVA followed by Fisher's PLSD tests. Results: H2O2 NaOCl solutions, and SS bonding agent were more cytotoxic than Ca(OH)2 or PBS. In the groups with H2O2 Or SS, only a few cells remained attached to the bottom of wells. The difference between these two groups was not statistically significant. H2O2, NaOCl and SS depressed the mitochondrial enzyme response by 97.7%, 97.3%, and 95.0%, respectively. on the other hand, Ca(OH)2 depressed the metabolic activity of cells by only 5%. While H2O2, NaOCl and SS caused extreme changes on the cell morphology, neither Ca(OH)2 nor PBS promoted dramatic changes in the cell morphology.

Effects of acetylsalicylic acid and acetic acid solutions in VX2 carcinoma cells. In vitro analysis

Purpose: To analyze, in vitro, the effects of acetylsalicylic acid (aspirin) and acetic acid solutions on VX2 carcinoma cells in suspension and to examine the correlation between these effects and neoplastic cell death. Methods: The VX2 tumor cells (107 cells/ml) were incubated in solutions containing differing concentrations (2.5% and 5%) of either acetylsalicylic acid or acetic acid, or in saline solution (controls). Every five minutes, cell viability was tested (using the trypan blue test) and analyzed under light microscopy. Results: Tumor cell viability (in %) decreased progressively and, by 30 minutes, neoplastic cell death had occurred in all solutions. Conclusion: Based on this experimental model and the methodology employed, we conclude that these solutions cause neoplastic cell death in vitro.

In vivo microbiological evaluation of the effect of biomechanical preparation of root canals using different irrigating solutions

The aim of this study was to evaluate the antimicrobial effect of biomechanical preparation using different irrigating solutions. Seventy-eight root canals from premolars of four dogs were used. After experimental induction of periapical lesions, the root canals were prepared using the following solutions for irrigation: Group 1) 2.5% sodium hypochlorite (NaOCl); Group 2) 2% chlorhexidine (CHX); Group 3) saline solution and Group 4) control group with no biomechanical preparation. The microbiological evaluation of the root canals was performed by counting the colony forming units (CFUs) using different culture mediums. Two absorbent paper cones were used in each root canal in order to collect the microbiological samples before, and thirty days after the biomechanical preparation. The culture plates were incubated in aerobic, anaerobic and microaerophilic environment. Statistical evaluation was carried out using analysis of variance, Tukey and Student tests. The results demonstrated that there was reduction in the number of microorganisms in the NaOCl and CHX groups (p<0.05). There was greater effectiveness in the chlorhexidine group. The group that used saline solution and the control group presented an increased number of microorganisms. It can be concluded that the use of antimicrobial irrigating solutions during biomechanical preparation promotes the reduction of endodontic microbiota. However, a considerable number of microorganisms were still observed.

Adhesion of real seal to human root dentin treated with different solutions

The aim of this study was to evaluate the effects of different irrigants on sealer-dentin bond strength when using Real Seal. Thirty single-rooted teeth were divided into 3 groups. In one group, the teeth were irrigated with 3 mL of 2.5% NaOCl after each file change, flushed with 17% EDTA for 3 min and finally rinsed with 3 mL of 2.5% NaOCl. In the other two groups, rinse with NaOCl was replaced with 2% chlorhexidine gluconate (CHX) and 0.9% saline, respectively. Each root was sectioned transversally into apical, middle and coronal thirds to obtain 2-mm-thick slices. Each slice was filled with Real Seal and Resilon. Push-out test was used to analyze bond strength and failure modes were classified as adhesive, cohesive or mixed, according to SEM observations. The push-out test did not reveal any statistically significant difference (p>0.05) between the irrigants. However, the groups exhibited significantly different (p<0.05) bond strengths in terms of the root canal third. Higher bond strength was observed at the apical third when compared with coronal third, while middle third presented intermediary values. Fifteen specimens were analyzed by SEM (5 per group). Eleven specimens exhibited adhesive failures (5 in saline, 4 in NaOCl and 2 in CHX group); 2 cohesive failures were observed in the CHX group, and 1 mixed failure each was observed in the CHX and NaOCl groups. The tested irrigants did not influence the bond strength of Resilon and Real Seal to dentin. The apical third exhibited higher mean bond strengths and adhesive failures were predominant.

Effectiveness of several solutions to prevent the formation of precipitate due to the interaction between sodium hypochlorite and chlorhexidine and its effect on bond strength of an epoxy-based sealer

To evaluate the effectiveness of isopropyl alcohol, saline or distilled water to prevent the precipitate formed between sodium hypochlorite (NaOCl) and chlorhexidine (CHX) and its effect on the bond strength of an epoxy-based sealer in radicular dentine. Methodology The root canals of 50 extracted human canines (n = 10) were instrumented. In G1, root canals were irrigated with 17% EDTA and 2.5% NaOCl; G2, as G1, except that 2% CHX was used as the final irrigant. In the other groups, intermediate flushes with isopropyl alcohol (G3), saline (G4) or distilled water (G5) were used between NaOCl and CHX. The specimens were submitted to SEM analysis to evaluate the presence of debris and smear layer, in the apical and cervical segments. In sequence, fifty extracted human canines were distributed into five groups (n = 10), similar to the SEM study. After root filling, the roots were sectioned transversally to obtain dentine slices, in the cervical, middle and apical thirds. The root filling was submitted to a push-out bond strength test using an electromechanical testing machine. Statistical analysis was performed using Kruskal–Wallis and Dunn's tests (α = 5%). Results All groups had similar amounts of residue precipitated on the canal walls (P > 0.05). The push-out bond strength values were similar for all groups, independently of the root third evaluated (P > 0.05). Conclusions Isopropyl alcohol, saline and distilled water failed to prevent the precipitation of residues on canal walls following the use of NaOCl and CHX. The residues did not interfere with the push-out bond strength of the root filling.

Effect of protective solutions and hydroxyethyl starch in the attenuation of the injuries of ischemia and reperfusion of splanchnic organs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparison of Celsior and Perfadex lung preservation solutions in rat lungs subjected to 6 and 12 hours of ischemia using an ex-vivo lung perfusion system

OBJECTIVE: This study evaluated the performance of lungs that were preserved with different solutions (Celsior, Perfadex or saline) in an ex vivo rat lung perfusion system. METHODS: Sixty Wistar rats were anesthetized, anticoagulated and randomized into three groups (n = 20). The rats were subjected to antegrade perfusion via the pulmonary artery with Perfadex, Celsior, or saline, followed by 6 or 12 hours of ischemia (4 degrees C, n = 10 in each group). Respiratory mechanics, gas exchange and hemodynamics were measured at 10-minute intervals during the reperfusion of heart-lung blocks in an ex vivo system (IL2-Isolated Perfused Rat or Guinea Pig Lung System, Harvard Apparatus, Holliston, Massachusetts, USA; Hugo Sachs Elektronik, Germany) for 60 minutes. The lungs were prepared for histopathology and evaluated for edema following reperfusion. Group comparisons were performed using ANOVA and the Kruskal-Wallis test with a 5% level of significance. RESULTS: Gas exchange was not significantly different between lungs perfused with either Perfadex or Celsior at the same ischemic times, but it was very low in lungs that were preserved with saline. Airway resistance was greater in the lungs that were preserved for 12 hours. Celsior lungs that were preserved for 6 and 12 hours exhibited lower airway resistance (p = 0.01) compared to Perfadex lungs. Pulmonary artery pressure was not different between the groups, and no significant differences in histopathology and apoptosis were observed between the groups. CONCLUSIONS: Lungs that were preserved with Celsior or Perfadex exhibited similar gas exchange and histopathological findings. Airway resistance was slightly lower in the Celsior-preserved lungs compared with the Perfadex-preserved lungs.

Intravenous Hypertonic Saline Solution (7.5%) and Oral Electrolytes to Treat of Calves with Noninfectious Diarrhea and Metabolic Acidosis

Objective The aim of this study was to compare the efficacy of treating osmotic diarrhea and dehydration in calves with hypertonic saline solution (HSS) IV, isotonic electrolyte solution (IES) PO, and a combination of these 2 solutions (HSS + IES). Experimental Design Eighteen male calves 830 days of age were used to evaluate the efficacy of 3 methods of fluid therapy after induction of osmotic diarrhea and dehydration. The diarrhea and dehydration were induced by administration of saccharose, spironolactone, and hydrochlorothiazide for 48 hours. The animals were randomly divided into 3 experimental groups: Group 1: 7.2% hypertonic saline solution-HSS (5 mL/kg IV); Group 2: oral isotonic electrolyte solution IES (60 mL/kg PO); or Group 3: HSS+IES. Clinical signs and laboratory finding observed 48 hours post-induction (Time 0) included diarrhea, dehydration, lethargy, and metabolic acidosis. Results Calves treated with HSS + IES experienced decreases in hematocrit, total protein concentration, albumin concentration, urea nitrogen concentration, and plasma volume as well as increases in blood pH, blood bicarbonate concentration, and central venous pressure between 1 and 3 hours post-treatment. These findings also were observed in animals treated with IES, however, at a slower rate than in the HSS + IES-treated animals. Animals treated with HSS continued to display signs of dehydration, lethargy, and metabolic acidosis 24 hours post-treatment. Conclusion Treatment with a combination of HSS and IES produced rapid and sustainable correction of hypovolemia and metabolic acidosis in calves with noninfections diarrhea and dehydration.

Comparison of the in vitro effects of saline, hypertonic hydroxyethyl starch, hypertonic saline, and two forms of hydroxyethyl starch on whole blood coagulation and platelet function in dogs.

OBJECTIVE To compare the in vitro effects of hypertonic solutions and colloids to saline on coagulation in dogs. DESIGN In vitro experimental study. SETTING Veterinary teaching hospital. ANIMALS Twenty-one adult dogs. INTERVENTIONS Blood samples were diluted with saline, 7.2% hypertonic saline solution with 6% hydroxyethylstarch with an average molecular weight of 200 kDa and a molar substitution of 0.4 (HH), 7.2% hypertonic saline (HTS), hydroxyethyl starch (HES) 130/0.4 or hydroxyethyl starch 600/0.75 at ratios of 1:22 and 1:9, and with saline and HES at a ratio of 1:3. MEASUREMENTS AND MAIN RESULTS Whole blood coagulation was analyzed using rotational thromboelastometry (extrinsic thromboelastometry-cloting time (ExTEM-CT), maximal clot firmness (MCF) and clot formation time (CFT) and fibrinogen function TEM-CT (FibTEM-CT) and MCF) and platelet function was analyzed using a platelet function analyzer (closure time, CTPFA ). All parameters measured were impaired by saline dilution. The CTPFA was prolonged by 7.2% hypertonic saline solution with 6% hydroxyethylstarch with an average molecular weight of 200 kDa and a molar substitution of 0.4 (HH) and HTS but not by HES solutions. At clinical dilutions equivalent to those generally administered for shock (saline 1:3, HES 1:9, and hypertonic solutions 1:22), CTPFA was more prolonged by HH and HTS than other solutions but more by saline than HES. No difference was found between the HES solutions or the hypertonic solutions. ExTEM-CFT and MCF were impaired by HH and HTS but only mildly by HES solutions. At clinically relevant dilutions, no difference was found in ExTEM-CFT between HTS and saline or in ExTEM-MCF between HH and saline. No consistent difference was found between the 2 HES solutions but HH impaired ExTEM-CFT and MCF more than HTS. At high dilutions, FibTEM-CT and -MCF and ExTEM-CT were impaired by HES. CONCLUSIONS Hypertonic solutions affect platelet function and whole blood coagulation to a greater extent than saline and HES. At clinically relevant dilutions, only CTPFA was markedly more affected by hypertonic solutions than by saline. At high dilutions, HES significantly affects coagulation but to no greater extent than saline at clinically relevant dilutions.

Graft preservation solutions in cardiovascular surgery.

Vein grafts are still the most commonly used graft material in cardiovascular surgery and much effort has been spent in recent years on investigating the optimal harvesting technique. One other related topic of similar importance remained more or less an incidental one. The storage solutions of vein grafts following procurement and prior to implantation are, despite their assumed impact, a relatively neglected theme. There is no doubt that the endothelium plays a key role in long-term patency of vein grafts, but the effects of the different storage solutions on the endothelium remain unclear : In a review of the literature, we could find 20 specific papers that addressed the question, of which the currently available preservation solutions are superior, harmless, damaging or ineffective. The focus lies on saline and autologous whole blood. Besides these two storage media, novel or alternative solutions have been investigated with surprising findings. In addition, a few words will be spent on potential alternatives and novel solutions on the market. As there is currently no randomized clinical trial regarding saline versus autologous whole blood available, this review compares all previous studies and methods of analysis to provide a certain level of evidence on this topic. In summary, saline has negative effects on the endothelial layers and therefore may compromise graft patency. Related factors, such as distension pressure, may outbalance the initial benefit of autologous whole blood or storage solutions and intensify the harmful effects of warm saline. In addition, there is no uniform consent on the superiority of autologous whole blood for vein graft storage. This may open the door to alternatives such as the University of Wisconsin solution or one of the specific designed storage solutions like TiProtec™ or Somaluthion™. Whether these preservation solutions are superior or advantageous remains the subject of further studies.