Mirian Marubayashi Hidalgo

Study of the effectiveness of propolis extract as a storage medium for avulsed teeth

The purpose of the present study was to evaluate the efficacy of propolis extract in maintaining the viability of human periodontal ligament (PDL) cells, and to radiographically analyze tooth replantation and the adjacent periodontium in dogs after storage in this extract. Human PDL cells were incubated with the experimental media propolis, milk, saliva, Hanks balanced salt solution (HBSS), and Dulbeccos modified Eagles medium (DMEM, positive controls), and distilled water (negative control). Cell viability was determined 0, 1, 3, 6, 12, and 24 h later by colorimetric MTT assay. Thirty incisors from dogs were divided into two storage time blocks (1 and 3 h) and were maintained in the experimental media. HBSS served as a positive control, and dry teeth (on gauze) as a negative control. The replanted teeth were radiographed once per month for 6 months. The radiographic images were standardized by the shortening/lengthening factor, and were both qualitatively and quantitatively analyzed. The in vitro results showed that the efficacy of propolis in maintaining functional viability of PDL cells was similar to that of milk. Propolis and milk were significantly better than controls from the 6-h time period. The in vivo results showed that teeth maintained in propolis medium exhibited replacement resorption with significant reduction in tooth length, similar to teeth maintained in saliva and dried teeth. This resorption was less intense with the 3-h storage time than the 1-h storage time. Conditions close to normal were found in teeth maintained in milk, similar to the HBSS control. Therefore, although propolis was effective in maintaining the viability of human PDL cells, resorption of the tooth replantation in dogs occurred under these experimental conditions.

Removal efficiency of propolis paste dressing from the root canal

Objetives The aim of this study was to investigate through scanning electron microscopy (SEM) the cleaning of root canal walls after the use of experimental propolis or calcium hydroxide root canal dressings. Material and Methods Twenty single-rooted teeth were used. After conventional cleaning and shaping procedures and removal of the smear layer with 17% EDTA, the teeth were divided into four groups according to the medication used (N=5): Group I (control) - No drug, Group II - Calcium hydroxide dressing, Group III - Propolis paste A70D and Group IV - Propolis paste D70D. The medications were introduced into the root canals and maintained for 7 days, then removed with a K-file and 5 mL of 1% sodium hypochlorite irrigation. Finally, the canals were flushed with 2 mL of 17% EDTA for 3 min. For SEM analysis, the roots were cleaved and microphotographs from the middle third of the root canal were taken at 750x. The cleaning of the root canal walls was determined by the number of open dentinal tubules as verified with the software Image Tool 3.1. The statistical analysis was performed by ANOVA and Tukeys test (p<0.05). Results The results showed no statistically significant difference between the calcium hydroxide and propolis groups. Conclusions The experimental propolis pastes presented acceptable physical characteristics to be used as intracanal medicaments.

Pharmacological Evaluation of Propolis Solutions for Endodontic Use

Abstract This study evaluated the anti-inflammatory and exudative activities of propolis solutions and their antimicrobial activity. The solutions were prepared and diluted in alcohol solution (PPE1, PPE2, PPE3, PPF18, and PPF19). Ear edema was previously induced in mice by the application of croton oil, and the irritative effect of the solutions was determined through the exudation test of Evans blue in rats. Antimicrobial activity by using a macrodilution method was determined. Eight aerobic bacteria, seven anaerobic bacteria, and two yeasts were tested. The PPE1, PPE2, and PPF18 solutions presented excellent anti-inflammatory activities. PPE1 solution showed the best antimicrobial effect. PPF18 showed an inhibitory effect for the majority of the aerobic bacteria in the dilution 1:8, inhibiting the growth of Klebsiella pneumoniae. and the yeasts in the dilution 1/16 and of Pseudomones aeruginosa. in the dilution 1/32. PPF19 was effective for inhibiting the growth of the aerobic bacteria and yeasts in the dilution 1/2. Our results suggest the possible application of PPE1, PPF18, and PPF19 solutions in endodontics.

Antibacterial activity of propolis-based toothpastes for endodontic treatment

This study evaluated the antibacterial activity of propolis-based toothpastes used as intracanal medication in endodontic treatment. The propolis-based toothpastes were prepared using an extract established in previous studies (identified as A70D and D70D). Calcium hydroxide paste was used as a control. The bacteria employed were Streptococcus mutans (ATCC 25175), Staphylococcus aureus (ATCC 6538), Staphylococcus aureus (ATCC 25923), Kocuria rhizophila (ATCC 9341), Escherichia coli (ATCC 10538), Pseudomonas aeruginosa (ATCC 27853), Enterococcus hirae (ATCC 10541), Streptococcus mutans (ATCC 25175). Five field strains isolated from saliva were used: Staphylococcus spp. (23.1 - coagulase positive), Staphylococcus spp. (23.5 - coagulase negative), Staphylococcus spp. (26.1 - coagulase positive), Staphylococcus spp. (26.5 - coagulase negative) and Staphylococcus epidermidis (6epi). The diffusion-well method on double-layer agar was used in a culture medium of Tryptic Soy Agar. The plates were kept at room temperature for two hours to allow the diffusion of pastes in the culture medium, and then incubated at 35o C for twenty-four hours in aerobiosis and in microaerophilia (S. mutans). After this period, the total diameter of the inhibition halo was measured. The results were analyzed by ANOVA analysis of variance, followed by the Tukey test at p<0.05. The propolis-based toothpastes presented antibacterial activity against 83.3% of the analyzed bacteria. For 66.7% of these bacteria, the propolis-based toothpastes exhibited greater antibacterial activity than calcium hydroxide. The present results allow us to conclude that the experimental pastes A70D and D70D showed good activity against aerobic bacteria, proving more effective than calcium hydroxide.

Dragon's Blood Sap (Croton Lechleri) As Storage Medium For Avulsed Teeth: In Vitro Study Of Cell Viability

Tooth replantation success depends on the condition of cementum periodontal ligament after tooth avulsion; which is influenced by storage medium. The dragons blood (Croton lechleri) sap has been suggested as a promising medium because it supports collagen formation and exhibits healing, anti-inflammatory and antimicrobial properties. Thus, the aim of this study was to evaluate the efficacy of dragons blood sap as a storage medium for avulsed teeth through evaluation of functional and metabolic cell viability. This in vitro study compared the efficacy of different storage media to maintain the viability of human peripheral blood mononuclear and periodontal ligament cells. A 10% dragons blood sap was tested while PBS was selected as its control. Ultra pasteurized whole milk was used for comparison as a commonly used storage medium. DMEM and distilled water were the positive and negative controls, respectively. The viability was assessed through trypan blue exclusion test and colorimetric MTT assay after 1, 3, 6, 10 and 24 h of incubation. The dragons blood sap showed promising results due to its considerable maintenance of cell viability. For trypan blue test, the dragons blood sap was similar to milk (p<0.05) and both presented the highest viability values. For MTT, the dragons blood sap showed better results than all storage media, even better than milk (p<0.05). It was concluded that the dragons blood sap was as effective as milk, the gold standard for storage medium. The experimental sap preserved the membrane of all cells and the functional viability of periodontal ligament cells.

Doença periodontal: estudo da resposta imune humoral

We compared levels of sera immunoglobulin IgG and IgA, and salivary IgG and secretory IgA (IgA-S) reactive to Actinobacillus actinomycetemcomitans (Aa). Doses of total sera IgM, IgG, and IgA of patients with early-onset (EOP) and adult (AP) periodontitis, and healthy controls without periodontal disease were assessed. Igs to Aa were determined by ELISA. Total sera Igs were quantified by simple radial immunodiffusion, using a mixture of sonicated extracts of five isolates obtained from EOP patients confirmed as Aa, and sonicated extract of Aa reference strain FDC Y4 as antigens. No significant differences were found between sera IgG and IgA and salivary IgG and IgA-S levels among groups EOP (n=9), AP (n=20), and control (n=20). These results suggest that there was no significant alteration of the Aa humoral immune response level in patients with periodontitis. By quantification of total sera Igs, we found no statistically significant differences between EOP (n=9) and AP (n=9) patients or healthy controls (n=9).

Avaliação da resposta imune celular em pacientes com periodontite

The purpose of this study was to compare the lymphoproliferative response to Actinobacillus actinomycetemcomitans (Aa) and to phytohemaglutinin (PHA) mitogen, in groups of patients with early-onset periodontitis (EOP, n = 9), adult periodontitis (AP, n = 20), and healthy controls without periodontal disease (n = 20). As antigens, a mixture of sonicated extracts of five isolates obtained from EOP patients, and sonicated extracts of Aa reference strains ATCC 29522, ATCC 29523, and FDC Y4 were used. No statistically significant difference was observed between the three groups as to the lymphoproliferative response to Aa. The use of previously heated and unheated Aa sonicated extracts demonstrated the existence of heat-labile factor or factors capable of depressing the specific lymphoproliferative response. This indicates the necessity of using a heated extract for a valid evaluation of lymphoproliferative response. As for the lymphoproliferative response to the PHA mitogen, no significant differences between groups were observed. This suggests that there was no induction of immunosuppression in the patients with periodontitis. In addition, in experiments with heated or unheated Aa extracts, the presence of heat-stable factor or factors capable of suppressing the PHA response was noticed. The presence of these factors could explain why no alteration of the cellular immune response was detected in the patients with periodontitis.

External apical root resorption diagnosis by using FII human dentine fraction and salivary IGg

BACKGROUND External apical root resorption as a consequence of orthodontic treatment is an inflammatory pathological process that results in permanent loss of tooth structure from the root apex. OBJECTIVES This study aimed to investigate the diagnostic potential of human dentine fractions and salivary IgG in external apical root resorption. PATIENTS AND METHODS Saliva samples were collected from 10 patients before (T0) and after 3 (T3), 6 (T6) and 12 (T12) months of orthodontic treatment. The total dentinal extract, obtained from human third molars, was fractioned by gel filtration chromatography in three fractions denominated FI, FII and FIII. The root resorption analysis of the upper central incisors was performed by digital image subtraction method. Reactivity of salivary IgG to antigenic fractions of dentine was determined by enzyme-linked immunosorbent assay (Elisa). RESULTS Regardless of treatment, FI dentin fraction with high MM (<300kDa) was the one that presented highest reactivity with salivary IgG. However, it was found higher salivary IgG reactivity for FII (69 to 45 kilodalton [kDa]) as compared to FIII (<45kDa) at (T6) and (T12), (P<0.05), the same periods in that the root resorptions were detected. CONCLUSION Our results suggest that FII human dentine fraction and salivary IgG have potential to be used in diagnosis and monitoring of external apical root resorption. The development of a practical and accessible biochemical test using saliva and FII dentine fraction may help in the prevention of severe root resorption.

Aqueous Extract of Psidium cattleianum as Intracanal Medication: An In Vitro Study of Cell Viability

Psidium cattleianum extract has shown activity against oral microorganisms and anti-inflammatory activity, and also tissue biocompatibility. These characteristics aroused interest in its use as an alternative to traditional intracanal medication. The aim of this study was to evaluate the aqueous extract of P. cattleianum cytotoxicity by structural and functional cell viability, in order to use as intracanal medication. The hypothesis is that aqueous extract of P. cattleianum should be used as intracanal medications due to better cytotoxicity response than cell culture medium. Positive control used was RPMI cell culture medium and negative control was water. Cell viability was analyzed after 1 h, 3 h, 6 h, 10 h and 24 h of incubation by exclusion method with trypan blue and MTT assay, using human mononuclear cells (PBMC) and human Periodontal Ligament Cells (PDL) in culture. By trypan blue assay, both PBMC and PDL cells showed an average viability higher (p>0.05) than RPMI when the cells were maintained in aqueous extract. Distilled water showed lowest average viability (p<0.05). By MTT assay, PDL cells showed increasing viability over time when maintained in aqueous extract culture. P. cattleianum was able to maintain the structural and functional cell viability, shoeing a higher performance than positive control. Also, this extract neutralized the detrimental effect of distillated water. The results about biocompatibility assays are promising to indicate the use of aqueous extract of P. cattleianum as an alternative to traditional intracanal medications.

Circulating dentinal antigen‐antibody immune complexes during orthodontic tooth movement in rats

BACKGROUND The immunogenic potential of dentin has been reported through dentin-reactive autoantibodies detection in human and animal model. This study aimed to investigate the formation and diagnostic value of immune complexes formation after autoantibodies production, and soluble dentin antigens levels associated to root resorption, in the course of orthodontic tooth movement, in rat experimental model. METHODS Forty Wistar rats (n = 8 for each group) were submitted to orthodontic tooth movement, in which the maxillary right first molar was mesially moved by applying of 55 g of force for 3, 7, 14, or 21 days. Untreated group was used as control. Circulating autoantibodies to rat dentinal extract, immune complexes, and soluble dentinal antigen levels were determined by immunoenzyme assays. Additionally, dentinal antigens were analyzed by immunoblot. RESULTS Higher serum dentin-reactive IgG and immune complex levels were detected in the 14- and 21-day groups (p < 0.05 and p < 0.001 respectively) but not in circulating dentinal antigen levels (p > 0.05), as compared to the control group. Reactivity was found to dentinal components with molecular mass (MM) ~120 and ~150 kDa, by immunoblot. CONCLUSION This work represents the first evidence of immune complexes formation and circulating soluble dentin antigens associated to root resorption in orthodontic tooth movement. Immune complexes formation could be used to early diagnosis of external root resorption.