Analysis of the reaction of subcutaneous tissues in rats and the antimicrobial activity of calcium hydroxide paste used in association with different substances

MIDENA, Raquel Zanin; GARCIA, Roberto Brandão; CAVENAGO, Bruno Cavalini; MARCIANO, Marina Angélica; MINOTTI, Paloma Gagliardi; ORDINOLA-ZAPATA, Ronald; WECKWERTH, Paulo Henrique; ANDRADE, Flaviana Bombarda de; DUARTE, Marco Antonio Hungaro

doi:10.1590/1678-775720140049

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Journal of Applied Oral Science

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ORIGINAL ARTICLES • J. Appl. Oral Sci. 23 (5) • Sep-Oct 2015 • https://doi.org/10.1590/1678-775720140049 copy

Analysis of the reaction of subcutaneous tissues in rats and the antimicrobial activity of calcium hydroxide paste used in association with different substances

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The aim of this study was to evaluate the subcutaneous tissue response in rats and the antimicrobial activity of intracanal calcium hydroxide dressings mixed with different substances against E. faecalis. Fifty four rats were divided into three experimental groups according to the vehicle in the calcium hydroxide treatment: 0.4% chlorohexidine in propylene glycol (PG), Casearia sylvestris Sw in PG and calcium hydroxide+PG (control group). The pastes were placed into polyethylene tubes and implanted into the subcutaneous tissue. After 7, 14 and 30 days, the samples were processed and histologically evaluated (hematoxylin and eosin). The tissue surface in contact with the material was analyzed, and the quantitative analysis determined the volume density occupied by the inflammatory infiltrate (giant cells, polymorphonuclear cells and mononuclear cells), fibroblasts, collagen fibers and blood vessels. For the antimicrobial analysis, 20 dentin blocks infected with E. faecalis were treated with calcium hydroxide pastes in different vehicles; 0.4% chlorhexidine in PG, PG, extract from Casearia sylvestris Sw in PG and a positive control (infection and without medication) for 7 days. The efficiency of the pastes was evaluated by the live/dead technique and confocal microscopy. The results showed that 0.4% chlorhexidine induced a higher inflammatory response than the other groups. The Casearia sylvestris Sw extract showed satisfactory results in relation to the intensity of the inflammatory response. In the microbiological test, there were no statistical differences between the evaluated intracanal dressings and the percentage of bacterial viability was between 33 and 42%. The control group showed an 86% viability. Antimicrobial components such as chlorhexidine or Casearia sylvestris Sw did not improve the antimicrobial activity against E. faecalis in comparison to the calcium hydroxide+PG treatment. In addition, the incorporation of chlorhexidine in the calcium hydroxide paste promoted the highest inflammatory response.

Calcium hydroxide; Chlorhexidine; Endodontics; Materials testing

Inflamatory cells
	Giant cells			Mononuclear cells			Polymorphonuclear cells
	7 days	14 days	30 days	7 days	14 days	30 days	7 days	14 days	30 days
CH+CLX 0.4 %	1.125^A;1,2 (0.0000-3.500)	1.563^A;1 (0.0000-8.250)	0.0000^A;2 (0.0000-1.250)	14.44^A;1 (6.250-19.50)	11.25^AB;1 (7.500-24.75)	5.000^A;2 (4.250-7.750)	0.6250^A;1 (0.0000-1.750)	0.2500^AB;1,2 (0.0000-1.000)	0.0000^A;2 (0.0000-0.2500)
CH+CS	0.1250^A;1 (0.0000-3.250)	0.0000^B;1,2 (0.0000-0.5000)	0.0000^A;2 (0.0000-0.0000)	14.32^A;1 (9.250-42.50)	9.875^A;1 (3.250-15.50)	1.563^B;2 (0.5000-4.750)	0.0000^AB;1 (0.0000-0.5000)	0.0000^A;1 (0.0000-0.2500)	0.0000^A;1 (0.0000-0.2500)
CH+PG	2.000^A;1 (0.0000-5.500)	0.6250^AB;1,2 (0.0000-2.000)	0.0000^A;2 (0.0000-0.2500)	17.25^A;1 (7.750-28.00)	13.75^B;1 (5.500-16.75)	1.188^B;2 (0.2500-4.000)	0.0000^B;1 (0.0000-1.500)	0.6875^B;1 (0.0000-1.750)	0.0000^A;1 (0.0000-0.5000)

Histological events
	Collagen fibers			Fibroblasts			Blood vessels
	7 days	14 days	30 days	7 days	14 days	30 days	7 days	14 days	30 days
CH + CLX 0,4 %	52.07^A;1 (43.50-58.00)	42.38^A;1 (17.50-54.75)	63.75^A;2 (57.75-71.75)	5.255^A;1 (3.500-6.750)	3.625^A;1 (1.000-7.500)	7.938^A;2 (4.500-11.00)	0.8750^A;1 (0.0000-5.130)	0.5000^A;1 (0.0000-3.500)	1.125^A;1 (0.5000-1.750)
CH+CS	46.50^A;1 (20.25-61.25)	56.63^B;1 (42.25-70.00)	53.06^B;1 (39.25-69.50)	7.875^B;1 (2.500-10.00)	7.500^B;1 (4.250-10.75)	7.563^A;1 (6.250-8.750)	1.250^A;1 (0.0000-2.750)	0.7500^A;2 (0.0000-1.250)	0.8125^A;1,2 (0.5000-1.500)
CH+PG	48.50^A;1 (36.25-52.25)	51.75^B;1 (45.50-67.50)	68.00^A;2 (53.50-79.00)	8.565^B;1 (4.000-10.75)	5.063^A;2 (2.000-8.750)	8.125^A;1 (4.500-10.50)	0.2500^B;1 (0.0000-1.500)	0.0625^A;1 (0.0000-1.250)	0.3750^B;1 (0.0000-0.5000)

Live cells (%)	CH + CLX 0,4 %	CH + CS	CH + PG	Positive control
Minimum	3.149	4.968	1.953	53.58
Median	42.45	34.58	33.59	86.62
Maximum	77.84	80.96	81.22	93.18
Mean	43.08	40.55	36.89	83.65

Faculdade De Odontologia De Bauru - USP Serviço de Biblioteca e Documentação FOB-USP, Alameda Dr. Octávio Pinheiro Brisolla 9-75, 17012-901 Bauru SP Brasil, Tel.: +55 14 3235-8373 - Bauru - SP - Brazil
E-mail: jaos@usp.br

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[1] Corresponding address: Raquel Zanin Midena - Alameda Octávio Pinheiro Brisolla, 9-75 - 17012-901 - Bauru - SP - Brazil - Phone/Fax number: 55 14-32358344 - e-mail: raquelmidena@gmail.com