Cytotoxicity, inflammation, biomineralization, and immunoexpression of IL-1β and TNF-α promoted by a new bioceramic cement

COSME-SILVA, Leopoldo; SANTOS, Amanda Ferreira dos; LOPES, Camila Soares; DAL-FABBRO, Renan; BENETTI, Francine; GOMES-FILHO, João Eduardo; QUEIROZ, India Olinta de Azevedo; ERVOLINO, Edilson; VIOLA, Naiana Viana

doi:10.1590/1678-7757-2020-0033

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Brasil

Journal of Applied Oral Science

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Original Article • J. Appl. Oral Sci. 28 • 2020 • https://doi.org/10.1590/1678-7757-2020-0033 copy

Cytotoxicity, inflammation, biomineralization, and immunoexpression of IL-1β and TNF-α promoted by a new bioceramic cement

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Aim

To evaluate the cytotoxicity, biocompatibility and mineralization capacity of BIO-C PULPO, and MTA.

Methodology

L929 fibroblasts were cultured and MTT assay was used to determine the material cytotoxicity on 6, 24, and 48 h. A total of 30 male rats (Wistar) aged between 4 and 6 months, weighing between 250 and 300 g were used. Polyethylene tubes containing BIO-C PULPO, MTA, and empty tubes were implanted into dorsal connective tissue. After the experimental periods (7, 15, 30, 60, and 90 days) the tubes were histologically analyzed using hematoxylin-eosin (H&E), immunolabeling of IL-1β and TNF-α, and von Kossa staining, or without staining for polarized light analysis. The average number of inflammatory cells was quantified; the mineralization assessment was determined by the area marked in μm2 and semiquantitative immunolabeling analyses of IL-1β and TNF-α were performed. Then, data underwent statistical analysis with a 5% significance level.

Results

It was observed that BIO-C PULPO and MTA presented cytocompatibility at 6, 24, and 48 similar or higher than control for all evaluated period. On periods 7 and 15 days, BIO-C PULPO was the material with the highest number of inflammatory cells (p<0.05). On periods 30, 60, and 90 days, BIO-C PULPO and MTA presented similar inflammatory reactions (p>0.05). No statistical differences were found between Control, BIO-C PULPO, and MTA for immunolabeling of IL-1β and TNF-α in the different periods of analysis (p<0.05). Positive von Kossa staining and birefringent structures under polarized light were observed in all analyzed periods in contact with both materials, but larger mineralization area was found with BIO-C PULPO on day 90 (p<0.05).

Conclusion

BIO-C PULPO was biocompatible and induced mineralization similar to MTA.

Biocompatibility; Biomaterials; Cytotoxicity; Pulpotomy; Dental Pulp

Time	Material	Inflammatory Cells	VK	POL	Fibrous capsule	Immunolabeling
		Mean ±SD*	Median (min-max values)^†	Median (min-max values)^†		Scores (median)^§ IL-1β TNF-α
7 days	Control	299 (±29)^aA	0 (0-0)^aA	0 (0-0)^aA	Thick	2^a	2.5^a
	MTA	214 (±50)^bA	51,365 (31,723-99,329)^bA	64,729 (38,748-89,105)^bA	Thick	2^a	2.5^a
	BIO-C PULPO	328 (±69)^aAB	15,538 (1,681-178,434)^bA	39,930 (2,980-128,260)^abA	Thick	3^a	3^a
15 days	Control	254 (±65)^aAB	0 (0-0)^aA	0 (0-0)^aA	Thick	2^a	2^a
	MTA	212 (±45)^aA	51,253 (32,558-73,778)^bA	45,671 (24,169-96,900)^bA	Thick	2^a	2^a
	BIO-C PULPO	396 (±84)^bA	24,142 (20,131-33,237)^abA	24,446 (12,515-34,351)^abA	Thick	2^a	2.5^a
30 days	Control	146 (±50)^aB	0 (0-0)^aA	0 (0-0)^aA	Thin	2^a	2^a
	MTA	206 (±88)^bA	27,652 (20,184-32,611)^bB	29,127 (17,423-43,998)^bAB	Thin	2^a	2^a
	BIO-C PULPO	294 (±107)^bAB	35,387 (25,120-999,70)^bA	21,538 (11,738-34,472)^bA	Thin	2^a	2^a
60 days	Control	134 (±21)^aB	0 (0-0)^aA	0 (0-0)^aA	Thin	2^a	2^a
	MTA	157 (±64)^aA	24,965 (16,840-31,919)^abAB	24,836 (21,956-31,085)^bAB	Thin	1.5^a	2^a
	BIO-C PULPO	204 (±57)^aBC	44,193 (30,507-67,682)^bA	29,396 (18,285-58,402)^bA	Thin	1.5^a	2^a
90 days	Control	144 (±8)^aB	0 (0-0)^aA	0 (0-0)^aA	Thin	1^a	1^a
	MTA	135 (±31)^aA	16,046 (3,889-16,979)^bB	15,458 (10,961-30,939)^bB	Thin	1^a	1^a
	BIO-C PULPO	138 (±24)^aC	27,992 (19,820-35,860)^cA	16,897 (12,563-17,232)^bA	Thin	1^a	1^a

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