Histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis of tissue response to sealing materials after furcation perforation

Abstract

Aim To evaluate in vivo tissue responses after sealing furcation perforations in dog's teeth with either Biodentine™, mineral trioxide aggregate (MTA) or gutta‐percha, by means of histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis. Methodology After root canal treatment, perforations were created in the central region of the pulp chamber floor using a round diamond bur and filled with one or other of the materials. The animals were euthanized after 120 days, and the teeth (n = 30) were processed for histopathological analysis of new mineralized tissue formation and collagen fibre reinsertion, immunohistochemical analysis of osteopontin (OPN) and alkaline phosphatase (ALP) and immunofluorescence analysis for bone morphogenetic protein (BMP‐2), cementum attachment protein (CAP), bone sialoprotein (BSP), osteocalcin (OCN) and cementum protein1 (CEMP1). Histoenzymology was performed for TRAP activity and osteoclast count. Data were analysed statistically (α = 0.05) using chi‐square and Kruskal–Wallis tests. Results Gutta‐percha did not induce mineralized tissue formation. MTA and BiodentineTM formed mineralized tissue in 88% and 92% of specimens, respectively, with no significant difference (P > 0.05). Gutta‐percha was associated with scattered collagen fibres parallel to the perforations. Groups treated with MTA or BiodentineTM had partial fibre reinsertion perpendicular to the newly formed mineralized tissue. All materials induced OPN and ALP expression, weakest for gutta‐percha and strongest for MTA (P < 0.05). Only MTA induced BMP‐2, BSP, OCN, CAP and CEMP1 expression. Osteoclast counts were similar in all groups (P = 0.97). Conclusions Mineral trioxide aggregate and BiodentineTM were biocompatible, with formation of mineralized tissue and partial reinsertion of collagen fibres. In addition, the participation of several molecules by which calcium silicate‐based materials induce the formation of mineralized tissue were noted, with expression of ALP and OPN mineralization markers, without interference in the number of osteoclasts. Only MTA stimulated the expression of proteins associated with the formation of a cementum‐like mineralized tissue.

Bone morphogenetic proteins in biomineralization of two endodontic restorative cements

“…This could suggest a reaction to the implantation of BD, indicating its mineralizing potential, as in cytokine measurements in the present study, this material significantly expressed IL‐8 at 30 days and also expressed mRNA of BMP2 in the same period. As for MTA and BMP2, our study confirmed the findings of Önay et al, 27 but Silva et al 28 showed that MTA stimulates BMP2 expression. This suggests that these affinities could result from the close relationship of BMPs with osteoblasts, where both BMP2 and BMP4 have effects on osteoblasts, 73 making it difficult to elucidate the specific roles during osteogenesis.…”

“…In addition, only one study to date has assessed the relationship between BMP4 and MTA, thus confirming its osteogenic‐odontogenic potential 26 . As for BMP2, a study has demonstrated no relationship between BMP2 and MTA, only in a special formulation known as MM‐MTA 27 ; however, another study has shown that MTA stimulates BMP2 expression 28 …”

“…26 As for BMP2, a study has demonstrated no relationship between BMP2 and MTA, only in a special formulation known as MM-MTA 27 ; however, another study has shown that MTA stimulates BMP2 expression. 28 Accordingly, the aim of the present study is to assess the role of MTA and BD in BMPs and inflammatory cytokines to elucidate the mechanisms whereby these materials induce mineralization and to confirm their biocompatibility.…”

Does MTA provide a more favourable histological response than other materials in the repair of furcal perforations? A systematic review

“…Their protocols for outcome assessment differed in, for example, observation time points and outcome assessment methods. Many studies assessed these characteristics by means of scores (Aladimi et al, 2020; Al‐Daafas & Al‐Nazhan, 2007; Bakhtiar et al, 2017; Cardoso et al, 2018b; Noetzel et al, 2006; Samiee et al, 2010; Silva et al, 2019; de Sousa Reis et al, 2019) or cell counting (da Fonseca et al, 2019; Hassanien et al, 2015; Neto et al, 2010, 2012; da Silva et al, 2011; Silva et al, 2017; Tawfik et al, 2016). Even those studies that analysed the outcomes by means of scores evaluated different characteristics in each score.…”

Present status and future directions – Managing perforations

“…The aim of perforation management is to maintain healthy periodontal tissues adjacent to the perforation without persistent inflammation or loss of periodontal attachment. In the case of established periodontal tissue breakdown, the aim is to re‐establish tissue attachment (Pitt Ford et al, 1995; Silva et al, 2019). Thus, successful perforation repair depends on the ability to seal the perforation and to re‐establish a healthy periodontal ligament (Seltzer et al, 1970).…”