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June 17, 2026Journal of Molecular Pathology0 citationsOpen Access

Comparative Cytotoxicity and Inflammatory Profiles of CeraSeal Versus AH Plus in Periodontal Tissue Repair: An In Vitro and In Vivo Study

GSGulnihol SharipovaBukhara State Medical Institute JRJasur RizaevSamarkand State University named after Sharof Rashidov SBShuxrat BoymuradovTashkent Pediatric Medical Institute

Key Points

To compare the cytotoxicity and inflammatory profiles of CeraSeal and AH Plus in periodontal tissue repair.
Exposed human periodontal ligament stem cells to sealer extracts for 120 hours and assessed cell death through various assays.
Utilized a rat model to evaluate inflammatory markers and osteogenic activity with two sealer types.
Conducted morphometric analysis to assess cellular preservation and inflammatory infiltration.
AH Plus showed greater necrotic cell death (357.6% LDH release) compared to CeraSeal (128.8%; p = 0.0079).
In vivo analysis showed higher IL-1β levels in AH Plus-treated tissues (52.25 cells/mm²) than in CeraSeal (24.88; p = 0.0002).
CeraSeal demonstrated superior cellular preservation (p = 0.0079) despite higher inflammatory infiltration compared to AH Plus.

Abstract

Background/Objectives: Endodontic perforation repair requires biomaterials that balance sealing ability with minimal cellular injury. AH Plus (epoxy resin-based) remains widely used despite cytotoxicity concerns. CeraSeal (calcium silicate-based bioceramic) is a potentially more biocompatible alternative. However, comparative data on sealer-induced cytotoxicity and inflammatory responses remain limited. This study compared the cytotoxicity and inflammatory profiles of CeraSeal and AH Plus using in vitro and in vivo approaches. Methods: Human periodontal ligament stem cells (hPDLSCs) were exposed to sealer extracts (1:4 AH Plus, 1:8 CeraSeal) for 120 h. Cell death was assessed by MTT, Live/Dead, LDH release, and Annexin V/PI flow cytometry. Oxidative stress was quantified via ROS generation (DCFH-DA). In a rat furcation perforation model (n = 8 teeth/group), inflammatory markers (TNF-α, IL-1β, CD68), osteogenic activity (ALP), and osteoclasts (TRAP) were evaluated. Results: AH Plus was associated with significantly greater necrotic cell death (357.6 ± 47.6% LDH release vs. CeraSeal 128.8 ± 37.5%; p = 0.0079) and reduced hPDLSC viability at all time points (p 0.05). In vivo, IL-1β was higher in AH Plus-treated tissues (52.25 vs. 24.88 cells/mm2; p = 0.0002), while TNF-α and CD68 were greater in CeraSeal (p ≤ 0.0011). ALP was higher in AH Plus (median 6.15 vs. 3.68; p = 0.0002), with no difference in TRAP-positive osteoclasts. Morphometric analysis showed superior cellular preservation with CeraSeal (p = 0.0079), while inflammatory infiltration was higher in CeraSeal (p = 0.0002). Conclusions: AH Plus was associated with a necrotic-inflammatory profile with elevated IL-1β and higher ALP expression. CeraSeal demonstrated better cellular preservation, lower LDH release, and a distinct inflammatory signature (higher TNF-α and CD68). These findings establish comparative response profiles for the two sealers and support CeraSeal as a potentially biocompatible alternative, though further mechanistic studies are warranted.

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