Efficacy of Chitosan as Final Irrigating Solution on TGF-β1 Release from Root Canal Dentin and its Effect on Dental Pulp Stem Cells Response

Abstract

Purpose: To study the effect of Chitosan final irrigating solution compared with EDTA using different irrigation techniques, on TGF-β1 release from root canal dentin, using enzyme-linked immunosorbent assay (ELISA), and to evaluate the effect of released TGF-β1 on Dental Pulp Stem Cells. Materials and Methods: Human DPSCs were isolated from impacted third molars. Forty eight human root segments were prepared to a standardized truncated cone-shaped canal with an open apex of 1 mm diameter, the volume of prepared segments were determined using CBCT.Samples were randomized into two experimental groups (n=24) regarding the final irrigating solution; Group I: 1.5% NaOCl followed by 17% EDTA, Group II: 1.5% NaOCl followed by 1% Chitosan, then subdivided into subgroups A and B (n = 12) according to the final irrigation technique: A: Needle Irrigation, B: XP-Endo Finisher agitation. The released TGF-β1 were quantified using ELISA at 4 hours, 1 day, and 3 days following irrigation. DPSCs response to1.5% NaOCl, 17% EDTA, 1% Chitosan, and different concentrations of released TGF-β1 was assessed using MTT assay. Results: No statistically significant difference in the quantity of TGF-β1 released by 17% EDTA and 1% Chitosan following needle irrigation or XP Endo Finisher agitation.1% Chitosan significantly increased DPSCs viability compared to17% EDTA (P < 0.05).TGF-β1 showed significant expansion of viable DPSCs in a positive correlation to its concentration (P <0.001). Conclusions: 1% Chitosan effectively released TGF-β1 from root canal dentin as17% EDTA. The released TGF-β1 promoted DPSCs proliferation at picogram levels in a concentration dependent manner.

Impact of using XP-endo finisher and nanobubble water during EDTA dentin conditioning on TGF-β1 release in regenerative endodontic procedures

“…TGF-β1 was quantified using ELISA following the canal irrigation procedure. This approach aligns with prior research, as measuring growth factors post-irrigation mirrors the clinical scenario where bleeding is induced at the apex after irrigation and drying [ 6 , 14 , 45 ]. This differs from the study by Galler et al, where growth factor release was measured during irrigation [ 10 ].…”

“…On the other hand, in the EDTA/DW/XP group, the positive effect of the EDTA activation using the XP-endo Finisher can be attributed to enhanced smear layer removal and thereby reaching inaccessible areas while preserving dentin. Moreover, the XP-endo Finisher has improved irrigation dynamics as the file exhibits increased flexibility and enhanced expansion capacity which leads to the improvement of the penetration of the irrigant into the dentinal tubules [ 45 ]. The increase in the TGF-β1 release in the EDTA/DW/XP group is in accordance with studies that showed that irrigation activation increases the growth factor release from dentin [ 7 , 13 , 34 , 47 ].…”

“…In this investigation, the root segment model was employed by decoronating the teeth to a 12 ± 1 standard length in accordance with several studies [13,15,39]. Using the root segment model is more closely aligned with the clinical scenario, since the release of growth factors occurs from the inner dentin surface exclusively [17].…”

“…Additionally, it mediates signaling the proliferation, differentiation and mineralization of odontoblasts [2,10,12]. Growth factors are stored in dentin, and their release is triggered by conditioning and demineralizing the dentin matrix with chelating agents like EDTA, citric acid, phosphoric acid, and chitosan [6,11,[13][14][15][16]. However, EDTA 17% has showed superior results regarding the growth factors' release [17].…”