The aim of this study was to evaluate the effect of different irrigating solutions as well as their combination and activation modes on root canal dentin microhardness. The protocol was registered in PROSPERO and PRISMA guidelines were followed. The structured question was as follows: “Which type of irrigating solution used in endodontic treatment causes more change in dentin microhardness?” The literature was screened via PubMed, Google Scholar, Scopus, and Science Direct. The last search was carried out in February 2023 with English language restriction. Two reviewers independently performed screening and evaluation of articles. A total of 470 articles were retrieved from all the databases, whereas only 114 articles were selected for full-text analysis. After applying eligibility criteria, 44 studies were evaluated and included in this review. The results showed that with increased contact time with irrigants, dentin microhardness decreases. Increased contact time with sodium hypochlorite (NaOCl) was associated with more reduction in dentin microhardness compared with other irrigants. Other irrigants, with the exception of distilled water, including EDTA, citric acid, herbal irrigants, glycolic acid, phytic acid, etc., in this study significantly decreased dentin microhardness. The maximum reduction in dentin microhardness was seen with 2.5% NaOCl after 15 min of contact time. The use of irrigating solutions alters the chemical composition of dentin, thereby decreasing its microhardness, which affects the clinical performance of endodontically treated teeth.