This systematic review aimed to determine how cellulose nanofiber (CNF) enhance the mechanical properties, biocompatibility, and functional versatility of dental materials, based on in vitro evidence. A systematic review protocol was developed according to PRISMA 2020. The literature search was conducted using MEDLINE/PubMed, Scopus, Web of Science, Scopus, and Embase for English-language publications with no date restrictions, yielding 180 publications. After removing duplicates, 150 remained, from which 23 were evaluated in full. 17 in vitro studies met the inclusion criteria, of which 13 had a low risk of bias, 4 had a moderate risk, and none had a high risk. Across all included investigations, CNF consistently improved mechanical performance, particularly flexural and compressive strength, while maintaining or enhancing biocompatibility in various cell culture models. CNF’s nanofibrillar structure and modifiable surface chemistry also expanded its functional versatility, enabling applications such as drug encapsulation and targeted antimicrobial delivery. Overall, CNF emerges as a promising biomaterial for modern dentistry, offering superior mechanical reinforcement, favorable cell responses, and wide-ranging functional modifications. Further in vivo research and clinical trials are necessary to confirm its long-term safety and efficacy, thereby facilitating the translation of these in vitro findings into standard dental practice.
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