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The layer\u2010by\u2010layer (LbL) assembly is a highly versatile methodology introduced 30 years ago, which consists of assembling complementary polyelectrolytes or biomolecules in a stepwise manner to form thin self\u2010assembled films. In view of its simplicity, compatibility with biological molecules, and adaptability to any kind of supporting material carrier, this technology has undergone major developments over the past decades. Specific applications have emerged in different biomedical fields owing to the possibility to load or immobilize biomolecules with preserved bioactivity, to use an extremely broad range of biomolecules and supporting carriers, and to modify the film's mechanical properties via crosslinking. In this review, the focus is on the recent developments regarding LbL films formed as 2D or 3D objects for applications in drug delivery and tissue engineering. Possible applications in the fields of vaccinology, 3D biomimetic tissue models, as well as bone and cardiovascular tissue engineering are highlighted. In addition, the most recent technological developments in the field of film construction, such as high\u2010content liquid handling or machine learning, which are expected to open new perspectives in the future developments of LbL, are presented.<\/jats:p>","DOI":"10.1002\/adhm.202302713","type":"journal-article","created":{"date-parts":[[2023,12,20]],"date-time":"2023-12-20T04:48:32Z","timestamp":1703047712000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":69,"title":["Recent Developments in Layer\u2010by\u2010Layer Assembly for Drug Delivery and Tissue Engineering Applications"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0126-8482","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Borges","sequence":"first","affiliation":[{"name":"CICECO \u2013 Aveiro Institute of Materials Department of Chemistry University of Aveiro Campus Universit\u00e1rio de Santiago  Aveiro 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