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. 2024 Apr 25;14:9497. doi: 10.1038/s41598-024-59713-0

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Histologic features of RNS variants. (A–C) Hematoxylin–eosin staining showed numerous tortuous blood vessels (bv) with an abnormally large diameter in the papillary layer and associated extensive inflammatory infiltrates in RNS-1 gingiva (B). Abnormally large and shredded collagen bundles running in all directions were found in RNS-2 gingiva (C). (D–F) Picrosirius red polarizing staining. (D) Fibers of collagen organized in bundles of fixed diameter running in perpendicular pathways. (E,F) In RNS-1 and RNS-2, collagen fibers did not organize in packed bundles, but appeared shredded (arrow) and organized in a mesh network. (G,H) Alizarin red staining identified calcium aggregates (arrow) in the reticular layer of the lamina propria of RNS-1 (G) and RNS-2 (H) mutants. Scale bars: (A–C), (G,H) = 200 μm; (D–F) = 50 μm.