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. 2022 Jun 30;7:35. doi: 10.1038/s41536-022-00229-4

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a ≥20 cm snout-to-tailtip axolotl femurs were exposed by dissecting upper hind limb skin and spreading the muscles above the femur. b A titanium plate (7.75 mm Plate, RISystem, Switzerland) was attached to the femur by 4 titanium screws. c A saw guide was used for osteotomy. d A Gigly wire (0.66 mm) was inserted under the bone and used for sawing. Green outline and arrow show the plastic foil used for soft tissue protection during the surgery. e Osteotomy resulted in 0.7 mm bone gap in axolotl (white arrow). f 0.7 mm fracture of mouse femur is shown (white arrow). Here, a 6-hole plate (10 mm MouseFix Plate XL, RISystem) was attached to the femur by 4 titanium screws to fixate the bone. Scale bars 1 mm. g Sketch of the three different osteotomy models used. Amputation of the limb at the mid-femur (left) leads to blastema formation in the region of the proximal femur end of the osteotomy gap of the plate-fixated fracture (middle) and the non-stabilized fracture (right). In the non-stabilized fracture the two bone ends do not stay aligned but converge in a random way. Dashed line shows the amputation plane.