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. 2021 May 28;6:29. doi: 10.1038/s41536-021-00139-x

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© The Author(s) 2021

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. 1 — a Histological section of tibial mono-cortical defects 5 days after injury, stained with Movat’s Pentachrome. The defect site is filled with soft tissue. b PCNA IHC shows active proliferation in the periosteum and defect site at this early time point. c Alkaline phosphatase staining demonstrates only a small region of osteogenic differentiation at the cortical edge. d After 14 days, the defects site is filled with woven bone, stained yellow-green with Pentachrome. e PCNA staining reveals absence of proliferation within the injury. f Alkaline phosphatase staining indicating osteogenic differentiation throughout the injury site. g Quantification of PCNA-positive cells in the injury site at POD 5 and 14. h Quantification of alkaline phosphatase staining at POD 5 and 14. i Frequency of bone-cartilage-stromal progenitor cells (BCSPs) was analyzed over the time course of fracture healing by flow cytometry using the following BCSP markers: CD45⁻ Ter119⁻ Tie2⁻ CD51⁺ CD90⁻ 6C3⁻ CD105⁺. Scale bar = 100 μm. ALP alkaline phosphatase, BCSP bone-cartilage-stromal progenitor cells, c cortical bone, PCNA proliferating cell nuclear antigen, POD postoperative day. **p < 0.01. Data were represented as mean ± s.e.m.