The objective here was to experimentally characterise the temporal evolution of the structural and mechanical properties of large volume immature regenerated tissues. We studied these evolving tissues from their genesis in controlled mechanical conditions. We developed an animal model based on the periosteal properties leading to unloaded regenerated skeletal tissue. To characterize the temporal evolution of mechanical properties, we carried out indentation tests coupled with macroscopic examinations and histological studies. This combined methodology yielded a range of information on osteogenesis at different scales: macroscopic by simple observation, mesoscopic by indentation test and microscopic by histological study. Results allowed us to identify different periods, providing a link between biological changes and material property evolution in bone tissue regeneration. The regenerated tissue evolves from a viscous, homogeneous, soft material to a heterogeneous stiffer material endowed with a lower viscosity. From a biological point of view, cell organization progresses from a proliferated cell clot to a mature structure closer to that of the bone. During the first seven days, mechanical and biological results revealed the same evolution: first, the regenerated tissue grew, then, differentiated into an osteochondral tissue and finally calcification began. While our biological results confirm those of other studies, our mechanical results provide the first experimental mechanical characterization by reduced Young's modulus of such tissue.Response to Reviewers: Comments for the Author:Editor's comment: A revised version of the manuscript should address all the points raised by the reviewers. In addition, please let the manuscript be revised by a native speaker of English and mention all changes made. In addition, change the title to: TEMPORAL EVOLUTION OF SKELETAL REGENERATED TISSUE: WHAT CAN MECHANICAL INVESTIGATION ADD TO BIOLOGICAL? The heading: Discussion and Conclusions should be changed to Discussion. There should be concluding statement of not more than 2-4 sentences at the end of the discussion. The references should be put in order according to the instructions to authorsWe take into account all these remarks Reviewer #1: The authors have attempted to accommodate most of the reviewer's concerns, and the paper is now more informative. For example, an engineering interpretation of the experiments has been added by developing a simple Finite Element Model of the biologic material. This reviewer truly appreciates the efforts done by the authors, and thinks that the manuscript results to be improved in its revised version.
Reviewer #3: CommentsThe authors should improve the flow and comprehensibility of language in their manuscript. Sections that have been added in the manuscript following the initial review comments should be quite more elaborate and thoroughly edited prior to publication.