DOI: 10.5281/zenodo.18508661
title: Supplementary materials for: "Layer-by-layer soft-tissue constraints on passive ex vivo limb joint range of motion in quadrupedal mammals: implications for palaeobiological inference"

description: This Zenodo record contains the supplementary materials supporting the study on layer-resolved passive ex vivo flexion–extension range of motion (ROM) in quadrupedal mammals. The package includes: (i) the full dataset integrating specimen metadata, biological reference variables, and joint ROM measurements across tissue-layer conditions; (ii) a repeatability pilot (juvenile alpaca hindlimb) quantifying within- and between-evaluator variability and inter-rater agreement; and (iii) a representative image set documenting end-range positions and digitisation references used for ROM quantification across the dissection sequence.Abbreviations: S, skin and subcutaneous tissue; M, myofascia; CL, capsulo-ligamentous tissues; O, osteology only.

Supplementary File 1. Dataset of joint range of motion and biological reference data

File description:

This dataset compiles the full experimental and contextual information underpinning the study “Layer-by-layer soft-tissue constraints on passive ex vivo limb joint range of motion in quadrupedal mammals: implications for palaeobiological inference”. It includes specimen metadata, anatomical descriptors, and layer-resolved flexion–extension range of motion (RoM) measurements obtained from four extant mammals representing contrasting postural categories. Each row corresponds to a unique combination of specimen, limb (fore/hind), joint, anatomical axis, and tissue-layer condition (skin and subcutaneous tissues, myofascia, capsulo-ligamentous tissues, and osteology). By convention, negative values denote extension and positive values denote flexion.For each joint and tissue-layer condition, the file also provides summary statistics (mean, standard deviation, and percentage change between conditions), together with complementary biological reference variables (body mass, estimated maximum terrestrial speed, and locomotor habit) compiled from published zoological and biomechanical sources. These auxiliary variables are included to support comparative interpretation and scaling context for interspecific differences in soft-tissue constraints, rather than to enable formal allometric modelling.

Supplementary File 2. Repeatability pilot for maximal extension endpoints in a juvenile alpaca hindlimb (ex vivo ROM)

File description:

This spreadsheet reports a repeatability pilot quantifying maximal extension ROM endpoints in a juvenile alpaca hindlimb across tissue-layer conditions (S+M+CL+O, M+CL+O, CL+O, O). Two evaluators (R1–R2) obtained three repeated measurements (M1–M3) per joint and condition. Within-evaluator variability is summarised as mean (X), standard deviation (SD), and intra-rater coefficient of variation (CV, %). Between-evaluator differences are reported as inter-rater CV (%) and absolute mean differences (Δ, °).Inter-rater agreement was assessed using a two-way mixed-effects model with absolute agreement, reporting ICC(3,1) with 95% confidence intervals (single measures); ICC(3,2) is also provided for the mean of two evaluators.

Supplementary File 3. Representative image set for layer-resolved passive flexion–extension ROM measurements

File description:

This PDF provides representative still images documenting the end-range positions used to quantify passive flexion and extension for each joint and tissue condition. For each species and joint, one example is shown for flexion and one for extension across the full dissection sequence: S+M+CL+O (intact), M+CL+O (skin and subcutaneous tissue removed), CL+O (myofascia removed; capsulo-ligamentous tissues preserved), and O (osteology only). Images include calibrated digital angle overlays and the proximal and distal segment reference lines used for digitisation in Tracker v6.1.6. Black boxes mask the head/face for ethical presentation and do not obscure anatomical landmarks used for digitisation. Abbreviations: S, skin and subcutaneous tissue; M, myofascia; CL, capsulo-ligamentous tissues; O, osteology only.

Supplementary Video. Representative ex vivo ROM measurement in the chilla fox shoulder under the S+M+CL condition.

File description:

This silent video documents a representative trial used to quantify passive sagittal-plane flexion–extension range of motion (ROM) at the shoulder of the chilla fox (Lycalopex griseus) under the S+M+CL tissue condition (skin and subcutaneous tissues, myofascia, and the capsulo-ligamentous envelope preserved; osteology intact). The footage illustrates limb handling and joint manipulation to reach end-range positions. The video is provided as methodological documentation to complement the protocol described in the Methods and the representative still images in Supplementary File 3.