Architecture mechanics mediated osteogenic progression in bone regeneration of artificial scaffolds

Long, Siyu 1 ; Yu, Peng1; Yang, Wei 1

Research facility: Sichuan University

Published Jun 25, 2025 on Dryad. https://doi.org/10.5061/dryad.dr7sqvb8w

Data files

Jun 25, 2025 version files 1.25 MB

Data_files.zip

1.25 MB
README.md

5.32 KB

Abstract

Scaffold architecture exerts a considerable influence on the osteogenic effect through stress transmission, as the deformation of scaffolds alters the mechanical microenvironment of cells adhering to scaffold surface. Despite extensive researches on bone regeneration influenced by scaffold architecture, present studies have not addressed the biological mechanism underlying scaffold architecture-induced stress stimulation (SASS) on cells yet, posing a great challenge in revealing the biomechanical cues between scaffold architecture and osteogenic progression. Therefore, Graphite (GP), Fullerene (FL), and Diamond (DM) scaffolds with gradient stress-stimulation to cells after deformation were prepared. Moreover, we analyzed the results of the finite element simulation of the compression and recovery process of the three types of scaffolds, and tested the macroscopic mechanical and viscoelastic properties. The cellular biomechanical mechanisms of SASS through single-cell RNA sequencing indicated that architectures providing SASS can induce the enrichment of focal adhesion and osteogenic differentiation pathways of bone mesenchymal stem cells, and balance bone resorption of osteoclasts and bone formation of osteoblasts. Besides, SASS enhances bone regeneration for repairing critical-sized defects in vivo. These results provide insights for artificial bone scaffold design and clarify the biomechanical cues between SASS and osteogenic progression.

Dataset DOI: 10.5061/dryad.dr7sqvb8w

Description of the data and file structure

We summarized the finite element simulation of the micro-strain distribution, macro-mechanical, and viscoelastic properties of GP, FL, and DM scaffolds during the compression and recovery process. One month after implantation in the femur of rats, the new bone at the defect was removed, and single-cell sequencing was performed. The KEGG, GO enrichment, and gene expression differences of mesenchymal stem cells (MSCs), osteoclasts (OCs), and osteoblasts (OBs) cell clusters in the three scaffolds were analyzed. In the in vivo experiments, the parameters of new bone, semi-quantitative analysis of IHC and IF were analyzed.

Files and variables

File: Data_files.zip

Description:

Porosity&surface-volume ratio.xlsx: Porosity and surface-volume ratio of GP, FL, and DM scaffolds.

Step1_Element proportion-strain curves.csv, Step1.5_Element proportion-strain curves.csv, Step2_Element proportion-strain curves.csv, Step3_Element proportion-strain curves.csv, Step3.5_Element proportion-strain curves.csv, Step4_Element proportion-strain curves.csv: Element proportion-strain curves at six time points (step 1, 1.5, 2, 3, 3.5, 4) for GP, FL, DM scaffolds and Bone (Step 1: Compress to 10N; Step 2: Maintain the gap for 10 minutes; Step 3: Return to the force of 0.05N; Step 3.5: Step 4: Maintain the gap for 5 minutes). "Start" and "end" refer to the range of strain, that is, the initial value and the final value. "Count" refers to the number of elements. "Ratio" refers to the proportion of the number of elements to the total number of elements. "Ratio(*100)" means "ratio" multiplied by 100. Step 1, 2, 3, and 4 refer to the end time points of each step, Step 1.5 is at the midpoint between Step 1 and Step 2, and Step 3.5 is at the midpoint between Step 3 and Step 4.

Mode, average, SD, peak width at half height.xlsx: The strain analysis on the elements proportion-strain curves of the mode, the average, the standard deviation, the peak width at half-height for GP, FL, DM scaffolds, and Bone.

Stress, total strain, elastic strain and creep strain curves for the elements with the mode&max strain in the process of step 1-4.xlsx: Stress, total strain, elastic strain and creep strain curves for the elements with the mode proportion of strain and the elements with the largest strain in the process of step 1-4.

Compressive curves&strength&modulus.xlsx: Compressive curves, strength, and modulus of the GP, FL, and DM scaffolds.

Macroscopic viscoelasticity.xlsx: The stress-time curves of the four-step process (Step 1: Compress to 10N; Step 2: Maintain the gap for 10 minutes; Step 3: Return to the force of 0.05N; Step 4: Maintain the gap for 5 minutes) of the GP, FL, DM scaffolds and the cancellous bones of pigs. The relaxation curves of step 2 and the recovery curves of step 4 are shown after normalizing the initial point. The creep curves of the GP, FL, DM scaffolds, and Bone. Compression cycle of the GP, FL, DM scaffolds, and Bone.

Gene expression.xlsx: The transcript levels of cell markers in the mesenchymal stem cell (MSCs), osteoclasts (OCs), osteoblasts (OBs), and osteocytes subtypes expressing these marker genes. The transcript levels of related genes in GP, FL, and DM scaffolds in MSCs, OCs, and OBs.

Cell ratio.xlsx: Cell ratio in the MSCs, OCs, OBs, and osteocytes subtypes from DM, FL, and GP scaffolds. The proportion of immune cells in the GP, FL, and DM groups.

KEGG enrichment.xlsx, GO enrichment.xlsx: KEGG pathway enrichment analyses of all differentially expressed genes by scRNA-seq in DM vs. GP scaffolds, DM vs. FL scaffolds, and FL vs. GP scaffolds in MSCs, OCs, OBs. GO pathway enrichment analyses of all differentially expressed genes by scRNA-seq in DM vs. GP scaffolds, DM vs. FL scaffolds, and FL vs. GP scaffolds in MSCs, OCs, and OBs.

RT PCR.xlsx: Real-time PCR of BMSCs, OBs, and OCs under dynamic compression in vitro (The expression levels of Fn1 and Ibsp in BMSCs, the expression levels of Fn1 and Ibsp in OBs, and the expression levels of Spp1 and Acp5 in OCs).

Routine blood tests.xlsx: Routine blood tests for white blood cell count (WBC), lymphocyte count (Lymph), Monocyte count (Mon), neutrophil granulocyte count (Gran), red blood cell count (RBC), hemoglobin (HGB), platelet count (PLT) and mean platelet volume (MPV) of control group and critical defect healing 2 days and 2 weeks after GP, FL, DM scaffolds implantation in rats.

BV_TV&BMD&Tb.Th.&Tb.N.&Bending strength.xlsx: Micro-CT quantification of critical defect femur in rats and Bamapigs healing 4 weeks and 12 weeks after surgery for GP, FL and DM groups. BV/TV, BMD, Tb.N., Tb.Th.. Bending strength of critical defect femurs in rats at 4 weeks and 12 weeks after implantation in the Bone, Blank, GP, FL, and DM groups.

Sirius red.xlsx: Semi-quantitative positive area results of Sirius red staining.

Statistics of IHC&IF.xlsx: Semi-quantitative mean optical density results of immunohistochemistry (IHC) staining. Semi-quantitative positive area results of immunofluorescence (IF) staining.