Influence of porosity and Young's modulus of gelatin methacrylate anhydride hydrogels on the biological behavior of mouse bone marrow mesenchymal stem cells

Kong Yue; Tian Feng; Liu Qinghua; Zhang Chao; Song Wei; Kong Yi; Li Zhao; Enhejirigala; Huang Sha

doi:10.3760/cma.j.cn501225-20250630-00286

Volume 41 Issue 10

Oct. 2025

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2025 > 41(10): 958-967

Kong Y,Tian F,Liu QH,et al.Influence of porosity and Young's modulus of gelatin methacrylate anhydride hydrogels on the biological behavior of mouse bone marrow mesenchymal stem cells[J].Chin J Burns Wounds,2025,41(10):958-967.DOI: 10.3760/cma.j.cn501225-20250630-00286.

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Influence of porosity and Young's modulus of gelatin methacrylate anhydride hydrogels on the biological behavior of mouse bone marrow mesenchymal stem cells

doi: 10.3760/cma.j.cn501225-20250630-00286

Kong Yue¹,
Tian Feng²,
Liu Qinghua²,
Zhang Chao²,
Song Wei²,
Kong Yi²,
Li Zhao²,
Enhejirigala^{1
,
,},
Huang Sha^{2
,
,}

1.
Department of Human Anatomy, School of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot 010010, China
2.
Research Center for Wound Repair and Tissue Regeneration, Medical Innovation Research Department, the PLA General Hospital, Beijing 100048, China

Funds:

General Program of National Natural Science Foundation of China 82472166, 32471432

Beijing Natural Science Foundation-Changping Innovation Joint Fund L234066

Natural Science Foundation Project of Inner Mongolia Autonomous Region 2025MS08143

Support Program for Young Science and Technology Talents in Higher Education Institutions of Inner Mongolia Autonomous Region NJYT24031

Doctoral Start-up Fund of Inner Mongolia Medical University YKD2023BSQD012

A University-level General Project of Inner Mongolia Medical University YKD2024MS006

Guiding Fund of Central Government for Supporting the Development of the Local Science and Technology 2024ZY0127

More Information

Corresponding author: Enhejirigala, Email: 572563157@qq.com; Huang Sha, Email: stellarahuang@sina.com
Received Date: 2025-06-30

Abstract

Abstract

Objective To investigate the influence of porosity and Young's modulus of gelatin methacrylate anhydride (GelMA) hydrogels on the biological behavior of mouse bone marrow mesenchymal stem cells (BMSCs), and to provide a theoretical basis for the development of tissue engineering scaffolds for skin wound repair. Methods This study was an experimental study. GelMA hydrogels that were prepared by photo-crosslinking 50 g/L GelMA solution for 30 and 60 seconds and 150 g/L GelMA solution for 30 and 60 seconds, were designated respectively as low-concentration short-time group, low-concentration long-time group, high-concentration short-time group, and high-concentration long-time group. The porosity of the 4 groups of hydrogels after freeze-drying for 48 hours was measured and calculated using Image J 1.54 software. The Young's modulus of the 4 groups of hydrogels was detected using a Young's modulus tester. The correlation between the porosity and Young's modulus was evaluated and a predictive model between them was established. The remaining mass percentages of the 4 groups of hydrogels after soaking in phosphate buffered saline for 7 days were calculated. BMSCs were isolated from the femurs of 10 C57BL/6 mice aged 1 day of unknown sex. Cell-loaded GelMA hydrogels were prepared by mixing BMSCs with 50 g/L GelMA solution and photo-crosslinking for 30 and 60 seconds, and by mixing BMSCs with 150 g/L GelMA solution and photo-crosslinking for 30 and 60 seconds, and were designated respectively as low-concentration short-time cell-loaded group, low-concentration long-time cell-loaded group, high-concentration short-time cell-loaded group, and high-concentration long-time cell-loaded group. The cell survival in the 4 cell-loaded groups of hydrogels after 7 days of culture was detected using a live/dead cell kit, and the cell survival rate was calculated. After 10 days of osteogenic induction culture of the hydrogels in the 4 cell-loaded groups, the mRNA expressions of osteogenic-related factors Runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP), and stemness-related factors SRY-box transcription factor 2 (Sox2) and Nanog in the cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction; the protein expressions of Runx2 and Nanog in the cells were detected by immunofluorescence method. Except for the protein expression (qualitative observation), the sample size of each group for the other indicators was three. Results The porosity of hydrogels in low-concentration short-time group, low-concentration long-time group, high-concentration short-time group, and high-concentration long-time group after freeze-drying for 48 hours decreased successively, being (92.7±0.9)%, (85.0±1.8)%, (68.6±1.2)%, and (56.3±5.8)%, respectively; the Young's modulus increased successively, being (5.933±0.020), (7.803±0.089), (20.772±0.106), and (22.498±0.060) kPa, respectively. Except for the Young's modulus of hydrogels between the high-concentration long-time group and high-concentration short-time group (P>0.05), the pairwise comparisons of hydrogel porosity or Young's modulus among the remaining groups showed statistically significant differences (P<0.05). The porosity and Young's modulus of the hydrogels were significantly negatively correlated (R²=0.91, P<0.05), and a linear equation predictive model between them was established accordingly. After soaking for 7 days, the remaining mass percentages of hydrogels in low-concentration long-time group, high-concentration short-time group, and high-concentration long-time group were significantly higher than that in low-concentration short-time group (P<0.05), and the remaining mass percentage of hydrogels in high-concentration long-time group was significantly higher than that in low-concentration long-time group (P<0.05). After 7 days of culture, a large number of live cells was observed in the 4 cell-loaded groups of hydrogels, and there was no statistically significant difference in the overall comparison of cell survival rate among the groups (P>0.05). After 10 days of osteogenic induction culture, the mRNA expressions of Runx2 and ALP in the cells of hydrogels in high-concentration long-time cell-loaded group were significantly higher than those in the other three cell-loaded groups (P<0.05), and the mRNA expressions of Sox2 and Nanog in the cells of hydrogels in low-concentration short-time cell-loaded group were significantly higher than those in the other three cell-loaded groups (P<0.05); the protein expression of Runx2 in the cells of hydrogels in high-concentration long-time cell-loaded group was the highest, and the protein expression of Nanog in the cells of hydrogels in low-concentration short-time cell-loaded group was the highest. Conclusions The porosity and Young's modulus of GelMA hydrogel synergistically regulate the biological behavior of mouse BMSCs, where high porosity and low Young's modulus are conducive to maintaining stemness, while low porosity and high Young's modulus drive osteogenic differentiation.
- Biocompatible materials,
- Hydrogels,
- Gelatin methacrylate anhydride,
- Porosity,
- Young's modulus,
- Bone marrow mesenchymal stem cells,
- Stemness,
- Osteogenic differentiation

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