ORIGINAL RESEARCH article
Front. Cell Dev. Biol.
Sec. Stem Cell Research
Volume 13 - 2025 | doi: 10.3389/fcell.2025.1669459
This article is part of the Research TopicMesenchymal Stromal Cells: Biology, Therapeutic Potential, and Translational ChallengesView all articles
In vitro and in vivo osteogenesis of rat adipose-derived stem cells combined with calcium alginate gel scaffold induced by calcitonin gene-related peptide
Provisionally accepted
- 1Ningde Municipal Hospital of Ningde Normal University, Ningde, China
- 2Department of Orthopaedics, Shanghai Sixth People’s Hospital Fujian, Quanzhou, China
- 3The First Hospital of Putian City, Putian, China
- 4Ningde Clinical Medical College of Fujian Medical University, Ningde, China
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Bone defect repair is clinically challenging due to the limitations of traditional treatments. Tissue engineering holds great potential for constructing bone substitutes. This study evaluates the osteogenic capability of calcitonin gene-related peptide (CGRP)-induced rat adipose-derived stem cells (ADSCs) combined with calcium alginate (CaAlg) scaffolds both in vitro and in vivo. ADSCs were isolated from rat inguinal fat pads, cultured, and characterized at passage 3. For in vitro experiments, cells were grouped and assessed over time using the CCK-8 assay for proliferation, alkaline phosphatase (ALP) activity assays, ALP staining, alizarin red staining (ARS), RT-PCR, and Western blotting for osteogenesis-related gene and protein expression. For in vivo experiments, constructs were evaluated after 12 weeks using X-ray, micro-CT, gross observation, and H&E staining. Results showed that ADSCs had clear surface antigen characteristics and displayed an "S"-shaped proliferation curve post-osteogenic induction. In vitro, CGRP and CaAlg scaffolds synergistically enhanced ADSC osteogenic differentiation, with higher early ALP activity and late-stage mineralization in the CGRP-ADSCs-CaAlg group. Additionally, osteogenesis-related gene and protein expressions were upregulated in CGRP-induced and scaffold-combined groups. In vivo, bone formation was observed in both ADSCs-CaAlg and CGRP-ADSCs-CaAlg groups, but not in the control group. These findings indicate that CGRP can induce ADSCs combined with CaAlg scaffolds to form tissue-engineered bone in vivo, with CGRP and CaAlg scaffolds showing a synergistic effect on promoting ADSCs osteogenic differentiation.
Keywords: adipose-derived stem cells, Calcitonin Gene-Related Peptide, Calcium alginateHydrogel, Tissue Engineering, three-dimensional culture, Osteogenic differentiation
Received: 19 Jul 2025; Accepted: 01 Sep 2025.
Copyright: © 2025 Huang, Liu, Lin, Zhang, Xu, Wang and Lin. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Xiaofeng Liu, Department of Orthopaedics, Shanghai Sixth People’s Hospital Fujian, Quanzhou, China
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