Mesenchymal stem/stromal cells-derived exosomal miRNA delivery enhances bone repair in osteoporotic conditions
Data files
Jul 27, 2026 version files 53.03 KB
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Figure_1.xlsx
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Figure_2.xlsx
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Figure_3.xlsx
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Figure_4.xlsx
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Figure_S1.xlsx
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README.md
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Abstract
Osteoporosis-associated bone fractures are a leading cause of disability in the elderly population. Developing effective therapeutic strategies to enhance bone repair under osteoporotic conditions remains a major clinical challenge. Increasing evidence indicates that aberrant lineage commitment of mesenchymal stem cells (MSCs) resident in bone marrow contributes to osteoporosis-related bone loss. However, incomplete understanding of the regulatory mechanisms governing MSC differentiation has limited the development of efficient therapeutic approaches. In this study, we identified microRNA-423 (miR-423) as a negative regulator of osteogenic differentiation, and demonstrated that inhibition of miR-423 significantly enhanced osteoblast differentiation of MSCs. To enable in vivo delivery of the miR-423 inhibitor for bone repair, MSC-derived exosomes (MSC-Exo) were employed as a delivery vehicle, generating Exo–miR-423 inhibitor construct. These exosomes were subsequently incorporated into an apatite-coated poly(lactic-co-glycolic acid) (PLGA) scaffold to form an Exo–miR-423 inhibitor/scaffold complex. Implantation of this complex significantly promoted bone healing in a calvarial defect model in ovariectomized (OVX) mice. Collectively, these findings demonstrate a promising miRNA modulated, exosome-based tissue engineering strategy for enhancing bone defect and fracture repair under osteoporotic conditions, and highlight its potential for further optimization and translational application.
Dataset DOI: 10.5061/dryad.d51c5b0j3
Description of the data and file structure
This dataset includes raw measurements of the osteogenic properties of scaffolds that integrate cell-derived exosomes loaded with miR-423 inhibitors in vitro and in a mouse calvarial defect model.
Files and variables
File: Figure_1.xlsx
Description: miR-423 inhibition promotes osteogenesis in OVX-derived MSCs. (B) Expression of osteogenic genes in OVX-derived MSCs treated with miR-423 inhibitor, miR-423 mimic, or control.
MSCs isolated from OVX mice were treated with miR-423 mimic, miR-423 inhibitor, or PBS control and subsequently induced toward osteogenic differentiation. The expression of osteogenic markers was measured by a real-time PCR assay.
File: Figure_2.xlsx
Description: miR-423 inhibition increases ALP expression and mineral deposition in OVX-derived MSCs. (B) ALP expression in OVX-derived MSCs treated with miR-423 inhibitor, miR-423 mimic, or PBS at day 7. (D) Mineral deposition in OVX-derived MSCs treated with miR-423 inhibitor, miR-423 mimic, or PBS at day 21, evaluated by Alizarin Red staining.
MSCs isolated from OVX mice were treated with miR-423 mimic, miR-423 inhibitor, or PBS control and subsequently induced toward osteogenic differentiation. To evaluate ALP activity, cells were lysed, incubated with ALP substrate solution, and absorbance was measured at 405 nm. Mineralization was evaluated using ARS staining. ARS-stained cells were dispersed in acetic acid, and absorbance was measured at 405 nm.
File: Figure_3.xlsx
Description: Exosome-based miR-423 inhibitor delivery increases osteogenic induction of OVX-MSCs in 3D. The apatite-PLGA scaffold was fabricated to deliver exosome-laden miR-423 inhibitor, exosomes alone, or control. Osteogenic induction of the constructs was evaluated in a 3D in vitro culture encapsulated with OVX-MSCs. (D) Gene expression of osteogenic markers (Runx2, OCN, Osterix) and the Wnt/ β-catenin signaling maker (CTNNB1) measured by a real-time PCR assay. (F) ALP expression at day 7. (H) Mineralization assessed by Alizarin Red staining at day 21.
Exosomes were isolated from the conditioned medium of cultured MSCs. Exosomes loaded with miR-423 inhibitor were generated by transfecting exosomes with miR-423 inhibitor using Exo-Fect™ siRNA/miRNA Transfection Kit. Exosomes loaded with miR-423 inhibitor were adsorbed onto the apatite layer of the PLGA scaffold. MSCs isolated from OVX mice were cultured on the scaffold and subsequently induced toward osteogenic differentiation. The expression of osteogenic markers was measured by a real-time PCR assay. ALP activity and mineral deposition were evaluated using ALP and ARS staining.
File: Figure_4.xlsx
Description: Exosome-laden miR-423 inhibitor enhances calvarial bone repair in OVX mice. Quantification of bone repair, including percent bone area (B), percent BV/TV (C), and Tb.N (D).
The in vivo bone regenerative effect of exosome/scaffold complex was evaluated using a calvarial defect model in OVX mice. A critical-sized calvarial defect was created in both Sham and OVX mice. OVX mice were implanted with blank scaffolds or scaffolds loaded with low-dose (10 µg) or high-dose (50 µg) Exosome-laden miR-423 inhibitor, while Sham mice received blank scaffolds. After eight weeks of surgery, all the animals were sacrificed and the calvarial samples were collected for μCT analysis and quantification.
File: Figure_S1.xlsx
Description: Expression of miR-423 in OVX-derived MSCs treated with miR-423 inhibitor, miR-423 mimic, or control, as measured by real-time PCR.
MSCs isolated from OVX mice were treated with miR-423 mimic, miR-423 inhibitor, or PBS control, and intracellular miR-423 levels were measured by a real-time PCR assay.
Abbreviations:
- 3D Three-dimensional
- ALP Alkaline phosphatase
- ARS Alizarin red S
- BV/TV Bone Volume / Tissue Volume
- CT Computed Tomography
- CTL Control
- COL1A1 Collagen type I alpha 1
- EXO Exosome
- MSC Mesenchymal stem/stromal cells
- OCN Osteocalcin
- OVX Ovariectomized
- PBS Phosphate-buffered saline
- PCR Polymerase chain reaction
- PLGA Poly(lactic-co-glycolic acid)
- RUNX2 Runt-related transcription factor 2
- SCAF Scaffold
- Tb.N Trabecular number