PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions
Abstract Background The bone tissue engineering (BTE) approach has been introduced as an alternative to conventional treatments for large non-healing bone defects. Magnetism promotes stem cells' adherence to biocompatible scaffolds toward osteoblast differentiation. Furthermore, osteogenic diff...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-04-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-02816-0 |
| _version_ | 1819061458494816256 |
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| author | Hadi Sadeghzadeh Ahmad Mehdipour Hassan Dianat-Moghadam Roya Salehi Ali Baradar Khoshfetrat Ayla Hassani Daryush Mohammadnejad |
| author_facet | Hadi Sadeghzadeh Ahmad Mehdipour Hassan Dianat-Moghadam Roya Salehi Ali Baradar Khoshfetrat Ayla Hassani Daryush Mohammadnejad |
| author_sort | Hadi Sadeghzadeh |
| collection | DOAJ |
| description | Abstract Background The bone tissue engineering (BTE) approach has been introduced as an alternative to conventional treatments for large non-healing bone defects. Magnetism promotes stem cells' adherence to biocompatible scaffolds toward osteoblast differentiation. Furthermore, osteogenic differentiation media are expensive and any changes in its composition affect stem cells differentiation. Moreover, media growth factors possess a short half-life resulting in the rapid loss of their functions in vivo. With the above in mind, we fabricated a multilayered nanocomposite scaffold containing the wild type of Type I collagen (Col I) with endogenous magnetic property to promote osteogenesis in rat ADSCs with the minimum requirement of osteogenic differentiation medium. Methods Fe3O4 NPs were synthesized by co-precipitation method and characterized using SEM, VSM, and FTIR. Then, a PCL/Col I nanocomposite scaffold entrapping Fe3O4 NPs was fabricated by electrospinning and characterized using SEM, TEM, AFM, VSM, Contact Angle, tensile stretching, and FTIR. ADSCs were isolated from rat adipose tissue and identified by flow cytometry. ADSCs were loaded onto PCL/Col I and PCL/Col I/Fe3O4-scaffolds for 1–3 weeks with/without osteogenic media conditions. The cell viability, cell adhesion, and osteogenic differentiation were evaluated using MTT assay, SEM, DAPI staining, ALP/ARS staining, RT-PCR, and western blotting, respectively. Results SEM, VSM, and FTIR results indicated that Fe3O4 was synthesized in nano-sized (15–30 nm) particles with spherical-shaped morphology and superparamagnetic properties with approved chemical structure as FTIR revealed. According to SEM images, the fabricated magnetic scaffolds consisted of nanofiber (500–700 nm). TEM images have shown the Fe3O4 NPs entrapped in the scaffold's fiber without bead formation. FTIR spectra analysis confirmed the maintenance of the natural structure of Col I, PCL, and Fe3O4 upon electrospinning. AFM data have shown that MNPs incorporation introduced stripe-like topography to nanofibers, while the depth of the grooves has decreased from 800 to 500 nm. Flow cytometry confirmed the phenotype of ADSCs according to their surface markers (i.e., CD29 and CD105). Additionally, Fe3O4 NP improved nanocomposite scaffold strength, wettability, porosity, biocompatibility and also facilitates the ALP activity, calcium-mineralization. Finally, magnetic nanocomposite scaffolds upregulated osteogenic-related genes or proteins’ expression (e.g., Col I, Runx2, OCN, ON, BMP2) in seeded ADSCs with/without osteo-differentiation media conditions. Conclusions Together, these results indicate that Fe3O4 NPs within the natural structure of Col I increase osteogenic differentiation in osteogenic cues-free media conditions. This effect could be translated in vivo toward bone defects healing. These findings support the use of natural ECM materials alongside magnetic particles as composite scaffolds to achieve their full therapeutic potential in BTE treatments. Graphical Abstract |
| first_indexed | 2024-12-21T14:43:12Z |
| format | Article |
| id | doaj.art-5f99ba1001224523bd9fa9cd1562c781 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-21T14:43:12Z |
| publishDate | 2022-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-5f99ba1001224523bd9fa9cd1562c7812022-12-21T19:00:07ZengBMCStem Cell Research & Therapy1757-65122022-04-0113111810.1186/s13287-022-02816-0PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditionsHadi Sadeghzadeh0Ahmad Mehdipour1Hassan Dianat-Moghadam2Roya Salehi3Ali Baradar Khoshfetrat4Ayla Hassani5Daryush Mohammadnejad6Department of Tissue Engineering, Faculty of Advanced Medical Sciences, Tabriz University of Medical SciencesDepartment of Tissue Engineering, Faculty of Advanced Medical Sciences, Tabriz University of Medical SciencesDepartment of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical SciencesDepartment of Medical Nanotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical ScienceChemical Engineering Faculty, Sahand University of TechnologyChemical Engineering Faculty, Sahand University of TechnologyDepartment of Tissue Engineering, Faculty of Advanced Medical Sciences, Tabriz University of Medical SciencesAbstract Background The bone tissue engineering (BTE) approach has been introduced as an alternative to conventional treatments for large non-healing bone defects. Magnetism promotes stem cells' adherence to biocompatible scaffolds toward osteoblast differentiation. Furthermore, osteogenic differentiation media are expensive and any changes in its composition affect stem cells differentiation. Moreover, media growth factors possess a short half-life resulting in the rapid loss of their functions in vivo. With the above in mind, we fabricated a multilayered nanocomposite scaffold containing the wild type of Type I collagen (Col I) with endogenous magnetic property to promote osteogenesis in rat ADSCs with the minimum requirement of osteogenic differentiation medium. Methods Fe3O4 NPs were synthesized by co-precipitation method and characterized using SEM, VSM, and FTIR. Then, a PCL/Col I nanocomposite scaffold entrapping Fe3O4 NPs was fabricated by electrospinning and characterized using SEM, TEM, AFM, VSM, Contact Angle, tensile stretching, and FTIR. ADSCs were isolated from rat adipose tissue and identified by flow cytometry. ADSCs were loaded onto PCL/Col I and PCL/Col I/Fe3O4-scaffolds for 1–3 weeks with/without osteogenic media conditions. The cell viability, cell adhesion, and osteogenic differentiation were evaluated using MTT assay, SEM, DAPI staining, ALP/ARS staining, RT-PCR, and western blotting, respectively. Results SEM, VSM, and FTIR results indicated that Fe3O4 was synthesized in nano-sized (15–30 nm) particles with spherical-shaped morphology and superparamagnetic properties with approved chemical structure as FTIR revealed. According to SEM images, the fabricated magnetic scaffolds consisted of nanofiber (500–700 nm). TEM images have shown the Fe3O4 NPs entrapped in the scaffold's fiber without bead formation. FTIR spectra analysis confirmed the maintenance of the natural structure of Col I, PCL, and Fe3O4 upon electrospinning. AFM data have shown that MNPs incorporation introduced stripe-like topography to nanofibers, while the depth of the grooves has decreased from 800 to 500 nm. Flow cytometry confirmed the phenotype of ADSCs according to their surface markers (i.e., CD29 and CD105). Additionally, Fe3O4 NP improved nanocomposite scaffold strength, wettability, porosity, biocompatibility and also facilitates the ALP activity, calcium-mineralization. Finally, magnetic nanocomposite scaffolds upregulated osteogenic-related genes or proteins’ expression (e.g., Col I, Runx2, OCN, ON, BMP2) in seeded ADSCs with/without osteo-differentiation media conditions. Conclusions Together, these results indicate that Fe3O4 NPs within the natural structure of Col I increase osteogenic differentiation in osteogenic cues-free media conditions. This effect could be translated in vivo toward bone defects healing. These findings support the use of natural ECM materials alongside magnetic particles as composite scaffolds to achieve their full therapeutic potential in BTE treatments. Graphical Abstracthttps://doi.org/10.1186/s13287-022-02816-0ScaffoldType I collagenMagnetic nanoparticlesAdipose-derived stem cellsDifferentiationOsteogenic cues-free media |
| spellingShingle | Hadi Sadeghzadeh Ahmad Mehdipour Hassan Dianat-Moghadam Roya Salehi Ali Baradar Khoshfetrat Ayla Hassani Daryush Mohammadnejad PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions Stem Cell Research & Therapy Scaffold Type I collagen Magnetic nanoparticles Adipose-derived stem cells Differentiation Osteogenic cues-free media |
| title | PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions |
| title_full | PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions |
| title_fullStr | PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions |
| title_full_unstemmed | PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions |
| title_short | PCL/Col I-based magnetic nanocomposite scaffold provides an osteoinductive environment for ADSCs in osteogenic cues-free media conditions |
| title_sort | pcl col i based magnetic nanocomposite scaffold provides an osteoinductive environment for adscs in osteogenic cues free media conditions |
| topic | Scaffold Type I collagen Magnetic nanoparticles Adipose-derived stem cells Differentiation Osteogenic cues-free media |
| url | https://doi.org/10.1186/s13287-022-02816-0 |
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