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Cited by 141Bone Tissue Engineering Materials3D Printing in Biomedical Researchbiodegradable polymer synthesis and properties

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Enhanced osteogenic differentiation of stem cells by 3D printed PCL scaffolds coated with collagen and hydroxyapatite

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Ebrahimi, Zahra, Irani, Shiva, Ardeshirylajimi, Abdolreza, Seyedjafari, Ehsan·2022·Scientific Reports
DOI 10.1038/s41598-022-15602-yPMC PMC9300684
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Abstract

Bone tissue engineering uses various methods and materials to find suitable scaffolds that regenerate lost bone due to disease or injury. Poly(ε-caprolactone) (PCL) can be used in 3D printing for producing biodegradable scaffolds by fused deposition modeling (FDM). However, the hydrophobic surfaces of PCL and its non-osteogenic nature reduces adhesion and cell bioactivity at the time of implantation. This work aims to enhance bone formation, osteogenic differentiation, and in vitro biocompatibility via PCL scaffolds modification with Hydroxyapatite (HA) and Collagen type I (COL). This study evaluated the osteosupportive capacity, biological behavior, and physicochemical properties of 3D-printed PCL, PCL/HA, PCL/COL, and PCL/HA/COL scaffolds. Biocompatibility and cells proliferation were investigated by seeding human adipose tissue-derived mesenchymal stem cells (hADSCs) onto the scaffolds, which were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and 6-diamidino-2-phenylindole (DAPI) staining. In addition, the bone differentiation potential of the hADSCs was assessed using calcium deposition, alkaline phosphatase (ALP) activity, and bone-related protein and genes. Although all constructed scaffolds support hADSCs proliferation and differentiation, the results showed that scaffold coating with HA and COL can boost these capacities in a synergistic manner. According to the findings, the tricomponent 3D-printed scaffold can be considered as a promising choice for bone tissue regeneration and rebuilding.

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Crossref · 10.1038/s41598-022-15602-y

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Figures

Figure 1 (a,b) ATR-FTIR spectra of powder HA, PCL scaffold, COL and HA-coated PCL scaffolds, and (c,d) The X-ray diffraction (XRD) spectra of PCL and composite scaffolds.
Figure 1 (a,b) ATR-FTIR spectra of powder HA, PCL scaffold, COL and HA-coated PCL scaffolds, and (c,d) The X-ray diffraction (XRD) spectra of PCL and composite scaffolds.CC BY · PubMed Central
Figure 2 SEM images and EDS spectra analysis of the morphology (a) PCL, (b) PCL/HA, (c) PCL/COL, and (d) PCL/HA/COL scaffold and the change of porosity of the 3D printed scaffolds.
Figure 2 SEM images and EDS spectra analysis of the morphology (a) PCL, (b) PCL/HA, (c) PCL/COL, and (d) PCL/HA/COL scaffold and the change of porosity of the 3D printed scaffolds.CC BY · PubMed Central
Figure 3 (a) Force-extension diagram, (b) Stress–strain diagram, and (c) compressive modulus for the different scaffolds. (d) Variation of swelling behavior of the flat-printed FDM scaffolds. (e) Weight loss of PCL, PCL/HA, PCL/COL and PCL/HA/COL scaffold during soaking in PBS and (f) PBS-Lipase for 28 days. (g) pH values of the scaffolds during the degradation in PBS and (h) lipase. Measurements were started after 1 day, day 0 control = pH 7,4. (n = 3 in each group, *p ≤ 0.05).
Figure 3 (a) Force-extension diagram, (b) Stress–strain diagram, and (c) compressive modulus for the different scaffolds. (d) Variation of swelling behavior of the flat-printed FDM scaffolds. (e) Weight loss of PCL, PCL/HA, PCL/COL and PCL/HA/COL scaffold during soaking in PBS and (f) PBS-Lipase for 28 days. (g) pH values of the scaffolds during the degradation in PBS and (h) lipase. Measurements were started after 1 day, day 0 control = pH 7,4. (n = 3 in each group, *p ≤ 0.05).CC BY · PubMed Central
Figure 4 The proliferation rate of hADSCs on the scaffolds after 1,3,7 to 14 days, and determined by MTT assay. (n = 3, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001).
Figure 4 The proliferation rate of hADSCs on the scaffolds after 1,3,7 to 14 days, and determined by MTT assay. (n = 3, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001).CC BY · PubMed Central
Figure 5 Bone mineralization capacity. In the following image, (a) Alizarin Red staining after a 14 days' culture. (b) After 21 days, Alizarin Red staining. After 21 days of culture, more staining was observed (Scale bar = 100 μm and 20 μm). (c) Calcium concentration of hADSCs on the scaffolds measured after 1,714 and 21 days' culture (n = 3, ***p ≤ 0.001).
Figure 5 Bone mineralization capacity. In the following image, (a) Alizarin Red staining after a 14 days' culture. (b) After 21 days, Alizarin Red staining. After 21 days of culture, more staining was observed (Scale bar = 100 μm and 20 μm). (c) Calcium concentration of hADSCs on the scaffolds measured after 1,714 and 21 days' culture (n = 3, ***p ≤ 0.001).CC BY · PubMed Central
Figure 6 (a) mRNA Expression of osteogenic related genes on scaffolds, Osteonectin (first column) and ALP (second column) after 21 days (n = 3, *p ≤ 0.05 vs. Control group). (b) ALP activity of hADSCs cells seeded on scaffolds in osteogenic medium.
Figure 6 (a) mRNA Expression of osteogenic related genes on scaffolds, Osteonectin (first column) and ALP (second column) after 21 days (n = 3, *p ≤ 0.05 vs. Control group). (b) ALP activity of hADSCs cells seeded on scaffolds in osteogenic medium.CC BY · PubMed Central
Figure 7 The expression pattern of osteogenic markers Osteonectin (first column), for hADSCs on PCL, PCL/HA, PCL/COL and PCL/HA/COL scaffold after 21 days’ incubation in osteogenic medium determined by ICC assay. Cell nuclei in the images are stained with 4′,6- diamidino-2-phenylindole (DAPI) (Scale bar represents 20 μm).
Figure 7 The expression pattern of osteogenic markers Osteonectin (first column), for hADSCs on PCL, PCL/HA, PCL/COL and PCL/HA/COL scaffold after 21 days’ incubation in osteogenic medium determined by ICC assay. Cell nuclei in the images are stained with 4′,6- diamidino-2-phenylindole (DAPI) (Scale bar represents 20 μm).CC BY · PubMed Central

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Cites

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Research works printed but not linked: 1

  1. Jayatissa, N. U., Unagolla, J. & Bhaduri, S. 3D printed polymer scaffolds for bone tissue engineering. Ohio J. Sci. 118, A15–A16 (2018).

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Metadata

Authors
Ebrahimi, Z., Irani, S., Ardeshirylajimi, A., & Seyedjafari, E.
Affiliations
  • Islamic Azad University, Tehran (IR)
  • Shahid Beheshti University (IR)
  • University of Tehran (IR)
Corresponding author
Seyedjafari, Ehsan
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2022-07-20
Volume
12
Issue
1
Pages
12359-12359
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-022-15602-y
PMC
PMC9300684
OpenAlex
W4286504968
Cited by
141
References
68
Citations by year
2026: 39 · 2025: 49 · 2024: 31 · 2023: 18 · 2022: 4
FWCI
7.41
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Topics
  • Physical Sciences › Engineering › Biomedical Engineering › Bone Tissue Engineering Materials
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
  • Physical Sciences › Materials Science › Biomaterials › biodegradable polymer synthesis and properties
Keywords
polycaprolactone, 3D-printed scaffolds, osteogenic differentiation, hydroxyapatite coating, human adipose-derived mesenchymal stem cells, bone tissue engineering, biocompatibility, cell proliferation, alkaline phosphatase activity, calcium deposition, fused deposition modeling
MeSH terms
Collagen, Humans, Osteogenesis, Polyesters, Stem Cells, Durapatite, Tissue Scaffolds, Printing, Three-Dimensional

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