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Cited by 208Bone Tissue Engineering Materials3D Printing in Biomedical ResearchAdditive Manufacturing and 3D Printing Technologies

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Three dimensional printed macroporous polylactic acid/hydroxyapatite composite scaffolds for promoting bone formation in a critical-size rat calvarial defect model

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Zhang, Haifeng, Mao, Xiyuan, Du, Zijing, Jiang, Wenbo, Han, Xiuguo, Zhao, Danyang, Han, Dong, Li, Qingfeng·2016·Science and Technology of Advanced Materials
DOI 10.1080/14686996.2016.1145532PMC PMC5101962
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Abstract

We have explored the applicability of printed scaffold by comparing osteogenic ability and biodegradation property of three resorbable biomaterials. A polylactic acid/hydroxyapatite (PLA/HA) composite with a pore size of 500 μm and 60% porosity was fabricated by three-dimensional printing. Three-dimensional printed PLA/HA, β-tricalcium phosphate (β-TCP) and partially demineralized bone matrix (DBM) seeded with bone marrow stromal cells (BMSCs) were evaluated by cell adhesion, proliferation, alkaline phosphatase activity and osteogenic gene expression of osteopontin (OPN) and collagen type I (COL-1). Moreover, the biocompatibility, bone repairing capacity and degradation in three different bone substitute materials were estimated using a critical-size rat calvarial defect model in vivo. The defects were evaluated by micro-computed tomography and histological analysis at four and eight weeks after surgery, respectively. The results showed that each of the studied scaffolds had its own specific merits and drawbacks. Three-dimensional printed PLA/HA scaffolds possessed good biocompatibility and stimulated BMSC cell proliferation and differentiation to osteogenic cells. The outcomes in vivo revealed that 3D printed PLA/HA scaffolds had good osteogenic capability and biodegradation activity with no difference in inflammation reaction. Therefore, 3D printed PLA/HA scaffolds have potential applications in bone tissue engineering and may be used as graft substitutes in reconstructive surgery.

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Graphical abstract CC BY · PubMed Central
Figure 1. 3D printing machine using the new mini-deposition system.
Figure 1. 3D printing machine using the new mini-deposition system.CC BY · PubMed Central
Figure 2. (A) Optical and (B) SEM images for three scaffolds: 3D printed PLA/HA (1), β-TCP (2) and DBM (3).
Figure 2. (A) Optical and (B) SEM images for three scaffolds: 3D printed PLA/HA (1), β-TCP (2) and DBM (3).CC BY · PubMed Central
Figure 3. BMSC responses to three scaffolds. (A) SEM images of the attachment of BMSCs on (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds after culturing for seven days. (B, C) The fluorescence images of proliferation of BMSCs on (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds after culturing for seven days. (← The state of proliferation of BMSCs; ▄ the selected areas in B corresponding to C. Length scales: (A) 50 μm, (B) 10 μm and (C) 8 μm).
Figure 3. BMSC responses to three scaffolds. (A) SEM images of the attachment of BMSCs on (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds after culturing for seven days. (B, C) The fluorescence images of proliferation of BMSCs on (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds after culturing for seven days. (← The state of proliferation of BMSCs; ▄ the selected areas in B corresponding to C. Length scales: (A) 50 μm, (B) 10 μm and (C) 8 μm).CC BY · PubMed Central
Figure 4. Quantitative analysis of three scaffolds: (A) cell adhesion rate, (B) cell proliferation, (C) ALP activity, (D) osteogenic gene expression and (E) COL-1 of BMSCs cultured on different scaffolds (*p＜0.05).
Figure 4. Quantitative analysis of three scaffolds: (A) cell adhesion rate, (B) cell proliferation, (C) ALP activity, (D) osteogenic gene expression and (E) COL-1 of BMSCs cultured on different scaffolds (*p＜0.05).CC BY · PubMed Central
Figure 5. Micro CT images for three scaffolds. (A) Plane and profile images of (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds before implanting. (B) Typical micro-CT images of (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) bone defect without scaffold at four weeks in vivo. (C) Micro-CT images of (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) bone defect without scaffold at eight weeks in vivo.
Figure 5. Micro CT images for three scaffolds. (A) Plane and profile images of (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds before implanting. (B) Typical micro-CT images of (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) bone defect without scaffold at four weeks in vivo. (C) Micro-CT images of (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) bone defect without scaffold at eight weeks in vivo.CC BY · PubMed Central
Figure 6. BV/TV and degradation rate in every group (A) Percentage of new bone formation at four and eight weeks. (B) Percentage of degradation of three scaffolds at four and eight weeks. (*p＜0.05)
Figure 6. BV/TV and degradation rate in every group (A) Percentage of new bone formation at four and eight weeks. (B) Percentage of degradation of three scaffolds at four and eight weeks. (*p＜0.05)CC BY · PubMed Central
Figure 7. HE images of implanted and control group after four and eight weeks. (A) Histological images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group four weeks after implantation. (B) Histological images of (1) implanted PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group eight weeks after implantation. Scale bars 10 μm.
Figure 7. HE images of implanted and control group after four and eight weeks. (A) Histological images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group four weeks after implantation. (B) Histological images of (1) implanted PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group eight weeks after implantation. Scale bars 10 μm.CC BY · PubMed Central
Figure 8. Immunohistochemical examination of osteocalcin and type I collagen were carried out in the margin of bone defect in different time (magnification ×100). (A) Osteocalcin expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group four weeks after implanting. (B) Osteocalcin expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group eight weeks after implanting. (C) Semi-quantitative scatter plot of osteocalcin expression. (D) Type I collagen expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group four weeks after implanting. (E) Type I collagen expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group eight weeks after implanting. (F) Semi-quantitative scatter plot of type I collagen expression (*p＜0.05).
Figure 8. Immunohistochemical examination of osteocalcin and type I collagen were carried out in the margin of bone defect in different time (magnification ×100). (A) Osteocalcin expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group four weeks after implanting. (B) Osteocalcin expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group eight weeks after implanting. (C) Semi-quantitative scatter plot of osteocalcin expression. (D) Type I collagen expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group four weeks after implanting. (E) Type I collagen expression images of implanted (1) PLA/HA, (2) β-TCP and (3) DBM scaffolds as well as (4) control group eight weeks after implanting. (F) Semi-quantitative scatter plot of type I collagen expression (*p＜0.05).CC BY · PubMed Central

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Research works printed but not linked: 10

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Metadata

Authors
Zhang, H., Mao, X., Du, Z., Jiang, W., Han, X., Zhao, D., Han, D., & Li, Q.
Affiliations
Shanghai Jiao Tong University (CN)
Corresponding author
Han, Dong
ORCID
Venue
Science and Technology of Advanced Materials
Publisher
Informa UK Limited
Published
2016-01-01
Volume
17
Issue
1
Pages
136-148
ISSN
  • 1468-6996 (ISSN-L)
  • 1878-5514
DOI
10.1080/14686996.2016.1145532
PMC
PMC5101962
OpenAlex
W2330522579
Cited by
208
References
61
Citations by year
2026: 11 · 2025: 18 · 2024: 14 · 2023: 24 · 2022: 31 · 2021: 32 · 2020: 27 · 2019: 23 · 2018: 13 · 2017: 14 · 2016: 1
FWCI
8.18
Type
journal article
Language
en
Open access
Licence
Locations
  • Science and Technology of Advanced Materials (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Natural Science Foundation of China (81272132)
Topics
  • Physical Sciences › Engineering › Biomedical Engineering › Bone Tissue Engineering Materials
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
  • Physical Sciences › Engineering › Industrial and Manufacturing Engineering › Additive Manufacturing and 3D Printing Technologies
Keywords
3D-printed scaffolds, macroporous scaffolds, bone tissue engineering, critical-size calvarial defect, mesenchymal stem cells, osteogenic differentiation, biodegradation, beta-tricalcium phosphate, demineralized bone matrix, bone substitutes, osteopontin, type I collagen, biocompatibility

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