In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and β-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells

doi:10.3390/ma13143057

. 2020 Jul 8;13(14):3057.

doi: 10.3390/ma13143057.

In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and β-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells

Shuaishuai Cao^{1

2}, Jonghyeuk Han³, Neha Sharma^{1

2}, Bilal Msallem^{1

2}, Wonwoo Jeong³, Jeonghyun Son³, Christoph Kunz¹, Hyun-Wook Kang³, Florian M Thieringer^{1

2}

Affiliations

¹ Department of Oral and Cranio-Maxillofacial Surgery, University Hospital Basel, Spitalstrasse 21, 4031 Basel, Switzerland.
² Medical Additive Manufacturing Research Group, Department of Biomedical Engineering, University of Basel, Gewerbestrasse 16, 4123 Allschwil, Switzerland.
³ Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), 50, UNIST-gil, Ulsan 44919, Korea.

PMID: 32650530
PMCID: PMC7412522
DOI: 10.3390/ma13143057

In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and β-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells

Shuaishuai Cao et al. Materials (Basel). 2020.

. 2020 Jul 8;13(14):3057.

doi: 10.3390/ma13143057.

Authors

Shuaishuai Cao^{1

2}, Jonghyeuk Han³, Neha Sharma^{1

2}, Bilal Msallem^{1

2}, Wonwoo Jeong³, Jeonghyun Son³, Christoph Kunz¹, Hyun-Wook Kang³, Florian M Thieringer^{1

2}

Affiliations

¹ Department of Oral and Cranio-Maxillofacial Surgery, University Hospital Basel, Spitalstrasse 21, 4031 Basel, Switzerland.
² Medical Additive Manufacturing Research Group, Department of Biomedical Engineering, University of Basel, Gewerbestrasse 16, 4123 Allschwil, Switzerland.
³ Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), 50, UNIST-gil, Ulsan 44919, Korea.

PMID: 32650530
PMCID: PMC7412522
DOI: 10.3390/ma13143057

Abstract

3D printed biomaterials have been extensively investigated and developed in the field of bone regeneration related to clinical issues. However, specific applications of 3D printed biomaterials in different dental areas have seldom been reported. In this study, we aimed to and successfully fabricated 3D poly (lactic-co-glycolic acid)/β-tricalcium phosphate (3D-PLGA/TCP) and 3D β-tricalcium phosphate (3D-TCP) scaffolds using two relatively distinct 3D printing (3DP) technologies. Conjunctively, we compared and investigated mechanical and biological responses on human dental pulp stem cells (hDPSCs). Physicochemical properties of the scaffolds, including pore structure, chemical elements, and compression modulus, were characterized. hDPSCs were cultured on scaffolds for subsequent investigations of biocompatibility and osteoconductivity. Our findings indicate that 3D printed PLGA/TCP and β-tricalcium phosphate (β-TCP) scaffolds possessed a highly interconnected and porous structure. 3D-TCP scaffolds exhibited better compressive strength than 3D-PLGA/TCP scaffolds, while the 3D-PLGA/TCP scaffolds revealed a flexible mechanical performance. The introduction of 3D structure and β-TCP components increased the adhesion and proliferation of hDPSCs and promoted osteogenic differentiation. In conclusion, 3D-PLGA/TCP and 3D-TCP scaffolds, with the incorporation of hDPSCs as a personalized restoration approach, has a prospective potential to repair minor and critical bone defects in oral and maxillofacial surgery, respectively.

Keywords: 3D printing; bone regeneration; ceramic printing; dental biomaterials; human dental pulp stem cell; in vitro research; polymer printing.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

**Figure 1**
Overview of the application of two types of 3DP technologies used to fabricate scaffold incorporated with hDPSCs for oral and maxillofacial bone reconstruction.

**Figure 2**
Characterizations of C-TCP, 3D-PLGA/TCP, and 3D-TCP scaffolds. SEM images showed the 3D interconnected structure and scaffold surface, and EDS-based assessments indicated the presence of β-TCP in these three scaffolds.

**Figure 3**
Mechanical properties of C-TCP, 3D-PLGA/TCP, and 3D-TCP scaffolds. (a) Compressive strain–stress curves indicated that 3D-PLGA/TCP possessed flexible mechanical properties. (b) Effects of 3DP and β-TCP amounts on the compressive modulus (* p < 0.050).

**Figure 4**
The effect of C-TCP, 3D-PLGA/TCP, and 3D-TCP scaffolds on hDPSCs viability. The live/dead assay results indicated that hDPSCs possessed excellent cell viability on the surfaces of all tested scaffolds (green: live cells; red: dead cells).

**Figure 5**
hDPSCs viability of C-TCP, 3D-PLGA/TCP, and 3D-TCP scaffolds. C-TCP and 3D-TCP scaffolds showed good cell viability over time, whereas 3D-PLGA/TCP scaffold had significantly lower cell viability than the other two groups (* p < 0.050).

**Figure 6**
hDPSCs proliferation on C-TCP, 3D-PLGA/TCP, and 3D-TCP scaffolds. AlamarBlue assays indicated that all scaffolds significantly promoted the proliferation of hDPSCs (* p < 0.050).

**Figure 7**
The effect of C-TCP, 3D-PLGA/TCP, and 3D-TCP scaffolds on ALP activity. The 3D interconnected structure and β-TCP amount enhanced the ALP activity on day 3, 7, and 14 (* p < 0.050).

See this image and copyright information in PMC

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References

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[1] Warnke P.H., Springer I.N.G., Wiltfang J., Acil Y., Eufinger H., Wehmöller M., Russo P.A.J., Bolte H., Sherry E., Behrens E., et al. Growth and transplantation of a custom vascularised bone graft in a man. Lancet. 2004;364:766–770. doi: 10.1016/S0140-6736(04)16935-3. - DOI - PubMed

[2] Warnke P.H., Springer I.N.G., Wiltfang J., Acil Y., Eufinger H., Wehmöller M., Russo P.A.J., Bolte H., Sherry E., Behrens E., et al. Growth and transplantation of a custom vascularised bone graft in a man. Lancet. 2004;364:766–770. doi: 10.1016/S0140-6736(04)16935-3. - DOI - PubMed

[3] Ciocca L., Lesci I.G., Mezini O., Parrilli A., Ragazzini S., Rinnovati R., Romagnoli N., Roveri N., Scotti R. Customized hybrid biomimetic hydroxyapatite scaffold for bone tissue regeneration. J. Biomed. Mater. Res. B Appl. Biomater. 2017;105:723–734. doi: 10.1002/jbm.b.33597. - DOI - PubMed

[4] Ciocca L., Lesci I.G., Mezini O., Parrilli A., Ragazzini S., Rinnovati R., Romagnoli N., Roveri N., Scotti R. Customized hybrid biomimetic hydroxyapatite scaffold for bone tissue regeneration. J. Biomed. Mater. Res. B Appl. Biomater. 2017;105:723–734. doi: 10.1002/jbm.b.33597. - DOI - PubMed

[5] Cao S., Zhang L., Chen Z., Li S., Jiang H., Tian Y., Gu W., Zhou M., Chen X. The Effect and Biocompatibility of Nanofibrous Nano-Hydroxyapatite/Polycaprolactone/Gelatin Scaffolds Multipurpose Membrane in Guiding Bone Regeneration. J. Biomater. Tissue Eng. 2017;7:721–729. doi: 10.1166/jbt.2017.1619. - DOI

[6] Cao S., Zhang L., Chen Z., Li S., Jiang H., Tian Y., Gu W., Zhou M., Chen X. The Effect and Biocompatibility of Nanofibrous Nano-Hydroxyapatite/Polycaprolactone/Gelatin Scaffolds Multipurpose Membrane in Guiding Bone Regeneration. J. Biomater. Tissue Eng. 2017;7:721–729. doi: 10.1166/jbt.2017.1619. - DOI

[7] Zhou M., Peng X., Mao C., Xu F., Hu M., Yu G.Y. Primate mandibular reconstruction with prefabricated, vascularized tissue-engineered bone flaps and recombinant human bone morphogenetic protein-2 implanted in situ. Biomaterials. 2010;31:4935–4943. doi: 10.1016/j.biomaterials.2010.02.072. - DOI - PubMed

[8] Zhou M., Peng X., Mao C., Xu F., Hu M., Yu G.Y. Primate mandibular reconstruction with prefabricated, vascularized tissue-engineered bone flaps and recombinant human bone morphogenetic protein-2 implanted in situ. Biomaterials. 2010;31:4935–4943. doi: 10.1016/j.biomaterials.2010.02.072. - DOI - PubMed

[9] Tang D., Tare R.S., Yang L.Y., Williams D.F., Ou K.L., Oreffo R.O. Biofabrication of bone tissue: Approaches, challenges and translation for bone regeneration. Biomaterials. 2016;83:363–382. doi: 10.1016/j.biomaterials.2016.01.024. - DOI - PubMed

[10] Tang D., Tare R.S., Yang L.Y., Williams D.F., Ou K.L., Oreffo R.O. Biofabrication of bone tissue: Approaches, challenges and translation for bone regeneration. Biomaterials. 2016;83:363–382. doi: 10.1016/j.biomaterials.2016.01.024. - DOI - PubMed

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In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and β-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells

Affiliations

In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and β-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells

Authors

Affiliations

Abstract

Conflict of interest statement

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