Poly(Dopamine)-Assisted Immobilization of Xu Duan on 3D Printed Poly(Lactic Acid) Scaffolds to Up-Regulate Osteogenic and Angiogenic Markers of Bone Marrow Stem Cells - PubMed (original) (raw)

Poly(Dopamine)-Assisted Immobilization of Xu Duan on 3D Printed Poly(Lactic Acid) Scaffolds to Up-Regulate Osteogenic and Angiogenic Markers of Bone Marrow Stem Cells

Chia-Hung Yeh et al. Materials (Basel). 2015.

Abstract

Three-dimensional printing is a versatile technique to generate large quantities of a wide variety of shapes and sizes of polymer. The aim of this study is to develop functionalized 3D printed poly(lactic acid) (PLA) scaffolds and use a mussel-inspired surface coating and Xu Duan (XD) immobilization to regulate cell adhesion, proliferation and differentiation of human bone-marrow mesenchymal stem cells (hBMSCs). We prepared PLA scaffolds and coated with polydopamine (PDA). The chemical composition and surface properties of PLA/PDA/XD were characterized by XPS. PLA/PDA/XD controlled hBMSCs' responses in several ways. Firstly, adhesion and proliferation of hBMSCs cultured on PLA/PDA/XD were significantly enhanced relative to those on PLA. In addition, the focal adhesion kinase (FAK) expression of cells was increased and promoted cell attachment depended on the XD content. In osteogenesis assay, the osteogenesis markers of hBMSCs cultured on PLA/PDA/XD were significantly higher than seen in those cultured on a pure PLA/PDA scaffolds. Moreover, hBMSCs cultured on PLA/PDA/XD showed up-regulation of the ang-1 and vWF proteins associated with angiogenic differentiation. Our results demonstrate that the bio-inspired coating synthetic PLA polymer can be used as a simple technique to render the surfaces of synthetic scaffolds active, thus enabling them to direct the specific responses of hBMSCs.

Keywords: 3D printed-scaffold; angiogenic; dopamine; osteogenic; poly (lactic acid); tissue engineering.

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Conflict of interest statement

The authors declare no conflict of interest. No benefit of any kind has been or will be received either directly or indirectly by the authors.

Figures

Figure 1

The (A) top view and side view of 3D printed poly(lactic acid) (PLA) scaffold; (B) Schematic illustration of dopamine-assisted immobilization of XD on surfaces.

Figure 2

XPS (A) N1s; (B) C1s; and (C) O1s high-resolution spectra obtained on PLA scaffolds after coating with dopamine.

Figure 3

SEM images of PLA scaffold coated with DA and XD.

Figure 4

Release profile of XD from PLA scaffolds in DMEM for (A) short and (B) long times. The values shown are means ± standard errors for all the assays.

Figure 5

The adhesion of hBMSCs cultured with various specimens for different time points. The values shown are means ± standard errors for all the assays. “#” indicates a significant difference (p < 0.05) compared to PLA; “*” indicates a significant difference (p < 0.05) compared to PLA/DA.

Figure 6

The morphology of hBMSCs adhered on PLA/PDA/XD scaffolds for 3 and 24 h.

Figure 7

The pFAK expression of hBMSCs cultured on various specimens for 3 h. The values shown are means ± standard errors for all the assays. “#” indicates a significant difference (p < 0.05) compared to PLA; “*” indicates a significant difference (p < 0.05) compared to PLA/DA.

Figure 8

(A) PrestoBlue® assay and (B) LDH assay of hBMSCs cultured on various specimens for different time points. The values shown are means ± standard errors for all the assays. “#” indicates a significant difference (p < 0.05) compared to PLA; “*” indicates a significant difference (p < 0.05) compared to PLA/DA.

Figure 9

(A) Col; (B) ALP; (C) BSP and (D) OC gene expression in the hBMSCs were cultured on the various specimens for 7 and 14 days. The values shown are means ± standard errors for all the assays. “@” indicates a significant difference (p < 0.05) compared to PLA; “*” indicates a significant difference (p < 0.05) compared to PLA/DA.

Figure 10

(A) Alizarin Red S staining and (B) quantification of calcium mineral deposits of hBMSCs cultured on various scaffolds for 7 and 14 days. The values shown are means ± standard errors for all the assays. “*” indicates a significant difference (p < 0.05) compared to PLA; “@” indicates a significant difference (p < 0.05) compared to PLA/DA.

Figure 11

The protein expression of (A) Ang-1 and (B) vWF of hBMSCs cultured on scaffolds for different days. The values shown are means ± standard errors for all the assays. “*” indicates a significant difference (p < 0.05) compared to PLA; “@” indicates a significant difference (p < 0.05) compared to PLA/DA.

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Poly(Dopamine)-Assisted Immobilization of Xu Duan on 3D Printed Poly(Lactic Acid) Scaffolds to Up-Regulate Osteogenic and Angiogenic Markers of Bone Marrow Stem Cells - PubMed (original) (raw)