Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Nov 25:23:100866.
doi: 10.1016/j.mtbio.2023.100866. eCollection 2023 Dec.

Synergistic effect of hierarchical topographic structure on 3D-printed Titanium scaffold for enhanced coupling of osteogenesis and angiogenesis

Leyi Liu  1   2 , Jie Wu  1   2 , Shiyu Lv  1   2 , Duoling Xu  1   2 , Shujun Li  3 , Wentao Hou  3 , Chao Wang  1   2 , Dongsheng Yu  1   2
Affiliations

Synergistic effect of hierarchical topographic structure on 3D-printed Titanium scaffold for enhanced coupling of osteogenesis and angiogenesis

Leyi Liu et al. Mater Today Bio. .

Abstract

The significance of the osteogenesis-angiogenesis relationship in the healing process of bone defects has been increasingly emphasized in recent academic research. Surface topography plays a crucial role in guiding cellular behaviors. Metal-organic framework (MOF) is an innovative biomaterial with nanoscale structural and topological features, enabling the modulation of scaffold physicochemical properties. This study involved the loading of varying quantities of UiO-66 nanocrystals onto alkali-heat treated 3D-printed titanium scaffolds, resulting in the formation of hierarchical micro/nano topography named UiO-66/AHTs. The physicochemical properties of these scaffolds were subsequently characterized. Furthermore, the impact of these scaffolds on the osteogenic potential of BMSCs, the angiogenic potential of HUVECs, and their intercellular communication were investigated. The findings of this study indicated that 1/2UiO-66/AHT outperformed other groups in terms of osteogenic and angiogenic induction, as well as in promoting intercellular crosstalk by enhancing paracrine effects. These results suggest a promising biomimetic hierarchical topography design that facilitates the coupling of osteogenesis and angiogenesis.

Keywords: Coupling of osteogenesis and angiogenesis; Hierarchical topography; UiO-66.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Image 1
Graphical abstract
Scheme 1
Scheme 1
A simplified illustration of manufacturing UiO-66 modified micro/nano structure on 3D printed titanium scaffold and general summary of the mechanism of the topographic structure regulating angiogenesis-osteogenesis relationship.
Fig. 1
Fig. 1
Morphological observation of different scaffolds: (a) The physical images of EBM printed scaffolds; (b) Micro-topography of EBM printed scaffolds; (c) Nano-topography of TI, AHT, 1/4UiO 66/AHT, 1/2UiO-66/AHT, UiO-66/AHT; (d) Three-dimensional reconstruction view of scaffolds surface by the CLSM (at microscale); (e) Three-dimensional reconstruction view of scaffolds surface by the AFM (at nanoscale).
Fig. 2
Fig. 2
Surface characteristics of different scaffolds: (a) FTIR spectra of synthetic UiO-66 nanocrystals; (b) XRD patterns of the simulated and synthetic UiO-66 nanocrystals; (c) Qualitative results of contact angle; (d) Fluorescent images of serum protein absorption on scaffolds; (e) Quantitative analysis of fluorescence density; (f) Quantitative analysis of absorbed BSA and FN on scaffolds; *p＜0.05,**p＜0.01, ***p＜0.005 compared to TI group; ##p ＜0.01, ###p ＜0.005 compared to AHT group; △△△p＜0.005.
Fig. 3
Fig. 3
Adhesion, morphology, viability, and proliferation of BMSCs on different titanium scaffolds: (a) Representative images of live/dead staining; (b) Quantitative counting of live cells on scaffolds; (c) Cell proliferative results examined with CCK-8 assays after 3, 5, and 7 days of seeding; (d) Representative immunofluorescence staining of ITGB1, Phalloidin, and DAPI; (e) SEM morphology of BMSCs incubated on the scaffolds for 3 days (white arrows, polygonal lamellipodium protrusions); **p＜0.01, ***p＜0.005 compared to TI group; #p＜0.05, ##p＜0.01, ###p ＜0.005 compared to AHT group; △p＜0.05, △△p＜0.01, △△△p＜0.005.
Fig. 4
Fig. 4
In vitro osteogenic differentiation of BMSCs incubated on different scaffolds: (a) Representative images of ALP staining results of BMSCs; (b) Representative results of mineralized nodules at Day 21 and Day 28; (c) Quantitative analysis of ALP activity; (d) Semi-quantitative analysis of Alizarin Red staining of BMSCs at Day 21 and Day 28; (e) Relative osteogenic-related mRNA expression of BMSCs; *p＜0.05, **p＜0.01, ***p＜0.005 compared to TI group, #p ＜0.05, ##p ＜0.01, ###p＜0.005 compared to AHT group, △p＜0.05, △△p＜0.01, △△△p＜0.005. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Fig. 5
Fig. 5
Adhesion, morphology, viability, and proliferation of HUVECs on different scaffolds: (a) Representative images of live/dead staining; (b) Quantitative caculation of live cells on scaffolds; (c) Cell proliferative results examined with CCK-8 assays after 3, 5, and 7 days of seeding; (d) Representative immunofluorescence staining of ITGB1, Phalloidin, and DAPI; (e) SEM morphology of HUVECs cultured on the scaffolds for 3 days (white arrows, polygonal lamellipodium protrusions); *p＜0.05, **p＜0.01, ***p＜0.005 compared to TI group, #p＜0.05, ##p＜0.01 compared to AHT group, △△p＜0.01.
Fig. 6
Fig. 6
In vitro angiogenic differentiation of HUVECs cultured on different scaffolds: (a) Representative wound healing migration images; (b) Quantitative analysis for the migrated HUVECs in scratch wound healing assay (0 h, 12 h); (c) Representative fluorescence images of tube formation; (d) Quantitative analysis of nodes and (e) junctions of tubular structures; (f) Relative angiogenic-related mRNA expression of HUVECs; *p＜0.05, **p＜0.01, ***p＜0.005 compared to TI group, #p ＜0.05, ##p ＜0.01, ###p ＜0.005 compared to AHT group, △p＜0.05, △△△p＜0.005.
Fig. 7
Fig. 7
In vitro osteogenic regulation of HUVECs on different scaffolds: (a) Relative osteogenesis-related mRNA expression of HUVECs; (b) Quantified analysis of BMP2 and BMP4 concentrations secreted by HUVECs in Elisa kit; (c) Schematic illustration of Scheme of BMSCs and HUVECs co-culture system; (d) Representative images of transmigrated BMSCs in co-culture system; (e) Quantitative analysis for transmigrated BMSCs; (f) Representative images of ALP staining in co-culture systems; (g) Quantitative analysis for ALP activity; *p＜0.05, **p＜0.01, ***p＜0.005 compared with TI group, #p ＜0.05, ##p ＜0.01, ###p ＜0.005 compared with AHT group, △p＜0.05, △△p＜0.01, △△△p＜0.005.
Fig. 8
Fig. 8
In vitro angiogenic regulation of BMSCs on different scaffolds: (a) Relative angiogenesis-related mRNA expression of BMSCs; (b) Quantified analysis of VEGF and bFGF concentrations secreted by BMSCs in Elisa kit; (c) Schematic illustration of Scheme of BMSCs and HUVECs co-culture system; (d) Representative images of transmigrated HUVECs in co-culture systems; (e) Quantitative analysis for transmigrated HUVECs; (f) Representative images of tube formation in co-culture systems; (g) Quantitative analysis for tube formation; *p＜0.05, **p＜0.01, ***p＜0.005 compared with TI group, #p ＜0.05, ##p ＜0.01, ###p ＜0.005 compared with AHT group, △p＜0.05, △△p＜0.01, △△△p＜0.005.
Fig. 9
Fig. 9
Histological observation of osseointegration in vivo: (a) Representative H&E staining images, red arrow: newly formed but unmineralized bone (osteoid), black arrow: mineralized and matured bone; (b) Goldner's Trichrome staining images, red arrow: newly formed but unmineralized bone (osteoid), black arrow: mineralized and matured bone; yellow arrow: new vessels. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Fig. 10
Fig. 10
Immunohistochemistry and immunofluorescence staining evaluation: (a) Representative immunohistochemistry staining images for OCN,CD31 and the colorimetric quantitative results, black triangle: blood vessel; (b) Representative immunofluorescence staining images for EMCN, CD31 and quantitative analysis of H type vessel, S:scaffold; *p＜0.05, **p＜0.01, ***p＜0.005 compared with TI group, #p ＜0.05, ##p ＜0.01, ###p ＜0.005 compared with AHT group.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Tian T., Zhang T., Lin Y., Cai X. Vascularization in craniofacial bone tissue engineering. J. Dent. Res. 2018;97:969–976. doi: 10.1177/0022034518767120. - DOI - PubMed
    1. Kumar B.P., Venkatesh V., Kumar K.A., Yadav B.Y., Mohan S.R. Mandibular reconstruction: overview. J. Maxillofac. Oral Surg. 2016;15:425–441. doi: 10.1007/s12663-015-0766-5. - DOI - PMC - PubMed
    1. Ferguson B.M., Entezari A., Fang J., Li Q. Optimal placement of fixation syste-m for scaffold-based mandibular reconstruction. J. Mech. Behav. Biomed. Mater. 2022;126 doi: 10.1016/j.jmbbm.2021.104855. - DOI - PubMed
    1. Palmquist A., Jolic M., Hryha E., Shah F.A. Complex geometry and integrated macro-porosity: clinical applications of electron beam melting to fabricate bespoke bone-anchored implants. Acta Biomater. 2022 doi: 10.1016/j.actbio.2022.06.002. - DOI - PubMed
    1. Spriano S., Yamaguchi S., Baino F., Ferraris S. A critical review of multifunctional titanium surfaces: new frontiers for improving osseointegration and host response, avoiding bacteria contamination. Acta Biomater. 2018;79:1–22. doi: 10.1016/j.actbio.2018.08.013. - DOI - PubMed

LinkOut - more resources