Polyetheretherketone with citrate potentiated influx of copper boosts osteogenesis, angiogenesis, and bacteria-triggered antibacterial abilities

doi:10.1016/j.jmst.2020.08.048

Abstract

Abstract:

A well designed coating for polyetheretherketone (PEEK) implants can provide enough support to overcome crucial medical challenges, which are insufficient osseointegration and high rate of infection. Herein, we utilize the co-deposition of polydopamine (PDA) and copper-citrate nanoclusters to construct a pH-responsive coating on porous PEEK for synergistic bone regeneration, vascular formation and anti-infection. Specifically, this PDA coating released high dose of copper and citrate at lower pH value, which increased intracellular copper content, boosted production of reactive oxygen species and severe damage of protein, leading to killing of 93 % planktonic bacterial and eradication of adherent bacteria. At pH of 7.4, the release of copper and citrate were in a slow and sustained behavior, synergistically enhanced vascular formation potential and osteodiffereration of Ad-MSC in vitro. After implanted in rabbit tibia for 6 and 12 weeks, the micro-CT evaluation and histological analysis consistently highlighted the ability of this PDA coating to increase new bone formation adjacent to coated PEEK implant and enhance bone-implant interfacial integration. These results were proven to be related to the synergistic effect that citrate facilitated a 2-fold influx of copper into cells, which not only enhanced the bacteria-killing ability but also encouraged bone regeneration of implants. This present work provides an effective method to control infections while promoting osseointegration simultaneously, which will show tremendous clinical application and can be a solution to current challenges facing orthopedics.

Key words: Citrate, Copper, Potentiated influx, Bone regeneration, Angiogenesis, Bacteria-triggered coating

Jianglong Yan, Dandan Xia, Pan Xiong, Yangyang Li, Wenhao Zhou, Qiyao Li, Pei Wang, Yufeng Zheng, Yan Cheng. Polyetheretherketone with citrate potentiated influx of copper boosts osteogenesis, angiogenesis, and bacteria-triggered antibacterial abilities[J]. J. Mater. Sci. Technol., 2021, 71: 31-43.

Figures/Tables 9

Fig. 1. (a) Scheme illustration of surface functionalization of PEEK. TEM images of (b1) DA capped CuNs and (b2) citrate capped CuNs with FFT images on the top right corner, and size distribution of (b3) DA capped CuNs and (b4) citrate capped CuNs analyzed with ImageJ software.

Fig. 2. (a) Surface morphology and (b) contact angle of different samples. Chemical composition of coatings: (c1) Survey spectra of XPS, and high resolution XPS spectra of (c2) Cu 2p from DCuN, (c3) Cu 2p from CCuN, (c4) C 1s from SP, (c5) C 1s from DCuN, and (c6) C 1s from CCuN.

Fig. 3. (a) The cumulative release profile of copper at different pH condition, and (b) pH controlled release of copper from DCuN and CCuN samples.

Fig. 4. Antibacterial evaluation: (a) antibacterial rate of different samples and (b) activity of bacteria adhered to different samples after treated with E. coli at different pH value for 6 h. After treated with different samples at pH 5.0 for 6 h, (c) relative PI intensity, (d) protein leakage, (e) relative ROS level, and (f) concentration of copper per bacteria. SEM observation of bacteria morphology after cultured with samples for (g) 6 h and (h) 7 d. *P < 0.05; **P < 0.01; ***P < 0.001.

Fig. 5. Schematic illustration of bacteria-triggered synergistic bactericidal mechanism of copper-citrate nanoclusters contained porous surface.

Fig. 6. In vitro proliferation of Ad-MSCs. (a) SEM morphology of Ad-MSCs and (b) cell activity evaluated by MTT. Osteodifferentiation of Ad-MSCs in monoculture or coculture system: (c, e, g) Qualitative and (d, f, h) quantitative of ALP expression, collagen secretion, and calcium deposition, respectively. *P < 0.05; **P < 0.01.

Fig. 7. In vitro proliferation of HUVECs. (a) SEM observation of HUVECs and (b) cell activity evaluated by MTT. (c) NO production of HUVECs in monoculture and coculture system. Gene expression of (d) HIF-1α, (e) VEGF, and (f) eNOS. *P < 0.05; **P < 0.01; ***P < 0.001.

Fig. 8. Micro-CT imaging and quantification of bone regeneration. (a) 3D reconstructed micro-CT images of the new bone formation and corresponding micro-CT images of the tibia bone defect in the axial plane at 6, and 12 weeks after implantation. Quantitative histomorphometry analyses of (b) BV/TV and (c) BMD. *P < 0.05.

Fig. 9. The methylene blue and acid fuchsine staining in each group at 6 and 12 weeks post-surgery.

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