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. 2023 Nov 3:11:1283526.
doi: 10.3389/fbioe.2023.1283526. eCollection 2023.

PRP coating on different modified surfaces promoting the osteointegration of polyetheretherketone implant

Xiaotong Shi  1 , Zongliang Wang  2 , Min Guo  2 , Yu Wang  2 , Zhiguo Bi  1 , Dongsong Li  1 , Peibiao Zhang  2 , Jianguo Liu  1
Affiliations

PRP coating on different modified surfaces promoting the osteointegration of polyetheretherketone implant

Xiaotong Shi et al. Front Bioeng Biotechnol. .

Abstract

Introduction: Polyetheretherketone (PEEK) material implants have been applied more and more clinically recently. In order to increase the osteogenic activity of PEEK material, the microstructure change of the material surface and the construction of functional microcoatings have become a hot research topic. This study investigated the ability of PEEK surfaces modified by different methods to carry Platelet-rich plasma (PRP) and the osteogenic ability of different PEEK microstructures after carrying PRP in vivo/in vitro. Methods: In this study, PEEK surfaces were modified by sulfuric acid, gaseous sulfur trioxide and sandpaper. Next, PRP from SD rats was prepared and incubated on PEEK material with different surface microstructures. Lactate dehydrogenase test, scanning electron microscope and Elisa assay was used to evaluate adhesion efficiency of PRP. Then in vitro tests such as CCK-8, ALP staining, ARS staining and RT-qPCR et al were used to further evaluate osteogenesis ability of the PRP coating on PEEK surface. Finally, The tibia defects of SD rats were established, and the new bone was evaluated by Micro-CT, HE staining, and immunofluorescence staining. Results: The sandpaper-polished PEEK with the strongest PRP carrying capacity showed the best osteogenesis. Our study found that the modified PEEK surface with PRP coating has excellent osteogenic ability and provided the basis for the interface selection of PRP for the further application of PEEK materials. Discussion: Among the three PEEK modified surfaces, due to the most PRP carrying and the strongest osteogenic ability in vitro/vivo, the frosted surface was considered to be the most suitable surface for the preparation of PRP coating.

Keywords: growth factor; osteointegration; platelet-rich plasma; polyetheretherketone; surface treatment.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

SCHEME 1
SCHEME 1
Concept illustration. PEEK samples with three different modified surfaces were fabricated and PRP coatings were prepared. This combination treatment promoted osteoblast proliferation and differentiation in vitro, while in vivo it could regulate immunity, promote angiogenesis, and ultimately promote osseointegration.
FIGURE 1
FIGURE 1
(A) The SEM morphology of the surface of PEEK samples from each group. (B) EDS results of elemental composition on the surface of PEEK samples in each group. (C) Results of water contact angles on PEEK surfaces in each group. (D) FT-IR spectral characteristics of PEEK surfaces in each group, n = 5.* indicate p < 0.05.
FIGURE 2
FIGURE 2
(A) AFM images of the surfaces of PEEK samples from each group. Yellow arrows: fully activated platelets; Red arrows: non-activated platelets. (B) Surface roughness of each group of PEEK samples analyzed based on AFM images, n = 3. (C) Compressive strength of PEEK samples in each group, n = 5. (D) Maximum compression load of PEEK samples in each group, n = 5.* indicate p < 0.05.
FIGURE 3
FIGURE 3
(A) SEM results of PEEK samples from each group after incubation with PRP. (B) Results of LDH assay for the number of platelets adhered to the surface of PEEK samples in each group, n = 3. (C) PDGF-BB release curves in each group, n = 3. (D) The initial release of PDGF-BB from the surface of each PEEK sample was measured, n = 3. (E) Final cumulative release of PDGF-BB from the surface of various PEEK samples, n = 3.* indicate p < 0.05.
FIGURE 4
FIGURE 4
(A) DAPI staining of nuclei of adherent cells on the surface of PEEK samples from each group. (B) Quantitative analysis of the nuclei of adherent cells on the surface of each PEEK sample, n = 3. (C) SEM images of adherent cells on the surface of PEEK samples from each group. (D) CCK-8 results of the proliferation of MC-3T3-E1 cells on the PEEK surface of each group for 1,3, and 7 days, n = 3.* indicate p < 0.05, # represents that compared with the same treatment method, the group incubated with PRP was significantly higher than the group not incubated, p < 0.05.
FIGURE 5
FIGURE 5
(A) ALP staining results of MC3T3-E1 cultured on PEEK surface for 7 days and 14 days in each group. (B) ALP quantitative analysis results of cells cultured on PEEK surface for 7 days and 14 days, n = 3. (C) ARS staining of cells cultured on PEEK surfaces for 21 days in each group. (D) Quantitative ARS analysis of cells cultured on PEEK surfaces for 21 days in each group, n = 3. (E–H) Osteogenesis-related gene (Col-1, Rux-2, OPN, OCN) expressions of MC3T3-E1 were evaluated by RT-qPCR (n = 3) 14 days after cultured on the surface of PEEK surface of each group, n = 3.* indicate p < 0.05, # represents that compared with the same treatment method, the group incubated with PRP was significantly higher than the group not incubated, p < 0.05.
FIGURE 6
FIGURE 6
(A) 2D images of in vivo osteogenesis in each group of samples. Yellow triangle: PEEK material location; Red arrow: new bone; Green arrow: new bone at the tibial burr site. (B–E) Quantitative analysis of in vivo samples of each group for bone volume fraction (BV/TV), trabecular bone number (Tb. N), trabecular bone thickness (Tb. Th) and trabecular bone separation (Tb. Sp), n = 3. (F) 3D reconstruction of PEEK samples and surrounding new bone in each group. PEEK samples are shown in gray, and new bone is shown in blue. * indicate p < 0.05, # represents that compared with the same treatment method, the group incubated with PRP was significantly higher than the group not incubated, p < 0.05.
FIGURE 7
FIGURE 7
(A) The HE staining results of the tibial specimens of each group. (B) Sirius staining results of tibial specimens from each group; Orange represented typeⅠcollagen, green represented type III collagen.
FIGURE 8
FIGURE 8
(A) OCN immunofluorescence staining of tibial samples from each group. Red represents the expression of OCN, and blue is the nucleus. (B) OPN immunofluorescence staining of tibial samples from each group. Red represents the expression of OPN, and blue is the nucleus.
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Grants and funding

The authors declare financial support was received for the research, authorship, and/or publication of this article. This work was financially supported by Jilin Province Development and Reform Commission (2020C030-2).