<i>In vitro</i> and <i>in vivo</i> evaluation of bone morphogenetic protein-2 (BMP-2) immobilized collagen-coated polyetheretherketone (PEEK)

doi:10.1007/s11706-015-0276-x

Front. Mater. Sci.

2015, Vol. 9

Issue (1) : 38-50 https://doi.org/10.1007/s11706-015-0276-x

RESEARCH ARTICLE

In vitro and in vivo evaluation of bone morphogenetic protein-2 (BMP-2) immobilized collagen-coated polyetheretherketone (PEEK)

Ya-Wei DU^1,²,Li-Nan ZHANG¹,Xin YE¹,He-Min NIE³,Zeng-Tao HOU¹,Teng-Hui ZENG⁴,Guo-Ping YAN²,Peng SHANG^1,^*(

)

1. Center for Translational Medicine Research and Development, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
2. School of Materials Science and Engineering, Wuhan Institute of Technology, Wuhan 430073, China
3. Department of Biomedical Engineering, College of Biology, Hunan University, Changsha 410082, China
4. Department of Spine Surgery, Shenzhen Second People’s Hospital, Shenzhen 518035, China

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Abstract

Polyetheretherketone (PEEK) is regarded as one of the most potential candidates of biomaterials in spinal implant applications. However, as a bioinert material, PEEK plays a limited role in osteoconduction and osseointegration. In this study, recombinant human bone morphogenetic protein-2 (rhBMP-2) was immobilized onto the surface of collagen-coated PEEK in order to prepare a multi-functional material. After adsorbed onto the PEEK surface by hydrophobic interaction, collagen was cross-linked with N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). EDC/NHS system also contributed to the immobilization of rhBMP-2. Water contact angle tests, XPS and SEM clearly demonstrated the surface changes. ELISA tests quantified the amount of rhBMP-2 immobilized and the release over a period of 30 d. In vitro evaluation proved that the osteogenesis differentiation rate was higher when cells were cultured on modified PEEK discs than on regular ones. In vivo tests were conducted and positive changes of major parameters were presented. This report demonstrates that the rhBMP-2 immobilized method for PEEK modification increase bioactivity in vitro and in vivo, suggesting its practicability in orthopedic and spinal clinical applications.

Keywords polyetheretherketone (PEEK) spinal implantation bone morphogenetic protein-2 (BMP-2) collagen surface modification

Corresponding Author(s): Peng SHANG

Online First Date: 21 January 2015 Issue Date: 02 March 2015

Cite this article:

Ya-Wei DU,Li-Nan ZHANG,Xin YE, et al. In vitro and in vivo evaluation of bone morphogenetic protein-2 (BMP-2) immobilized collagen-coated polyetheretherketone (PEEK)[J]. Front. Mater. Sci., 2015, 9(1): 38-50.

URL:

https://academic.hep.com.cn/foms/EN/10.1007/s11706-015-0276-x
https://academic.hep.com.cn/foms/EN/Y2015/V9/I1/38

Fig.1 The dimension of intervertebral disc space was measured by microCT (a) cross-section image and (b) side image.

Fig.2 The red midline was the center mark which was set according to the highest point of L6 vertebra and the non-fluorescent area was the scanning area containing 100 cross-section images above and below the red line.

Fig.3 The ROI was circled carefully with the gray threshold value set from 100 to 250: (a) the cross-section image above the center mark; (b) the cross-section image on the center mark; (c) the cross-section image below the center mark.

Fig.4 (a) The geometry design of simplified PEEK intervertebral disc prostheses by SolidWorks 2013 software. (b) A wire was glued to the polymer prostheses and elongated in horizontal direction.

Fig.5 SEM images of (a) unmodified PEEK and (b)(c)(d) BIPEEK. Collagen fibrils were obvious with the diameter of about 50–200 nm in (b)–(d).

Tab.1 Contact angles of different samples

Fig.6 The survey XPS spectra of PEEK, CCPEEK and BIPEEK.

Fig.7 C 1s spectra of (a) PEEK and (b) BIPEEK; O 1s spectra of (c) PEEK and (d) BIPEEK.

Fig.8 The residual rate of rhBMP-2 on the surface of BIPEEK.

Fig.9 Cell proliferation of MC3T3-E1 cells cultured on PEEK, CCPEEK and BIPEEK after 1, 3, 5 and 7 d. (_* denotes significant difference: P<0.05)

Fig.10 ALP activity of MC3T3-E1 cells cultured on PEEK, CCPEEK and BIPEEK after 7 and 14 d. (_* denotes significant difference: P<0.05)

Fig.11 Alizarin red S staining of PEEK, CCPEEK and BIPEEK cultured with MC3T3-E1 cells after 14 d.

Fig.12 Semi-quantitative analysis of alizarin red S staining of PEEK, CCPEEK and BIPEEK cultured with MC3T3-E1 cells after 14 d. (_* denotes significant difference: P<0.05)

Fig.13 Calluses in specimens in (a) the control group and (b) the experimental group after implanting prostheses for 4 weeks.

Tab.2 Differences between control and experimental groups at the time of 4 and 8 weeks after implantation

Tab.1

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