Bioactivity of Titanium Metal Hybridized with Inactivated Bacterial Biofilm

doi:10.11900/0412.1961.2017.00290

Acta Metall Sin

2017, Vol. 53

Issue (10): 1402-1412 DOI: 10.11900/0412.1961.2017.00290

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Bioactivity of Titanium Metal Hybridized with Inactivated Bacterial Biofilm

Xiaofeng LU^1,², Ming XIAO^1,², Yangmei CHEN^1,², Bangcheng YANG^1,²(

)

1 Engineering Research Center in Biomaterials, Sichuan University, Chengdu 610064, China
2 National Engineering Research Center for Biomaterials, Sichuan University, Chengdu 610064, China

Cite this article:

Xiaofeng LU, Ming XIAO, Yangmei CHEN, Bangcheng YANG. Bioactivity of Titanium Metal Hybridized with Inactivated Bacterial Biofilm. Acta Metall Sin, 2017, 53(10): 1402-1412.

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Abstract

Bacterial biofilm contained with proteins and polysaccharides, which made it have the potential to mimic extracellular matrix for tissue repair. It also has the properties to attach the substrate firmly. In order to regulate the bioactivity of titanium metal, inactivated bacterial biofilm was hybridized on the metal surfaces. Staphylococcus aureus (S.aureus) was cultured for 5 d on pure titanium metal (P-Ti) and that subjected to acid-alkali treatment (AA-Ti), and then the bacteria was inactivated at 121 ℃ and 0.13 MPa for 30 min to get hybridized titanium metals BP-Ti and BAA-Ti respectively. BCA (bicinchoninic acid) assay and phenol-sulfuric acid analysis showed the extracellular matrix (ECM) in the biofilm on BAA-Ti contented less polysaccharide and more protein than that on BP-Ti. Enzyme immunoassay showed the staphylococcal enterotoxins in both biofilms were lower than the threshold value. SBF (simulated body fluid) soaking experiments showed both BAA-Ti and BP-Ti could induce apatite formation after 5 d, and BAA-Ti induced less apatite than the BP-Ti did. The AA-Ti could induce apatite formation at 1 d, but there is no apatite formation on P-Ti even after 5 d. It indicated that the biofilm decreased the bioactivity of BAA-Ti, but increased the bioactivity of BP-Ti. This effect might depend on the roles of proteins and polysaccharide in the biofilms which could restrain the apatite formation for the former, and accelerate the apatite formation for the later. When osteosarcoma cell MG-63 was cultured on the metals for 3 d, the cell proliferation ability on the metals followed by the order of BAA-Ti

Key words: titanium biofilm hybridization polysaccharide protein bioactivity

Received: 12 July 2017

ZTFLH:

TG178

Fund: Supported by National Key Research and Development Program of China (No.2016YFC1102700), National Nature Science Foundation of China (Nos.31570966 and 31771035), Key Program of Science & Technology Development of Sichuan Province (No.2012FZ0122), Key Program of Science & Technology Development of Chengdu (No.2015-HM01-00142-SF) and the Jiangsu Collaborative Innovation Center of Biomedical Functional Materials

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	Xiaofeng LU
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https://www.ams.org.cn/EN/10.11900/0412.1961.2017.00290 OR https://www.ams.org.cn/EN/Y2017/V53/I10/1402

Fig.1 SEM images (a, b) and XRD spectra (c, d) of P-Ti (a, c) and AA-Ti (b, d) without biofilm

Fig.2 SEM images of P-Ti (a~c) and AA-Ti (d~f) cultured with S.aureus for 1 d (a, d), 3 d (b, e) and 5 d (c, f)

Fig.3 XRD spectra of BP-Ti (a) and BAA-Ti (b) cultured with S.aureus for 5 d

Fig.4 AFM images of P-Ti (a), AA-Ti (b), BP-Ti (c) and BAA-Ti (d)

Fig.5 AFM images of the surface potential of P-Ti (a), AA-Ti (b), BP-Ti (c) and BAA-Ti (d)

Fig.6 Optical density (OD) of staphylococcal enterotoxins (A, B, C, D and E) on BAA-Ti and BP-Ti(**—p<0.05)

Fig.7 Surface SEM images of P-Ti (a1~a3), BP-Ti (b1~b3), AA-Ti (c1~c3) and BAA-Ti (d1~d3) after soaked in SBF for 1 d (a1~d1), 3 d (a2~d2) and 5 d (a3~d3)

Fig.8 XRD spectra of P-Ti, BP-Ti, AA-Ti and BAA-Ti after soaked in SBF for 1 d (a1, a2), 3 d (b1, b2) and 5 d (c1, c2)

Fig.9 FTIR spectra of P-Ti, BP-Ti, AA-Ti and BAA-Ti after soaked in SBF for 1 d (a1, a2), 3 d (b1, b2) and 5 d (c1, c2)

Fig.10 MTT assay of MG-63 cultured on the specimens for different days

Fig.11 CLSM of MG-63 cultured on P-Ti (a1~a3), BP-Ti (b1~b3), AA-Ti (c1~c3) and BAA-Ti (d1~d3) for 1 d (a1~d1), 2 d (a2~d2) and 3 d (a3~d3)

Fig.12 SEM images of MG-63 cultured on P-Ti (a), BP-Ti (b), AA-Ti (c) and BAA-Ti (d) for 3 d

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