滅活細(xì)菌生物膜雜化生物活性鈦金屬材料研究

滅活細(xì)菌生物膜雜化生物活性鈦金屬材料研究Bioactivity of Titanium Metal Hybridized with Inactivated Bacterial Biofilm

為調(diào)控生物活性鈦金屬材料的生物活性，使用滅活的細(xì)菌生物膜對其進(jìn)行雜化。金黃色葡萄球菌(S.aureus)在純Ti (P-Ti)和酸堿處理Ti (AA-Ti)表面培養(yǎng)5 d后在121 ℃、0.13 MPa條件下滅活30 min，獲得細(xì)菌生物膜雜化的純Ti (BP-Ti)和酸堿處理Ti (BAA-Ti)。BCA蛋白分析和苯酚-硫酸分析表明，在BAA-Ti表面的細(xì)菌生物膜的細(xì)胞外基質(zhì)較BP-Ti表面細(xì)菌生物膜的細(xì)胞外基質(zhì)含較多蛋白，而后者含有較多的多糖。酶免疫分析表明，在BP-Ti和BAA-Ti表面的腸毒素都低于閾值。模擬體液(SBF)浸泡實(shí)驗(yàn)表明，BAA-Ti和BP-Ti在5 d內(nèi)均可誘導(dǎo)磷灰石形成，但BAA-Ti誘導(dǎo)磷灰石能力低于BP-Ti。AA-Ti在1 d內(nèi)可誘導(dǎo)磷灰石生成，但在5 d內(nèi)P-Ti表面無磷灰石生成。表明生物膜提高了BP-Ti的生物活性，但降低了BAA-Ti的生物活性。這可能源于生物膜中多糖有利于促進(jìn)磷灰石的形成，而其中的蛋白抑制磷灰石的形成。MG-63細(xì)胞培養(yǎng)實(shí)驗(yàn)表明，細(xì)胞增殖能力順序?yàn)椋築AA-Ti <BP-Ti <AA-Ti<P-Ti，表明生物膜中的蛋白抑制細(xì)胞的增殖。Ti表面雜化的生物膜顯著影響鈦金屬材料的生物學(xué)性能，可通過表面雜化生物膜的方式實(shí)現(xiàn)對鈦的生物活性的調(diào)控。

Bacterial biofilm contained with proteins and polysaccharides, which made it have thepotential to mimic extracellular matrix for tissue repair. It also has the properties to attach the substratefirmly. In order to regulate the bioactivity of titanium metal, inactivated bacterial biofilm was hybridized onthe metal surfaces. Staphylococcus aureus (S.aureus) was cultured for 5 d on pure titanium metal (P-Ti) andthat subjected to acid-alkali treatment (AA-Ti), and then the bacteria was inactivated at 121 ℃ and 0.13MPa for 30 min to get hybridized titanium metals BP-Ti and BAA-Ti respectively. BCA assay andphenol-sulfuric acid analysis showed the extracellular matrix (ECM) in the biofilm on BAA-Ti contentedless 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 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<BP-Ti<AA-Ti<P-Ti, which indicated that the proteins might inhibit the cell proliferation. The biofilms hybridized on titanium metal could influence the biological response of titanium metal. It might be possible to regulate the bioactivity of titanium by biofilm hybridization.

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