Department of Chemistry and Biochemistry, Bioscience Institute, State University of São Paulo - UNESP, campus Botucatu, Botucatu, São Paulo, Brazil.
Electron Microscopy Center, IBB, UNESP, Botucatu, São Paulo, Brazil.
J Biomed Mater Res A. 2019 Aug;107(8):1597-1604. doi: 10.1002/jbm.a.36673. Epub 2019 Apr 9.
Molecular mechanism governing inflammatory scenario in response to titanium (Ti)-nanotexturing surfaces needs to be better addressed. Thus, we subjected pre-osteoblast to different Ti-texturing surfaces, as follows: machined (Mac), double acid-etching (DAE), and nanoscaled hydroxyapatite-blasted titanium surface (nHA), considering the cells chronically responding either directly (when the cells were cultured onto the surfaces) or indirectly (when the cells were challenged with the conditioned medium by the surfaces), up to 10 days. Our results showed that there is a dynamic requirement of inflammatory-related genes activation in response to nHA by up expressing IL1ß, IL6, IL10, and IL33 (direct condition) and IL6, IL10, IL18 (indirect condition). Importantly, our data show that there is inflammasome involvement, once NLRP3, ASC1, and CASP1 genes were also required. As we found a strong signal of IL10, an anti-inflammatory cytokine, we further investigated Sonic Hedgehog (Shh) signaling cascade. Surprisingly, Shh ligand and Smoothened (Smo) genes were up-modulated in response to nHA, while Patched (Ptc) was down-modulated. Finally, an interactome was built using bioinformatics reinforcing Shh signaling cascade on modulating IL10 transcripts by Src mediating this process and this prevalence of anti-inflammatory picture might explain the low profile of RANKL transcripts in response to nHA, compromising the osteoclastogenesis surrounding the implants. Taking our results into account, our data show that the inflammatory landscape promoted by nHA is strictly modulated by Shh signaling promoted anti-inflammatory pathways. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1597-1604, 2019.
调控钛（Ti）纳米纹理表面炎症反应的分子机制尚需进一步阐明。因此，我们将前成骨细胞分别置于不同的 Ti 纹理表面，包括机械加工（Mac）、双酸蚀刻（DAE）和纳米羟基磷灰石喷钛表面（nHA），考虑细胞的慢性直接（当细胞在表面上培养时）或间接（当细胞被表面的条件培养基所挑战时）反应，时间长达 10 天。我们的结果表明，nHA 可通过上调白细胞介素 1β（IL1β）、白细胞介素 6（IL6）、白细胞介素 10（IL10）和白细胞介素 33（IL33）（直接条件）以及白细胞介素 6（IL6）、白细胞介素 10（IL10）和白细胞介素 18（IL18）（间接条件）来满足炎症相关基因激活的动态需求。重要的是，我们的数据表明炎症小体的参与，因为 NLRP3、ASC1 和 CASP1 基因也需要。由于我们发现了强烈的抗炎细胞因子 IL10 信号，我们进一步研究了 Sonic Hedgehog（Shh）信号级联。令人惊讶的是，nHA 反应上调了 Shh 配体和 Smoothened（Smo）基因，而 Patched（Ptc）基因下调。最后，我们使用生物信息学构建了一个相互作用网络，强化了 Shh 信号级联通过 Src 介导的这个过程来调节 IL10 转录，这种抗炎信号的优势可能解释了 nHA 反应中 RANKL 转录物水平低的现象，从而影响了植入物周围破骨细胞的形成。考虑到我们的结果，我们的数据表明，nHA 促进的炎症景观受到 Shh 信号促进的抗炎途径的严格调控。
