Transfection system of amino-functionalized calcium phosphate nanoparticles: in vitro efficacy, biodegradability, and immunogenicity study.
Transfection system of amino-functionalized calcium phosphate nanoparticles: in vitro efficacy, biodegradability, and immunogenicity study.
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DOI：
10.1021/am507193a
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发表时间：
2015-03
影响因子：
9.5
通讯作者：
Babak Mostaghaci;Julia Susewind;G. Kickelbick;C. Lehr;B. Loretz
中科院分区：
文献类型：
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作者：
Babak Mostaghaci;Julia Susewind;G. Kickelbick;C. Lehr;B. Loretz
已经开发了许多方法以使用磷酸钙（CaP）将核苷酸递送到活细胞中。最近显示用N-（2-氨基乙基）-3-氨基丙基三甲氧基硅烷对CaP纳米颗粒（CaP NP）进行表面官能化以获得具有正表面电荷的分散的NP，其能够转染（Chem.Mater. 2013，25（18），3667）。在这项研究中，不同的晶体结构的氨基改性的CaP纳米粒子（透钙磷石和羟基磷灰石）进行了研究，他们在细胞培养系统中的相互作用更详细。定性（共聚焦激光扫描显微镜）和定量（流式细胞术）转染实验与两个细胞系显示较高的转染效率透钙磷石与羟基磷灰石。转染还揭示了细胞类型依赖性。HEK 293细胞比A549细胞更容易被转染。该结果得到细胞毒性结果的支持。A549细胞对CaP NPs表现出更高程度的耐受性。此外，考虑了表面修饰对与巨噬细胞和补体相互作用的影响，巨噬细胞和补体是先天免疫系统的两个重要组成部分。胺表面功能化具有减少促炎细胞因子释放的作用。通过C3 a补体活化测定研究的补体相互作用确实显示在没有或具有胺修饰的CaP NP之间没有显著差异，并且总体上弱相互作用。最后，研究了CaP NPs在生物介质中的两种晶体结构以及在酸性和中性pH下的降解。两种氨基修饰的CaP NPs在中性pH下均在数天内崩解，透钙磷石NPs在酸性pH下崩解明显更快。总之，这种氨基官能化的CaP NPs的良好转染能力以及优异的生物相容性、生物降解性，和低免疫原性使它们成为进一步评价的感兴趣的候选物。
Many methods have been developed in order to use calcium phosphate (CaP) for delivering nucleotides into living cells. Surface functionalization of CaP nanoparticles (CaP NPs) with N-(2-aminoethyl)-3-aminopropyltrimethoxysilane was shown recently to achieve dispersed NPs with a positive surface charge, capable of transfection (Chem. Mater. 2013, 25 (18), 3667). In this study, different crystal structures of amino-modified CaP NPs (brushite and hydroxyapatite) were investigated for their interaction in cell culture systems in more detail. Qualitative (confocal laser scanning microscopy) and quantitative (flow cytometry) transfection experiments with two cell lines showed the higher transfection efficacy of brushite versus hydroxyapatite. The transfection also revealed a cell type dependency. HEK293 cells were easier to transfect compared to A549 cells. This result was supported by the cytotoxicity results. A549 cells showed a higher degree of tolerance toward the CaP NPs. Further, the impact of the surface modification on the interaction with macrophages and complement as two important components of the innate immune system were considered. The amine surface functionalization had an effect of decreasing the release of proinflammatory cytokines. The complement interaction investigated by a C3a complement activation assay did show no significant differences between CaP NPs without or with amine modification and overall weak interaction. Finally, the degradation of CaP NPs in biological media was studied with respect to the two crystal structures and at acidic and neutral pH. Both amino-modified CaP NPs disintegrate within days at neutral pH, with a notable faster disintegration of brushite NPs at acidic pH. In summary, the fair transfection capability of this amino functionalized CaP NPs together with the excellent biocompatibility, biodegradability, and low immunogenicity make them interesting candidates for further evaluation.