多功能纳米金刚石在神经发育和超分辨率成像中的应用系统研究。

IF 11.3 1区 医学 Q1 Medicine Biomaterials Research Pub Date : 2023-04-27 DOI:10.1186/s40824-023-00384-9
Jaeheung Kim, Moon Sung Kang, Seung Won Jun, Hyo Jung Jo, Dong-Wook Han, Chang-Seok Kim
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引用次数: 1

摘要

背景:中枢神经系统缺陷神经元的再生是神经退行性疾病治疗的热点问题。由于受损的神经元往往无法实现新生神经突的自发修复,各种组织工程方法都集中在神经突发生上,以实现受损神经元细胞的再生。同时,由于对更好的诊断的需求,荧光显微镜超分辨率成像技术的研究引发了技术的发展,超越了由光学衍射极限决定的经典分辨率,以精确观察神经元的行为。本文研究了多功能纳米金刚石作为神经生成促进剂和超分辨率成像探针的作用。方法:在HT-22海马神经元细胞中加入含nd的生长培养基和分化培养基,培养10 d,研究nd诱导神经发生的能力。利用nd的光闪烁特性,利用定制的双光子显微镜观察nd的离体和离体图像,并进行直接随机光学重建显微镜(dSTORM)超分辨率重建。此外,在静脉注射NDs后24 h进行小鼠脑离体成像。结果:NDs能被细胞内吞,促进神经细胞的自发发生,无任何分化因子,具有良好的生物相容性,无明显毒性。通过dSTORM将nd -内吞噬细胞的图像重建为超分辨率图像,从而解决了纳米颗粒造成的图像失真问题,包括尺寸膨胀和难以区分附近位置的颗粒。此外,NDs在小鼠脑内的离体图像证实了NDs可以穿透血脑屏障(BBB)并保留其用于dSTORM的光闪烁特性。结论:NDs具有dSTORM超分辨率成像、神经生成促进和血脑屏障穿透等功能,具有显著的生物学应用潜力。
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A systematic study on the use of multifunctional nanodiamonds for neuritogenesis and super-resolution imaging.

Background: Regeneration of defective neurons in central nervous system is a highlighted issue for neurodegenerative disease treatment. Various tissue engineering approaches have focused on neuritogenesis to achieve the regeneration of damaged neuronal cells because damaged neurons often fail to achieve spontaneous restoration of neonatal neurites. Meanwhile, owing to the demand for a better diagnosis, studies of super-resolution imaging techniques in fluorescence microscopy have triggered the technological development to surpass the classical resolution dictated by the optical diffraction limit for precise observations of neuronal behaviors. Herein, the multifunctional nanodiamonds (NDs) as neuritogenesis promoters and super-resolution imaging probes were studied.

Methods: To investigate the neuritogenesis-inducing capability of NDs, ND-containing growing medium and differentiation medium were added to the HT-22 hippocampal neuronal cells and incubated for 10 d. In vitro and ex vivo images were visualized through custom-built two-photon microscopy using NDs as imaging probes and the direct stochastic optical reconstruction microscopy (dSTORM) process was performed for the super-resolution reconstruction owing to the photoblinking properties of NDs. Moreover, ex vivo imaging of the mouse brain was performed 24 h after the intravenous injection of NDs.

Results: NDs were endocytosed by the cells and promoted spontaneous neuritogenesis without any differentiation factors, where NDs exhibited no significant toxicity with their outstanding biocompatibility. The images of ND-endocytosed cells were reconstructed into super-resolution images through dSTORM, thereby addressing the problem of image distortion due to nano-sized particles, including size expansion and the challenge in distinguishing the nearby located particles. Furthermore, the ex vivo images of NDs in mouse brain confirmed that NDs could penetrate the blood-brain barrier (BBB) and retain their photoblinking property for dSTORM application.

Conclusions: It was demonstrated that the NDs are capable of dSTORM super-resolution imaging, neuritogenic facilitation, and BBB penetration, suggesting their remarkable potential in biological applications.

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来源期刊
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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