Involvement of miR-199a-5p-loaded mesoporous silica nanoparticle-polyethyleneimine-KALA in osteogenic differentiation

IF 3.4 3区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Dental Sciences Pub Date : 2024-07-01 DOI:10.1016/j.jds.2024.01.007

Tianyue Wang , Hidemi Nakata , Bing Shen , Ziying Jiao , Kaori Yokota , Shinji Kuroda , Shohei Kasugai , Eriko Marukawa

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引用次数: 0

Abstract

Background/purpose

While there are numerous reports on surgical techniques and materials for bone grafting, limited methods are available to enhance the body's inherent capacity to heal bones. Here we investigated microRNA-199a (miR-199a), a molecular that promotes osteoblast differentiation and bone healing.

Materials and methods

To construct a miR-199a delivery complex, miR-199a-5p mimics were coated with mesoporous silica nanoparticles (MSNs) following modified with polyethyleneimine (PEI) and peptide WEAKLAKALAKALAKHLAKALAKALKACEA (KALA) to obtain 199a-5p-loaded MSN-PEI-KALA. Nanoparticle complexes are assessed for particle size and zeta potential using transmission electron microscopy and dynamic light scattering. Then MC3T3-E1 cells are exposed to MSN_miR-199a-5p @PEI-KALA. The impact of MSN_miR-199a-5p@PEI-KALA at varying concentrations on cell viability is assessed using Cell Counting Kit-8. Cell uptake and distribution were analyzed by double fluorescent staining with fluorescein amidite-labeled MSN_miR-199a@PEI-KALA and lysosome labeling. On day 7 after osteogenic induction, alkaline phosphatase (ALP) staining was conducted.

Results

The findings indicated that the nanoparticle complexes encapsulating PEI and peptide exhibited an augmentation in both particle size and zeta potential. At a dosage of 10 μg/mL, MSN_miR-199a@PEI-KALA displayed the lowest cytotoxicity compared to the control group. MC3T3-E1 cells treated with MSN_miR-199a-5p@PEI-KALA exhibited intensified ALP staining and elevated mRNA expression levels of ALP, runt-related transcription factor 2, and osteopontin, suggesting the involvement of miR-199a-5p-loaded MSN-PEI-KALA in osteogenic differentiation.

Conclusion

The successful construction of the delivering complex MSN_miR-199a@PEI-KALA in present research highlights the promise of this biomaterial carrier for the application of miRNAs in treating bone defects.

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miR-199a-5p负载介孔二氧化硅纳米粒子-聚乙烯亚胺-KALA参与成骨分化过程

背景/目的尽管有关骨移植手术技术和材料的报道不胜枚举，但用于增强人体固有的骨骼愈合能力的方法却十分有限。材料与方法为了构建 miR-199a 递送复合物，miR-199a-5p 模拟物被包覆在介孔二氧化硅纳米颗粒（MSNs）上，然后用聚乙烯亚胺（PEI）和肽 WEAKLAKALAKALAKHLAKALAKALKACEA（KALA）修饰，得到 199a-5p 负载的 MSN-PEI-KALA。利用透射电子显微镜和动态光散射评估纳米颗粒复合物的粒度和 ZETA 电位。然后将 MC3T3-E1 细胞暴露于 MSN_miR-199a-5p @PEI-KALA 中。使用细胞计数试剂盒-8 评估不同浓度的 MSN_miR-199a-5p@PEI-KALA 对细胞活力的影响。通过亚氨酸荧光素标记的MSN_miR-199a@PEI-KALA和溶酶体标记的双重荧光染色分析细胞摄取和分布情况。结果研究结果表明，包裹 PEI 和多肽的纳米颗粒复合物的粒径和 ZETA 电位都有所增大。与对照组相比，当剂量为 10 μg/mL 时，MSN_miR-199a@PEI-KALA 的细胞毒性最低。用MSN_miR-199a-5p@PEI-KALA处理的MC3T3-E1细胞的ALP染色增强，ALP、runt相关转录因子2和骨素的mRNA表达水平升高，表明miR-199a-5p负载的MSN-PEI-KALA参与了成骨分化。结论本研究成功构建了MSN_miR-199a@PEI-KALA递送复合物，凸显了这种生物材料载体在应用miRNA治疗骨缺损方面的前景。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Dental Sciences 医学-牙科与口腔外科

CiteScore

5.10

自引率

14.30%

发文量

348

审稿时长

6 days

期刊介绍： he Journal of Dental Sciences (JDS), published quarterly, is the official and open access publication of the Association for Dental Sciences of the Republic of China (ADS-ROC). The precedent journal of the JDS is the Chinese Dental Journal (CDJ) which had already been covered by MEDLINE in 1988. As the CDJ continued to prove its importance in the region, the ADS-ROC decided to move to the international community by publishing an English journal. Hence, the birth of the JDS in 2006. The JDS is indexed in the SCI Expanded since 2008. It is also indexed in Scopus, and EMCare, ScienceDirect, SIIC Data Bases. The topics covered by the JDS include all fields of basic and clinical dentistry. Some manuscripts focusing on the study of certain endemic diseases such as dental caries and periodontal diseases in particular regions of any country as well as oral pre-cancers, oral cancers, and oral submucous fibrosis related to betel nut chewing habit are also considered for publication. Besides, the JDS also publishes articles about the efficacy of a new treatment modality on oral verrucous hyperplasia or early oral squamous cell carcinoma.