Ahn Guk Young, Kim Sung-Eun, Yun Tae Hoon, Choi Inseong, Park Daewon, Choi Sung-Wook
Biomedical and Chemical Engineering, Department of Biotechnology, The Catholic University of Korea, 43 Jibong-ro Wonmi-gu, Gyeonggi-do, 14662, Bucheon-si, Republic of Korea.
Department of Orthopaedic Surgery and Rare Diseases Institute, Korea University Medical Centre, Guro Hospital, 80 Guro-dong, Guro-gu, Seoul, 152-703, Republic of Korea.
Biomater Res. 2021 Sep 23;25(1):28. doi: 10.1186/s40824-021-00231-9.
Alendronate (Alen) is promising material used for bone-targeted drug delivery due to its high bone affinity and therapeutic effects on bone diseases. In addition, Alen can enhance the osteogenic differentiation of osteoblastic cell. Recently, nanodiamonds (NDs) with hardness, non-toxicity, and excellent biocompatibility are employed as promising materials for carrier systems and osteogenic differentiation. Therefore, we prepared Alen-conjugated NDs (Alen-NDs) and evaluated their osteogenic differentiation performances.
Alen-NDs were synthesized using DMTMM as a coupling reagent. Morphological change of Mouse calvaria-derived preosteoblast (MC3T3-E1) treated with Alen-NDs was observed using the confocal microscope. The osteogenic differentiation was confirmed by cell proliferation, alkaline phosphatase (ALP), calcium deposition, and real-time polymerase chain reaction assay.
Alen-NDs were prepared to evaluate their effect on the proliferation and differentiation of osteoblastic MC3T3-E1 cells. The Alen-NDs had a size of about 100 nm, and no cytotoxicity at less than 100 μg/mL of concentration. The treatment of NDs and Alen-NDs reduced the proliferation rate of MC3T3-E1 cells without cell death. Confocal microscopy images confirmed that the treatment of NDs and Alen-NDs changed the cellular morphology from a fibroblastic shape to a cuboidal shape. Flow cytometry, alkaline phosphatase (ALP) activity, calcium deposition, and real-time polymerase chain reaction (RT-PCR) confirmed the higher differentiation of MC3T3-E1 cells treated by Alen-NDs, compared to the groups treated by osteogenic medium and NDs. The higher concentration of Alen-ND treated in MC3T3-E1 resulted in a higher differentiation level.
Alen-NDs can be used as potential therapeutic agents for osteoporosis treatment by inducing osteogenic differentiation.
阿仑膦酸盐（Alen）因其对骨的高亲和力及对骨疾病的治疗作用，是用于骨靶向给药的有前景的材料。此外，Alen可增强成骨细胞的成骨分化。近来，具有硬度、无毒性及优异生物相容性的纳米金刚石（NDs）被用作载体系统及成骨分化的有前景材料。因此，我们制备了阿仑膦酸盐共轭纳米金刚石（Alen-NDs）并评估了它们的成骨分化性能。
使用DMTMM作为偶联剂合成Alen-NDs。用共聚焦显微镜观察经Alen-NDs处理的小鼠颅骨来源的前成骨细胞（MC3T3-E1）的形态变化。通过细胞增殖、碱性磷酸酶（ALP）、钙沉积及实时聚合酶链反应测定法确认成骨分化。
制备Alen-NDs以评估其对成骨细胞MC3T3-E1细胞增殖和分化的影响。Alen-NDs尺寸约为100nm，浓度低于100μg/mL时无细胞毒性。NDs和Alen-NDs处理降低了MC3T3-E1细胞的增殖率但无细胞死亡。共聚焦显微镜图像证实，NDs和Alen-NDs处理使细胞形态从成纤维细胞形状变为立方形。流式细胞术、碱性磷酸酶（ALP）活性、钙沉积及实时聚合酶链反应（RT-PCR）证实，与成骨培养基和NDs处理组相比，Alen-NDs处理的MC3T3-E1细胞分化更高。在MC3T3-E1中处理的Alen-ND浓度越高，分化水平越高。
Alen-NDs可通过诱导成骨分化用作骨质疏松症治疗的潜在治疗剂。
