MOE Key Laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China.
J Mater Chem B. 2020 Apr 21;8(15):3016-3027. doi: 10.1039/c9tb02899d. Epub 2020 Mar 24.
Controlling and detecting cell differentiation in human mesenchymal stem cells (hMSCs) for regenerative medicine remain challenging at present. Here, we developed multifunctional gold nanoparticles (AuNPs) to control and detect osteogenic differentiation in human mesenchymal stem cells (hMSCs) in real-time. The polyethyleneimine (PEI) capped AuNPs (Au-PEI) were synthesized first and were then conjugated with a matrix metalloproteinase 13 (MMP13)-sensitive peptide-FITC group (EGPLGVRGKG-FITC) to obtain multifunctional AuNPs (AuNP-PEI-peptide-FITC). AuNP-PEI-peptide-FITC could bind siRNA with PEI by electrostatic interactions to form the AuNP-PEI-peptide-FITC/siRNA nanocomplexes, allowing efficient siRNA delivery in hMSCs. The adipogenic-related gene peroxisome proliferator-activated receptor γ (PPARγ) was targeted for silencing by the AuNP-PEI-peptide-FITC/siRNA nanocomplexes to control osteogenic differentiation in hMSCs. After demonstrating that the AuNP nanocomplexes could control cell differentiation, the versatility of this tool was illustrated by showing that it can be used as a nanoprobe for real time detection of the osteogenic differentiation of hMSCs. This was done by measuring the activity of the MMP13 enzyme (produced during osteogenic differentiation) through the recovery of FITC fluorescence. This multifunctional AuNP showed a robust new methodology for controlling cell fate and simultaneously detecting cell differentiation in real-time for hMSCs, which is promising for multiple applications in regenerative medicine.
目前，控制和检测人骨髓间充质干细胞（hMSCs）的细胞分化在再生医学领域仍然具有挑战性。在这里，我们开发了多功能金纳米粒子（AuNPs），以实时控制和检测人骨髓间充质干细胞（hMSCs）的成骨分化。首先合成了聚乙烯亚胺（PEI）封端的金纳米粒子（Au-PEI），然后将其与基质金属蛋白酶 13（MMP13）敏感肽-FITC 基团（EGPLGVRGKG-FITC）缀合，得到多功能金纳米粒子（AuNP-PEI-肽-FITC）。AuNP-PEI-肽-FITC 可以通过静电相互作用与 PEI 结合形成 AuNP-PEI-肽-FITC/siRNA 纳米复合物，从而允许在 hMSCs 中高效递送 siRNA。通过 AuNP-PEI-肽-FITC/siRNA 纳米复合物靶向沉默脂肪形成相关基因过氧化物酶体增殖物激活受体 γ（PPARγ），以控制 hMSCs 的成骨分化。在证明 AuNP 纳米复合物可以控制细胞分化后，通过测量基质金属蛋白酶 13（MMP13）酶的活性（在成骨分化过程中产生）来实时检测 hMSCs 的成骨分化，说明了该工具的多功能性。通过恢复 FITC 荧光来实现。这种多功能 AuNP 为控制细胞命运并同时实时检测 hMSCs 的细胞分化提供了一种强大的新方法，有望在再生医学的多个应用中得到广泛应用。
