Construction of a Microfluidic Platform With Core-Shell CdSSe@ZnS Quantum Dot-Encoded Superparamagnetic Iron Oxide Microspheres for Screening and Locating Matrix Metalloproteinase-2 Inhibitors From Fruits of Rosa roxburghii

Abstract

The microfluidic platform is a versatile tool for screening and locating bioactive molecules from functional foods. Here, a layer-by-layer assembly approach was used to fabricate core-shell CdSSe@ZnS quantum dot encoded superparamagnetic iron oxide microspheres, which served as a carrier for matrix metalloproteinase-2. The matrix metalloproteinase-2 camouflaged magnetic microspheres was further incorporated into a homemade microfluidic platform and incubated with extracts of fruits of Rosa roxburghii. The flow rate of the microfluidic platform was tuned. The major influencing parameters on ligand binding, such as dissociate solvents, incubation pH, ion strength, temperature, and incubation time were also optimized by using ellagic acid as a model compound. The specific binding ligands were sent for structure elucidation by mass spectrometry. The absolute recovery of ellagic acid ranged from 101.14 to 102.40% in the extract of R. roxburghii under the optimal extraction conditions. The linearity was pretty well in the range of 0.009–1.00 mg·ml−1 (R2 = 0.9995). The limit of detection was 0.003 mg·ml−1. The relative SDs of within-day and between-day precision were <1.91%. A total of thirteen ligands were screened out from fruits of R. roxburghii, which were validated for their inhibitory effect by enzyme assay. Of note, eleven new matrix metalloproteinase-2 inhibitors were identified, which may account for the antitumor effect of fruits of R. roxburghii.

Ligand fishing as a tool to screen natural products with anticancer potential

“…In this context, Tao et al. [172] constructed a very interesting ligand fishing system that combines immobilization on MPs with a microfluidic chip system. In it, the authors prepared and characterized a quantum dot magnetic particle with immobilized MMP‐2 before submitting it to a microfluidic chip.…”

“…Finally, the eluted ligands were pumped into a UPLC‐Q‐TOF/MS system to be analyzed. Among the twelve ligands fished and identified, ellagic acid stands out as it is a well‐known MMP‐2 inhibitor [172, 173]. This polyphenolic compound possesses antiproliferative activity in various types of cancer cells, such as glioblastoma (U87 and U118), human osteogenic sarcoma (CRL1343), human colorectal adenocarcinoma (Caco‐2), human prostate cancer (DU 145) and breast cancer (Hs 578T and MCF‐7) [174–176].…”

“…This polyphenolic compound possesses antiproliferative activity in various types of cancer cells, such as glioblastoma (U87 and U118), human osteogenic sarcoma (CRL1343), human colorectal adenocarcinoma (Caco‐2), human prostate cancer (DU 145) and breast cancer (Hs 578T and MCF‐7) [174–176]. Furthermore, the other eleven ligands fished (niga‐ichigoside F 1 , rosamultin, kajiichigoside F 1 , zebirioside O, peduncloside, potentillanoside A, quadranoside VIII, corilagin, oreganol A, medicagenic acid, and valerenic acid) were described as MMP‐2 inhibitors for the first time, evidencing the high throughput potential of this approach to isolate new molecules that could be useful for cancer treatment [172].…”

“…There are more than 20 isoforms of MMPs, each having specific substrates, tissue distribution, molecular mass, and gene coding. Among them, gelatinases (MMP-2 and 9) are particularly involved in cancer invasion and metastasis as their increased expression has been related to the invasiveness of gliomas [168], malignant transformations after human papilloma virus infections [169], reduced survival in patients with primary breast carcinoma [170] and esophageal cancer [171].…”