Mmp 2

We extracted blood samples from the heart of the test mice on the final day of experiments. The plasma levels of MMP-1, MMP-2, MMP-9, histamine, hyaluronidase, hyaluronic acid, IL-6, TNF-α, and AGEs were determined using commercial ELISA kits (MMP-1: MyBioSource, San Diego, CA, USA; MMP-2, MMP-9, hyaluronic acid, and TNF-α: R&D Systems, Minneapolis, MN, USA; histamine: Bertin Pharm., Montigny-le-Bretonneux, France; hyaluronidase: AB Clonal Inc., Tokyo, Japan; IL-6: Proteintech, Rosemont, IL, USA; and AGEs: OxiSelect AGE connection ELISA kit, Cell Biolabs Inc., San Diego, CA, USA), according to the instructions of the respective manufacturers.

Supernatants obtained from cell cultures treated with IC₅₀ conditions of the respective NPs were treated according to the manufacturer’s protocols for the assessment of the expression of different extracellular proteins and soluble mediators by ELISA. The following mouse kits from MyBiosourse, USA, were utilized: collagen1 (MBS724458), TGF-β (MBS160136), MMP2 (MBS824667), MMP9 (MBS175917), IL-6 (MBS824703), and TNF-α (MBS825075).
Mouse alpha-smooth muscle actin (α-SMA) ELISA kit (MBS267551, MyBioSource, USA) was also used to determine the expression level of the intracellular α-SMA in Fbls and TAFs according to the manufacturer’s protocol.