All animal protocols and experiments were in accordance with the Purdue Animal Care and Use Committee (protocol number: 1612001512) and conducted in compliance to applicable regulations. As a model system, athymic nude mice (n = 10, 6 weeks old, NCr-Fox1nu, Charles River Laboratories, USA) were used in the in vivo animal tests. The Si NN-patch was sterilized with 70% ethanol, exposed to UV light for 30 min, and then immersed in a DyLight 800 dye (1 μl/ml; Thermo Fisher Scientific, USA) for 5 min. For the transdermal nanoinjection, the Si NN-patch was gently placed on the skin of mice awake with a medical-grade adhesive (Roll-On Adhesive, JOBST, USA) and pressed firmly for ~10 s. The behavior of the mice was monitored in real time and recorded with a high-resolution video camera (EOS 700D, Canon Inc.). For the intramuscular nanoinjection, the mice were anesthetized with avertin (Sigma-Aldrich, USA) by intraperitoneal nanoinjection at a dose of 250 mg/kg, followed by careful incision of the skin with surgical scissors to expose the gluteal and lumbar muscles. In these tests, the incision was made on the upper back of the mice, where the Si NN-patch was inserted. The incisional site was then sutured using surgical needle and thread. To induce acute inflammation, PMA (100 μM, 40 μl; Sigma-Aldrich, USA) as a positive control was rubbed on the skin, muscle, and ear. After ~5 hours of implementations, 100 μl of luminol (body weight, 200 mg/kg; Sigma-Aldrich, USA) was administered by intraperitoneal nanoinjection, and then the mice were imaged (exposure time, 3 min; binning, 4). For the histological examination, approximately 4 μm of tissue section was cut, fixed in 10% formalin (Sigma-Aldrich, USA) for 24 hours, and then stained with H&E (Sigma-Aldrich, USA) using a light microscope (Eclipse 90i, Nikon Inc.).

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