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. 2021 Nov 8;11(11):1160. doi: 10.3390/jpm11111160

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Pulverization of sCA-miRNA34a with PEG blending. (A) During sCA-miRNA34a production, methoxy-PEG-CO(CH₂)₂COO-NHS (Mw 10,000) targeting the OH group of sCA ([Ca₁₀(PO₄)^6−X(CO₃)^X(OH)₂]) was initially mixed with the constituents and three kinds of pulverization, bath sonication (38 kHz), wet jet-milling (30 pass), and AFA (1 MHz), performed after generation of the particles. The solution prepared for intravenous injection was directly (without dilution) analyzed by DLS, and both particle size distribution and autocorrelation function are shown for each sample. (B) Degradation of miRNA34a after the pulverization. (C) Proliferation assay of HCT116 human colon cancer cells at 48 and 72 h. (D) The mRNA expression of Survivin, Bcl-2, and E2F1 in HCT116 at 36 h after treatment. Data represent mean ± SEM (n = 4). p-values were obtained using the two-tailed t test.

Pulverization of sCA-miRNA34a with PEG blending. (A) During sCA-miRNA34a production, methoxy-PEG-CO(CH₂)₂COO-NHS (Mw 10,000) targeting the OH group of sCA ([Ca₁₀(PO₄)^6−X(CO₃)^X(OH)₂]) was initially mixed with the constituents and three kinds of pulverization, bath sonication (38 kHz), wet jet-milling (30 pass), and AFA (1 MHz), performed after generation of the particles. The solution prepared for intravenous injection was directly (without dilution) analyzed by DLS, and both particle size distribution and autocorrelation function are shown for each sample. (B) Degradation of miRNA34a after the pulverization. (C) Proliferation assay of HCT116 human colon cancer cells at 48 and 72 h. (D) The mRNA expression of Survivin, Bcl-2, and E2F1 in HCT116 at 36 h after treatment. Data represent mean ± SEM (n = 4). p-values were obtained using the two-tailed t test.