Figure 1.
Evaluation of cytotoxicity after tissue adhesives treatments for short-time using four conventional assays.
Mouse L929 fibroblasts (1×105 cells) were seeded in a 12 well plates. The cells were incubated control and treated with two tissue adhesives (PACA and Dermabond®) and then further incubated for 24 hrs. Cell viability was analyzed using the (A) WST assay, (B) NR assay, (C) LIVE/DEAD viability/cytotoxicity assay, Scale bars = 100 µm, (D) TUNEL assay. *P<0.05 vs. control (n = 6 pairs). **P>0.05 vs. PACA (n = 6 pairs). Evaluation of cytotoxicity after tissue adhesives for time course on fibroblast cell culture. Two tissue adhesive were treated with (E) direct contact and (F) indirect contact for 24 hr, 48 hr, and 72 hr. Cell viability was analyzed using the WST assay. *P<0.05, #P>0.05 vs. control (n = 4 pairs). **P>0.05 vs. PACA (n = 4 pairs).
Figure 2.
Evaluation of in vivo biocompatibility from incision rat model.
Histological analysis of Normal (negative), PACA-treated, Dermabond®–treated and untreated (positive) Sprague–Dawley rats at day 7 by (A) H&E staining and (B) MT staining. Scale bars = 10 µm.
Figure 3.
Evaluation of cytotoxicity after tissue adhesives treatments using Raman spectroscopy.
(A) Averaged Raman spectra of L929 cells (a: control, b: PACA, c: Dermabond®) and the spectral differences of control and treated cells (d: control-PACA, e: control-Dermabond®, f: PACA-Dermabond®). (B) Relative intensities of the Raman peaks for control and treated cells with two tissue adhesives (PACA and Dermabond®).
Table 1.
Peak assignment for Raman spectra of L929 cells.
Figure 4.
Raman spectroscopy to evaluate cytotoxic changes in response to effects of tissue adhesive on the DNA.
(A) Averaged Raman spectra of DNA (a: control, b: PACA, c: Dermabond®) and the spectral differences of control and treated DNA (d: control-PACA, e: control-Dermabond®, f: PACA-Dermabond®). (B) Relative intensities of the Raman peaks for control DNA and treated DNA with two tissue adhesive (PACA and Dermabond®). (C) PCR amplification of the 776 bp β-actin using same dose of DNA samples (Control, PACA and Dermabond®).
Figure 5.
PCA-SVM scores of the control and two tissue adhesive-treated (A) cells and (B) DNA.
The SVM classifier was used with the linear discriminant function.
Table 2.
Classification rate of the proposed PCA-SVM classifiers for evaluating biocompatibility of the PACA and Dermabond® tissue adhesives on the Raman spectral of cells and DNA.*