Ation for 24 hours to induce quiescence. Quiescent cells were incubated with CT-1 for 24 hours. Following this stimulation, CD-NP was added into HCF every 24 hours. Next, cells were labelled with the BrdU label for 24 hours and measured at absorbance 370 nm. Relative DNA synthesis of tested groups was normalized against the Title Loaded From File control (no addition of CD-NP) groups, where the control groups were set as the reference.10. Statistical AnalysisResults are presented in mean 6 standard deviation. The oneway analysis of variance (ANOVA) was used to compare significant difference and p,0.05 is denoted as statistically significant.Results 1. In vitro Release from FilmsFrom figure 1a, film 1 and film 3 had the lowest and highest initial (burst) release of 13 and 65 respectively. Subsequently, film 1 and film 3 released 60 and 99 CD-NP by 30 days. Film 2 had an intermediate initial burst release of 31 and released 93 of CD-NP at 16574785 the end of 30 days. In figure 1b, the concentration of the CD-NP (following the burst release) from allCenderitide-Eluting Filmfilms were more or less similar from 1 to 30 days (in the range of 1?6 mg/mL).investigated. In figure 4b, the cGMP production levels after the addition of released CD-NP from all three films were elevated significantly compared to the control group (p,0.05).2. In vitro Degradation and Mass LossThe degradation of the films was determined by measuring the molecular mass (figure 2a) and total mass (figure 2b) changes. There was no significant molecular mass change and mass loss in all three tested films, indicating the slow degradation of PCL.5. Effects of CD-NP on Human Cardiac Fibroblast (HCF) Cell ViabilityIn figure 5, the graphs of cell index (CI) of HCF against time is presented, where CI increment denotes increase in cell proliferation or cell spreading. Figure 5a shows the cell viability of HCF after daily dose of 37 mg/mL CD-NP compared to control. It can be seen that in the first 48 hours, there was no distinct difference between the CD-NP group and control, however, a downward trend started to develop after the 3rd dose was administered. By the addition of the 4th dose (figure 5a), it was clear that daily dose of CD-NP at concentration of 37 mg/mL resulted in lower CI compared to control. Figure 5b, c, d shows the cell viability study of HCF of films 1, 2 and 3 respectively against the control group. Both film 1 and 3 showed immediate decline of CI compared to control, whilst, film 2 only saw decline in CI compared to control on the 4th day. The relative cell index (RCI) is used to describe the cell viability in a comparative manner, where lower the RCI value denotes greater extent of inhibition. Figure 6a b, c and d shows the correlation between the RCI (primary y-axis) and peptide concentration (Title Loaded From File secondary y-axis) with respect to time. From figure 6a, we can see that daily dosing of CD-NP results in “spikes” of CD-NP to 37 mg/mL daily (secondary y-axis), but the RCI was only less than 1 on the 2nd day onwards. Films 1 and 3 had RCI value less than 1 from 0 to 5 days. Film 2 however only saw RCI less than 1 after the 1st day. By the 5th day, all three films had RCI 23977191 less than 1.3. Surface MorphologyFigure 3a, b and c present the initial surface morphology of films 1, 2 and 3 respectively. Both film 1 and film 3 appear to be more porous compared to film 2, which may be due to the use of an immiscible co-solvent system. And by using a longer period of emulsification, film 3 appears to be mo.Ation for 24 hours to induce quiescence. Quiescent cells were incubated with CT-1 for 24 hours. Following this stimulation, CD-NP was added into HCF every 24 hours. Next, cells were labelled with the BrdU label for 24 hours and measured at absorbance 370 nm. Relative DNA synthesis of tested groups was normalized against the control (no addition of CD-NP) groups, where the control groups were set as the reference.10. Statistical AnalysisResults are presented in mean 6 standard deviation. The oneway analysis of variance (ANOVA) was used to compare significant difference and p,0.05 is denoted as statistically significant.Results 1. In vitro Release from FilmsFrom figure 1a, film 1 and film 3 had the lowest and highest initial (burst) release of 13 and 65 respectively. Subsequently, film 1 and film 3 released 60 and 99 CD-NP by 30 days. Film 2 had an intermediate initial burst release of 31 and released 93 of CD-NP at 16574785 the end of 30 days. In figure 1b, the concentration of the CD-NP (following the burst release) from allCenderitide-Eluting Filmfilms were more or less similar from 1 to 30 days (in the range of 1?6 mg/mL).investigated. In figure 4b, the cGMP production levels after the addition of released CD-NP from all three films were elevated significantly compared to the control group (p,0.05).2. In vitro Degradation and Mass LossThe degradation of the films was determined by measuring the molecular mass (figure 2a) and total mass (figure 2b) changes. There was no significant molecular mass change and mass loss in all three tested films, indicating the slow degradation of PCL.5. Effects of CD-NP on Human Cardiac Fibroblast (HCF) Cell ViabilityIn figure 5, the graphs of cell index (CI) of HCF against time is presented, where CI increment denotes increase in cell proliferation or cell spreading. Figure 5a shows the cell viability of HCF after daily dose of 37 mg/mL CD-NP compared to control. It can be seen that in the first 48 hours, there was no distinct difference between the CD-NP group and control, however, a downward trend started to develop after the 3rd dose was administered. By the addition of the 4th dose (figure 5a), it was clear that daily dose of CD-NP at concentration of 37 mg/mL resulted in lower CI compared to control. Figure 5b, c, d shows the cell viability study of HCF of films 1, 2 and 3 respectively against the control group. Both film 1 and 3 showed immediate decline of CI compared to control, whilst, film 2 only saw decline in CI compared to control on the 4th day. The relative cell index (RCI) is used to describe the cell viability in a comparative manner, where lower the RCI value denotes greater extent of inhibition. Figure 6a b, c and d shows the correlation between the RCI (primary y-axis) and peptide concentration (secondary y-axis) with respect to time. From figure 6a, we can see that daily dosing of CD-NP results in “spikes” of CD-NP to 37 mg/mL daily (secondary y-axis), but the RCI was only less than 1 on the 2nd day onwards. Films 1 and 3 had RCI value less than 1 from 0 to 5 days. Film 2 however only saw RCI less than 1 after the 1st day. By the 5th day, all three films had RCI 23977191 less than 1.3. Surface MorphologyFigure 3a, b and c present the initial surface morphology of films 1, 2 and 3 respectively. Both film 1 and film 3 appear to be more porous compared to film 2, which may be due to the use of an immiscible co-solvent system. And by using a longer period of emulsification, film 3 appears to be mo.