pNIPMAm MP characterization Sample Clauses

pNIPMAm MP characterization. (A) Phase image of MPs demonstrates the even polydispersity and spherical shape of particles. (B,C) Colloidal-phase mediated heteroaggregation was scaled up to produce uniformly coated MPs as indicated by SEM and confocal imaging (D) Xxxxxxx counter size analysis of MPs demonstrates a narrow size distribution with an average diameter of 4.6 ± 1.0 μm. (E) MPs demonstrated increased growth factor binding capacity with increasing initial growth factor loading concentrations. Average loading efficiency for BMP4 was 55.5 ± 6.2%. (F, G) Release of BMP4 from MPs was directly dependent on the initial loading amount of BMP4 and demonstrated increased release for 7 days with less than 60% of BMP4 was released at both loading amounts. Delivery of BMP4 via pNIPMAm MPs to skeletal myoblasts Bioactivity of BMP-4 loaded MPs was evaluated using an in vitro alkaline phosphatase (ALP) assay to quantify ALP activity of skeletal myoblasts (C2C12s), following treatment with soluble BMP4, unloaded MPs, unloaded MPs and soluble BMP4, or BMP4 loaded MPs. Treatment with BMP4 loaded MPs induced ALP activity similar to delivery of soluble BMP-4, suggesting that loaded MPs maintain BMP-4 bioactivity and ability to initiate a functional response, even with less than 25% release of BMP4 (Figure 3.3A). Overall, treatment of cells with BMP-4 (soluble or loaded) demonstrated increased ALP activity as compared to unloaded MP and untreated groups. Interestingly, simultaneous treatment of unloaded MPs and soluble BMP4 induced significantly less ALP activity in comparison to soluble BMP4 treatment, suggesting that MPs may sequester free BMP-4 and prevent subsequent interactions with cells. No differences in DNA content were observed between groups (Figure 3.3B) Figure 3.3. BMP4 bioactivity was evaluated by quantifying ALP activity of C2C12 cells after 3 days of treatment. BMP4 (soluble or loaded) increased ALP activity compared to unloaded MP and No MP. BMP4 MPs induced similar ALP activity to soluble BMP4, indicating comparable bioactivity. Unloaded MPs with soluble BMP4 induced less ALP activity compared to soluble BMP4 alone, suggesting that MPs actively sequester free BMP4 and prevent interactions with cells. ( n =3; p<0.05 compared to: * No MP; $ Unloaded MP) Incorporation of pNIPMAm MPs in EBs MPs were incorporated within ESC aggregates using forced aggregation via centrifugation within agarose microwells at 1:10, 1:3, 1:1, and 3:1 (MP:cell) seeding ratios. Rhodamine B-labeled MPs incor...
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