Catalyst-Free Plasma-Assisted Copolymerization of Poly(ε-caprolactone)-poly(ethylene glycol) for Biomedical Applications

Catalyst-free ring-opening polymerization (ROP) strategy was developed to overcome the disadvantage of incomplete and expensive removal of catalyst used during the multistep wet chemical processes. Nano-sized biocompatible and low molecular weight poly­(ε-carolactone)-poly­(ethylene glycol) (PCL-PEG) copolymer coatings were deposited via a single-step, low-pressure, pulsed-plasma polymerization process. Experiments were performed at different monomer feed ratio and effective plasma power. The coatings were analyzed by XPS, as well as MALDI ToF. Ellipsometric measurement showed deposition rates ranging from 1.3 to 3 nm/min, depending on the ratio of the PCL/PEG precursors introduced in the reactor. Our results have demonstrated that plasma copolymerized PCL-PEG coatings can be tailored in such a way to be cell adherent, convenient for biomedical implants such as artificial skin substrates, or cell repellent, which can be used as antibiofouling surfaces for urethral catheters, cardiac stents, and so on. The global objective of this study is to tailor the surface properties of PCL by copolymerizing it with PEG in the pulsed plasma environment to improve their applicability in tissue engineering and biomedical science

MCs increase OCC metastasis biological function: adherence, migration and invasion.

<p><b>A.</b> OCC-eGFP (NIH:OVCAR3, left chart and SKOV3, right chart) were seeded on a Matrigel (BD Biosciences)-coated well for 10 min, 15 min, 30 min and 1 hour. Increased adherence to the ECM is observed when OCC were preemptively cultivated with MC (light grey bars) compared to the control (dark grey bars). <b>B.</b> Scheme representing the invasion and migration assay. <b>C.</b> Sorted OCC were seeded on (un) coated transwells and GFP signal of each well under the coated membrane was acquired after 24 h. Increased migration and invasion through the ECM is observed when OCC were preemptively cultivated with MC (light grey bars) compared to the control (dark grey bars). SEM are represented, n = 3, | p<0.05 t-Student Test.</p

MC protects OCC from chemotherapy induced cell death.

<p><b>A.</b> Explanative scheme of FACS analysis. <b>B</b>. OCC or OCC co-cultivated with MSC were treated for 48 hours with 90 µM of Cisplatin and 6 nm of Paclitaxel. Cell types were discriminated by FACS using GFP as a cancer cell marker, and CD73 as an MC marker. Cell death analysis was carried out using the dual Calcein/Live Dead staining. <b>C</b>. Quantitation of cell death analysis experiments. OCC display a resistance to conventional chemotherapy treatment when co-cultivated with MC compared to the control. SEM are represented, n = 3, | p<0.05 t-Student Test.</p

Specificity of the test: Absence of interference by fibrin degradation products.

<p>(*) 210 ng/mL is the detection limit of the STA Liatest.</p><p>(**) the levels of D-dimer in untreated plasmas and in t-PA treated plasmas being beneath the detection limit of D-dimer (i.e., 210 ng/mL), we conclude only that the SF concentration in this group was ≤200 ng/mL.</p><p>Fibrin degradation products (D-dimer) were added to normal plasma at the indicated final concentrations. SF was then determined.</p><p>For each sample, D-dimer measurement was performed in duplicate (but this is not strictly necessary).</p

MC contact enriches biological function gene clusters.

<p>Table showing the biological clusters identified by IPA analysis. p-values for the different clusters are: “Metastasis” (NIH:OVCAR3, p = 6.48*10⁻⁶; SKOV3, p = 5.42*10⁻⁵), “Proliferation of cell lines” (NIH:OVCAR3, p = 4.36.48*10⁻⁷; SKOV3, p = 6.79*10⁻⁶) and “Cell death of tumor cell line” (NIH:OVCAR3, p = 5.68*10⁻⁸; SKOV3, p = 7.62*10⁻⁵). These clusters were shared between NIH:OVCAR3-eGFP and SKOV3-eGFP.</p

MC sustains OCC proliferation in a serum free cytokine free context. A.

<p>OCC were cultivated on a MC expressing mOrange or direclty on the plastic dish. Proliferation assay was carried out in a serum free-cytokine free media. Pictures were taken every two days, and GFP cells were quantified. <b>B.</b> Detail of the well on day 14, we can notice OCC display a normal morphology. <b>C.</b> Chart displaying cell count carried over 14 days. OVCAR3 eGFP when seeded on a MC are able to sustain cell cycle in a serum free cytokine free media. Similar results were obtained with SKOV3 (data not shown). SEM are represented, n = 3, | p<0.05 t-Student Test.</p

Evolution of SF and D-dimer levels in patients under anticoagulant therapy.

<p>SF and D-dimer (D-Di) concentrations were determined daily in patients before and after starting anticoagulant therapy.</p

Evaluation of the area under the ROC (Receiving Operating Characteristic) curve for SF and D-dimer derived from patients with suspected Pulmonary Embolism or Deep Vein Thrombosis.

<p>Evaluation of the area under the ROC (Receiving Operating Characteristic) curve for SF and D-dimer derived from patients with suspected Pulmonary Embolism or Deep Vein Thrombosis.</p

Sensitivity, Specificity, Positive- (PPV) and Negative-Predictive Values (NPV) (95% Confidence Intervals) for Soluble Fibrin (SF) and D-dimer concentrations in Pulmonary Embolism and Deep Vein Thrombosis.

<p>Sensitivity, Specificity, Positive- (PPV) and Negative-Predictive Values (NPV) (95% Confidence Intervals) for Soluble Fibrin (SF) and D-dimer concentrations in Pulmonary Embolism and Deep Vein Thrombosis.</p

Plasma Soluble Fibrin (SF) and D-dimer levels in Patients with suspected Pulmonary Embolism (PE) or Deep Vein Thrombosis (DVT).

<p>SF threshold value  =  300 ng/mL; D-dimer threshold value  =  500 ng/mL.</p