Liposome Formulation and In Vitro Testing in Non-Physiological Conditions Addressed to Ex Vivo Kidney Perfusion.
Silvia PisaniEnrica ChiesaIda GentaRossella DoratiMarilena GregoriniMaria Antonietta GrignanoMarina RamusGabriele CeccarelliStefania CroceChiara ValsecchiManuela MontiTeresa RampinoBice ContiPublished in: International journal of molecular sciences (2022)
This work focuses on formulating liposomes to be used in isolated kidney dynamic machine perfusion in hypothermic conditions as drug delivery systems to improve preservation of transplantable organs. The need mainly arises from use of kidneys from marginal donors for transplantation that are more exposed to ischemic/reperfusion injury compared to those from standard donors. Two liposome preparation techniques, thin film hydration and microfluidic techniques, are explored for formulating liposomes loaded with two model proteins, myoglobin and bovine serum albumin. The protein-loaded liposomes are characterized for their size by DLS and morphology by TEM. Protein releases from the liposomes are tested in PERF-GEN perfusion fluid, 4 °C, and compared to the in vitro protein release in PBS, 37 °C. Fluorescent liposome uptake is analyzed by fluorescent microscope in vitro on epithelial tubular renal cell cultures and ex vivo on isolated pig kidney in hypothermic perfusion conditions. The results show that microfluidics are a superior technique for obtaining reproducible spherical liposomes with suitable size below 200 nm. Protein encapsulation efficiency is affected by its molecular weight and isoelectric point. Lowering incubation temperature slows down the proteins release; the perfusion fluid significantly affects the release of proteins sensitive to ionic media (such as BSA). Liposomes are taken up by epithelial tubular renal cells in two hours' incubation time.
Keyphrases
- drug delivery
- drug release
- contrast enhanced
- cancer therapy
- protein protein
- quantum dots
- single cell
- induced apoptosis
- binding protein
- stem cells
- magnetic resonance imaging
- cell therapy
- heart failure
- computed tomography
- acute myocardial infarction
- magnetic resonance
- photodynamic therapy
- mass spectrometry
- signaling pathway
- high glucose
- cell proliferation
- cerebral ischemia
- label free
- mesenchymal stem cells
- acute coronary syndrome
- circulating tumor cells
- high resolution
- bone marrow
- oxidative stress
- cell cycle arrest
- left ventricular
- molecularly imprinted
- liquid chromatography
- wound healing
- percutaneous coronary intervention
- pi k akt