PMID- 37686348
OWN - NLM
STAT- MEDLINE
DCOM- 20230911
LR  - 20230911
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 24
IP  - 17
DP  - 2023 Aug 31
TI  - Investigation and Comparison of Active and Passive Encapsulation Methods for 
      Loading Proteins into Liposomes.
LID - 10.3390/ijms241713542 [doi]
LID - 13542
AB  - In this work, four different active encapsulation methods, microfluidic (MF), 
      sonication (SC), freeze-thawing (FT), and electroporation (EP), were investigated 
      to load a model protein (bovine serum albumin-BSA) into neutral liposomes made 
      from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC):cholesterol (Chol) and 
      charged liposomes made from DSPC:Chol:Dioleoyl-3-trimethylammonium propane 
      (DOTAP), DSPC:Chol:1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), and 
      DSPC:Chol:phosphatidylethanolamine (PE). The aim was to increase the protein 
      encapsulation efficiency (EE%) by keeping the liposome size below 200 nm and the 
      PDI value below 0.7, which warrants a nearly monodisperse preparation. 
      Electroporation (100 V) yielded the best results in terms of EE%, with a dramatic 
      increase in liposome size (>600 nm). The FT active-loading method, either applied 
      to neutral or charged liposomes, allowed for obtaining suitable EE%, keeping the 
      liposome size range below 200 nm with a suitable PDI index. Cationic liposomes 
      (DSPC:Chol:DOTAP) loaded with the FT active method showed the best results in 
      terms of EE% (7.2 +/- 0.8%) and size (131.2 +/- 11.4 nm, 0.140 PDI). In vitro release 
      of BSA from AM neutral and charged liposomes resulted slower compared to PM 
      liposomes and was affected by incubation temperature (37  degrees C, 4  degrees C). The empty 
      charged liposomes tested for cell viability on Human Normal Dermal Fibroblast 
      (HNDF) confirmed their cytocompatibility also at high concentrations (10(10) 
      particles/mL) and cellular uptake at 4  degrees C and 37  degrees C. It can be concluded that 
      even if both microfluidic passive and active methods are more easily transferable 
      to an industrial scale, the FT active-loading method turned out to be the best in 
      terms of BSA encapsulation efficiencies, keeping liposome size below 200 nm.
FAU - Pisani, Silvia
AU  - Pisani S
AUID- ORCID: 0000-0002-5396-1601
AD  - Department of Drug Sciences, University of Pavia, Viale T. Taramelli 12, 27100 
      Pavia, Italy.
FAU - Di Martino, Deborah
AU  - Di Martino D
AUID- ORCID: 0009-0007-8445-8535
AD  - Unit of Cellular and Molecular Neurobiology, IRCCS Mondino Foundation, Via 
      Mondino 2, 27100 Pavia, Italy.
FAU - Cerri, Silvia
AU  - Cerri S
AUID- ORCID: 0000-0002-6466-5381
AD  - Unit of Cellular and Molecular Neurobiology, IRCCS Mondino Foundation, Via 
      Mondino 2, 27100 Pavia, Italy.
FAU - Genta, Ida
AU  - Genta I
AUID- ORCID: 0000-0001-5710-0588
AD  - Department of Drug Sciences, University of Pavia, Viale T. Taramelli 12, 27100 
      Pavia, Italy.
FAU - Dorati, Rossella
AU  - Dorati R
AUID- ORCID: 0000-0001-5774-9547
AD  - Department of Drug Sciences, University of Pavia, Viale T. Taramelli 12, 27100 
      Pavia, Italy.
FAU - Bertino, Giulia
AU  - Bertino G
AD  - Otorhinolaryngology Unit, Department of Surgical Sciences, Fondazione IRCCS 
      Policlinico San Matteo, 27100 Pavia, Italy.
FAU - Benazzo, Marco
AU  - Benazzo M
AD  - Otorhinolaryngology Unit, Department of Surgical Sciences, Fondazione IRCCS 
      Policlinico San Matteo, 27100 Pavia, Italy.
FAU - Conti, Bice
AU  - Conti B
AUID- ORCID: 0000-0002-0034-2815
AD  - Department of Drug Sciences, University of Pavia, Viale T. Taramelli 12, 27100 
      Pavia, Italy.
LA  - eng
GR  - grant #08053922/Italian Ministry of Health, RC-2023/
GR  - . PE00000007 INF-ACT/NextGeneration EU-MUR PNRR Extended Partnership initiative 
      on Emerging Infectious Diseases/
PT  - Journal Article
DEP - 20230831
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - MR86K0XRQP (1,2-dioleoyloxy-3-(trimethylammonium)propane)
RN  - 0 (Liposomes)
RN  - 27432CM55Q (Serum Albumin, Bovine)
SB  - IM
MH  - Humans
MH  - *Liposomes
MH  - *Serum Albumin, Bovine
MH  - Electroporation
MH  - Electroporation Therapies
PMC - PMC10487800
OTO - NOTNLM
OT  - bovine serum albumin
OT  - electroporation
OT  - freeze-thawing
OT  - liposomes
OT  - microfluidic
COIS- The authors declare no conflict of interest.
EDAT- 2023/09/09 11:44
MHDA- 2023/09/11 06:42
PMCR- 2023/08/31
CRDT- 2023/09/09 01:13
PHST- 2023/07/26 00:00 [received]
PHST- 2023/08/26 00:00 [revised]
PHST- 2023/08/29 00:00 [accepted]
PHST- 2023/09/11 06:42 [medline]
PHST- 2023/09/09 11:44 [pubmed]
PHST- 2023/09/09 01:13 [entrez]
PHST- 2023/08/31 00:00 [pmc-release]
AID - ijms241713542 [pii]
AID - ijms-24-13542 [pii]
AID - 10.3390/ijms241713542 [doi]
PST - epublish
SO  - Int J Mol Sci. 2023 Aug 31;24(17):13542. doi: 10.3390/ijms241713542.