Poly (lactide-co-glycolide)-polymethacrylate nanoparticles for intramuscular delivery of plasmid encoding interleukin-10 to prevent autoimmune diabetes in mice.
OBJECTIVE
Determine the efficiency of cationic nanoparticles prepared by blending poly (lactide-co-glycolide; PLGA) and methacrylate copolymer (Eudragit(R) E100) to deliver a therapeutic gene encoding mouse interleukin-10, in vitro and in vivo.


METHODS
Nanoparticles prepared with PLGA and E100 were evaluated for delivery of plasmid DNA encoding mouse interleukin-10 in vitro and in vivo in mice upon intramuscular injection. Blood-glucose, serum interferon-gamma levels and histology of pancreas were studied to determine therapeutic efficacy. Histological evaluation of skeletal muscle from the injection site was performed to assess the biocompatibility of nanoparticles.


RESULTS
PLGA/E100 nanoparticles showed endosomal escape evidenced by confocal microscopy and buffering ability. Transfecting HEK293 cells with plasmid-loaded PLGA/E100 nanoparticles resulted in significantly (p < 0.05) greater expression of interleukin-10 compared to PLGA nanoparticles. Mice treated with PLGA/E100 nanoparticles displayed higher serum levels of interleukin-10 and lower blood glucose levels compared to those treated with interleukin-10 plasmid alone or PLGA nanoparticles. High expression of interleukin-10 facilitated suppression of interferon-gamma levels and reduced islet infiltration. Histology of muscle showed that nanoparticles were biocompatible and did not cause chronic inflammatory response.


CONCLUSIONS
Nanoparticles prepared by blending PLGA with methacrylate can efficiently and safely deliver plasmid DNA encoding mouse interleukin-10 leading to prevention of autoimmune diabetes.
Basarkar
Ashwin
A
Department of Pharmaceutical Sciences, College of Pharmacy, Nursing, and Allied Sciences, North Dakota State University, Fargo, North Dakota 58105, USA.
Singh
Jagdish
J
D000818:Animals; D001327:Autoimmune Diseases / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control*; D001786:Blood Glucose / Q000378:metabolism; D002021:Buffers; D003920:Diabetes Mellitus / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control*; D016503:Drug Delivery Systems; D005079:Excipients / Q000737:chemistry*; D006801:Humans; D007273:Injections, Intramuscular; D050417:Insulin-Secreting Cells / Q000473:pathology; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics*; D019344:Lactic Acid / Q000737:chemistry*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D053758:Nanoparticles; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry*; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer
D001786:Blood Glucose; D002021:Buffers; D005079:Excipients; D016753:Interleukin-10; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D007371:Interferon-gamma
J Gene Med. 2002 Jan-Feb;4(1):84-91
11828391
Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10876-80
8855275
Int J Pharm. 2005 Sep 14;301(1-2):277-85
16019173
Drug Deliv. 2004 Mar-Apr;11(2):107-12
15200009
Mol Ther. 2001 May;3(5 Pt 1):658-64
11356070
Proc Natl Acad Sci U S A. 2000 Jan 18;97(2):811-6
10639162
Am J Ther. 2005 Nov-Dec;12(6):580-91
16280652
Diabetes. 1996 Jun;45(6):812-7
8635658
Immunol Res. 2002;25(3):229-45
12018462
Biochim Biophys Acta. 1999 Feb 16;1444(2):171-90
10023051
Mol Ther. 2001 Oct;4(4):313-6
11592833
Gene Ther. 2000 Dec;7(24):2099-104
11223991
Eur J Immunol. 1998 May;28(5):1719-26
9603479
J Control Release. 2005 May 18;104(2):359-77
15907586
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13844-9
9391115
Biomacromolecules. 2004 Mar-Apr;5(2):379-88
15002997
Mol Ther. 2006 Jul;14(1):88-96
16488666
Gene Ther. 1997 Aug;4(8):823-32
9338011
Int J Pharm. 2005 Jul 14;298(1):255-62
15941631
Gene Ther. 2003 Jan;10(2):131-42
12571642
Science. 1990 Mar 23;247(4949 Pt 1):1465-8
1690918
Nature. 2001 Dec 13;414(6865):792-8
11742411
Biochem Biophys Res Commun. 2002 Nov 22;299(1):29-34
12435385
Diabetes Res Clin Pract. 2004 Dec;66 Suppl 1:S27-32
15563975
Eur J Pharm Sci. 2003 Mar;18(3-4):211-20
12659932
Mol Ther. 2006 Oct;14(4):476-84
16829204
J Control Release. 2003 Mar 7;88(2):333-42
12628339
Mol Ther. 2005 Jun;11(6):990-5
15922971
Acta Pharmacol Sin. 2003 Aug;24(8):751-6
12904273