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Liposomal targeting of prednisolone phosphate to synovial lining macrophages during experimental arthritis inhibits M1 activation but does not favor M2 differentiation.

Hofkens W, Schelbergen R, Storm G, van den Berg WB, van Lent PL - PLoS ONE (2013)

Bottom Line: In vitro studies showed that Lip-PLP phagocytosed by M1 macrophages resulted in a suppression of the M1 phenotype and induction of M2 markers (IL-10, TGF-β, IL-1RII, CD163, CD206 and Ym1).This study clearly shows that systemic treatment with PLP-liposomes selectively targets synovial lining macrophages and inhibits M1 activation.In contrast to in vitro findings, PLP-liposomes do not cause a shift of synovial lining macrophages towards M2.

View Article: PubMed Central - PubMed

Affiliation: Rheumatology Research and Advanced Therapeutics, Department of Rheumatology, Radboud University Medical Centre, Nijmegen, The Netherlands.

ABSTRACT

Background: To determine the effects of liposomal targeting of prednisolone phosphate (Lip-PLP) to synovial lining macrophages on M1 and M2 polarization in vitro and during experimental arthritis.

Material and methods: Experimental arthritis (antigen and immune complex induced) was elicited in mice and prednisolone containing liposomes were given systemically. Synovium was investigated using microarray analysis, RT-PCR and histology. Bone-marrow macrophages were stimulated towards M1 using LPS and IFNγ before treatment by PLP-liposomes. M1 and M2 markers were determined using RT-PCR.

Results: Microarray analysis of biopsies of inflamed synovium during antigen induced arthritis (AIA) showed an increased M1 signature characterized by upregulation of IL-1β, IL-6 and FcγRI starting from day 1 and lasting up until day 7 after arthritis induction. The M2 signature remained low throughout the 7 day course of arthritis. Treatment of AIA with intravenously delivered Lip-PLP strongly suppressed joint swelling and synovial infiltration whereas colloidal gold containing liposomes exclusively targeted the macrophages within the inflamed synovial intima layer. In vitro studies showed that Lip-PLP phagocytosed by M1 macrophages resulted in a suppression of the M1 phenotype and induction of M2 markers (IL-10, TGF-β, IL-1RII, CD163, CD206 and Ym1). In vivo, Lip-PLP treatment strongly suppressed M1 markers (TNF-α, IL-1β, IL-6, IL-12p40, iNOS, FcγRI, Ciita and CD86) after local M1 activation of lining macrophages with LPS and IFN-γ and during experimental AIA and immune complex arthritis (ICA). In contrast, M2 markers were not significantly upregulated in antigen-induced arthritis and down regulated in immune complex arthritis.

Conclusion: This study clearly shows that systemic treatment with PLP-liposomes selectively targets synovial lining macrophages and inhibits M1 activation. In contrast to in vitro findings, PLP-liposomes do not cause a shift of synovial lining macrophages towards M2.

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Effect of Lip-PLP on M1 and M2 marker expression within the synovium during ICA.A: Photomicrographs of frontal knee joint sections of mice with ICA at day 1 after treatment. B: Histological scoring of synovial infiltration mice with ICA at day 1 after treatment with saline or Lip-PLP. C+D: Expression of M1 (C) and M2 (D) markers in the synovium during ICA. RE = Relative Expression compared to values of GAPDH. Mice were treated at day 1 after induction of ICA and biopsies were obtained at day 1 after treatment. Data are expressed as mean +/− SD of eight animals. UD = undetectable. Statistical significance was determined by Student's t-test. * = P<0.05, ** = P<0.01 compared to saline treatment.
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pone-0054016-g006: Effect of Lip-PLP on M1 and M2 marker expression within the synovium during ICA.A: Photomicrographs of frontal knee joint sections of mice with ICA at day 1 after treatment. B: Histological scoring of synovial infiltration mice with ICA at day 1 after treatment with saline or Lip-PLP. C+D: Expression of M1 (C) and M2 (D) markers in the synovium during ICA. RE = Relative Expression compared to values of GAPDH. Mice were treated at day 1 after induction of ICA and biopsies were obtained at day 1 after treatment. Data are expressed as mean +/− SD of eight animals. UD = undetectable. Statistical significance was determined by Student's t-test. * = P<0.05, ** = P<0.01 compared to saline treatment.

Mentions: Similar to the AIA, systemic delivery of Lip-PLP inhibited synovial infiltration at day 1 after injection (Fig. 6A) and significantly suppressed M1 factors TNF-α (30-fold), IL-1β (230-fold), IL-6 (116-fold), IL-12p40 (not detected anymore), FcγRI (32-fold), Ciita (18-fold) and CD86 (7-fold) (Fig. 6B). Treatment with Lip-PLP even suppressed M2 factors and only CD163 expression was somewhat upregulated by Lip-PLP (4-fold), suggesting that Lip-PLP inhibits joint inflammation in ICA largely through suppression of M1 macrophages.


Liposomal targeting of prednisolone phosphate to synovial lining macrophages during experimental arthritis inhibits M1 activation but does not favor M2 differentiation.

Hofkens W, Schelbergen R, Storm G, van den Berg WB, van Lent PL - PLoS ONE (2013)

Effect of Lip-PLP on M1 and M2 marker expression within the synovium during ICA.A: Photomicrographs of frontal knee joint sections of mice with ICA at day 1 after treatment. B: Histological scoring of synovial infiltration mice with ICA at day 1 after treatment with saline or Lip-PLP. C+D: Expression of M1 (C) and M2 (D) markers in the synovium during ICA. RE = Relative Expression compared to values of GAPDH. Mice were treated at day 1 after induction of ICA and biopsies were obtained at day 1 after treatment. Data are expressed as mean +/− SD of eight animals. UD = undetectable. Statistical significance was determined by Student's t-test. * = P<0.05, ** = P<0.01 compared to saline treatment.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585322&req=5

pone-0054016-g006: Effect of Lip-PLP on M1 and M2 marker expression within the synovium during ICA.A: Photomicrographs of frontal knee joint sections of mice with ICA at day 1 after treatment. B: Histological scoring of synovial infiltration mice with ICA at day 1 after treatment with saline or Lip-PLP. C+D: Expression of M1 (C) and M2 (D) markers in the synovium during ICA. RE = Relative Expression compared to values of GAPDH. Mice were treated at day 1 after induction of ICA and biopsies were obtained at day 1 after treatment. Data are expressed as mean +/− SD of eight animals. UD = undetectable. Statistical significance was determined by Student's t-test. * = P<0.05, ** = P<0.01 compared to saline treatment.
Mentions: Similar to the AIA, systemic delivery of Lip-PLP inhibited synovial infiltration at day 1 after injection (Fig. 6A) and significantly suppressed M1 factors TNF-α (30-fold), IL-1β (230-fold), IL-6 (116-fold), IL-12p40 (not detected anymore), FcγRI (32-fold), Ciita (18-fold) and CD86 (7-fold) (Fig. 6B). Treatment with Lip-PLP even suppressed M2 factors and only CD163 expression was somewhat upregulated by Lip-PLP (4-fold), suggesting that Lip-PLP inhibits joint inflammation in ICA largely through suppression of M1 macrophages.

Bottom Line: In vitro studies showed that Lip-PLP phagocytosed by M1 macrophages resulted in a suppression of the M1 phenotype and induction of M2 markers (IL-10, TGF-β, IL-1RII, CD163, CD206 and Ym1).This study clearly shows that systemic treatment with PLP-liposomes selectively targets synovial lining macrophages and inhibits M1 activation.In contrast to in vitro findings, PLP-liposomes do not cause a shift of synovial lining macrophages towards M2.

View Article: PubMed Central - PubMed

Affiliation: Rheumatology Research and Advanced Therapeutics, Department of Rheumatology, Radboud University Medical Centre, Nijmegen, The Netherlands.

ABSTRACT

Background: To determine the effects of liposomal targeting of prednisolone phosphate (Lip-PLP) to synovial lining macrophages on M1 and M2 polarization in vitro and during experimental arthritis.

Material and methods: Experimental arthritis (antigen and immune complex induced) was elicited in mice and prednisolone containing liposomes were given systemically. Synovium was investigated using microarray analysis, RT-PCR and histology. Bone-marrow macrophages were stimulated towards M1 using LPS and IFNγ before treatment by PLP-liposomes. M1 and M2 markers were determined using RT-PCR.

Results: Microarray analysis of biopsies of inflamed synovium during antigen induced arthritis (AIA) showed an increased M1 signature characterized by upregulation of IL-1β, IL-6 and FcγRI starting from day 1 and lasting up until day 7 after arthritis induction. The M2 signature remained low throughout the 7 day course of arthritis. Treatment of AIA with intravenously delivered Lip-PLP strongly suppressed joint swelling and synovial infiltration whereas colloidal gold containing liposomes exclusively targeted the macrophages within the inflamed synovial intima layer. In vitro studies showed that Lip-PLP phagocytosed by M1 macrophages resulted in a suppression of the M1 phenotype and induction of M2 markers (IL-10, TGF-β, IL-1RII, CD163, CD206 and Ym1). In vivo, Lip-PLP treatment strongly suppressed M1 markers (TNF-α, IL-1β, IL-6, IL-12p40, iNOS, FcγRI, Ciita and CD86) after local M1 activation of lining macrophages with LPS and IFN-γ and during experimental AIA and immune complex arthritis (ICA). In contrast, M2 markers were not significantly upregulated in antigen-induced arthritis and down regulated in immune complex arthritis.

Conclusion: This study clearly shows that systemic treatment with PLP-liposomes selectively targets synovial lining macrophages and inhibits M1 activation. In contrast to in vitro findings, PLP-liposomes do not cause a shift of synovial lining macrophages towards M2.

Show MeSH
Related in: MedlinePlus