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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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Liposomal targeting of PLP to the inflamed synovial lining strongly suppresses joint inflammation during AIA.A: Knee joint swelling as measured by 99MTc-uptake is strongly suppressed after a single injection of Lip-PLP. B: Photomicrographs of frontal knee joint sections of mice with AIA at day 5 after treatment and naïve mice. Note that the inflammatory infiltrate is reduced in mice treated with Lip-PLP. Original magnification ×100, Asterisks points to synovial infiltrate, hash sign points to inflammatory exudates. C: Histological scoring of synovial infiltration at day 1 and day 5 after systemic treatment with Lip-PLP or saline. D: Silver staining of frontal knee joint sections of mice with AIA, treated by intravenous injection with gold-containing liposomes. Note that the silver staining of the gold particles is mostly observed within the synovial lining cells (arrows). Mice were treated at day 3 after induction of AIA. Values are the mean of 8 mice per group. Original magnification ×100; insert ×400. F = femur, JS = joint space. Statistical significance was determined by Student's t-test. * = P<0.05 compared to saline treatment.
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pone-0054016-g002: Liposomal targeting of PLP to the inflamed synovial lining strongly suppresses joint inflammation during AIA.A: Knee joint swelling as measured by 99MTc-uptake is strongly suppressed after a single injection of Lip-PLP. B: Photomicrographs of frontal knee joint sections of mice with AIA at day 5 after treatment and naïve mice. Note that the inflammatory infiltrate is reduced in mice treated with Lip-PLP. Original magnification ×100, Asterisks points to synovial infiltrate, hash sign points to inflammatory exudates. C: Histological scoring of synovial infiltration at day 1 and day 5 after systemic treatment with Lip-PLP or saline. D: Silver staining of frontal knee joint sections of mice with AIA, treated by intravenous injection with gold-containing liposomes. Note that the silver staining of the gold particles is mostly observed within the synovial lining cells (arrows). Mice were treated at day 3 after induction of AIA. Values are the mean of 8 mice per group. Original magnification ×100; insert ×400. F = femur, JS = joint space. Statistical significance was determined by Student's t-test. * = P<0.05 compared to saline treatment.

Mentions: Mice with established antigen-induced arthritis (AIA) expressing a strong M1 signature at day 3, were treated with a single intravenous injection of Lip-PLP (10 mg/kg) and showed a strong suppression of joint swelling as measured by 99MTc-uptake by 74% within 1 day when compared to saline controls and by 61% when compared to free PLP treatment (Fig. 2A). At day 5 after treatment, Lip-PLP had almost completely suppressed joint swelling and histological examination of frontal knee joint sections showed a mean suppression of the synovial infiltrate of 29% at day 1 and of 80% at day 5 after treatment (Fig. 2B+C).


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)

Liposomal targeting of PLP to the inflamed synovial lining strongly suppresses joint inflammation during AIA.A: Knee joint swelling as measured by 99MTc-uptake is strongly suppressed after a single injection of Lip-PLP. B: Photomicrographs of frontal knee joint sections of mice with AIA at day 5 after treatment and naïve mice. Note that the inflammatory infiltrate is reduced in mice treated with Lip-PLP. Original magnification ×100, Asterisks points to synovial infiltrate, hash sign points to inflammatory exudates. C: Histological scoring of synovial infiltration at day 1 and day 5 after systemic treatment with Lip-PLP or saline. D: Silver staining of frontal knee joint sections of mice with AIA, treated by intravenous injection with gold-containing liposomes. Note that the silver staining of the gold particles is mostly observed within the synovial lining cells (arrows). Mice were treated at day 3 after induction of AIA. Values are the mean of 8 mice per group. Original magnification ×100; insert ×400. F = femur, JS = joint space. Statistical significance was determined by Student's t-test. * = P<0.05 compared to saline treatment.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0054016-g002: Liposomal targeting of PLP to the inflamed synovial lining strongly suppresses joint inflammation during AIA.A: Knee joint swelling as measured by 99MTc-uptake is strongly suppressed after a single injection of Lip-PLP. B: Photomicrographs of frontal knee joint sections of mice with AIA at day 5 after treatment and naïve mice. Note that the inflammatory infiltrate is reduced in mice treated with Lip-PLP. Original magnification ×100, Asterisks points to synovial infiltrate, hash sign points to inflammatory exudates. C: Histological scoring of synovial infiltration at day 1 and day 5 after systemic treatment with Lip-PLP or saline. D: Silver staining of frontal knee joint sections of mice with AIA, treated by intravenous injection with gold-containing liposomes. Note that the silver staining of the gold particles is mostly observed within the synovial lining cells (arrows). Mice were treated at day 3 after induction of AIA. Values are the mean of 8 mice per group. Original magnification ×100; insert ×400. F = femur, JS = joint space. Statistical significance was determined by Student's t-test. * = P<0.05 compared to saline treatment.
Mentions: Mice with established antigen-induced arthritis (AIA) expressing a strong M1 signature at day 3, were treated with a single intravenous injection of Lip-PLP (10 mg/kg) and showed a strong suppression of joint swelling as measured by 99MTc-uptake by 74% within 1 day when compared to saline controls and by 61% when compared to free PLP treatment (Fig. 2A). At day 5 after treatment, Lip-PLP had almost completely suppressed joint swelling and histological examination of frontal knee joint sections showed a mean suppression of the synovial infiltrate of 29% at day 1 and of 80% at day 5 after treatment (Fig. 2B+C).

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