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Celastrol nanoparticles inhibit corneal neovascularization induced by suturing in rats.

Li Z, Yao L, Li J, Zhang W, Wu X, Liu Y, Lin M, Su W, Li Y, Liang D - Int J Nanomedicine (2012)

Bottom Line: In vivo, suture-induced CNV was chosen to evaluate the effect of CNPs on CNV in rats.After treatment with CNPs, the length and area of CNV reduced from 1.16 ± 0.18 mm to 0.49 ± 0.12 mm and from 7.71 ± 0.94 mm(2) to 2.29 ± 0.61 mm(2), respectively.Macrophage infiltration decreased significantly in the CNP-treated corneas.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, People's Republic of China.

ABSTRACT

Purpose: Celastrol, a traditional Chinese medicine, is widely used in anti-inflammation and anti-angiogenesis research. However, the poor water solubility of celastrol restricts its further application. This paper aims to study the effect of celastrol nanoparticles (CNPs) on corneal neovascularization (CNV) and determine the possible mechanism.

Methods: To improve the hydrophilicity of celastrol, celastrol-loaded poly(ethylene glycol)-block-poly(ɛ-caprolactone) nanopolymeric micelles were developed. The characterization of CNPs was measured by dynamic light scattering and transmission electron microscopy analysis. Celastrol loading content and release were assessed by ultraviolet-visible analysis and high performance liquid chromatography, respectively. In vitro, human umbilical vein endothelial cell proliferation and capillary-like tube formation were assayed. In vivo, suture-induced CNV was chosen to evaluate the effect of CNPs on CNV in rats. Immunohistochemistry for CD68 assessed the macrophage infiltration of the cornea on day 6 after surgery. Real-time quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay were used to evaluate the messenger ribonucleic acid and protein levels, respectively, of vascular endothelial growth factor, matrix metalloproteinase 9, and monocyte chemoattractant protein 1 in the cornea.

Results: The mean diameter of CNPs with spherical shape was 48 nm. The celastrol loading content was 7.36%. The release behavior of CNPs in buffered solution (pH 7.4) showed a typical two-phase release profile. CNPs inhibited the proliferation of human umbilical vein endothelial cells in a dose-independent manner and suppressed the capillary structure formation. After treatment with CNPs, the length and area of CNV reduced from 1.16 ± 0.18 mm to 0.49 ± 0.12 mm and from 7.71 ± 0.94 mm(2) to 2.29 ± 0.61 mm(2), respectively. Macrophage infiltration decreased significantly in the CNP-treated corneas. CNPs reduced the expression of vascular endothelial growth factor, matrix metalloproteinase 9, and monocyte chemoattractant protein 1 in the cornea on day 6 after suturing.

Conclusion: CNPs significantly inhibited suture-induced CNV by suppressing macrophage infiltration and the expression of vascular endothelial growth factor and matrix metalloproteinase 9 in the rat cornea.

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Celastrol nanoparticles inhibit neovascularization and macrophage infiltration in rat cornea. Rats were treated with celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Corneal tissue was fixed in 10% neutralized buffered formaldehyde, and 5 μm paraffin sections were stained with hematoxylin and eosin. (A) Celastrol nanoparticle-treated group showed significantly inhibited neovascularization and inflammatory cell infiltration in cornea on day 6 after surgery when compared with (B) the control group. Red arrows show new blood vessels. Corneal sections were processed by anti-rat CD68 staining in (C) celastrol nanoparticle-treated group and (D) the control group. Celastrol nanoparticles suppressed macrophage infiltration in the cornea on day 6 after surgery. The number of infiltrated macrophage (CD68+) cells in the corneal stroma was counted in five randomly selected fields (×400) of the immunofluorescence-stained slides.Notes: The number of infiltrating macrophages was analyzed by IBM SPSS (v 16.0; SPSS Inc, Chicago, IL). The data are presented as mean ± standard deviation (magnification: ×100). Statistical significance is based on the difference when compared with the control; *P < 0.001.Abbreviation: CI, confidence interval.
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f6-ijn-7-1163: Celastrol nanoparticles inhibit neovascularization and macrophage infiltration in rat cornea. Rats were treated with celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Corneal tissue was fixed in 10% neutralized buffered formaldehyde, and 5 μm paraffin sections were stained with hematoxylin and eosin. (A) Celastrol nanoparticle-treated group showed significantly inhibited neovascularization and inflammatory cell infiltration in cornea on day 6 after surgery when compared with (B) the control group. Red arrows show new blood vessels. Corneal sections were processed by anti-rat CD68 staining in (C) celastrol nanoparticle-treated group and (D) the control group. Celastrol nanoparticles suppressed macrophage infiltration in the cornea on day 6 after surgery. The number of infiltrated macrophage (CD68+) cells in the corneal stroma was counted in five randomly selected fields (×400) of the immunofluorescence-stained slides.Notes: The number of infiltrating macrophages was analyzed by IBM SPSS (v 16.0; SPSS Inc, Chicago, IL). The data are presented as mean ± standard deviation (magnification: ×100). Statistical significance is based on the difference when compared with the control; *P < 0.001.Abbreviation: CI, confidence interval.

Mentions: Histologic analysis of the cornea revealed that CNP treatment led to a significant reduction in the number of infiltrating inflammatory cells in the treated group when compared with the control group. New blood vessels were inhibited and corneal edema was also distinctly mitigated by celastrol ( Figure 6A and B). The number of infiltrating macrophages was significantly reduced in the cornea on day 6 after surgery in the celastrol group compared to the control group (P < 0.001; Figure 6C–E). Histopathological evaluation was performed three times for six sections of each group.


Celastrol nanoparticles inhibit corneal neovascularization induced by suturing in rats.

Li Z, Yao L, Li J, Zhang W, Wu X, Liu Y, Lin M, Su W, Li Y, Liang D - Int J Nanomedicine (2012)

Celastrol nanoparticles inhibit neovascularization and macrophage infiltration in rat cornea. Rats were treated with celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Corneal tissue was fixed in 10% neutralized buffered formaldehyde, and 5 μm paraffin sections were stained with hematoxylin and eosin. (A) Celastrol nanoparticle-treated group showed significantly inhibited neovascularization and inflammatory cell infiltration in cornea on day 6 after surgery when compared with (B) the control group. Red arrows show new blood vessels. Corneal sections were processed by anti-rat CD68 staining in (C) celastrol nanoparticle-treated group and (D) the control group. Celastrol nanoparticles suppressed macrophage infiltration in the cornea on day 6 after surgery. The number of infiltrated macrophage (CD68+) cells in the corneal stroma was counted in five randomly selected fields (×400) of the immunofluorescence-stained slides.Notes: The number of infiltrating macrophages was analyzed by IBM SPSS (v 16.0; SPSS Inc, Chicago, IL). The data are presented as mean ± standard deviation (magnification: ×100). Statistical significance is based on the difference when compared with the control; *P < 0.001.Abbreviation: CI, confidence interval.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3298384&req=5

f6-ijn-7-1163: Celastrol nanoparticles inhibit neovascularization and macrophage infiltration in rat cornea. Rats were treated with celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Corneal tissue was fixed in 10% neutralized buffered formaldehyde, and 5 μm paraffin sections were stained with hematoxylin and eosin. (A) Celastrol nanoparticle-treated group showed significantly inhibited neovascularization and inflammatory cell infiltration in cornea on day 6 after surgery when compared with (B) the control group. Red arrows show new blood vessels. Corneal sections were processed by anti-rat CD68 staining in (C) celastrol nanoparticle-treated group and (D) the control group. Celastrol nanoparticles suppressed macrophage infiltration in the cornea on day 6 after surgery. The number of infiltrated macrophage (CD68+) cells in the corneal stroma was counted in five randomly selected fields (×400) of the immunofluorescence-stained slides.Notes: The number of infiltrating macrophages was analyzed by IBM SPSS (v 16.0; SPSS Inc, Chicago, IL). The data are presented as mean ± standard deviation (magnification: ×100). Statistical significance is based on the difference when compared with the control; *P < 0.001.Abbreviation: CI, confidence interval.
Mentions: Histologic analysis of the cornea revealed that CNP treatment led to a significant reduction in the number of infiltrating inflammatory cells in the treated group when compared with the control group. New blood vessels were inhibited and corneal edema was also distinctly mitigated by celastrol ( Figure 6A and B). The number of infiltrating macrophages was significantly reduced in the cornea on day 6 after surgery in the celastrol group compared to the control group (P < 0.001; Figure 6C–E). Histopathological evaluation was performed three times for six sections of each group.

Bottom Line: In vivo, suture-induced CNV was chosen to evaluate the effect of CNPs on CNV in rats.After treatment with CNPs, the length and area of CNV reduced from 1.16 ± 0.18 mm to 0.49 ± 0.12 mm and from 7.71 ± 0.94 mm(2) to 2.29 ± 0.61 mm(2), respectively.Macrophage infiltration decreased significantly in the CNP-treated corneas.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, People's Republic of China.

ABSTRACT

Purpose: Celastrol, a traditional Chinese medicine, is widely used in anti-inflammation and anti-angiogenesis research. However, the poor water solubility of celastrol restricts its further application. This paper aims to study the effect of celastrol nanoparticles (CNPs) on corneal neovascularization (CNV) and determine the possible mechanism.

Methods: To improve the hydrophilicity of celastrol, celastrol-loaded poly(ethylene glycol)-block-poly(ɛ-caprolactone) nanopolymeric micelles were developed. The characterization of CNPs was measured by dynamic light scattering and transmission electron microscopy analysis. Celastrol loading content and release were assessed by ultraviolet-visible analysis and high performance liquid chromatography, respectively. In vitro, human umbilical vein endothelial cell proliferation and capillary-like tube formation were assayed. In vivo, suture-induced CNV was chosen to evaluate the effect of CNPs on CNV in rats. Immunohistochemistry for CD68 assessed the macrophage infiltration of the cornea on day 6 after surgery. Real-time quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay were used to evaluate the messenger ribonucleic acid and protein levels, respectively, of vascular endothelial growth factor, matrix metalloproteinase 9, and monocyte chemoattractant protein 1 in the cornea.

Results: The mean diameter of CNPs with spherical shape was 48 nm. The celastrol loading content was 7.36%. The release behavior of CNPs in buffered solution (pH 7.4) showed a typical two-phase release profile. CNPs inhibited the proliferation of human umbilical vein endothelial cells in a dose-independent manner and suppressed the capillary structure formation. After treatment with CNPs, the length and area of CNV reduced from 1.16 ± 0.18 mm to 0.49 ± 0.12 mm and from 7.71 ± 0.94 mm(2) to 2.29 ± 0.61 mm(2), respectively. Macrophage infiltration decreased significantly in the CNP-treated corneas. CNPs reduced the expression of vascular endothelial growth factor, matrix metalloproteinase 9, and monocyte chemoattractant protein 1 in the cornea on day 6 after suturing.

Conclusion: CNPs significantly inhibited suture-induced CNV by suppressing macrophage infiltration and the expression of vascular endothelial growth factor and matrix metalloproteinase 9 in the rat cornea.

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Related in: MedlinePlus