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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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Inhibitory effects of celastrol nanoparticles on suture-induced corneal neovascularization in rats. Animals were sutured (two stitches) in the temporal cornea, and then treated with subconjunctival injection of celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Slit-lamp microphotographs of corneas were taken after mydriasis on day 6 after surgery. The pictures show (A) an intensive corneal neovascularization in response to suturing and (B) a marked inhibition of neovascularization by celastrol nanoparticles. The images were taken by perfusing the cornea with India ink on day 6 after suturing. Note the distinct suppression of corneal neovascularization by celastrol nanoparticles. (C) Abundant corneal neovascularization reached the sutures. (D) Shorter and fewer areas of corneal neovascularization were found in the celastrol-treated group. (E and F) show the inhibition effect of celastrol nanoparticles on the length and area of corneal neovascularization, respectively.Notes: The length and area of corneal neovascularization were measured with Image-Pro® Plus 5.1 (Media Cybernetics Inc, Bethesda, MD) in triplicate for each cornea. The data are presented as mean ± standard deviation of five rats. *P < 0.001 versus the control group (magnification: ×25).Abbreviations: CI, confidence interval; CNPs, celastrol nanoparticles.
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f5-ijn-7-1163: Inhibitory effects of celastrol nanoparticles on suture-induced corneal neovascularization in rats. Animals were sutured (two stitches) in the temporal cornea, and then treated with subconjunctival injection of celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Slit-lamp microphotographs of corneas were taken after mydriasis on day 6 after surgery. The pictures show (A) an intensive corneal neovascularization in response to suturing and (B) a marked inhibition of neovascularization by celastrol nanoparticles. The images were taken by perfusing the cornea with India ink on day 6 after suturing. Note the distinct suppression of corneal neovascularization by celastrol nanoparticles. (C) Abundant corneal neovascularization reached the sutures. (D) Shorter and fewer areas of corneal neovascularization were found in the celastrol-treated group. (E and F) show the inhibition effect of celastrol nanoparticles on the length and area of corneal neovascularization, respectively.Notes: The length and area of corneal neovascularization were measured with Image-Pro® Plus 5.1 (Media Cybernetics Inc, Bethesda, MD) in triplicate for each cornea. The data are presented as mean ± standard deviation of five rats. *P < 0.001 versus the control group (magnification: ×25).Abbreviations: CI, confidence interval; CNPs, celastrol nanoparticles.

Mentions: To investigate the effect of CNPs on CNV in vivo, 0.1 mL of CNPs by subconjunctival injection was administered at a concentration of 5.44 mg/mL on day 0 and day 3 after suture surgery in a rat model. New blood vessel buds appeared and began to develop into the corneal suture on day 1 or 2 after surgery. Abundant CNV reached its peak on day 6 and began to regress after day 8 in the control group. Corneal edema was observed during neovascularization (Figure 5A). However, shorter and fewer new corneal vessels and substantially less edema were found in the celastrol-treated group (Figure 5B). Corneal angiogenesis produced a black color after ink perfusion on day 6 in both groups (Figure 5C and D). Administration of 0.544 mg of CNPs reduced the length and area of CNV. The average length and area were 1.16 ± 0.18 mm and 7.71 ± 0.94 mm2, respectively, in the control group on day 6 after surgery, whereas the average length and area in the celastrol group decreased to 0.49 ± 0.12 mm and 2.29 ± 0.61 mm2, respectively (both P values <0.001) (Figure 5E and F). No eye irritation, corneal epithelial defect, or other adverse complications related to the topical CNPs were observed in any of the tested animals.


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)

Inhibitory effects of celastrol nanoparticles on suture-induced corneal neovascularization in rats. Animals were sutured (two stitches) in the temporal cornea, and then treated with subconjunctival injection of celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Slit-lamp microphotographs of corneas were taken after mydriasis on day 6 after surgery. The pictures show (A) an intensive corneal neovascularization in response to suturing and (B) a marked inhibition of neovascularization by celastrol nanoparticles. The images were taken by perfusing the cornea with India ink on day 6 after suturing. Note the distinct suppression of corneal neovascularization by celastrol nanoparticles. (C) Abundant corneal neovascularization reached the sutures. (D) Shorter and fewer areas of corneal neovascularization were found in the celastrol-treated group. (E and F) show the inhibition effect of celastrol nanoparticles on the length and area of corneal neovascularization, respectively.Notes: The length and area of corneal neovascularization were measured with Image-Pro® Plus 5.1 (Media Cybernetics Inc, Bethesda, MD) in triplicate for each cornea. The data are presented as mean ± standard deviation of five rats. *P < 0.001 versus the control group (magnification: ×25).Abbreviations: CI, confidence interval; CNPs, celastrol nanoparticles.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3298384&req=5

f5-ijn-7-1163: Inhibitory effects of celastrol nanoparticles on suture-induced corneal neovascularization in rats. Animals were sutured (two stitches) in the temporal cornea, and then treated with subconjunctival injection of celastrol nanoparticles (0.544 mg per rat) on day 0 and day 3 after surgery. Slit-lamp microphotographs of corneas were taken after mydriasis on day 6 after surgery. The pictures show (A) an intensive corneal neovascularization in response to suturing and (B) a marked inhibition of neovascularization by celastrol nanoparticles. The images were taken by perfusing the cornea with India ink on day 6 after suturing. Note the distinct suppression of corneal neovascularization by celastrol nanoparticles. (C) Abundant corneal neovascularization reached the sutures. (D) Shorter and fewer areas of corneal neovascularization were found in the celastrol-treated group. (E and F) show the inhibition effect of celastrol nanoparticles on the length and area of corneal neovascularization, respectively.Notes: The length and area of corneal neovascularization were measured with Image-Pro® Plus 5.1 (Media Cybernetics Inc, Bethesda, MD) in triplicate for each cornea. The data are presented as mean ± standard deviation of five rats. *P < 0.001 versus the control group (magnification: ×25).Abbreviations: CI, confidence interval; CNPs, celastrol nanoparticles.
Mentions: To investigate the effect of CNPs on CNV in vivo, 0.1 mL of CNPs by subconjunctival injection was administered at a concentration of 5.44 mg/mL on day 0 and day 3 after suture surgery in a rat model. New blood vessel buds appeared and began to develop into the corneal suture on day 1 or 2 after surgery. Abundant CNV reached its peak on day 6 and began to regress after day 8 in the control group. Corneal edema was observed during neovascularization (Figure 5A). However, shorter and fewer new corneal vessels and substantially less edema were found in the celastrol-treated group (Figure 5B). Corneal angiogenesis produced a black color after ink perfusion on day 6 in both groups (Figure 5C and D). Administration of 0.544 mg of CNPs reduced the length and area of CNV. The average length and area were 1.16 ± 0.18 mm and 7.71 ± 0.94 mm2, respectively, in the control group on day 6 after surgery, whereas the average length and area in the celastrol group decreased to 0.49 ± 0.12 mm and 2.29 ± 0.61 mm2, respectively (both P values <0.001) (Figure 5E and F). No eye irritation, corneal epithelial defect, or other adverse complications related to the topical CNPs were observed in any of the tested animals.

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.

Show MeSH
Related in: MedlinePlus