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. 2012;5(5):555-61.
doi: 10.3980/j.issn.2222-3959.2012.05.03. Epub 2012 Oct 18.

Homemade lyophilized cross linking amniotic sustained-release drug membrane with anti-scarring role after filtering surgery in rabbit eyes

Wan Li  1 , Wen-Jian Chen, Wei Liu, Liang Liang, Ming-Chang Zhang
Affiliations

Homemade lyophilized cross linking amniotic sustained-release drug membrane with anti-scarring role after filtering surgery in rabbit eyes

Wan Li et al. Int J Ophthalmol. 2012.

Abstract

Aim: To investigate the antifibrotic effect of the freeze-dried bilayered fibrin-binding amniotic membrane as a drug delivery system on glaucoma surgery in rabbit model. The aim of this study was to prepare a novel local delivery system for the sustained and controllable release of 5-Fu.

Methods: Twenty-four Japanese white rabbits were randomized into three groups: the experimental group (ocular trabeculectomy in combination with 5-Fu loaded freeze-dried bilayered fibrin-binding amniotic membrane transplantation), the control group (ocular trabeculectomy in combination with 5-Fu) and the blank group (single trabeculectomy). HE staining, massion staining and immunohistochemistry for α-SMA were performed on days 7, 14, 21 and 30 following surgery. The concentration of 5-Fu in rabbit aqueous humor was examined by high performance liquid chromatography (HPLC) 3 days after the surgery.

Results: Statistical differences were noted in intraocular pressure among groups on day 7, 14, 21 and 30 following surgery. Histology further demonstrated that trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane yielded well wound healing and no scar formation and was beneficial for long term effect.

Conclusion: HPLC showed a good slow-release effect with freeze-dried bilayered fibrin-binding amniotic membrane.

Keywords: drug delivery system; filtering surgery; freeze-dried bilayered fibrin-binding amniotic membrane; glaucoma; inhibition of scarring formation.

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Figures

Figure 1
Figure 1. Staining of rabbit corneoscleral region 14 days
A, B: after trabeculectomy and implantation of lyophilized crosslinking amniotic sustained-release drug membrane under the conjunctival flap; C, D: after trabeculectomy and implantation of 5-Fu cotton under the conjunctival flap; E, F: after trabeculectomy. (The existence of amniotic membranes and the good function of filtration tracks were shown in A, B; the good function of filtration tracks was shown in C, D; the existence of fibroblasts in filtration tracks was show in E, F).
Figure 2
Figure 2. HE staining of rabbit corneoscleral region 30 days
A, B: after trabeculectomy and implantation of lyophilized crosslinking amniotic sustained-release drug membrane under the conjunctival flap; C, D: after trabeculectomy and implantation of 5-Fu cotton under the conjunctival flap; E, F: after trabeculectomy. (The existence of amniotic membranes and the good function of filtration tracks were shown in A, B; most of the filtration tracks were blocked in C, D; the filtration tracks were blocked completely, and large amount of fibroblasts in E, F).
Figure 3
Figure 3. Massion staining of rabbit corneoscleral region 14 days
A, B: after trabeculectomy and implantation of lyophilized crosslinking amniotic sustained-release drug membrane under the conjunctival flap; C, D: after trabeculectomy and implantation of 5-Fu cotton under the conjunctival flap; E, F: after trabeculectomy. (The existence of filtration tracks and a small amount of collagen in A-D; the existence of amniotic membranes in A, B; a great amount of intensive and parallel collagen in filtration tracks and conjunctiva in E, F)
Figure 4
Figure 4. Massion staining of rabbit corneoscleral region 30 days
A, B: after trabeculectomy and implantation of lyophilized crosslinking amniotic sustained-release drug membrane under the conjunctival flap; C, D: after trabeculectomy and implantation of 5-Fu cotton under the conjunctival flap; E, F: after trabeculectomy. (The existence of amniotic membranes, filtration bleb and fistulas in A, B; a great amount of collagen in filtration fistulas and a small amount of in C, D; a great amount of collagen in conjunctiva and filtration fistulas in E, F)
Figure 5
Figure 5. Immunohistochemical staining of α-actin protein: expression of α-SMA 14 days after operation.
A: without obvious expression. Experimental group, trabeculectomy surgery with lyophilized crosslinking amniotic release membrane under conjunctival flap; B: small amount of expression. Control group, trabeculectomy surgery with 5-Fu cotton under conjunctival flap; C: highly expressed ofα-SMA. Blank group, trabeculectomy.
Figure 6
Figure 6. Experimental group, control group and blank group was statistically significant using 2-way ANOVA. The experimental group and control group was statistically significant using the LSD method. The control group and blank group were statistically significant.
Figure 7
Figure 7. Standard chromatograms by the detection of high performance liquid chromatography (HPLC).
Figure 8
Figure 8. HPLC chromatograms of rabbit aqueous humor 5-Fu 3 days post operation
A: in the experimental group; B: in the control group. Absorbance: A> B. 5-Fu release was higher in group A than group B after three days post operation.
See this image and copyright information in PMC

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