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. 2011;4(6):582-9.
doi: 10.3980/j.issn.2222-3959.2011.06.02. Epub 2011 Dec 18.

Study of self-made freeze-dried bilayered fibrin-binding amniotic membrane in ocular trabeculectomy in rabbits

Wan Li  1 , Wen-Jian Che, Ming-Chang Zhang
Affiliations

Study of self-made freeze-dried bilayered fibrin-binding amniotic membrane in ocular trabeculectomy in rabbits

Wan Li et al. Int J Ophthalmol. 2011.

Abstract

Aim: To investigate the antifibrotic effect of freeze-dried bilayered fibrin-binding amniotic membrane on trabeculectomy in a rabbit model.

Methods: Twenty-four Japanese white rabbits were randomized into three groups: the experimental group (ocular trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation), the control group (ocular trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane) and the blank group (single trabeculectomy). Clinical observation, hematoxylin-eosin staining, Massion staining, real-time PCR and immunohistochemistry for α-SMA were performed on days 7, 14, 21 and 30 following surgery.

Results: Statistical differences were noted in survival analysis and intraocular pressure (IOP) among groups on days 7, 14, 21 and 30 following surgery. Histology, immunohistochemistry and real-time PCR further demonstrated that trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane resulted in good wound healing and no scar formation.

Conclusion: Self-made freeze-dried bilayered fibrin-binding amniotic membrane may inhibit the formation of scarring in glaucoma after trabeculectomy.

Keywords: amniotic membrane; fibrin-binding; freeze-dried; trabeculectomy.

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Figures

Figure 1
Figure 1. Freeze-dried bilayered fibrin-binding amniotic membrane
Figure 2
Figure 2. Survival analysis in three groups
Blue: trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; Yellow: trabeculectomy and natural bilayered fibrin-binding amniotic membrane; Grey: single trabeculectomy
Figure 3
Figure 3. IOP measurement in three groups (pairwise comparison using the LSD method)
Red: trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; Blue: trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane; Green: single trabeculectomy
Figure 4
Figure 4. HE staining on day 14 post-surgery
A and B: HE staining on pathological slices of surgical corneosclera on day 14 following trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; C and D: HE staining on pathological slices of surgical corneosclera on day 14 following trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane transplantation; E and F: HE staining on pathological slices of surgical corneosclera on day 14 following single trabeculectomy (significant filtration blebs and passageway in images A, B, C and D with the amniotic membrane present; E and F depict scar formation in the filtration passageway).
Figure 5
Figure 5. Massion staining on day 14 post-surgery
A and B: Massion trichrome staining on pathological slices of surgical corneosclera on day 14 following trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; C and D: Massion trichrome staining on pathological slices of surgical corneosclera on day 14 following trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane transplantation; E and F: Massion trichrome staining on pathological slices of surgical corneosclera on day 14 following single trabeculectomy (a small quantity of collagen is seen in filtration blebs and passageways in integrated amniotic membrane in images A, B, C and D; abundant collagen is observed in the filtration passageway in images E and F)
Figure 6
Figure 6. Massion staining on day 30 post-surgery
A and B: Massion trichrome staining on pathological slices of surgical corneosclera on day 30 following trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; C and D: Massion trichrome staining on pathological slices of surgical corneosclera on day 30 following trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane transplantation; E and F: Massion trichrome staining on pathological slices of surgical corneosclera on day 30 following single trabeculectomy (the amniotic membrane and significant filtration blebs and passageways in images A and B, lack of an amniotic membrane and significant filtration blebs and passageways in images C and D and abundant collagen deposited in the subconjunctival filtration passageway in images E and F)
Figure 7
Figure 7. Immunohistochemical staining : expression of α-SMA on day 14 post-surgery
A: Weak α-SMA expression after trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; B: Limited α-SMA expression after trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane; C: Abundant α-SMA expression after single trabeculectomy
Figure 8
Figure 8. mRNA expression of CTGF in the ocular surgical area of rabbits in three groups on day 14 post-surgery
Red: trabeculectomy in combination with freeze-dried bilayered fibrin-binding amniotic membrane transplantation; Blue: trabeculectomy in combination with natural bilayered fibrin-binding amniotic membrane transplantation; Green: single trabeculectomy
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