Aqueous Vascular Endothelial Growth Factor (VEGF) Levels in Type 3 Neovascularization

Age-related Macular Degeneration Clinical Trial

Official title:

Aqueous Humor Levels of Vascular Endothelial Growth Factor Before and After Intravitreal Bevacizumab in Type 3 Versus Type 1 and 2 Neovascularization. A Prospective, Case-control Study.

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT01350323
• Other study ID #: UM-O-1
• Status: Completed
• Phase: N/A
• First received: May 2, 2011
• Last updated: May 6, 2011
• Start date: May 2009
• Est. completion date: September 2010

Study information

• Verified date: March 2009
• Source: University of Molise
• Contact: n/a
• Is FDA regulated: No
• Health authority: Italy: Ethics Committee
• Study type: Interventional

Clinical Trial Summary

This is a pilot, prospective, interventional, case-control study investigating aqueous levels of vascular endothelial growth factor (VEGF) in eyes with AMD-related neovascularization treated with intravitreal bevacizumab at the Medical Retina Department, University of Molise, Campobasso.

Description:

This was a pilot, prospective, interventional, case-control study investigating aqueous levels of VEGF in eyes with AMD-related NV treated with intravitreal bevacizumab.

Classification of neovascularization was based on its anatomic localization as ascertained by multimodal imaging employing FA (fluorescein angiography), ICGA (indocyanine green angiography) and OCT (optical coherence tomography) as recently proposed by Freund et al.

Diagnostic procedures and follow-up At baseline all the patients underwent BCVA measurement using early treatment diabetic retinopathy study (ETDRS) chart at 4 meters, fundus biomicroscopy, FA, ICGA and sd-OCT. All examinations were repeated in occasion of the follow-up appointments with exception of angiographic tests.

Angiographic tests and OCTs scans were recorded using Spectralis sd-OCT (Spectralis SD-OCT, Heidelberg Engineering, Heidelberg, Germany).

CMT was calculated after acquiring a sequence of 145 horizontal sections recorded in the high-resolution mode (1024 A-scans/30 degrees) and covering an area of 20 degrees (horizontal) × 15 degrees (vertical) with a distance of ~30 µm between individual sections. On follow-up examinations, the imaging processing software allowed reevaluation at exactly the same location.

The images were then processed by the "Thickness Map" analysis program. Field 1 of the map analysis protocol (central 1 mm) was used for central retinal thickness calculations.

Aqueous Sampling and Bevacizumab Injections All patients with AMD-related neovascularization received three intravitreal injections of 1.25 mg/0.05 ml of bevacizumab (Avastin; Genentech Inc, South San Francisco, California, USA) at baseline, month 1 and month 2.

Before injection, topical anesthesia was induced by tetracaine (1%) eye drops. Povidone-iodine was applied to the eyelid margins, the lashes, the conjunctiva bulbi and the fornices. After application of a sterile drape, a lid speculum was inserted.

Immediately before each intravitreal injection, aqueous sampling was performed by aspirating 0.05-0.1 ml of aqueous using a 30-gauge needle connected to a tuberculin syringe at the temporal limbus. All the samples were collected in the operating theatre under operating microscope control.

Intravitreal bevacizumab (IVB) injection was then performed using a 30-gauge needle in the inferotemporal quadrant at 3.5 mm to 4 mm posterior to the limbus.

The undiluted aqueous samples were transferred into sterile containers and immediately stored in a -80°C freezer until analysis.

Control Group Reference samples were obtained from 14 age-matched patients undergoing cataract surgery.

Aqueous humor samples were obtained in the same fashion described above for eyes with wet-AMD. The undiluted aqueous samples were transferred into sterile containers and immediately stored in a -80°C freezer until analysis.

Vascular Endothelial Growth Factor Assay Collected samples were gradually equilibrated to room temperature before beginning the assay and diluted 1:1 with the sample diluent provided by the manufacturer.

The VEGF content was determined on 50 µl of diluted sample with a human VEGF ELISA kit (EHVEGF, Pierce Biotechnology, Rockford, Illinois, USA) according to the manufacturer's instruction and using an extended standard curve including a 16, 8 and 4 pg/ml standard point. All assays were performed in duplicate. The minimum detectable concentration of VEGF was 3.5 pg/ml. Values inferior to 3.5pg/ml were considered equal to 1 for statistical analysis.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 33
• Est. completion date: September 2010
• Est. primary completion date: September 2010
• Accepts healthy volunteers: No
• Gender: Both
• Age group: 55 Years to 95 Years

Eligibility

Inclusion Criteria:

• wet age-related macular degeneration type 1, 2 or 3

Exclusion Criteria:

• BCVA at baseline less than 1.0 logMAR

• any previous treatment of the neovascular lesion

• previous vitrectomy

• laser coagulation within the last 3 months

• previous participation in any studies using investigational drugs within 3months preceding day 0

• intraocular surgery (including cataract surgery) in the study eye within 3 months preceding day 0

• glaucoma in the study eye; diabetes mellitus

• use of immunosuppressive drugs

• malignant tumors of any location

Study Design

Allocation: Non-Randomized, Intervention Model: Parallel Assignment, Masking: Open Label

Related Conditions & MeSH terms

• Age-related Macular Degeneration
• Macular Degeneration
• Neovascularization, Pathologic

Intervention

Procedure:
• Aqueous sample
Aqueous sample (0.1ml of aqueous)+ intravitreal injection of bevacizumab (1.25mg/0.05ml)
• Aqueous sample
Aqueous sample (0.1ml)+intravitreal injection of bevacizumab (1.25mg/0.05ml)
• Aqueous sample
Aqueous sample (0.1ml)+ intravitreal injection of bevacizumab (1.25mg/0.05ml)
• Aqueous sample
Aqueous sample (0.1ml)

Locations

Country	Name	City	State
Italy	University of Molise	Campobasso

Sponsors (1)

Lead Sponsor	Collaborator
University of Molise

Country where clinical trial is conducted

Italy, 

References & Publications (3)

Bottoni F, Massacesi A, Cigada M, Viola F, Musicco I, Staurenghi G. Treatment of retinal angiomatous proliferation in age-related macular degeneration: a series of 104 cases of retinal angiomatous proliferation. Arch Ophthalmol. 2005 Dec;123(12):1644-50. — View Citation

Wells JA, Murthy R, Chibber R, Nunn A, Molinatti PA, Kohner EM, Gregor ZJ. Levels of vascular endothelial growth factor are elevated in the vitreous of patients with subretinal neovascularisation. Br J Ophthalmol. 1996 Apr;80(4):363-6. — View Citation

Yannuzzi LA, Negrão S, Iida T, Carvalho C, Rodriguez-Coleman H, Slakter J, Freund KB, Sorenson J, Orlock D, Borodoker N. Retinal angiomatous proliferation in age-related macular degeneration. Retina. 2001;21(5):416-34. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Aqueous VEGF concentration	To establish if different types of wet-AMD were associated to different concentrations of VEGF at baseline	Change from baseline of VEGF concentration at month 1	No
Secondary	Macular thickness assessment	to compare central macular thickness before and after treatment respectively.	Change from baseline of macular thickness at month 1	No