1. The authors have no financial interests to disclose
Autologous Serum Eyedrops Diluted with Methylcellulose for the Treatment of Ocular Surface Diseases Unresponsive to Conventional Therapy
Felipe Fernandes Nicola (Presenting Author); Diane R. Marinho, MD; Samuel Rymer, MD, PhD; Sergio Kwitko, MD; Karla Linck Scheid, MD; Claudete Inês Locatelli.
The authors have no financial interests to disclose

2. Introduction
Since the first descriptions of autologous serum as treatment of dry eye at 1984 by Fox,RI et al 1, studies2,3 have been analyzing autologous serum eyedrops diluted mostly with saline solution as a vehicle.
It is known that saline solution is not comfortable to ocular surface use and can cause some degree of irritability, specially in patients with ocular surface diseases.
Recent studies 4,5 used sodium hialuronidase as a vehicle for autologous serum eyedrops analysis.
Methylcellulose is well tolerated and is widely used as vehicle to the eyedrops development.
1. Fox RI, Chan R, Michelson JB, Belmont JB, Michelson PE. Beneficial effect of artificial tears made with autologous serum in patients with keratoconjunctivitis sicca. Arthritis and Rheumatism 1984;27(2):459–61.
2. Autologe Serum-Augentropfen zur Therapie der Augenoberfläche. G. Geerling, D. Hartwig. Der Ophthalmologe.December 2002, Volume 99, Issue 12, pp
3. Proposal of standardized guidelines for the production and quality control of autologous serum eye drops in Korea: based on a nationwide survey. Hye Ryun Lee. 11 MAR 2014; DOI:  /trf.12558
4. Autologous serum eye drops diluted with sodium hyaluronate: clinical and experimental comparative study. López-García JS, Acta Ophthalmic Feb;92(1):e22-9. doi: /aos Epub 2013 Jul 26.
5. Comparison of autologous serum eye drops with different diluents. Cho YK,Curr Eye Res Jan;38(1):9-17. doi: / Epub 2012 Aug 28.

3. Purpose
To evaluate clinical effects and safety of 50% autologous serum eyedrops (AS) diluted with methylcellulose 0,5% in patients with severe dry eye resistant to conventional therapy.

4. Methods Study Design: prospective longitudinal intervention study.
Inclusion Criteria: patients with persistent epithelial defects or severe dry eye unresponsive to conventional therapy (use of lubrificant eyedrops, punctal oclusion or use of therapeutic contact lenses).
Exclusion Criteria: no adesion to the treatment; sistemic infection or puncture site infection; imminent risk of corneal perforation.
Intervantion: lubrificant treatment with AS diluted with methylcellulose 0,5% (1:1).

5. Methods Autologous Serum Preparation*
120ml peripheral venous blood was withdrawn from each patient, distributed in forty 4mL tubes and immediately centrifuged by rotations per minute during 10 minutes at room temperature.
In a laminar flow cabinet, 50mL of autologous serum were withdrawn from tubes and added to 50mL of methylcellulose 0,5%.
2mL were taken from the sterile tubes for microbiologic analisis
*AS was produced as recommended by the Infection Committee of Hospital de Clínicas de Porto Alegre.

6. Methods
All included patients were evaluated for subjective and objetive criteria before and after 6 months of AS use.
Subjetive Criteria: Sulisbury Eye Evaluation Questionnaire (five questions with high sensibility for dry eye6,7).
Objective Criteria: Biomicroscopy evaluation including break up time (BUT), fluorescein staining, rose bengal staining, Schirmer test and tear meniscus height.
Statistical Analysis: McNemar chi-square test or Mann- Whittney test using the IBM SPSS 21.
6. Schein OD. Relation between signs and symtoms of dry eye in the elderly. A population-based perspective.
7. Bandeen-Roche K. Self-reported assessment of dry eye in a population based-setting.

7. Results
A total of 44 eyes were analyzed before and after 6 months of AS use.
Subjective criteria showed improvement in symptoms in 26 patients (59%) (p <0.001).
BUT improved in 15 patients (34%) (p=0.002).
Fluorescein Staining improved in 25 eyes (57%) (p> 0.001).
Schirmer Test increased in 13 eyes (30%) (p=0.021).
Rose Bengal staining pattern improved in 22 eyes (50%) (p=0.004).
Tear Meniscus increased in 13 eyes (29%) (p=0.049).
No complications were reported.

8. Table 1. Autologous Serum Eyedrops Diluted with Methylcellulose for the Treatment of Ocular Surface Diseases Unresponsive to Conventional Therapy. RESULTS
Subjective Criteria
BUT
Fluorescein
Schirmer Test
Rose Bengal Staining
Tear Meniscus
Height
Worsened 2 11 8 4
Stable 18 27 17 30 14
Improved 26 15 25 13 22
p>0.001
p=0.002
p=0.021
p=0.004
p=0.049

10. Conclusion
AS is a safe and effective therapeutic option for ocular surface diseases.
Subjective improvement is not always related to objective improvement in eye examination.

11. Conclusion
Methylcellulose is an excellent vehicle for dilution and well tolerated by patients.
Further studies using traditional dilutions with saline are necessary for comparative results.