Material and Methods
Eighteen patients with VKC were included in this study: (i) during
active inflammation (group A), (ii) during quiescent phase (group Q) and
after recovery of VKC (group R). Ten healthy subjects were included as
control group. The study was performed at the Department of Sense Organs
and
Department of Pediatrics at Sapienza University of Rome, following the
tenants of the Declaration of Helsinky. Informed consent was signed by
all participants and caregiver, and the study was approved by the Ethic
Committee of University Sapienza of Rome.
Diagnosis of VKC was based on clinical history and clinical
presentation.4 Patients with VKC were included in
different groups: (i) Active group: presence of persistent ocular
symptoms associated with moderate to severe conjunctival hyperemia and
secretion and presence of corneal involvement; (ii) quiescent group:
absence or mild and intermittent ocular symptoms, absence of
conjunctival hyperemia and of corneal involvement; (iii) recovery group:
history of VKC with absence of symptoms and signs of inflammation from
at least 1 year.4, 20 All patients did not use
antiallergic topical and systemic treatments from at least 7 days.
At baseline visit, clinical history including skin prick test results
was collected and the presence of ocular signs and symptoms of VKC was
investigated. Specifically, the presence and severity of conjunctival
hyperemia and secretion, limbal and upper tarsal papillary reaction was
evaluated by slit lamp examination and scored from 0 to 3 (0=absence, 1=
mild, 2=moderate and 3=severe).3, 4 Corneal epithelial
damage was evaluated by fluorescein vital staining and scored by Oxford
scale.21 Patients were asked for the presence of
symptoms of such as itching, tearing and photophobia. Symptoms were
scored from 0 to 3 (0=absence, 1= mild, 2=moderate and 3=severe).
Conjunctival impression cytology and tear samples were collected in both
eyes of VKC and healthy subjects.22, 23 Patients
included in group A which received treatment with Hydrocortisone
(Cortivis eye drop, Medivis S.r.l., Catania, Italy) eye drops 4 times
daily for 7 days. At the end of treatment period patients were evaluated
as described above and tear and conjunctival samples were collected.
Tear samples were collected by imbibition of sterile Sharp-tip
Microsponges (Alcon Lab. Inc., Fort Worth, TX) gently placed at the
inferior conjunctival fornix and removed after 30 seconds. Tears were
collected by centrifugation at 13000 rpm for 3 minutes and stored at
-20°C. 22 Conjunctival impression cytology was
performed after tear collection in both eyes by applying a sterile cell
culture insert (Millicell-CM, 0.4 μm, Ø 12 mm, Millipore) to the nasal
and temporal bulbar conjunctiva, and then stored at -20°C.24
Western blot analysis was performed on biological samples to evaluate
tear sCD14 and conjunctival epithelium CD14, TLR-4 and 9.