LOSAT Medical Device Research NVP

Summary

• Service
• In vitro cell models
• Animal models
• Analyses on biological samples from clinical trials

• Rates

Service

Cellular cytotoxicity testing and analysis on biological specimens from clinical studies performed at the Laboratory of Ocular Surface Analysis and Translational Research, head Prof. Piera Versura and the Laboratory of Nephro-vascular Pathology, head Prof. Gianandrea Pasquinelli

Specifically, the laboratories involved intend to offer the test for in vitro cytotoxicity (basic test and integrated test with hypoxic stress, hyperosmolarity stress, oxidative stress and inflammatory stress). This test provides the general requirements for evaluating the cytotoxic potential of a medical device with the goal of assessing the compatibility of a device or material with a biological system.

They also make themselves available to perform analysis on biological samples as this type of activity may be commissioned as part of clinical trials, to evaluate biological markers in cells and tears sampled from subjects enrolled in the study. In particular, we talk about conjunctival cytology examinations, tear analysis (total protein amount and quantitative levels of lysozyme, lactotransferrin, lipocalin A, albumin) and ultrastructural analysis (transmission and scanning electron microscopy).

In vitro cell models

The following specific cell lines are in use:

• endothelial cell line (human vascular vessel) (HUVEC),

• human aortic smooth muscle cells (HAOSMC)

• glial cell line (human retina) (MIO-M1),

• human corneal epithelial cells (ATCC CRL-11516)

• human scleral fibroblasts – line in-house

• human conjunctival epithelial cells (ATCC CCL-20.2)

• retinal pigment epithelial cell line (human retina) (ARPE-19)

At request, for specific research purposes, co-culture models (direct and indirect contact) can be set up. Estimation of costs is available upon request.

In vitro cell models

• Cytotoxicity – direct and eluted contactISO 10993-5:2009 "Biological evaluation of medical devices Part 5: Tests for in vitro cytotoxicity" provides the general requirements for evaluating the cytotoxic potential of a medical device with the aim to evaluating how compatible a device or material is with a biological system.
• Hypoxic stress – performed by CoCl₂ adding or in a hypoxic chamber

• Hyperosmolarity stress – performed by NaCl adding to reach variable degree of mOsm/L medium

• Oxidative stress – performed by H₂O₂ adding

• Inflammatory stress – performed by TNF- and IL-1 adding

Qualitative * and quantitative ^§ methods of evaluation are available for each model applied, and are estimated upon the complexity of experiments:

• *morphological evaluation in light, fluorescent, and electron microscopy, immunocyto/histochemistry,

• *^§Cytofluorimetry for phenotype control

• ^§ MTT, MTS, and crystal violet assays,

• Molecular biology RT-qPCR for biomarkers identified on specific pathways

• ELISA assays on culture media

• Incucyte assays on wound closure, apoptotic signalling

Animal models

Dry Eye Disease – induced by a Controlled-Environment Chamber (CEC) that modulates specific conditions of airflow, humidity and temperature to produce an hyperevaporative stress

Analyses on biological samples from clinical trials

Lab for animal or human samples collection and detection.
The activity can be commissioned in the frame of Clinical trials, to evaluate biological markers in cells and tears sampled from subjects enrolled in the study.

In particular:

Conjunctival cytology

• goblet cells analysis (morphology and immunohistochemistry on mucins)
• cytofluorimetry on membrane target (HLA-DR)

Tear analysis

• major protein profile (Total protein amount, and quantitative levels of Lysozyme, Lactotransferrin,Lipocalin A, Albumin )
• inflammatory cytokines , to detect the panel IL-1, IL-6, IL-8 (CXC-L8), and TNF
• NGS on pelleted cells to evaluate targets of interest (miRNA, microbiome profiles)

Ultrastructural analysis

• Transmission electron microscopy
  
  

Rates

Rates are differentiated according to the type of test or analysis required and have been determined by taking into account the cost of consumables used (reagents), the cost of equipment maintenance and use, and the man time devoted to the activity.

To receive the updated fee schedule, email the managers.