CERES Programme - Project 3-103
CERES Programme (National Plan for Research-Development and Innovation) Project 3-103 (2003-2005): In Vitro Models for Study of Molecular Mechanisms Involved in Cutaneous Wound Healing
Co-ordinator: University of Bucharest
Project director: prof. Dana Iordăchescu, PhD
Team members: prof. Otilia Zărnescu, PhD, lect. Anișoara Cîmpean, PhD; PhD students: Adela Bonoiu, Gabriela Bucată, Raluca Ciubăr, Laura Homeghiu, Cristina Iancu, Cristina Stan; Master student: Valentina Mitran
Partners: University of Medicine and Pharmacy Carol Davila, Bucharest, Department of Dermatology II (dr. Daniel Boda), National Institute for Research and Development in Physics and Nuclear Engineering Horia Hulubei (dr. Ioan Dorobanțu)
The project takes part in Subprogramme S1 «Fundamental and pre-competitive research» having as general objects: stimulation of high performance scientific activity in the field of the biomedical sciences, of consolidation of a research team previously constituted in the frame of a World Bank project, of increase of the Romanian science visibility and of promotion of the international collaborations.
The specific objectiv of the project:
«Advanced researches at the cellular and molecular level of a fundamental biologic process – the maintenance of the human tegument homeostasis».
The precise objectives of the project:
1. Realization of some experimental models for study of wound repair;
2. Comparative studies to choice an optimal model for research of the molecular mechanisms involved in skin lesion repair;
3. Simulation of some lesions on the chosen experimental model and study of the induced morphological and biochemical changes;
4. Estimation of the dynamics of cell – matrix interactions during the lesion repair.
Activities for fulfilment of the project objectives:
1. Obtaining of keratinocyte cultures from human epidermis, in monolayer and in the presence of some cell matrix components;
2. Obtaining of fibroblast cultures from human dermis, in monolayer and in the presence of some cell matrix components;
3. Obtaining of a keratinocyte-fibroblast co-culture;
4. Comparative studies on cell morphology, viability and proliferation;
5. Estimation of the reproduction grade of differentiation markers and of the keratinocyte growth stopping;
6. Study of the action of some compounds with hyperproliferative effect;
7. Realization of mechanical, hyperthermic and oxidant lesions, by gamma irradiation, exposure to irritant substances; estimation of morphologic changes induced by lesions, estimation of some pro-inflammatory cytokines, study of the post-lesional evolution of some matrix metalloproteinases;
8. Study of the efficiency of some compound’s action: insulin, hydrocortisone and vitamin A upon the lesion repair.
The material base:
Animal cell culture laboratory with sterile areas (laminar-flow hood, CO2 incubator, inverted microscope + photo-documentation system + epi-fluorescence system, refrigerated centrifuge and micro-centrifuge, balance, vortex), Laboratory for material processing (systems of humidified and dry sterilization, balances), Station for water purification (equipment for obtaining the ultrapure water Milli Q Synthesis), Laboratory for cell cryopreservation (deep freezer at –96oC, liquid nitrogen cryostorage containers) and freezing room, Laboratory of Biochemistry (UV/VIS spectrophotometers, spectrofluorimeter, refrigerated centrifuge, electrophoresis and western-blot systems, UV/VIS transiluminator, thermostated water baths, analytical balances, magnetic stirrers) and Multi-user Research Center of Molecular Biology (micro-plate reader TECAN, image computerized densitometer, spectrophotometer).
