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KERATINOCYTE GROWTH FACTOR AND CYTOKINES IN SKIN BURNS.
INTRODUCTION: Intense inflammatory responses are activated by burns that affect a large total body surface area. Changes in plasma levels of cytokines after burns occur before metabolic abnormalities unsettle the patient. So it may be possible to develop therapeutic interventions that may attenuate the acute inflammatory response by decreasing the expression of these cytokines. The importance of growth factors in the healing process was demonstrated in cultured keratinocytes and fibroblasts. The keratinocyte growth factor (KGF) is a growth factor active in the repair of wounds, being the most potent stimulator of mitotic cells.
PURPOSE: To assess the level of keratinocyte growth factor (KGF) and IL-1ß, IL-6, IL-8, IL-10, IL-12 and TNF-alfa of patients with burns produced by cultured primary dermal fibroblasts and the gene expression.
METHODS: 10 patients will be include (05 patients in the study group and 05 patients in the control group) admitted to the Burns Care Unit of the Discipline of Plastic Surgery, Federal University of São Paulo (UNIFESP) between 25% and 50% of total body surface area (TBSA), deep second-degree or third degree, with need to perform surgical debridement. The control group will be constituted by patients with less than 5% of TBSA, deep second-degree or third degree, with need to perform surgical debridement. The authors will evaluate the levels of IL-1ß, IL-6, IL-8, IL-10, IL-12p70 and tumor necrosis factor alpha (TNF-alfa) in samples of the culture media of primary dermal fibroblasts of patients selected using flow cytometry. The level of keratinocyte growth factor (KGF), in the same samples will be evaluated by ELISA. The keratinocyte growth factor (KGF) and TNF-alfa gene expression will be evaluated in the culture of primary dermal fibroblasts from the same patients. The gene expression of KGF and cytokines will be done by qRT-PCR and RT-PCR array. The experiments will be done in duplicate.
KERATINOCYTE GROWTH FACTOR AND CYTOKINES IN SKIN BURNS.
INTRODUCTION: Intense inflammatory responses are activated by burns that affect a large total body surface area. Changes in plasma levels of cytokines after burns occur before metabolic abnormalities unsettle the patient. So it may be possible to develop therapeutic interventions that may attenuate the acute inflammatory response by decreasing the expression of these cytokines. The importance of growth factors in the healing process was demonstrated in cultured keratinocytes and fibroblasts. The keratinocyte growth factor (KGF) is a growth factor active in the repair of wounds, being the most potent stimulator of mitotic cells.
PURPOSE: To assess the level of keratinocyte growth factor (KGF) and IL-1ß, IL-6, IL-8, IL-10, IL-12 and TNF-alfa of patients with burns produced by cultured primary dermal fibroblasts and the gene expression.
METHODS: 10 patients will be include (05 patients in the study group and 05 patients in the control group) admitted to the Burns Care Unit of the Discipline of Plastic Surgery, Federal University of São Paulo (UNIFESP) between 25% and 50% of total body surface area (TBSA), deep second-degree or third degree, with need to perform surgical debridement. The control group will be constituted by patients with less than 5% of TBSA, deep second-degree or third degree, with need to perform surgical debridement. The authors will evaluate the levels of IL-1ß, IL-6, IL-8, IL-10, IL-12p70 and tumor necrosis factor alpha (TNF-alfa) in samples of the culture media of primary dermal fibroblasts of patients selected using flow cytometry. The level of keratinocyte growth factor (KGF), in the same samples will be evaluated by ELISA. The keratinocyte growth factor (KGF) and TNF-alfa gene expression will be evaluated in the culture of primary dermal fibroblasts from the same patients. The gene expression of KGF and cytokines will be done by qRT-PCR and RT-PCR array. The experiments will be done in duplicate.
Key words: burns, cytokines, inflammation, keratinocyte growth factor, fibroblasts.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Minor Burns | Total burn surface area less than 5% of second and third degree. | ||
| Major Burns. | Total Burn Surface Area more than 25% of second and third degree, and less than 50%. |
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| Measure | Description | Time Frame |
|---|---|---|
| Keratinocyte Growth Factor | Level of KGF in cultured fibroblasts at second passage in media culture and qRT-PCR and RT-PCR array. | cultured cells at second passage |
| Measure | Description | Time Frame |
|---|---|---|
| Interleucin 1 beta, 6, 8, 10, 12 and tumor necrosis factor alfa | Level of cytokines in cultured fibroblasts at second passage in media culture and qRT-PCR and RT-PCR array. | cultured fibroblasts at second passage |
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Inclusion Criteria:
Exclusion Criteria:
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ACUTE BURNED PATIENTS
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| Name | Affiliation | Role |
|---|---|---|
| ALFREDO GRAGNANI FILHO, MD, PhD | UFSaoPaulo | Principal Investigator |
| LYDIA M FERREIRA, MD Full Prof | UFSaoPaulo | Study Chair |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Burn Center of Federal University of Sao Paulo | São Paulo | São Paulo | Brazil |
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| ID | Term |
|---|---|
| D002056 | Burns |
| C565529 | Growth Factors, Combined Defect of |
| D007249 | Inflammation |
| D000080424 | Cytokine Release Syndrome |
| ID | Term |
|---|---|
| D014947 | Wounds and Injuries |
| D010335 | Pathologic Processes |
| D013568 | Pathological Conditions, Signs and Symptoms |
| D018746 | Systemic Inflammatory Response Syndrome |
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Normal skin will be collected at first burned patient debridment and send to Cell Culture Laboratory to start fibroblast and queratinocyte culture.
| D012769 | Shock |