Plastic Surgery Research Council
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CHRYSIN (5,7-DIHYDROXYFLAVONE) ACCELERATES DIFFERENTIATION OF PRIMARY HUMAN KERATINOCYTES IN VITRO
Presenter: Taylor J Jaraczewski
Co-Authors: Pathak PR; Roy M; King TW
Univ of Wisconsin School of Medicine & Public Health

Introduction: Wound healing affects millions of people and the impact on the US economy is over $25B annually. We are interested in discovering novel strategies to enhance keratinocyte migration, proliferation, and differentiation in order to improve wound healing. Chrysin (5,7-dihydroxyflavone), a natural flavonoid found in the blue passion flower, has recently been shown to protect keratinocytes from UV-induced damage. We investigated the effect of chrysin on cell differentiation in primary human keratinocytes.

Methods: Primary human keratinocytes were isolated from neonatal foreskin. The basal (60 µM calcium) EpiLifeĆ medium (Invitrogen, Carlsbad, CA) supplemented with human keratinocyte growth supplements (Invitrogen) was used for experiments. Chrysin was purchased from MP Biomedicals (Santa Ana, CA). Effetcs of chrysin on keratinocyte differentiation was investigated the presence of low (60 µM) and high calcium (1 mM). Light microscopy was utilized to observe cell phenotype. Western blots were performed for differentiation markers keratin 1 and keratin 10.

Results: We observed an increased number of enlarged and flat cells in the presence of chrysin compared to controls under basal (low calcium) conditions. The cells also expressed differentiation markers keratin 1 and keratin 10, while these markers were not detected in the absence of chrysin. Under differentiating conditions, (high calcium), chrysin notably accelerated cell differentiation and stratification compared to controls, which also resulted in higher levels of keratin 1 & keratin 10.

Conclusions: Calcium is known to promote terminal keratinocyte differentiation in vitro. The addition of chrysin stimulated terminal differentiation and stratification to an even greater extent. This observation was further supported by expression of higher levels of keratin 1 and keratin 10 in the presence of chrysin. Therefore, we conclude that chrysin is a potent stimulator of keratinocyte differentiation. Chrysin could potentially be utilized to accentuate differentiation and accelerate the wound healing process in patients.


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