Protective Effect of Tropical Highland Blackberry Juice (Rubus adenotrichos Schltdl.) Against UVB‐Mediated Damage in Human Epidermal Keratinocytes and in a Reconstituted Skin Equivalent Model

Abstract

Solar ultraviolet (UV) radiation, particularly its UVB (280–320 nm) spectrum, is the primary environmental stimulus leading to skin carcinogenesis. Several botanical species with antioxidant properties have shown photochemopreventive effects against UVB damage. Costa Rica’s tropical highland blackberry (Rubus adenotrichos) contains important levels of phenolic compounds, mainly ellagitannins and anthocyanins, with strong antioxidant properties. In this study, we examined the photochemopreventive effect of R. adenotrichos blackberry juice (BBJ) on UVB-mediated responses in human epidermal keratinocytes and in a three-dimensional (3D) reconstituted normal human skin equivalent (SE). Pretreatment (2 h) and posttreatment (24 h) of normal human epidermal keratinocytes (NHEKs) with BBJ reduced UVB (25 mJ cm−2)-mediated (1) cyclobutane pyrimidine dimers (CPDs) and (2) 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) formation. Furthermore, treatment of NHEKs with BBJ increased UVB-mediated (1) poly(ADP-ribose) polymerase cleavage and (2) activation of caspases 3, 8 and 9. Thus, BBJ seems to alleviate UVB-induced effects by reducing DNA damage and increasing apoptosis of damaged cells. To establish the in vivo significance of these findings to human skin, immunohistochemistry studies were performed in a 3D SE model, where BBJ was also found to decrease CPDs formation. These data suggest that BBJ may be developed as an agent to ameliorate UV-induced skin damage.

Prodifferentiation, Anti-Inflammatory and Antiproliferative Effects of Delphinidin, a Dietary Anthocyanidin, in a Full-Thickness Three-Dimensional Reconstituted Human Skin Model of Psoriasis

“…Primary normal human epidermal keratinocytes established in low calcium Epi-Life growth medium supplemented with HKGS (Invitrogen), were seeded at 20% confluence and synchronically switched and maintained in progenitor cell-targeted cell culture media (CnT-02–07 CELLnTEC, ZenBio, Research Triangle Park, N.C., USA) prior to establishment of 3D cultures. The dermal and epidermal components were prepared as previously described [17]. The full-thickness human PSE model (SOR-300-FT) consisted of human psoriatic keratinocytes and collagen-contracted fibroblasts cultured to form a multilayered, highly differentiated epidermis (MatTek Corporation, Ashland, Mass., USA).…”

Rhus coriaria L. Fruit Extract Prevents UV-A-Induced Genotoxicity and Oxidative Injury in Human Microvascular Endothelial Cells

“…Polyphenol-rich extracts from Rubus spp. and grapevine (Vitis vinifera L.) exhibit pro-apoptotic effect after UVR exposure in vitro and in vivo, as reported in the literature [66][67][68]. More studies are needed to clarify the mechanism of apoptosis' induction from the extract pre-treatment along with UV-A induced damage.…”

Cultivo Celular e Ingeniería de Tejidos: Aplicaciones en Biomedicina

“…En el LAINTEC también se cuenta con un pequeño banco de líneas celulares de distintos tipos de cáncer y algunas líneas de tejidos normales, y se han establecido modelos de cultivos tisulares tridimensionales de piel humana, los cuales se utilizan como modelos de estudio para realizar ensayos de citotoxicidad y de determinación de la actividad biológica (basado en marcadores moleculares de proliferación, viabilidad y muerte celular) de sustancias de origen natural. En estos modelos se ha evaluado -mayoritariamente-el potencial anticancerígeno de diversos extractos de plantas, incluyendo plantas medicinales (moringa, tempate, chilillo) y especies frutales (mora, mango, manzana, guayaba) [10]- [17]. En el caso de los modelos tisulares, al tratarse de reconstrucciones 3D, los constructos de piel recrean de forma más cercana el tejido in vivo, por lo que permiten generar información pre-clínica de alta calidad y de mayor potencial predictivo.…”

“…Esto permite proporcionar herramientas clave para facilitar la medicina personalizada, al tiempo que ahorra dinero y reduce la cantidad de animales utilizados para la investigación. A la fecha, estos estudios han permitido confirmar el potencial biomédico de sustancias bioactivas presentes en extractos de diversas plantas en Costa Rica, tales como la mora [10], manzana, anona y ciruelo [13]- [15], [30], guayaba [11] y varias plantas medicinales (Phyllanthus, Senna reticulata, Pettiveria alliaceae, Moringa) [12], [16], [17], [31], [32].…”