Optimization and Characterization of an Engineered Human Skin Equivalent

Black, Annie; Bouez, Charbel; Perrier, Éric; Schlotmann, Kordula; Chapuis, François; Damour, O.

doi:10.1089/ten.2005.11.723

Cited by 119 publications

(108 citation statements)

References 32 publications

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“…Since epithelial BMs are composed of an intricate network of proteins at the epithelial-stromal interface (Yurchenko and O'Rear, 1994;Christiano and Uitto 1996;Li et al 2001), BM assembly needs to be studied in biological systems in which a high degree of tissue complexity can be achieved, such as human skin equivalents (HSEs) (Andriani et al 2003;Berking and Herlyn 2001). However, HSEs have shown limited success forming structured BM (Black et al 2005;Yang et al 2006) due to MMP-mediated degradation of BM components after their synthesis and secretion (Amano et al 2001;Amano et al 2005). These studies have shown that Type IV Collagen can suppress this MMP-9 expression and stabilize the BM interface during the early stages of BM assembly.…”

Section: Discussionmentioning

confidence: 99%

The basement membrane microenvironment directs the normalization and survival of bioengineered human skin equivalents

Segal¹,

Andriani²,

Pfeiffer³

et al. 2008

Matrix Biology

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Epithelial-mesenchymal interactions promote the morphogenesis and homeostasis of human skin. However, the role of the basement membrane (BM) during this process is not well-understood. To directly study how BM proteins influence epidermal differentiation, survival and growth, we developed novel 3D human skin equivalents (HSEs). These tissues were generated by growing keratinocytes at an air-liquid interface on polycarbonate membranes coated with individual matrix proteins (Type I Collagen, Type IV Collagen or fibronectin) that were placed on contracted Type I Collagen gels populated with dermal fibroblasts. We found that only keratinocytes grown on membranes coated with the BM protein Type IV Collagen showed optimal tissue architecture that was similar to control tissues grown on de-epidermalized dermis (AlloDerm) that contained intact BM. In contrast, tissues grown on proteins not found in BM, such as fibronectin and Type I Collagen, demonstrated aberrant tissue architecture that was linked to a significant elevation in apoptosis and lower levels of proliferation of basal keratinocytes. While all tissues demonstrated a normalized, linear pattern of deposition of laminin 5, tissues grown on Type IV Collagen showed elevated expression of α6 integrin, Type IV Collagen and Type VII Collagen, suggesting induction of BM organization. Keratinocyte differentiation (Keratin 1 and filaggrin) was not dependent on the presence of BM proteins. Thus, Type IV Collagen acts as a critical microenvironmental factor in the BM that is needed to sustain keratinocyte growth and survival and to optimize epithelial architecture.

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Section: Discussionmentioning

confidence: 99%

The basement membrane microenvironment directs the normalization and survival of bioengineered human skin equivalents

Segal¹,

Andriani²,

Pfeiffer³

et al. 2008

Matrix Biology

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“…The current models developed by using advanced materials science and engineering methods are capable of mimicking the entire epidermis; but models mimicking also the complete organization of the dermis layer have not been developed yet. These cases are considered as important clinical issues that have not been fully solved [11,12].…”

Section: Introductionmentioning

confidence: 99%

Electrospinning of nanofibrous polycaprolactone (PCL) and collagen-blended polycaprolactone for wound dressing and tissue engineering

Zeybek¹,

Duman²,

Ürkmez³

2014

uujms

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Fabrication of nanofibrous biomaterials based on natural materials through various techniques is a popular research topic, particularly for biomedical applications. Electrospinning, a well-established technique for nanofiber production has also been extended for producing nanofibrous structures of natural materials that mimic natural extracellular matrix of mammalian tissues. Collagen nanofiber production utilizes hexafluoro propanol (HFP) as a solvent for electrospinning. A novel cost-effective electrospun nanofibrous membrane is established for wound dressing and allogeneic cultured epidermal substitute through the cultivation of human dermal keratinocytes for skin defects. Several synthetic polymers such as polycaprolactone (PCL) are generally electrospun for tissue engineering applications because of their remarkable mechanical stability and slow degradation rates. The large surface area of the polymer nanofibers with specific modifications facilitates cell adhesion and control of their cellular functions. The objectives of this study were to optimize fabrication parameters of electrospun nanofibrous membranes from biodegradable PCL and collagen-blended nanofibrous membranes to combine mechanical integrity and spinnability of PCL with high biocompatibility of collagen, and to examine keratinocyte attachment, morphology, proliferation, and cell-matrix interactions. Results prove that the porous nanofibrous PCL and modified PCL-blended collagen nanofibrous membranes are suitable for the attachment and proliferation of keratinocytes, and might have the potential to be applied as wound dressing as well as in tissue engineering as an epidermal substitute for the treatment of skin defects and burn wounds.

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“…, Antonio Manuel Lizana Moreno , Olga Espinosa Ibáñez 1,3 , Álvaro Sierra Sánchez 1,3 , Alexandra Ordoñez Luque 1,3 , Salvador Arias Santiago 1,3…”

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Pilot study of cutaneous tolerability of fibrin-agarose substitutes in healthy volunteers

González¹,

Moreno²,

Porcel³

et al. 2016

Actual. Med.

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Estudio piloto de tolerabilidad cutánea de láminas de fibrina-agarosa en voluntarios sanos ResumenObjetivos: En el presente estudio se persigue comprobar posibles reacciones adversas, derivadas del uso tópico de láminas de fibrina-agarosa en el antebrazo de voluntarios sanos. Metodología: Se llevó a cabo un estudio experimental en siete voluntarios sanos, cinco varones y dos mujeres, que no presentaban ningún tipo de lesión cutánea visible. En el antebrazo de cada voluntario se colocaron dos láminas de fibrina-agarosa de 4 cm 2 . Cada lámina se cubrió con un apósito impregnado y sobre una de las láminas se aplicó pomada antibiótica con mupirocina. Ambas láminas se cubrieron finalmente con un apósito protector y se mantuvieron en contacto directo sobre la piel durante 48 horas. Resultados: Los resultados determinaron que no se detectaron reacciones adversas después de 48 horas de evolución ni en los siguientes 7 días en ningún voluntario. Se observaron diferencias entre las dos láminas implantadas en cada voluntario, ya que al retirar el apósito cubierto con pomada antibiótica, la lámina presentaba un aspecto más hidratado que la que no llevaba pomada antibiótica. Conclusiones: El uso tópico de las láminas de fibrina-agarosa en voluntarios sanos no presenta reacciones adversas del tipo irritación o alergia al aplicarse directamente por vía tópica. Aunque el tamaño muestral del estudio es limitado, sugiere que la combinación de fibrina-agarosa se presenta como el biomaterial idóneo para el desarrollo de un modelo de piel artificial humana. AbstrasctPurpose: This study aims to analyse possible adverse reactions resulting from the topical use of fibrin-agarose substitutes in the forearm of healthy volunteers. Methods: An experimental study was carried out in seven healthy volunteers, five males and two females, who did not have any cutaneous lesion. Two fibrin-agarose substitutes of 4 cm2 were placed in the forearm of each volunteer. Each substitute was covered with an impregnated dressing and one of the substitutes was covered with antibiotic ointment (mupirocin). Both substitutes were finally covered with a protective dressing. The substitutes were maintained for 48 hours. Results:The results determined that no adverse reactions were detected in any volunteer after 48 hours and a week of evolution. Differences were observed between the two substitutes implanted in each volunteer, since when removing the covered dressing with antibiotic ointment, the substitute presented a more hydrated appearance than the one without antibiotic cream. Conclusions: The implant of fibrin-agarose substitutes in healthy volunteers does not present irritation or allergic type adverse reactions when they applied directly topically on the skin. Although the sample size is low, the fibrin-agarose combination is presented as the biomaterial suitable for the development of an artificial human skin model. Enviado: 01-12-2016 Revisado:14-12-2016 Aceptado: 23-12-2016 Palabras clave: Láminas de fibrina-agarosa, Piel artificial, Tolerabilidad...

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Optimization and Characterization of an Engineered Human Skin Equivalent

Cited by 119 publications

References 32 publications

The basement membrane microenvironment directs the normalization and survival of bioengineered human skin equivalents

The basement membrane microenvironment directs the normalization and survival of bioengineered human skin equivalents

Electrospinning of nanofibrous polycaprolactone (PCL) and collagen-blended polycaprolactone for wound dressing and tissue engineering

Pilot study of cutaneous tolerability of fibrin-agarose substitutes in healthy volunteers

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