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Producción Científica, Dermo-epidermal equivalents based on plasma-derived fibrin hydrogels have been ex tensively studied for skin engineering. However, they showed rapid degradation and contraction over time and low mechanical properties which limit their reproducibility and lifespan. In order to achieve better mechanical properties, elasticity and biological properties, we incorporated a elastin-like recombinamer (ELR) network, based on two types of ELR, one modified with azide (SKS-N3 ) and other with cyclooctyne (SKS-Cyclo) chemical groups at molar ratio 1:1 at three different SKS (serine-lysine-serine sequence) concentrations (1, 3, and 5 wt.%), into plasma-derived fibrin hydrogels. Our results showed a decrease in gelation time and contraction, both in the absence and presence of the encapsulated human primary fibroblasts (hFBs), higher mechanical properties and increase in elasticity when SKSs content is equal or higher than 3%. However, hFBs proliferation showed an improvement when the lowest SKS content (1 wt.%) was used but started decreasing when increasing SKS concentration at day 14 with respect to the plasma control. Proliferation of human primary keratinocytes (hKCs) seeded on top of the hybrid-plasma hydrogels containing 1 and 3% of SKS showed no differences to plasma control and an increase in hKCs proliferation was observed for hybrid-plasma hydrogels containing 5 wt.% of SKS. These promising results showed the need to achieve a balance between the reduced contraction, the better mechanical properties and biological properties and indicate the potential of using this type of hydrogel as a testing platform for pharmaceutical products and cosmetics, and future work will elucidate their potential., Gobierno Español, (Grant/Award Numbers: FPU15-00448, PID2019-110709RB-100, RED2018-102417-T), Junta de Castilla y León (VA317P18, Infrared2018-UVA06), Interreg. V España-Portugal POCTEP (0624_2IQBIONEURO_6_E), Programa de Actividades de I + D entre Grupos de Investigación de la Comunidad de Madrid (S2018/BAA-4480), Programa Estatal de I + D + i Orientada a los Retos de la So-ciedad (RTI2018-101627-B-I00)

Producción Científica, Dermo-epidermal equivalents based on plasma-derived fibrin hydrogels have been ex tensively studied for skin engineering. However, they showed rapid degradation and contraction over time and low mechanical properties which limit their reproducibility and lifespan. In order to achieve better mechanical properties, elasticity and biological properties, we incorporated a elastin-like recombinamer (ELR) network, based on two types of ELR, one modified with azide (SKS-N3 ) and other with cyclooctyne (SKS-Cyclo) chemical groups at molar ratio 1:1 at three different SKS (serine-lysine-serine sequence) concentrations (1, 3, and 5 wt.%), into plasma-derived fibrin hydrogels. Our results showed a decrease in gelation time and contraction, both in the absence and presence of the encapsulated human primary fibroblasts (hFBs), higher mechanical properties and increase in elasticity when SKSs content is equal or higher than 3%. However, hFBs proliferation showed an improvement when the lowest SKS content (1 wt.%) was used but started decreasing when increasing SKS concentration at day 14 with respect to the plasma control. Proliferation of human primary keratinocytes (hKCs) seeded on top of the hybrid-plasma hydrogels containing 1 and 3% of SKS showed no differences to plasma control and an increase in hKCs proliferation was observed for hybrid-plasma hydrogels containing 5 wt.% of SKS. These promising results showed the need to achieve a balance between the reduced contraction, the better mechanical properties and biological properties and indicate the potential of using this type of hydrogel as a testing platform for pharmaceutical products and cosmetics, and future work will elucidate their potential., Gobierno Español, (Grant/Award Numbers: FPU15-00448, PID2019-110709RB-100, RED2018-102417-T), Junta de Castilla y León (VA317P18, Infrared2018-UVA06), Interreg. V España-Portugal POCTEP (0624_2IQBIONEURO_6_E), Programa de Actividades de I + D entre Grupos de Investigación de la Comunidad de Madrid (S2018/BAA-4480), Programa Estatal de I + D + i Orientada a los Retos de la So-ciedad (RTI2018-101627-B-I00)

Producción Científica, Dermo-epidermal equivalents based on plasma-derived fibrin hydrogels have been ex tensively studied for skin engineering. However, they showed rapid degradation and contraction over time and low mechanical properties which limit their reproducibility and lifespan. In order to achieve better mechanical properties, elasticity and biological properties, we incorporated a elastin-like recombinamer (ELR) network, based on two types of ELR, one modified with azide (SKS-N3 ) and other with cyclooctyne (SKS-Cyclo) chemical groups at molar ratio 1:1 at three different SKS (serine-lysine-serine sequence) concentrations (1, 3, and 5 wt.%), into plasma-derived fibrin hydrogels. Our results showed a decrease in gelation time and contraction, both in the absence and presence of the encapsulated human primary fibroblasts (hFBs), higher mechanical properties and increase in elasticity when SKSs content is equal or higher than 3%. However, hFBs proliferation showed an improvement when the lowest SKS content (1 wt.%) was used but started decreasing when increasing SKS concentration at day 14 with respect to the plasma control. Proliferation of human primary keratinocytes (hKCs) seeded on top of the hybrid-plasma hydrogels containing 1 and 3% of SKS showed no differences to plasma control and an increase in hKCs proliferation was observed for hybrid-plasma hydrogels containing 5 wt.% of SKS. These promising results showed the need to achieve a balance between the reduced contraction, the better mechanical properties and biological properties and indicate the potential of using this type of hydrogel as a testing platform for pharmaceutical products and cosmetics, and future work will elucidate their potential., Gobierno Español, (Grant/Award Numbers: FPU15-00448, PID2019-110709RB-100, RED2018-102417-T), Junta de Castilla y León (VA317P18, Infrared2018-UVA06), Interreg. V España-Portugal POCTEP (0624_2IQBIONEURO_6_E), Programa de Actividades de I + D entre Grupos de Investigación de la Comunidad de Madrid (S2018/BAA-4480), Programa Estatal de I + D + i Orientada a los Retos de la So-ciedad (RTI2018-101627-B-I00)

Producción Científica, Dermo-epidermal equivalents based on plasma-derived fibrin hydrogels have been ex tensively studied for skin engineering. However, they showed rapid degradation and contraction over time and low mechanical properties which limit their reproducibility and lifespan. In order to achieve better mechanical properties, elasticity and biological properties, we incorporated a elastin-like recombinamer (ELR) network, based on two types of ELR, one modified with azide (SKS-N3 ) and other with cyclooctyne (SKS-Cyclo) chemical groups at molar ratio 1:1 at three different SKS (serine-lysine-serine sequence) concentrations (1, 3, and 5 wt.%), into plasma-derived fibrin hydrogels. Our results showed a decrease in gelation time and contraction, both in the absence and presence of the encapsulated human primary fibroblasts (hFBs), higher mechanical properties and increase in elasticity when SKSs content is equal or higher than 3%. However, hFBs proliferation showed an improvement when the lowest SKS content (1 wt.%) was used but started decreasing when increasing SKS concentration at day 14 with respect to the plasma control. Proliferation of human primary keratinocytes (hKCs) seeded on top of the hybrid-plasma hydrogels containing 1 and 3% of SKS showed no differences to plasma control and an increase in hKCs proliferation was observed for hybrid-plasma hydrogels containing 5 wt.% of SKS. These promising results showed the need to achieve a balance between the reduced contraction, the better mechanical properties and biological properties and indicate the potential of using this type of hydrogel as a testing platform for pharmaceutical products and cosmetics, and future work will elucidate their potential., Gobierno Español, (Grant/Award Numbers: FPU15-00448, PID2019-110709RB-100, RED2018-102417-T), Junta de Castilla y León (VA317P18, Infrared2018-UVA06), Interreg. V España-Portugal POCTEP (0624_2IQBIONEURO_6_E), Programa de Actividades de I + D entre Grupos de Investigación de la Comunidad de Madrid (S2018/BAA-4480), Programa Estatal de I + D + i Orientada a los Retos de la So-ciedad (RTI2018-101627-B-I00)

Clinical use of bioengineered skin in reconstructive surgery has been established for more than 30 years. The limitations and ethical considerations regarding the use of animal models have expanded the application of bioengineered skin in the areas of disease modeling and drug screening. These skin models should represent the anatomical and physiological traits of native skin for the efficient replication of normal and pathological skin conditions. In addition, reliability of such models is essential for the conduction of faithful, rapid, and large-scale studies. Therefore, research efforts are focused on automated fabrication methods to replace the traditional manual approaches. This report presents an overview of the skin models applicable to skin disease modeling along with their fabrication methods, and discusses the potential of the currently available options to conform and satisfy the demands for disease modeling and drug screening., Biomaterials & Tissue Biomechanics

Clinical use of bioengineered skin in reconstructive surgery has been established for more than 30 years. The limitations and ethical considerations regarding the use of animal models have expanded the application of bioengineered skin in the areas of disease modeling and drug screening. These skin models should represent the anatomical and physiological traits of native skin for the efficient replication of normal and pathological skin conditions. In addition, reliability of such models is essential for the conduction of faithful, rapid, and large-scale studies. Therefore, research efforts are focused on automated fabrication methods to replace the traditional manual approaches. This report presents an overview of the skin models applicable to skin disease modeling along with their fabrication methods, and discusses the potential of the currently available options to conform and satisfy the demands for disease modeling and drug screening., Biomaterials & Tissue Biomechanics

Bioengineered skin was initially developed for clinical application in reconstructive surgery, but nowadays it is also considered as a suitable skin model for research. Research applications range from assessment of cosmetic and pharmaceutical products to the modeling of pathological skin conditions. For example, psoriasis is a complicated skin disease that strongly concerns the scientific community because of its unclear pathogenesis and the current inability to cure it. The Pharmacogenomics Lab of ETH has been working on the development of psoriatic skin models in order to investigate the driving mechanisms of the disease and to later implement high-throughput drug screening to explore treatment possibilities. However, the manual production into transwell inserts results in unstable, self-contracting and inconsistent skin models, pain points that the Pharmacogenomics Lab would like to eliminate. A recent collaboration between the Product Development Group and the Pharmacogenomics Lab has been recently initiated in an attempt to standardize the fabrication of the psoriatic skin models. For this purpose, this Master Thesis focused on the investigation of the potentials and limitations of automated injection molding, a method developed by the Product Development Group, in the generation of consistent and representative dermo-epidermal models for psoriatic skin. After a series of experiments, it was seen that in-mold fabrication achieves homogeneous and consistent dermal hydrogels that maintain their dimensions throughout the whole cultivation period regardless of the collagen concentration employed for the dermal matrix, while automated injection molding allows the utilization of collagen concentrations higher than the unstable 5mg/ml, traditionally used in the manual fabrication method. All collagen concentrations exhibited similar biological performance, but the optimum fibroblasts viability was achieved in case of 5mg/mL, meaning that the whole injection mold, Biomedical Engineering | Biomaterials and Tissue Biomechanics

Severe exposure to HD induces blistering skin reactions and significant loss of stem cell keratinocytes that are required for a continuous renewal of the epidermal cell layer. Therefore, HD injuries require long healing periods leaving significant cosmetic and/or functional deficits. We are developing bioengineered skin from embryonic stem cells for improved therapy for HD-induced skin damage. Since stem cell keratinocytes lack major histocompatibility class II antigens, they exhibit little immunogenicity and are suitable for allograft use. The multipotential nature of stem cells may be particularly valuable for skin damage where several cell types and growth factors are necessary for proper repair and regeneration. We differentiated embryonic stem cells to skin keratinocytes in a cellular matrix cultured in an air/liquid system to form the bioengineered skin, mimicking normal skin. The efficacy of bioengineered skin for HD-induced skin lesions was evaluated using a C57BL/6 mouse model and 2-chloroethylethyl sulfide (half-mustard or CEES). Mice were exposed to 3 microliters neat CEES for 10 minutes, and at 48 h post exposure, the injured skin was excised and the site was cleaned with debridase. Next, the bioengineered skin was transferred directly to the wound and affixed with a non-adherent sterile gauze pad. The affixed with a non-adherent sterile gauze pad. The embryonic stem cell derived bioengineered skin exhibited growth and healing in 1 to 3 weeks. In contrast, the CEES exposed animals not treated with bioengineered skin had a) no skin growth and b) obvious contraction of the skin in the injured area. Our results demonstrate that for the first time, topically applied fresh or frozen bioengineered skin or skin keratinocytes from embryonic stem cells produce improved healing when applied 48 h after HD exposure., See also ADM002075. Presented at the Army Science Conference (25th) held in Orlando, Florida on 27-30 November 2006. The original document contains color images.