Álvaro Sierra-Sánchez, Jorge Cabañas-Penagos, Sandra Igual-Roger, Luis Martínez-Heredia, Olga Espinosa-Ibáñez, Raquel Sanabria-de la Torre, María I Quiñones-Vico, Ana Ubago-Rodríguez, Antonio Lizana-Moreno, Ana Fernández-González, Jorge Guerrero-Calvo, Natividad Fernández-Porcel, Arena Ramírez-Muñoz, Salvador Arias-Santiago
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Herein, the biological properties (cell viability, cell metabolic activity, protein secretion profile and histology) of several variations of a clinical HPSS model, regarding the biomaterial composition (alone or combined with six secondary biomaterials - serine, fibronectin, collagen, two types of laminins and hyaluronic acid), the cellular structure (trilayer, bilayer, monolayer and control without cells) and their skin tissue of origin (abdominal or foreskin cells) and the manufacturing process [effect of partial dehydration process in cell viability and comparison between submerged (SUB) and air/liquid interface (ALI) methodologies] have been evaluated and compared. Results reveal that the use of human plasma as a main biomaterial determines the <i>in vitro</i> properties, rather than the secondary biomaterials added. Moreover, the characteristics are similar regardless of the skin cells used (from abdomen or foreskin). However, the manufacture of more complex cellular substitutes (trilayer and bilayer) has been demonstrated to be better in terms of cell viability, metabolic activity and wound healing protein secretion (bFGF, EGF, VEGF-A, CCL5) than monolayer HPSSs, especially when ALI culture methodology is applied. Moreover, the application of the dehydration, although required to achieve an appropriate clinical structure, reduce cell viability in all cases. 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引用次数: 0
摘要
人血浆是一种天然生物材料,由于其蛋白质成分,被广泛用于临床产品的开发,尤其是在皮肤科领域。在这种情况下,这种生物材料被单独用作支架或与其他材料结合,用于开发细胞人血浆皮肤替代品(HPSS)。在此,我们将根据生物材料的组成(单独或与六种辅助生物材料--丝氨酸、纤连蛋白、胶原蛋白、两种层粘连蛋白和透明质酸--结合使用)、细胞结构(三层或四层)、细胞的存活率、细胞代谢活性、蛋白质分泌情况和组织学,对临床 HPSS 模型的几种变体的生物特性(细胞存活率、细胞代谢活性、蛋白质分泌情况和组织学)进行分析、对细胞结构(三层、双层、单层和无细胞对照组)及其来源的皮肤组织(腹部细胞或包皮细胞)和制造工艺[部分脱水工艺对细胞活力的影响以及浸没(SUB)和空气/液体界面(ALI)方法的比较]进行了评估和比较。结果表明,使用人血浆作为主要生物材料比添加辅助生物材料更能决定体外特性。此外,无论使用哪种皮肤细胞(腹部或包皮),其特性都是相似的。不过,事实证明,与单层 HPSSs 相比,制造更复杂的细胞替代物(三层和双层)在细胞活力、代谢活性和伤口愈合蛋白分泌(bFGF、EGF、VEGF-A、CCL5)方面更胜一筹,尤其是在采用 ALI 培养方法时。此外,脱水虽然是实现适当临床结构的必要条件,但在所有情况下都会降低细胞活力。这些数据表明,这种 HPSS 模型既稳健又可靠,而且根据目标皮肤病的不同,这里分析的几种亚型可能是很有前景的临床方法。
Biological properties and characterization of several variations of a clinical human plasma-based skin substitute model and its manufacturing process.
Human plasma is a natural biomaterial that due to their protein composition is widely used for the development of clinical products, especially in the field of dermatology. In this context, this biomaterial has been used as a scaffold alone or combined with others for the development of cellular human plasma-based skin substitutes (HPSSs). Herein, the biological properties (cell viability, cell metabolic activity, protein secretion profile and histology) of several variations of a clinical HPSS model, regarding the biomaterial composition (alone or combined with six secondary biomaterials - serine, fibronectin, collagen, two types of laminins and hyaluronic acid), the cellular structure (trilayer, bilayer, monolayer and control without cells) and their skin tissue of origin (abdominal or foreskin cells) and the manufacturing process [effect of partial dehydration process in cell viability and comparison between submerged (SUB) and air/liquid interface (ALI) methodologies] have been evaluated and compared. Results reveal that the use of human plasma as a main biomaterial determines the in vitro properties, rather than the secondary biomaterials added. Moreover, the characteristics are similar regardless of the skin cells used (from abdomen or foreskin). However, the manufacture of more complex cellular substitutes (trilayer and bilayer) has been demonstrated to be better in terms of cell viability, metabolic activity and wound healing protein secretion (bFGF, EGF, VEGF-A, CCL5) than monolayer HPSSs, especially when ALI culture methodology is applied. Moreover, the application of the dehydration, although required to achieve an appropriate clinical structure, reduce cell viability in all cases. These data indicate that this HPSS model is robust and reliable and that the several subtypes here analysed could be promising clinical approaches depending on the target dermatological disease.
期刊介绍:
Regenerative Biomaterials is an international, interdisciplinary, peer-reviewed journal publishing the latest advances in biomaterials and regenerative medicine. The journal provides a forum for the publication of original research papers, reviews, clinical case reports, and commentaries on the topics relevant to the development of advanced regenerative biomaterials concerning novel regenerative technologies and therapeutic approaches for the regeneration and repair of damaged tissues and organs. The interactions of biomaterials with cells and tissue, especially with stem cells, will be of particular focus.