Abstract
Significant progress has been made over the past 25 years in the development of in vitro-engineered substitutes that mimic human skin, either to be used as grafts for the replacement of lost skin, or for the establishment of in vitro human skin models. In this sense, laboratory-grown skin substitutes containing dermal and epidermal components offer a promising approach to skin engineering. In particular, a human plasma-based bilayered skin generated by our group, has been applied successfully to treat burns as well as traumatic and surgical wounds in a large number of patients in Spain. There are some aspects requiring improvements in the production process of this skin; for example, the relatively long time (three weeks) needed to produce the surface required to cover an extensive burn or a large wound, and the necessity to automatize and standardize a process currently performed manually. 3D bioprinting has emerged as a flexible tool in regenerative medicine and it provides a platform to address these challenges. In the present study, we have used this technique to print a human bilayered skin using bioinks containing human plasma as well as primary human fibroblasts and keratinocytes that were obtained from skin biopsies. We were able to generate 100 cm2, a standard P100 tissue culture plate, of printed skin in less than 35 min (including the 30 min required for fibrin gelation). We have analysed the structure and function of the printed skin using histological and immunohistochemical methods, both in 3D in vitro cultures and after long-term transplantation to immunodeficient mice. In both cases, the generated skin was very similar to human skin and, furthermore, it was indistinguishable from bilayered dermo-epidermal equivalents, handmade in our laboratories. These results demonstrate that 3D bioprinting is a suitable technology to generate bioengineered skin for therapeutical and industrial applications in an automatized manner.
Highlights
Skin injuries caused by burns, chronic ulcers from different etiology, infections, cancer surgery, and other genetic and somatic diseases require effective treatment to prevent morbidity or mortality
Several approaches have been explored for skin replacement therapy, such as cultured autologous epithelial autografts (CEA), but their results are far from ideal, since they are limited by their fragility and the difficulty of handling, unpredictable take rate and sensitivity to mechanical shearing forces for at least two months post grafting [6,7,8]
As shown in the histological staining (Fig. 3B), printed equivalents generated a tissue with a structure similar to that obtained differentiating handmade skin equivalents (Fig 3A) and similar to normal human skin (Fig 4 C)
Summary
Skin injuries caused by burns, chronic ulcers from different etiology, infections, cancer surgery, and other genetic and somatic diseases require effective treatment to prevent morbidity or mortality. Several approaches have been explored for skin replacement therapy, such as cultured autologous epithelial autografts (CEA) (for a review see [5]), but their results are far from ideal, since they are limited by their fragility and the difficulty of handling, unpredictable take rate and sensitivity to mechanical shearing forces for at least two months post grafting [6,7,8] In response to these limitations, new approaches for skin engineering have been tested and developed in recent years. A human plasma-derived bilayered (including dermis and epidermis) skin model was generated by our group and applied successfully to treat burns and traumatic and surgical wounds [16, 17]
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