Bioprinted Amniotic Fluid-Derived Stem Cells Accelerate Healing of Large Skin Wounds

Abstract

Stem cells obtained from amniotic fluid show high proliferative capacity in culture and multilineage differentiation potential. Because of the lack of significant immunogenicity and the ability of the amniotic fluid-derived stem (AFS) cells to modulate the inflammatory response, we investigated whether they could augment wound healing in a mouse model of skin regeneration. We used bioprinting technology to treat full-thickness skin wounds in nu/nu mice. AFS cells and bone marrow-derived mesenchymal stem cells (MSCs) were resuspended in fibrin-collagen gel and “printed” over the wound site. At days 0, 7, and 14, AFS cell- and MSC-driven wound closure and re-epithelialization were significantly greater than closure and re-epithelalization in wounds treated by fibrin-collagen gel only. Histological examination showed increased microvessel density and capillary diameters in the AFS cell-treated wounds compared with the MSC-treated wounds, whereas the skin treated only with gel showed the lowest amount of microvessels. However, tracking of fluorescently labeled AFS ceils and MSCs revealed that the cells remained transiently and did not permanently integrate in the tissue. These observations suggest that the increased wound closure rates and angiogenesis may be due to delivery of secreted trophic factors, rather than direct cell-cell interactions. Accordingly, we performed proteomic analysis, which showed that AFS cells secreted a number of growth factors at concentrations higher than those of MSCs. In parallel, we showed that AFS cell-conditioned media induced endothelial cell migration in vitro. Taken together our results indicate that bioprinting AFS cells could be an effective treatment for large-scale wounds and burns.

Impression 3D en médecine régénératrice et ingénierie tissulaire

“…La limite actuelle de la bio-impression in situ est qu'elle ne peut être utilisée que pour des pertes tissulaires localisées en superficie comme la peau ou la calvaria. Cette approche a été utilisée avec des imprimantes jet d'encre pour imprimer des cellules souches issues du liquide amniotique pour le traitement de brûlures [35]. Notre groupe a utilisé une imprimante laser pour imprimer de l'hydroxyapatite dans des défauts de calvaria [25].…”

Advances in bioprinted cell-laden hydrogels for skin tissue engineering

“…Histological analyses also revealed that the newly formed skin contained organized dermal collagen and a fully developed epidermis. The same research group used a similar procedure and bioprinting technique to print amniotic fluidderived stem cells (AFSC) and bone marrow-derived mesenchymal stem cells (MSC) suspended in a fibrinogencollagen gel into full-thickness skin wounds in mice [184]. The cell-biomaterial solution was dispensed to the skin wounds (2.0 9 2.0 cm 2 ) with uniform cell distribution.…”

“…In situ skin printing is challenging as it requires the development of dedicated bioprinting devices, the integration with imaging systems to obtain data from the wound site, and the design of bioinks capable of instructing printed cells to perform Cell liver spheroids Extrusion Liver [185] their native functions. Binder et al [18,184] developed a device for the in situ skin printing composed of a cartridge delivery system containing a series of inkjet nozzles and a laser scanning system, both mounted on a portable XYZ plotting system. The data obtained from the laser was used to reconstruct a 3D model of the wound, which was subsequently employed to determine the skin area that was missing from the wound.…”

“…2) [90,95], with potential to address the limitations of current clinical therapies, including the vascularization, regeneration of skin appendages and integration with the host tissue [52,149]. Bioprinting processes also enable the integration with medical imaging techniques (e.g., microcomputed tomography, magnetic resonance imaging) and path-planning devices, allowing the design of patient specific implants for direct printing either in vitro or in situ [184,200].…”

“…Bioprinting provides a powerful tool to arrange cells, biomaterials and soluble factors within a 3D environment on a length scale comparable to the complex heterogeneity found in natural tissue (10-100 lm), enabling new perspectives as well as unmatched possibilities in the design of biomimetic substitutes for tissue regeneration [10,80,125]. In the context of skin tissue engineering, bioprinting has been explored for the fabrication of 3D constructs containing skin cells positioned in distinct layers to resemble the anatomy of native skin [117,184].…”