Abstract
Engineered tissue-like structures often instigate an inflammatory response in the host that can inhibit wound healing and ultimately lead to the rejection of the implant. In our previous study, we have characterized the properties and biocompatibility of novel multiparticulate drug delivery systems (MDDS), based on collagen matrix with gradual release of anti-inflammatory drug flufenamic acid, we evaluated their anti-inflammatory potential and demonstrated their efficiency against burns and soft tissue lesions. In addition to these results, FA was previously described as a stimulant for adipogenesis, therefore we hypothesized that MDDS might also be appropriate for adipose tissue engineering. After the cell-scaffold constructs were obtained, cell morphology, adhesion and spreading on the systems were highlighted by scanning electron microscopy, immunostaining and confocal microscopy. The effect of FA-enriched materials on adipogenesis was evaluated at gene and protein level, by RT-qPCR, confocal microscopy and immunohistochemistry. Our current work indicates that flufenamic acid plays a beneficial role in adipocyte differentiation, with a direct effect upon the gene and protein expression of important early and late markers of adipogenesis, such as PPARγ2 and perilipin.
Highlights
The field of tissue engineering (TE) combines principles of engineering with notions from natural sciences in order to develop complex structures capable of mimicking natural tissue, in both aspect and function, fit for the purpose of regeneration [1]
Our results showed the successful formation of the cell-scaffold constructs, as well as the successful differentiation of human adipose-derived stem cells (hASCs), with better accumulation of intracellular lipid droplets (Oil Red O staining) and increased PPARγ2 and perilipin expression levels (RT-qPCR, immunohistochemistry and immunofluorescence coupled with confocal microscopy) in multiparticulate drug delivery systems (MDDS) compared to a collagen composite matrix without flufenamic acid (FA)
(30%), but one of them enriched with FA in the composite gel (M4), against a simple composite gel without FA or microcapsules (COL), as potential candidates for adipose tissue engineering
Summary
The field of tissue engineering (TE) combines principles of engineering with notions from natural sciences in order to develop complex structures capable of mimicking natural tissue, in both aspect and function, fit for the purpose of regeneration [1]. In vitro an ATE construct can be generated by seeding a biomaterial, made out of natural/synthetic compounds or a combination of the two, with cells capable of differentiating in mature adipocytes in the presence of a proper differentiation. The best choice for cellular component would seem to be human adipose-derived stem cells (hASCs), hypo-immunogenic cells suitable for designing biocompatible tissue constructs, which can be harvested in abundance from autologous fat tissue through minimal invasive procedures [4,5]. Preclinical studies on the use of hASCs in vitro and in vivo have been performed, and their efficacy has been established in clinical trials [13,14,15]
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