Regeneration of full-thickness skin defects by differentiated adipose-derived stem cells into fibroblast-like cells by fibroblast-conditioned medium

Background Fibroblasts are ubiquitous cells in the human body and are absolutely necessary for wound healing such as for injured skin. This role of fibroblasts was the reason why we aimed to differentiate human adipose-derived stem cells (hADSCs) into fibroblasts and to test their wound healing potency. Recent reports on hADSC-derived conditioned medium have indicated stimulation of collagen synthesis as well as migration of dermal fibroblasts in wound sites with these cells. Similarly, human fibroblast-derived conditioned medium (F-CM) was reported to contain a variety of factors known to be important for growth of skin. However, it remains unknown whether and how F-CM can stimulate hADSCs to secrete type I collagen. Methods In this study, we obtained F-CM from the culture of human skin fibroblast HS27 cells in DMEM media. For an in-vivo wound healing assay using cell transplantation, balb/c nude mice with full-thickness skin wound were used. Results Our data showed that levels of type I pro-collagen secreted by hADSCs cultured in F-CM increased significantly compared with hADSCs kept in normal medium for 72 h. In addition, from a Sircol collagen assay, the amount of collagen in F-CM-treated hADSC conditioned media (72 h) was markedly higher than both the normal medium-treated hADSC conditioned media (72 h) and the F-CM (24 h). We aimed to confirm that hADSCs in F-CM would differentiate into fibroblast cells in order to stimulate wound healing in a skin defect model. To investigate whether F-CM induced hADSCs into fibroblast-like cells, we performed FACS analysis and verified that both F-CM-treated hADSCs and HS27 cells contained similar expression patterns for CD13, CD54, and CD105, whereas normal medium-treated hADSCs were significantly different. mRNA level  analysis for Nanog, Oct4A, and Sox2 as undifferentiation markers and vimentin, HSP47, and desmin as matured fibroblast markers supported the characterization that hADSCs in F-CM were highly differentiated into fibroblast-like cells. To discover the mechanism of type I pro-collagen expression in hADSCs in F-CM, we observed that phospho-smad 2/3 levels were increased in the TGF-β/Smad signaling pathway. For in-vivo analysis, we injected various cell types into balb/c nude mouse skin carrying a 10-mm punch wound, and observed a significantly positive wound healing effect in this full-thickness excision model with F-CM-treated hADSCs rather than with untreated hADSCs or the PBS injected group. Conclusions We differentiated F-CM-treated hADSCs into fibroblast-like cells and demonstrated their efficiency in wound healing in a skin wound model

Estimating the Additional Greenhouse Gas Emissions in Korea: Focused on Demolition of Asbestos Containing Materials in Building

When asbestos containing materials (ACM) must be removed from the building before demolition, additional greenhouse gas (GHG) emissions are generated. However, precedent studies have not considered the removal of ACM from the building. The present study aimed to develop a model for estimating GHG emissions created by the ACM removal processes, specifically the removal of asbestos cement slates (ACS). The second objective was to use the new model to predict the total GHG emission produced by ACM removal in the entire country of Korea. First, an input-equipment inventory was established for each step of the ACS removal process. Second, an energy consumption database for each equipment type was established. Third, the total GHG emission contributed by each step of the process was calculated. The GHG emissions generated from the 1,142,688 ACS-containing buildings in Korea was estimated to total 23,778 tonCO2eq to 132,141 tonCO2eq. This study was meaningful in that the emissions generated by ACS removal have not been studied before. Furthermore, the study deals with additional problems that can be triggered by the presence of asbestos in building materials. The method provided in this study is expected to contribute greatly to the calculation of GHG emissions caused by ACM worldwide