We would like to acknowledge K

We would like to acknowledge K.W. of inflammatory cells (e.g., neutrophils, macrophages, CD4+ and?CD8+?lymphocytes) compared to STSG, acellular NVM (Acell-NVM) and NVM+AF at post-surgery days 7 and/or 14. Wounds treated with NVM+pFF offered only an increase in CD8+ lymphocyte influx. Furthermore, reduced alpha-smooth muscle mass actin (SMA) expression in wound areas and reduced contraction of the wounds was observed with NVM+AF compared to Acell-NVM. Xenogeneic transplantation of NVM+hFF increased SMA expression in wounds compared to NVM+AF. An improved scar quality was observed for wounds treated with NVM+AF compared to Acell-NVM, NVM+hFF and NVM+pFF at day 56. In conclusion, application of autologous fibroblasts improved the overall end result of wound healing in comparison to fetal dermal cells and Acell-NVM, whereas application of fetal dermal fibroblasts in NVM did not improve wound healing of full-thickness wounds in a porcine model. Although human fetal dermal cells exhibited an increased immune response, this did not seem to impact scar quality. test was used as a post hoc test. A value of <0.05 was considered statistically significant. Graph bars symbolize the mean and the standard deviation per treatment. Results Mesenchymal stem cell characteristics of fetal dermal cells The presence of mesenchymal stem cells in a fetal dermal cell populace and their characteristics were assessed by circulation cytometric analysis and differentiation capacity. Human fetal dermal cells were positive for CD105, Dicer1 CD73 and CD90, and were negative for CD14, CD31, CD34, CD45, CD79a and HLA-DR (data not shown). Both human and porcine fetal dermal cells were able to differentiate into osteoblasts, chondrocytes and adipocytes (data not shown). These results meet the criteria as defined by Dominici et al. (2006)Phenotypical characterization of adult MSCs derived from different sources has been explained by van den Bogaerdt et JNJ-39758979 al. (2009). AF, hFF and pFF cultured on glass slides showed comparable staining of the cytoplasm for HSP47 (Fig. ?(Fig.1d1d-?-f).f). SMA expression in these cells was also comparable for AF and pFF; however, the SMA expression in hFF seemed slightly lower (Fig. ?(Fig.1a1a-?-c).c). SMA expression by these cells was represented as a stress fiber phenotype. Open in a separate window Fig. 1 Cell characterization by SMA and HSP47 expression in hFF, pFF and AF (porcine), cells cultured on glass for 2C3?days and stained for DAPI (50?m Autologous fibroblasts in collagen-based dermal substitutes improved scar quality Scars of the differently treated wounds were macroscopically evaluated 56?days post-surgery using an adapted POSAS scar assessment level (Table ?(Table2).2). A lower score represents a better scar quality, indicating closer similarity to unaffected skin. Figure ?Physique2a2a illustrates a macroscopic view of the scars at 56?days post-surgery. Wounds transplanted with NVM+hFF, NVM+pFF, Acell-NVM or STSG exhibited a more star-shaped scar (distortion) compared to wounds treated with JNJ-39758979 NVM+AF. The overall observer scores (scar quality) of the NVM+hFF and NVM+pFF group were much like STSG and Acell-NVM treatment (Fig.?2b). Wounds treated with NVM+AF showed an improved scar quality compared to Acell-NVM, NVM+hFF and NVM+pFF at post-surgery day 56 (Fig.?2b). No differences of other macroscopic (Table ?(Table2)2) and microscopic (Table ?(Table3)3) evaluation scores were observed between the different treatments. Open in a separate windows Fig. 2 Macroscopic evaluation of treated wounds at day 56. a Representative photographs of wounds per treatment. b Scar quality (overall observer score) as determined by scoring parameters of the POSAS level. Scar quality was improved (represented by lower scores) in wounds transplanted with NVM+AF ((MannCWhitney test, <0.05). Split thickness skin graft, Novomaix+STSG, Novomaix JNJ-39758979 made up of human fetal fibroblasts + STSG, Novomaix made up of porcine fetal fibroblasts + STSG, Novomaix made up of autologous fibroblasts +STSG Collagen-based dermal substitutes seeded with autologous fibroblasts reduced scar contraction and SMA expression Excessive wound contraction is usually a frequently encountered characteristic of burn wound scars. Contraction of the wound was macroscopically evaluated (see description above), but also determined by planimetric analysis at days 21 and 56 post-surgery (Fig.?3a). Increased contraction was noted for wounds treated with NVM+hFF versus Acell-NVM at day 21. Full-thickness wounds transplanted with NVM+hFF or NVM+pFF showed statistically significantly more contraction compared to NVM+AF at JNJ-39758979 post-wounding days 21 and 56 (Fig.?3b). A lower wound contraction was only observed in wounds transplanted with NVM+hFF compared to NVM+pFF at day 21. Furthermore, addition of AFs to a dermal substitute reduced contraction of full-thickness wounds in comparison Acell-NVM (day 21 and 56). Overall, contraction increased over time. Open in a separate window Fig. 3 Wound contraction and SMA+ myofibroblasts in the wound area. a Planimetric analysis of wound area to determine wound contraction at day 21 and day 56. Contraction is usually expressed as percentage contraction of total wound area. NVM+AF ((MannCWhitney test, (MannCWhitney test, (MannCWhitney test, p?