Instead, Ad-MSCs-SF and D-Ad-MSCs-SF were equally effective and needed 36 hours to complete the coverage of the scratch area by DFs (Figure?4E)

Instead, Ad-MSCs-SF and D-Ad-MSCs-SF were equally effective and needed 36 hours to complete the coverage of the scratch area by DFs (Figure?4E). sterilized with ethanol 70 vol% and exposed to UV light for one hour (C.I. = 0.67). (c) Decellularized SF patch stored in water at 4C (C.I. = 0.69). (d) Decellularized SF patch stored under dry conditions at 4C (C.I. = 0.69). (e) Decellularized SF patch frozen stored in water at -20C (C.I. = 0.69). (f) Decellularized SF patch frozen stored under dry conditions at -20C (C.I. = 0.69). (A I = amide I; A II = amide II; A III = amide III). The intrinsic crystalline structure of SF patches was not affected by any of the treatments carried out on them, from sterilization to decellularization, freezing and storing under dry or wet conditions at +4C or -20C, as demonstrated by the closely similar profiles and by the values of crystallinity. (E) DSC thermograms of SF patches. (a) Untreated control sample (C.I. = 0.69). (b) SF patch sterilized with ethanol 70?vol% and exposed to UV light for six hours. (c) Decellularized SF patch stored in water at 4C. (d) Decellularized SF patch stored under dry conditions at 4C. (e) Decellularized SF patch frozen stored in water at -20C. (f) Decellularized SF patch frozen stored under dry conditions at -20C. Sterilization caused a slight low-temperature broadening of the melting/degradation endotherm, but the main peak still remained at high temperature (284C) and the -sheet crystalline regions retained their thermal stability, as indicated by the FTIR results. scrt396-S2.tiff (6.4M) GUID:?3E470038-CE6E-44C0-8100-71E57B00BE28 Additional file 3: Figure S3 Histological analysis of skin wounds upon treatment with SF, Ad-MSCs-SF and D-Ad-MSCs-SF patches. On day 14 after treatments, some mice were sacrificed and wounds were investigated by histology. Control wounds treated with SF patches alone showed a dermis displaying important hypercellularity, scanty collagen fiber alignment and continuous epidermis with Rabbit polyclonal to ACADM evident signs of dysplasia determined by the immature status (A, B). Wounds treated with D-Ad-MSCs-SF patches showed a more advanced epidermal organization and a dermis very rich in cells and microvessels (C, D). The wound treated with Ad-MSCs-SF showed the highest degree of tissue organization (E, F); the multilayer structure of epidermis was formed, the dermis still showed hypercellularity with the presence of numerous neoformed small vessels. It was also possible to observe early pilo-sebaceous units (arrowheads). In B, D and F are shown, at higher magnifications, the skin of mice treated with SF, D-Ad-MSCs-SF and Ad-MSCs-SF patches, respectively. In the figure, wound edges are Cardiolipin indicated by arrows; e = epidermis; d = dermis. scrt396-S3.tiff (13M) GUID:?7CFE8998-B7BD-4E62-B4F7-65350A01C481 Additional file 4: Figure S4 Wound healing process in mouse tissue by Ad-MSCs-SF and D-Ad-MSCs-SF. In mouse tissues that received SF, Ad-MSCs-SF and D-Ad-MSCs-SF patches, Col41 (A,B,C) and Vegf (D,E,F) were investigated by immunohistochemistry. Expression of Col41was observed in every sample. Basal membrane was continuously and sharply stained in Ad-MSCs-SF as well as in D-Ad-MSCs-SF demonstrating that the epidermal-dermal junction had been restored. Cardiolipin An average of 10 to 12 spindle shaped Vegf positive cells per field (100 magnification) were observed in the dermal layer of Ad-MSCs-SF. Conversely, reactive cells in D-Ad-MSCs-SF treated samples were less numerous (two to four per field, at 100 Cardiolipin magnification) and were characterized by a less intense staining. A similar number of Vegf positive cells was detected in SF treated samples. Immunohistochemical staining with anti-HuNu was additionally performed to demonstrate the fate of human transplanted Ad-MSCs in host tissues. The anti-HuNu antibody reacted with some cells located in.