Efficacy of gelatin hydrogels incorporating triamcinolone acetonide for prevention of fibrosis in a mouse model

Abstract

Introduction Triamcinolone acetonide (TA), a steroid, is often used clinically to prevent dysfunctions associated with fibrosis. The objective of this study was to examine whether TA can be suspended in a gelatin sheet for tissue engineering using a mouse skin wound model. Methods TA was suspended in biodegradable gelatin and freeze-dried in a sheet form. The sheet was analyzed for homogeneity and controlled release of TA by high-performance liquid chromatography. We made two skin wounds on the dorsal side of mice. Gelatin sheets with TA (TA sheet) and without TA (control sheet) were attached to each skin wound. To determine the efficacy of the prepared TA sheet on the skin wounds, TA-sheet versus TA-injection experiments were conducted. Hematoxylin and eosin staining was performed to assess the grade of epithelialization and alpha smooth muscle actin (α-SMA) immunohistochemical staining was conducted to evaluate myofibroblast infiltration. Results In the TA-release test in vitro , 7.7 ± 2.3% of TA was released from the sheet by 24 h. After replacing the initial phosphate-buffered saline (PBS) with collagenase PBS, the amount of released TA increased over time. The wound area/original skin wound area after 15 days with the TA sheet was significantly larger than that with the control sheet (26.9 ± 5.5% vs 10.7 ± 2.6%, p = 0.023). The α-SMA positive area/whole area with the TA sheet was significantly lower than that with the control sheet (4.65 ± 0.66% vs 7.24 ± 0.7%, p = 0.023). Furthermore, the α-SMA positive area/whole area with the TA sheet was significantly lower than that with TA injection (5.32 ± 0.45% vs 7.93 ± 0.75%, p = 0.013). Conclusions We developed a TA sheet and confirmed both the homogeneity of the suspended TA and controlled-release of the TA in the presence of collagenase in vitro . The TA sheet caused less myofibroblast infiltration into the tissue than the control sheet or TA injection did.

Triamcinolone acetonide induces the autophagy of Ag85B-treated WI-38 cells via SIRT1/FOXO3 pathway

“…In mice with skin wound healing, TA inhibited myofibroblast tissue infiltration. 13 TA is widely used in the treatment of fibrosis-related diseases via TGF pathway, such as keloids. 16,17 Consistently, this study found that TA could inhibit fibrosis by the downregulation of TGF-β1 and VEGF.…”

“…12 It is often used clinically to prevent fibrosis-related dysfunction. 13 Chen et al suggested that intra-injection of TA significantly induced keloid cell apoptosis and inhibited keloid formation. 14 In addition, TA combined with 5-fluorouracil inhibited autophagy and fibrosis in urethral scar fibroblasts by increasing the expression of miR-192-5p.…”

Combined therapy of platelet-rich plasma and basic fibroblast growth factor using gelatin-hydrogel sheet for rotator cuff healing in rat models

“…After the solution was frozen, freeze-drying was performed for 48 h. Freeze-drying gelatin-hydrogel was cut into 2 mm × 2 mm segments. The segments were cross-linked by dehydrothermal treatment at 140 °C for 48 h in a vacuum oven [ 28 ].…”

Bioactive gelatin-sheets as novel biopapers to support prevascularization organized by laser-assisted bioprinting for bone tissue engineering

“…After freezing, gelatin-sheets were transferred to a freeze-drier for 3 d (Takara freeze-dryer, Takara ATM Co., Tokyo, Japan) and underwent a dehydrothermal crosslinking (CL) procedure using the following parameters: 120 • C 24 h, 140 • C 24 h or 160 • C 24 h (Vacuum drying device, SATO VAC Inc., Saitama, Japan). The range of CL temperatures was chosen based on previous experiments [21,22]. The crosslinked gelatin sheets were punched out to obtain gelatin disks 8 mm in diameter using an 8 mm punch biopsy tool (biopsy punch, Kai Medical, Seki, Japan) (figure 1).…”