Abstract
BackgroundThere is a lack of proper animal models to study keloid formation. AimTo create three-dimensional poly lactic-co-glycolic acid (PLGA) scaffolds containing autologous platelet-rich plasma (PRP) as an in vitro culture environment for keloid fibroblasts (KL), and to study their implantation into nude mice to mimic the process of keloid formation. MethodsNormal fibroblasts (FB) and KL cells were isolated from surgical specimens and transduced with lentivirus loaded with green fluorescent protein (GFP) and luciferase genes. The FB and KL cells were three-dimensionally cultured for 14–18days in PLGA scaffolds containing PRP. Ten mice were implanted with KL cells in their left forelimbs(KL), and FB-scaffolds (FB+PLGA) in their right forelimbs. An additional ten mice were implanted with PLGA scaffolds without cells (PLGA) in their left forelimbs, and KL-scaffolds (KL+PLGA) in their right forelimbs. Graft volume and collagen content were analyzed 120days after the implantation. Resultsin vivo luminescence cell imaging showed that the FB cells proliferated in the PLGA scaffolds within 60days after implantation, and reached a plateau afterwards until 120days after implantation. The KL cells continuously proliferated in the PLGA scaffolds until 120days after implantation. The KL+PLGA group showed higher graft volumes than the FB+PLGA group 120days after the implantation (median volume, 166.95 vs. 63.34mm3); however, the difference is not statistically significant (P=0.743), due to a large variation of the graft volume within each group. Furthermore, Sirius red staining revealed increased collagen I deposition, and immunohistochemistry showed large-scale accumulation of α-smooth muscle actin (α-SMA), collagen I, and collagen III in the KL+PLGA grafts. ConclusionThe three-dimensional PLGA scaffold containing PRP supports keloid fibroblast growth and contributes to keloid formation in a nude mouse model.
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