Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing

Abstract

Background: Adipose tissue has been proven to play a crucial role in wound healing, while kindlin-2, an integrin-associated protein, has been shown to regulate cell adhesion, migration, and differentiation. This study aimed to explore its involvement in the cell differentiation of 3T3-L1 preadipocytes and its role in wound healing.Methods: Cell adhesion, Cell Counting Kit-8 (CCK-8), Transwell, and in vitro wound healing assays, along with adipogenic and osteogenic differentiation induction were performed in 3T3-L1 preadipocytes in which kindlin-2 was knocked down or overexpressed. In vivo, kindlin-2 (+/−) transgenic mice were constructed, and wound healing was analyzed by immunohistochemistry (IHC) in a mouse dorsal wound model. Realtime polymerase chain reaction (RT-PCR) and western blotting were performed to analyze the expression of adipokines and adipogenic markers in mouse wound tissues. Adipogenic differentiation induction of adipose tissue stromal vascular fraction (SVF) were performed, and the expression of adipogenic markers in SVF was detected by western blotting. The target signaling pathway highly related to adipogenic differentiation was explored by computational biology and verified by western blotting.Results: Knockdown of kindlin-2 was found to inhibit the adhesion, migration, and adipogenic differentiation of 3T3-L1 preadipocytes while promoting their osteogenic differentiation. In contrast, kindlin-2 overexpression resulted in increased adhesion, migration, and adipogenic differentiation of 3T3-L1 preadipocytes while reducing osteogenic differentiation. In vivo, downregulation of kindlin-2 inhibited adipogenesis in kindlin-2 transgenic mice, resulting in delayed wound healing by inhibiting inflammation, angiogenesis, collagen remodeling, and wound contraction. Mechanistically, we found that kindlin-2 could regulate adipogenic differentiation through PI3K/AKT/mTOR signaling pathway.Conclusions: Our study revealed the essential role that kindlin-2 has in the differentiation and wound healing of 3T3-L1 preadipocytes, which offers a theoretical basis for further research and a novel strategy for wound healing.

Benefits of negative pressure wound therapy with fat migration during revisional total hip arthroplasty in an obese patient: A case report

“…Studies have shown that preadipocytes, which are highly migratory fibroblast-like stem cells, facilitate wound healing. [ 10 ] Previous studies on transgenic mice have shown this effect can be explained by the role of adiponectin, which is a potent mediator of wound healing through re-epithelialization and angiogenesis. [ 11 ] In addition, adiponectin promotes preadipocyte proliferation and migration, serving as a potential therapeutic approach to treating surgical wounds in complex cases such as this patient.…”

SENP3 deletion promotes M2 macrophage polarization and accelerates wound healing through smad6/IκB/p65 signaling pathway

“…Skin tissue and cellular protein extraction and Western blotting were performed according to our previous protocols [ 45 ]. All the antibodies are presented in Supplementary Table 2 .…”

“…Immunohistochemistry performed on paraffin sections and immunofluorescence staining performed on cells were described in our previous study [ 45 ]. For tissue immunofluorescence staining, sections were permeabilized using 0.2% Triton X-100 before blocking using 20% goat serum and then incubated with primary antibodies at 4 °C overnight.…”