Functional Cellular Networks Laboratory, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon 21999, Republic of Korea.
SeoAh Song Dermatologic Clinic, Seoul 05557, Republic of Korea.
Int J Mol Sci. 2023 Apr 28;24(9):7986. doi: 10.3390/ijms24097986.
Angiogenesis promotes rejuvenation in multiple organs, including the skin. Heat shock protein 90 (HSP90), hypoxia-inducible factor-1 alpha (HIF-1α), and vascular endothelial growth factor (VEGF) are proangiogenic factors that stimulate the activities of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and extracellular signal-regulated kinase 1/2 (ERK1/2). Poly-D,L-lactic acid (PDLLA), polynucleotide (PN), and calcium hydroxyapatite (CaHA) are dermal fillers that stimulate the synthesis of dermal collagen. However, it is not yet known whether these compounds promote angiogenesis, which leads to skin rejuvenation. Here, we evaluated whether PDLLA, PN, and CaHA stimulate angiogenesis and skin rejuvenation using HO-treated senescent macrophages and endothelial cells as an in vitro model for skin aging, and we used young and aged C57BL/6 mice as an in vivo model. Angiogenesis was evaluated via endothelial cell migration length, proliferation, and tube formation after conditioned media (CM) from senescent macrophages was treated with PDLLA, PN, or CaHA. Western blot showed decreased expression levels of HSP90, HIF-1α, and VEGF in senescent macrophages, but higher expression levels of these factors were found after treatment with PDLLA, PN, or CaHA. In addition, after exposure to CM from senescent macrophages treated with PDLLA, PN, or CaHA, senescent endothelial cells expressed higher levels of VEGF receptor 2 (VEGFR2), PI3K, phosphorylated AKT (pAKT), and phosphorylated ERK1/2 (pERK1/2) and demonstrated greater capacities for cell migration, cell proliferation, and tube formation. Based on the levels of 4-hydroxy-2-nonenal, the oxidative stress level was lower in the skin of aged mice injected with PDLLA, PN, or CaHA, while the tumor growth factor (TGF)-β1, TGF-β2, and TGF-β3 expression levels; the density of collagen fibers; and the skin elasticity were higher in the skin of aged mice injected with PDLLA, PN, or CaHA. These effects were greater in PDLLA than in PN or CaHA. In conclusion, our results are consistent with the hypothesis that PDLLA stimulates angiogenesis, leading to the rejuvenation of aged skin. Our study is the first to show that PDLLA, PN, or CaHA can result in angiogenesis in the aged skin, possibly by increasing the levels of HSP90, HIF-1α, and VEGF and increasing collagen synthesis.
血管生成促进包括皮肤在内的多种器官的年轻化。热休克蛋白 90（HSP90）、缺氧诱导因子-1α（HIF-1α）和血管内皮生长因子（VEGF）是促进血管生成的因子，可刺激磷酸肌醇 3-激酶（PI3K）、蛋白激酶 B（AKT）和细胞外信号调节激酶 1/2（ERK1/2）的活性。聚-D,L-乳酸（PDLLA）、多核苷酸（PN）和钙羟磷灰石（CaHA）是刺激真皮胶原合成的真皮填充剂。然而，目前尚不清楚这些化合物是否能促进血管生成，从而导致皮肤年轻化。在这里，我们使用 HO 处理的衰老巨噬细胞和内皮细胞作为皮肤衰老的体外模型，评估了 PDLLA、PN 和 CaHA 是否能刺激血管生成和皮肤年轻化，并用年轻和年老的 C57BL/6 小鼠作为体内模型。用 PDLLA、PN 或 CaHA 处理衰老巨噬细胞的条件培养基（CM）后，通过内皮细胞迁移长度、增殖和管形成评估血管生成。Western blot 显示，衰老巨噬细胞中 HSP90、HIF-1α 和 VEGF 的表达水平降低，但用 PDLLA、PN 或 CaHA 处理后这些因子的表达水平升高。此外，用 PDLLA、PN 或 CaHA 处理衰老巨噬细胞的 CM 处理后，衰老的内皮细胞表达更高水平的血管内皮生长因子受体 2（VEGFR2）、PI3K、磷酸化 AKT（pAKT）和磷酸化 ERK1/2（pERK1/2），并具有更强的细胞迁移、细胞增殖和管形成能力。基于 4-羟基-2-壬烯醛的水平，在注射 PDLLA、PN 或 CaHA 的老年小鼠皮肤中，氧化应激水平较低，而肿瘤生长因子（TGF）-β1、TGF-β2 和 TGF-β3 的表达水平；胶原纤维密度；以及皮肤弹性在注射 PDLLA、PN 或 CaHA 的老年小鼠皮肤中较高。PDLLA 的这些作用大于 PN 或 CaHA。总之，我们的结果与 PDLLA 刺激血管生成，导致老年皮肤年轻化的假设一致。我们的研究首次表明，PDLLA、PN 或 CaHA 可导致老年皮肤中的血管生成，可能是通过增加 HSP90、HIF-1α 和 VEGF 的水平和增加胶原合成。
