Accelerated Wound Healing and Keratinocyte Proliferation through PI3K/Akt/pS6 and VEGFR2 Signaling by Topical Use of Pleural Fluid
Chen-Liang Tsai,
Chih-Ying Changchien,
Ying Chen,
Hsin-Han Chang,
Wen-Chiuan Tsai,
Yi-Wen Wang,
Kai-Chieh Chou,
Ming-Hsien Chiang,
Yu-Ling Tsai,
Hao-Chung Tsai,
Chieh-Yung Wang,
Ming-Sheng Shen,
Li-Ting Cheng,
Hung-Yi Lin,
Tse-Bin Yang,
Chih-Feng Chian
Affiliations
Chen-Liang Tsai
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Chih-Ying Changchien
Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Ying Chen
Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan
Hsin-Han Chang
Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan
Wen-Chiuan Tsai
Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Yi-Wen Wang
Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan
Kai-Chieh Chou
Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan
Ming-Hsien Chiang
Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan
Yu-Ling Tsai
Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Hao-Chung Tsai
Department of Internal Medicine, Division of Chest Medicine, Tri-Service General Hospital Songshan Branch, Taipei 105, Taiwan
Chieh-Yung Wang
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Ming-Sheng Shen
Department of Internal Medicine, Taichung Armed Force General Hospital, Taichung 411, Taiwan
Li-Ting Cheng
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Hung-Yi Lin
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Tse-Bin Yang
Department of Internal Medicine, Taipei City Hospital Renai Branch, Taipei 106, Taiwan
Chih-Feng Chian
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Impaired wound healing is an ongoing issue that cancer patients undergoing chemotherapy or radiotherapy face. Our previous study regarding lung-cancer-associated pleural fluid (LCPF) demonstrated its propensity to promote endothelial proliferation, migration, and angiogenesis, which are crucial features during cutaneous wound healing. Therefore, the current study aimed to investigate the effect of pleural fluid on cutaneous wound closure in vitro and in vivo using HaCaT keratinocytes and a full-thickness skin wound model, respectively. Both heart-failure-associated pleural fluid (HFPF) and LCPF were sequentially centrifuged and filtered to obtain a cell-free status. Treatment with HFPF and LCPF homogeneously induced HaCaT proliferation with cell cycle progression, migration, and MMP2 upregulation. Western blotting revealed increased PI3K/Akt phosphorylation and VEGFR2/VEGFA expression in HaCaT cells. When treated with the PI3K inhibitor, LCPF-induced keratinocyte proliferation was attenuated with decreased pS6 levels. By applying the VEGFR2 inhibitor, LCPF-induced keratinocyte proliferation was ameliorated by pS6 and MMP2 downregulation. The effect of LCPF-induced cell junction rearrangement was disrupted by co-treatment with a VEGFR2 inhibitor. Compared with a 0.9% saline dressing, LCPF significantly accelerated wound closure and re-epithelization when used as a dressing material in a full-thickness wound model. Histological analysis revealed increased neo-epidermis thickness and dermis collagen synthesis in the LCPF-treated group. Furthermore, LCPF treatment activated basal keratinocytes at the wound edge with the upregulation of Ki-67, VEGFA, and MMP2. Our preliminaries provided the benefit of wet dressing with pleural fluid to improve cutaneous wound closure through enhanced re-epithelization and disclosed future autologous application in cancer wound treatment.
