Restoring Prohealing/Remodeling-Associated M2a/c Macrophages Using ON101 Accelerates Diabetic Wound Healing

Ching-Wen Lin; Chih-Chiang Chen; Wen-Yen Huang; Yen-Yu Chen; Shiou-Ting Chen; Hung-Wen Chou; Chien-Ming Hung; Wan-Jiun Chen; Chia-Sing Lu; Shi-Xin Nian; Shyi-Gen Chen; Hsuen-Wen Chang; Vincent H S Chang; Li-Ying Liu; Ming-Liang Kuo; Shun-Cheng Chang

doi:10.1016/j.xjidi.2022.100138

Restoring Prohealing/Remodeling-Associated M2a/c Macrophages Using ON101 Accelerates Diabetic Wound Healing

JID Innov. 2022 Jun 2;2(5):100138. doi: 10.1016/j.xjidi.2022.100138. eCollection 2022 Sep.

Authors

Ching-Wen Lin¹, Chih-Chiang Chen^{2

3}, Wen-Yen Huang⁴, Yen-Yu Chen¹, Shiou-Ting Chen⁴, Hung-Wen Chou¹, Chien-Ming Hung⁴, Wan-Jiun Chen¹, Chia-Sing Lu⁵, Shi-Xin Nian², Shyi-Gen Chen^{1

6}, Hsuen-Wen Chang⁷, Vincent H S Chang^{7

8

9}, Li-Ying Liu², Ming-Liang Kuo¹⁰, Shun-Cheng Chang¹¹

Affiliations

¹ Oneness Biotech Co., Ltd., Taipei, Taiwan.
² Department of Dermatology, Taipei Veterans General Hospital, Taipei, Taiwan.
³ Department of Dermatology, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
⁴ Microbio Co., Ltd, Taipei, Taiwan.
⁵ NTU YongLin Institute of Health, National Taiwan University, Taipei, Taiwan.
⁶ Division of Plastic and Reconstructive Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
⁷ TMU Laboratory Animal Center, Office of Research and Development, Taipei Medical University, Taipei, Taiwan.
⁸ Department of Physiology, School of Medicine, Taipei Medical University, Taipei, Taiwan.
⁹ The PhD Program for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.
¹⁰ Microbio Shanghai Co., Ltd., Shanghai, China.
¹¹ Division of Plastic Surgery, Integrated Burn & Wound Care Center, Department of Surgery, Shuang-Ho Hospital; Department of Surgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.

Abstract

Diabetic wounds exhibit chronic inflammation and delayed tissue proliferation or remodeling, mainly owing to prolonged proinflammatory (M1) macrophage activity and defects in transition to prohealing/proremodeling (M2a/M2c; CD206⁺ and/or CD163⁺) macrophages. We found that topical treatment with ON101, a plant-based potential therapeutic for diabetic foot ulcers, increased M2c-like (CD163⁺ and CD206⁺) cells and suppressed M1-like cells, altering the inflammatory gene profile in a diabetic mouse model compared with that in the controls. An in vitro macrophage-polarizing model revealed that ON101 directly suppressed CD80⁺ and CD86⁺ M1-macrophage polarization and M1-associated proinflammatory cytokines at both protein and transcriptional levels. Notably, conditioned medium collected from ON101-treated M1 macrophages reversed the M1-conditioned medium‒mediated suppression of CD206⁺ macrophages. Furthermore, conditioned medium from ON101-treated adipocyte progenitor cells significantly promoted CD206⁺ and CD163⁺ macrophages but strongly inhibited M1-like cells. ON101 treatment also stimulated the expression of GCSF and CXCL3 genes in human adipocyte progenitor cells. Interestingly, treatment with recombinant GCSF protein enhanced both CD206⁺ and CD163⁺ M2 markers, whereas CXCL3 treatment only stimulated CD163⁺ M2 macrophages. Depletion of cutaneous M2 macrophages inhibited ON101-induced diabetic wound healing. Thus, ON101 directly suppressed M1 macrophages and facilitated the GCSF- and CXCL3-mediated transition from M1 to M2 macrophages, lowering inflammation and leading to faster diabetic wound healing.

Keywords: ADPC, adipocyte progenitor cell; CM, conditioned medium; HFD, high-fat diet; HG, high-glucose; IHC, immunohistochemistry; M1-CM, M1-associated conditioned medium; STAT3, signal transducer and activator of transcription 3; iNOS, inducible nitric oxide synthase; m-Clo, mannosylated clodronate.