. 2025 Nov 19;23(11):444.

doi: 10.3390/md23110444.

Botryocladia leptopoda Extracts Promote Wound Healing Ability via Antioxidant and Anti-Inflammatory Activities and Regulation of MMP/TIMP Expression

Shin-Ping Lin^{1

2

3}, Tsung-Kai Yi⁴, Yi-Feng Kao⁴, Ming-Chieh Tu⁴, Chen-Che Hsieh⁵, Yu-Chieh Chou², Jheng-Jhe Lu⁶, Shella Permatasari Santoso^{7

8

9}, Huey-Jine Chai⁴, Kuan-Chen Cheng^{6

10

11

12

13}

Affiliations

¹ School of Food Safety, Taipei Medical University, Taipei 11031, Taiwan.
² Ph.D. Program in Drug Discovery and Development Industry, Taipei Medical University, Taipei 11031, Taiwan.
³ Research Center of Biomedical Device, Taipei Medical University, Taipei 11031, Taiwan.
⁴ Seafood Technology Division, Fisheries Research Institute, Ministry of Agriculture, Keelung 20246, Taiwan.
⁵ Department of Seafood Science, College of Hydrosphere, National Kaohsiung University of Science and Technology, Kaohsiung 81157, Taiwan.
⁶ Institute of Biotechnology, National Taiwan University, Taipei 10617, Taiwan.
⁷ Department of Chemical Engineering, Faculty of Engineering, Widya Mandala Surabaya Catholic University, Surabaya 60114, Indonesia.
⁸ Chemical Engineering Master Program, Widya Mandala Surabaya Catholic University, Surabaya 60114, Indonesia.
⁹ Collaborative Research Center for Zero Waste and Sustainability, Widya Mandala Surabaya Catholic University, Surabaya 60114, Indonesia.
¹⁰ Institute of Food Science and Technology, National Taiwan University, Taipei 10617, Taiwan.
¹¹ Department of Medical Research, China Medical University Hospital, China Medical University, Taichung 40402, Taiwan.
¹² Department of Optometry, Asia University, Taichung 41354, Taiwan.
¹³ Department of Food Science, Fu Jen Catholic University, New Taipei City 242062, Taiwan.

PMID: 41295412
PMCID: PMC12653739
DOI: 10.3390/md23110444

Botryocladia leptopoda Extracts Promote Wound Healing Ability via Antioxidant and Anti-Inflammatory Activities and Regulation of MMP/TIMP Expression

Shin-Ping Lin et al. Mar Drugs. 2025.

. 2025 Nov 19;23(11):444.

doi: 10.3390/md23110444.

Authors

Affiliations

¹ School of Food Safety, Taipei Medical University, Taipei 11031, Taiwan.
² Ph.D. Program in Drug Discovery and Development Industry, Taipei Medical University, Taipei 11031, Taiwan.
³ Research Center of Biomedical Device, Taipei Medical University, Taipei 11031, Taiwan.
⁴ Seafood Technology Division, Fisheries Research Institute, Ministry of Agriculture, Keelung 20246, Taiwan.
⁵ Department of Seafood Science, College of Hydrosphere, National Kaohsiung University of Science and Technology, Kaohsiung 81157, Taiwan.
⁶ Institute of Biotechnology, National Taiwan University, Taipei 10617, Taiwan.
⁷ Department of Chemical Engineering, Faculty of Engineering, Widya Mandala Surabaya Catholic University, Surabaya 60114, Indonesia.
⁸ Chemical Engineering Master Program, Widya Mandala Surabaya Catholic University, Surabaya 60114, Indonesia.
⁹ Collaborative Research Center for Zero Waste and Sustainability, Widya Mandala Surabaya Catholic University, Surabaya 60114, Indonesia.
¹⁰ Institute of Food Science and Technology, National Taiwan University, Taipei 10617, Taiwan.
¹¹ Department of Medical Research, China Medical University Hospital, China Medical University, Taichung 40402, Taiwan.
¹² Department of Optometry, Asia University, Taichung 41354, Taiwan.
¹³ Department of Food Science, Fu Jen Catholic University, New Taipei City 242062, Taiwan.

PMID: 41295412
PMCID: PMC12653739
DOI: 10.3390/md23110444

Abstract

Wound healing is a complex process involving coordinated actions of multiple cell types. Therefore, when developing therapeutics to promote wound healing, it is essential to consider the synergistic contributions of various cells at different stages of the healing process. In this study, we evaluated the potential of different extracts of Botryocladia leptopoda as wound-healing agents by examining their effects on antioxidant activity, cytotoxicity, cell migration, anti-inflammatory properties, and expressions of specific biomarkers associated with wound healing. Results indicated that the ethanol extract (FE) and hexane extract (HE) exhibited the highest DPPH radical scavenging activity, reaching up to 94%. The alkaline extract (AE) showed the strongest antioxidant ability in the FICA assay, with a maximum of 99%. In addition, the FE and AE provided anti-inflammatory actions that inhibited tumor necrosis factor (TNF)-α and interleukin (IL)-6 in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Further analyses suggested that the FE and AE enhanced cell proliferation (210% and 112%) and migration (442.2% and 535.6%) and regulated wound healing-related genes, including matrix metalloproteinase 2, MMP9, and tissue inhibitor of metalloproteinase 2 (TIMP2) to avoid scar formation and accelerate wound healing. Lastly, the identification of potential compounds within the extract using the UHPLC system further supports its prospective medical applications. Taken together, these findings indicated that the FE and AE from B. leptopoda exhibited remarkable in vitro wound-healing properties, highlighting their potential for applications in pharmaceutical industries and health food development.

Keywords: Botryocladia leptopoda; anti-inflammation; antioxidant; cell proliferation; wound healing.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

**Figure 1**
Effects of different solvent extracts on total phenolic contents of *Botryocladia leptopoda* (a, b) extraction yields. Different letters on the error bars indicate significant differences among the extraction groups and treatments (p < 0.05). AE: alkaline extract obtained at 25 °C; FE: ethanol extract obtained at 25 °C; HE: ethanol extract obtained at 70 °C; HW: hot water extract obtained at 100 °C.

**Figure 2**
Antioxidant effects of various extracts (a) against DPPH radicals and (b) in a FICA assay. Different letters on the error bars indicate significant differences among the extraction groups and treatments (p < 0.05). AE: alkaline extract obtained at 25 °C; FE: ethanol extract obtained at 25 °C; HE: ethanol extract obtained at 70 °C; HW: hot water extract obtained at 100 °C.

**Figure 3**
Effects of *Botryocladia leptopoda* extracts on expression levels of proinflammatory-associated genes in LPS-stimulated RAW 264.7 macrophages. (a) *TNF-α* and (b) *IL-6*. Different letters on the error bars indicate significant differences among the different concentrations of the extract (p < 0.05). AE: alkaline extract obtained at 25 °C; FE: ethanol extract obtained at 25 °C.

**Figure 4**
Cytotoxic effects on L929 fibroblast cells under different concentrations of (a) FE and (b) AE treatment. Different letters on the error bars indicate significant differences among the different concentrations of the extract (p < 0.05). FE: ethanol extract obtained at 25 °C; AE: alkaline extract obtained at 25 °C.

**Figure 5**
Cell proliferation on L929 fibroblast cells under different concentrations of (a) FE and (b) AE treatment. Different letters on the error bars indicate significant differences among the different concentrations of the extract (p < 0.05). FE: ethanol extract obtained at 25 °C; AE: alkaline extract obtained at 25 °C.

**Figure 6**
Cell migration of L929 cells under different concentrations of *Botryocladia leptopoda* extracts. (a) In vitro scratch assay of cell diffusion and (b) quantitative analysis of cell diffusion. Original magnification ×40. Different letters on the error bars indicate significant differences among the different concentrations of the extract (p < 0.05). AE: alkaline extract obtained at 25 °C; FE: ethanol extract obtained at 25 °C.

**Figure 7**
Effects of different *Botryocladia leptopoda* extracts on H₂O₂-induced changes in an intracellular reduced ratio of (a) *MMP-2*, (b) *MMP-9* and (c) *TIMP-2*, mRNA expressions. Different letters on the error bars indicate significant differences among the different concentrations of the extract (p < 0.05). AE: alkaline extract obtained at 25 °C; FE: ethanol extract obtained at 25 °C.

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Botryocladia leptopoda Extracts Promote Wound Healing Ability via Antioxidant and Anti-Inflammatory Activities and Regulation of MMP/TIMP Expression

Affiliations

Botryocladia leptopoda Extracts Promote Wound Healing Ability via Antioxidant and Anti-Inflammatory Activities and Regulation of MMP/TIMP Expression

Authors

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Abstract

Conflict of interest statement

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References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Medical

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