Maqui berry cystatin inhibits cathepsin k activity and stimulates osteogenic differentiation of human dental pulp cells

Authors

  • Luana Raphael da Silva Universidade Estadual Paulista (UNESP). Faculdade de Odontologia de Araraquara. Departamento de Odontologia Restauradora.
  • Eduardo Pereira de Universidade Federal de São Carlos (UFSCar., Departamento de Genética e Evolução.
  • Bárbara Roma Mendes Universidade Estadual Paulista (UNESP). Faculdade de Odontologia de Araraquara. Departamento de Odontologia Restauradora.
  • Breno Henrique Amancio Universidade Estadual Paulista (UNESP). Faculdade de Odontologia de Araraquara. Departamento de Odontologia Restauradora.
  • Flávio Henrique-Silva Universidade Federal de São Carlos (UFSCar). Departamento de Genética e Evolução.
  • Gisele Faria Universidade Estadual Paulista (UNESP). Faculdade de Odontologia de Araraquara. Departamento de Odontologia Restauradora.

DOI:

https://doi.org/10.1590/1678-7765-2025-0661

Keywords:

Cathepsin K, Cystatins, Dental pulp

Abstract

Objectives  Cystatins, endogenous inhibitors of cysteine proteases, regulate extracellular matrix degradation. Their plant-derived homologs (phytocystatins) include MaquiCPI-3, a recombinant protein obtained from Aristotelia chilensis (maqui berry). This study aimed to investigate the inhibitory effect of MaquiCPI-3 on human cathepsin K (CTSK) activity and its cytotoxicity and impact on the proliferation, migration, and osteogenic differentiation of human dental pulp cells (hDPCs). Methodology  The inhibitory activity of MaquiCPI-3 against CTSK was measured using a spectrofluorometer with the fluorogenic substrate Z-Phe-Arg-AMC. The hDPCs from third molars were characterized by flow cytometry for mesenchymal (CD90, CD73, CD105) and hematopoietic (CD34, CD45) markers. The hDPCs, either exposed to MaquiCPI-3 or left untreated (control), were assessed for viability (MTT assay), proliferation (bromodeoxyuridine incorporation), chemotaxis (Transwell assay), mineralized nodule formation (Alizarin Red S staining), alkaline phosphatase activity (thymolphthalein release), and expression of mineralization-related genes (qPCR). Data were analyzed using one- or two-way ANOVA with appropriate post hoc tests or nonparametric alternatives (α=0.05). Results  MaquiCPI-3 potently inhibited CTSK (Ki=1.72 nM, Ki,app=2.08 nM), showed no cytotoxicity, and significantly enhanced ALP activity, mineralized nodule formation, and expression of BMP-2 and osteocalcin, stimulating no hDPC proliferation or migration when compared with the control. Conclusions  MaquiCPI-3 increased no cell proliferation or migration, its ability to inhibit CTSK activity and induce an osteogenic phenotype shows promising potential therapeutic strategies aimed at repairing and regenerating pulp and periapical tissues and controlling bone resorption.

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Published

2026-02-16

Issue

Vol. 34 (2026)

Section

Original Articles

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How to Cite

Silva, L. R. da, Eduardo Pereira de, Mendes, B. R., Amancio, B. H., Henrique-Silva, F., & Faria, G. (2026). Maqui berry cystatin inhibits cathepsin k activity and stimulates osteogenic differentiation of human dental pulp cells. Journal of Applied Oral Science, 34, e20250661. https://doi.org/10.1590/1678-7765-2025-0661