Levels of low-molecular-weight hyaluronan in periodontitis-treated patients and its immunostimulatory effects on CD4+ T lymphocytes



During periodontitis, chronic inflammation triggers soft tissue breakdown, and hyaluronan is degraded into fragments of low molecular weight (LMW-HA). This investigation aimed to elucidate whether LMW-HA fragments with immunogenic potential on T lymphocytes remain in periodontal tissues after periodontal treatment.

Materials and methods

GCF samples were obtained from 15 periodontitis-affected patients and the LMW-HA, RANKL, and OPG levels were analyzed before and after 6 months of periodontal treatment by ELISA. Eight healthy individuals were analyzed as controls. Besides, human T lymphocytes were purified, exposed to infected dendritic cells, and pulsed with LMW-HA. Non-treated T lymphocytes were used as control. The expression levels of the transcription factors and cytokines that determine the Th1, Th17, and Th22 lymphocyte differentiation and function were analyzed by RT-qPCR. Similarly, the expression levels of RANKL and CD44 were analyzed.


In the GCF samples of periodontitis-affected patients, higher levels of LMW-HA were detected when compared with those of healthy individuals (52.1 ± 15.4 vs. 21.4 ± 12.2, p < 0.001), and these increased levels did not decrease after periodontal therapy (52.1 ± 15.4 vs. 45.7 ± 15.9, p = 0.158). Similarly, the RANKL levels and RANKL/OPG ratios did not change after periodontal therapy. Furthermore, in human T lymphocytes, LMW-HA induced higher expression levels of the Th1, Th17, and Th22-related transcription factors and cytokines, as well as CD44 and RANKL, as compared with non-treated cells.


In some patients, increased levels of LMW-HA persist in periodontal tissues after conventional periodontal therapy, and these remaining LMW-HA fragments with immunostimulatory potential could induce the polarization of a pathologic Th1/Th17/Th22-pattern of immune response on T lymphocytes.

Clinical relevance

The persistence of increased levels of LMW-HA in periodontal tissues after periodontal therapy could favor the recurrence of the disease and further breakdown of periodontal supporting tissues.

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Data availability

All data generated and analyzed during this study are included in this article.


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We are grateful to Darna Venegas (Microbiology Laboratory, Faculty of Dentistry, Universidad de Chile) for sharing her expertise on bacterial cultures.


This investigation has been financially supported by grants FONDECYT 1181780 (RV) and FONDECYT 11190073 (CC), from the Chilean Governmental Agencia Nacional de Investigación y Desarrollo (ANID). GM was a recipient of the Ph.D. Scholarship CONICYT 21170297 from ANID. EAC was a recipient of the Ph.D. scholarship from the School of Graduates of the Faculty of Dentistry, Universidad de Chile.

Author information




All authors made substantial contributions to the conception and design of the study. FC and GM organized the study, processed the blood samples, performed the cell cultures, and obtained the RNA samples. FC and GM carried out the ELISA experiments and performed the data analysis. JPI, JG, and GM carried out the RT-qPCR experiments and performed the data analysis. FC, EAC, and EV were involved in drafting the manuscript and figures. PC examined the study individuals, performed the periodontal diagnosis and treatment, and critically evaluated and supplemented the manuscript. RV carried out the gingival crevicular fluid sampling. CC and RV designed and implemented the research protocol, performed the data analysis, and prepared the final version of the figures and manuscript for submission. All the authors read and approved the final manuscript.

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Correspondence to Rolando Vernal.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The performed study design was reviewed and approved by the Ethics Committee for Human Research of Faculty of Dentistry from Universidad de Chile (Protocol #2010/14). Informed consent was obtained from all individual participants included in the study.

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Castillo, F., Monasterio, G., Ibarra, J.P. et al. Levels of low-molecular-weight hyaluronan in periodontitis-treated patients and its immunostimulatory effects on CD4+ T lymphocytes. Clin Oral Invest (2021). https://doi.org/10.1007/s00784-021-03808-9

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  • LMW-HA
  • Hyaluronic acid
  • T lymphocytes
  • Cytokines
  • CD44