The effects of LPS and IL-1β on the expression of HBD-2 and HBD-3 in tissue engineered oral mucosa

doi:10.19438/j.cjoms.2024.06.003

Abstract

Abstract: PURPOSE: To investigate the expression of human beta-defensin 2(HBD-2) and human beta-defensin 3 (HBD-3) in tissue engineered oral mucosa equivalents under the stimulation of Escherichia coli lipopolysaccharide (LPS) and recombinant human interleukin-1(IL-1β), and to evaluate the possibility of using tissue engineered oral mucosa in the treatment of infectious oral diseases. METHODS: Real-time fluorescent quantitative reverse transcription PCR(RT-qPCR) and Western blot were used to evaluate the mRNA and protein expression of HBD-2 and HBD-3 in tissue engineered oral mucosa substitutes under the stimulation of LPS and IL-1β. SPSS 26.0 software package was used for data analysis. RESULTS: LPS and IL-1β stimulated the mRNA and protein expression of HBD-2 and HBD-3 in tissue-engineered oral mucosa, and showed a dose-dependent increase after 24 hours of culture(P<0.05). At 100 ng/mL, LPS induced HBD-2 mRNA by about 2.7-fold and HBD-3 mRNA by about 5-fold, respectively. The maximal production of HBD-2 mRNA was about 1.6-fold and HBD-3 about 4.8-fold by 20 ng/mL IL-1β. As to the results of western blot analysis, both LPS and IL-1β induced HBD-2 protein secretion at the maximal concentration in this study. HBD-3 protein was hardly observed without any treatment. After incubation with LPS and IL-1β, HBD-3 protein was significantly up-regulated(P<0.05). CONCLUSIONS: Tissue engineered oral mucosa equivalents are armed with defense activity against invading microorganisms in a sense, which may provide a therapy substitute for infectious oral diseases in the future.

Key words: Tissue engineered oral mucosa, HBD-2, HBD-3, Immune defense

CLC Number:

R739.8

WANG Yang, XUAN Jing, WANG Hai-yan, HUANG Yi, XIANG Mei-juan. The effects of LPS and IL-1β on the expression of HBD-2 and HBD-3 in tissue engineered oral mucosa[J]. China Journal of Oral and Maxillofacial Surgery, 2024, 22(6): 536-540.

References

[1] Xiong X, Jia J, He S, et al.Cryopreserved lip mucosa tissue derived keratinocytes can fabricate tissue engineered palatal mucosa equivalent[J]. J Biomed Mater Res B Appl Biomater, 2010, 94(1): 165-170.
[2] Sroussi HY, Epstein JB, Bensadoun RJ, et al.Common oral complications of head and neck cancer radiation therapy: mucositis, infections, saliva change, fibrosis, sensory dysfunctions, dental caries, periodontal disease, and osteoradionecrosis[J]. Cancer Med, 2017, 6(12): 2918-2931.
[3] Gao X, Ding J, Liao C, et al.Defensins: the natural peptide antibiotic[J]. Adv Drug Deliv Rev, 2021, 179: 114008.
[4] Fu J, Zong X, Jin M, et al.Mechanisms and regulation of defensins in host defense[J]. Signal Transduct Target Ther, 2023, 8(1): 300.
[5] Bals R.Epithelial antimicrobial peptides in host defense against infection[J]. Respir Res, 2000, 1(3): 141-150.
[6] Eggerstedt M, Torres-maldonado S, Danielian A, et al. Impact of defensins-containing body cream on skin composition[J]. J Cosmet Dermatol, 2023, 22(2): 620-627.
[7] Pazgier M, Hoover DM, Yang D, et al.Human beta-defensins[J]. Cell Mol Life Sci, 2006, 63(11): 1294-1313.
[8] Senel S.An overview of physical, microbiological and immune barriers of oral mucosa[J]. Int J Mol Sci, 2021, 22(15): 7821.
[9] Ting DSJ, Mohammed I, Lakshminarayanan R, et al.Host defense peptides at the ocular surface: roles in health and major diseases, and therapeutic potentials[J]. Front Med (Lausanne), 2022, 9: 835843.
[10] Lai J, Rigas Y, Kantor N, et al.Living with your biome: how the bacterial microbiome impacts ocular surface health and disease[J]. Expert Rev Ophthalmol, 2024, 19(2): 89-103.
[11] Mohammed AI, Sangha S, Nguyen H, et al.Assessment of oxidative stress-induced oral epithelial toxicity[J]. Biomolecules, 2023, 13(8): 1239.
[12] Zhai YJ, Feng Y, Ma X, et al.Defensins: defenders of human reproductive health[J]. Hum Reprod Update, 2023, 29(1): 126-154.
[13] Nazeer N, Rodriguez-lecompte JC, Ahmed M. Bacterial-specific aggregation and killing of immunomodulatory host defense peptides[J]. Pharmaceuticals (Basel), 2021, 14(9): 839.
[14] Chong KT, Xiang L, Wang X, et al.High level expression of human epithelial beta-defensins (hBD-1, 2 and 3) in papillomavirus induced lesions[J]. Virol J, 2006, 3: 75.
[15] Niemiec MJ, Grumaz C, Ermert D, et al.Dual transcriptome of the immediate neutrophil and Candida albicans interplay[J]. BMC Genomics, 2017, 18(1): 696.
[16] Casaroto AR, Da Silva RA, Salmeron S, et al.Candida albicans-cell interactions activate innate immune defense in human palate epithelial primary cells via nitric oxide(NO) and β-defensin 2 (hBD-2)[J]. Cells, 2019, 8(7): 707.
[17] Kim MJ, Lee WY, Choe YH.Expression of TIM-3, human β-defensin-2, and FOXP3 and correlation with disease activity in pediatric Crohn's disease with infliximab therapy[J]. Gut Liver, 2015, 9(3): 370-380.
[18] Wang P, Li G, Gao L, et al.Human β-defensin 2 enhances IL-1β production and pyroptosis through P2X7-mediated NLRP3 expression in macrophages[J]. Biocell, 2022, 46(5): 1197-1207.
[19] Zaga-Clavellina V, Martha RV, Flores-Espinosa P.In vitro secretion profile of pro-inflammatory cytokines IL-1β, TNF-α, IL-6, and of human beta-defensins(HBD)-1, HBD-2, and HBD-3 from human chorioamniotic membranes after selective stimulation with Gardnerella vaginalis[J]. Am J Reprod Immunol, 2012, 67(1): 34-43.
[20] Leeyaphan C, Hau C, Takeoka S, et al.Immune response in human chromoblastomycosis and eumycetoma - focusing on human interleukin-17A, interferon-gamma, tumour necrosis factor-alpha, interleukin-1 beta and human beta-defensin-2[J]. Mycoses, 2016, 59(12): 751-756.
[21] Lee SH, Jun HK, Lee HR, et al.Antibacterial and lipopolysaccharide (LPS)-neutralising activity of human cationic antimicrobial peptides against periodontopathogens[J]. Int J Antimicrob Agents, 2010, 35(2): 138-145.
[22] Saqib Z, De Palma G, Lu J, et al.Alterations in fecal β-defensin-3 secretion as a marker of instability of the gut microbiota[J]. Gut Microbes, 2023, 15(1): 2233679.