Acquired pellicle and biofilm engineering with CaneCPI-5: Insights from proteomic and microbiomics analysis

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Araujo, Tamara Teodoro ; Dionizio, Aline ; Carvalho, Thamyris de Souza ; Debortolli, Ana Luiza Bogaz ; Vertuan, Mariele ; de Souza, Beatriz Martines ; Camara, Joao Victor Fraza ; Henrique-Silva, Flavio ; Chiaratti, Marcos ; Santos, Angelica ; Alves, Lindomar ; Ferro, Milene ; Magalhaes, Ana Carolina ; Buzalaf, Marilia Afonso Rabelo

• Document type: Journal article
• Source: ARCHIVES OF ORAL BIOLOGY; v. 166, p. 8-pg., 2024-06-28.

Abstract

Objective: In this in vivo proof-of-concept study, acquired pellicle engineering was implemented to promote alterations in the protein composition of the acquired enamel pellicle (AEP) and the bacterial composition of the dental biofilm after treatment with Sugarcane cystatin (CaneCPI-5). Design: After prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with the following solutions: 1) deionized water (H2O- negative control or 2) 0.1 mg/mL CaneCPI-5. The AEP and biofilm were formed along 2 or 3 h, respectively. The AEP was collected with electrode filter papers soaked in 3 % citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. The biofilm microbiome was collected with a dental curette. The DNA was extracted, amplified, and analyzed by 16S-rRNA Next Generation Sequencing (NGS). Results: Treatment with CaneCPI-5 increased several proteins with antimicrobial, acid-resistance, affinity for hydroxyapatite, structural and calcium binding properties, such as Cysteine-rich-3 (6-fold-p = 0.03), Cystatin-B (5.5-fold-p < 0.01), Neutrophil-defensin 1 (4.7-fold-p < 0.01), Mucin (3.9-fold-p < 0.01), Immunoglobulinheavy-constant (3.8-fold-p < 0.01) and Lactotransferrin (2.8-fold-p < 0.01). Microbiome revealed that several commensal bacteria had their abundance increased after rinsing with CaneCPI-5, such as Corynebacterium and Neisseria, while Streptococcus and Prevotella nigrescens were decreased. The results indicate the efficiency of CaneCPI-5 in promoting beneficial changes in the AEP and biofilm, making this phytocystatin a potential target for incorporation into dental products. Conclusion: Cane demonstrated the capability to alter the protein composition of the acquired enamel pellicle (AEP) and the initial colonizers of the biofilm, enhancing the presence of proteins and bacteria crucial for dental protection. (AU)

• FAPESP's process: 19/26070-1 - Modulation of acquired enamel pellicle to control dental mineral loss: unravelling mechanisms to make therapies possible
• Grantee: Marília Afonso Rabelo Buzalaf
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 19/08032-5 - Acquired pellicle engineering for protection against caries and tooth erosion: effect of cystatin, statherin and hemoglobin on the proteomic profile of the acquired pellicle, bacterial biofilm and dental enamel
• Grantee: Tamara Teodoro Araujo
• Support Opportunities: Scholarships in Brazil - Doctorate