Inclusion of a recombinant sugarcane cistatin in dental products for the protection of dental enamel

• Grant number: 17/22771-0
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Duration: February 01, 2019 - October 31, 2019
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Adelita Carolina Santiago
• Grantee: Adelita Carolina Santiago
• Host Company: Ravenna Indústria e Comércio de Cosméticos Ltda. - ME
• CNAE: Fabricação de cosméticos, produtos de perfumaria e de higiene pessoal
• City: Bauru
• Pesquisadores principais: Flavio Henrique da Silva ; Marília Afonso Rabelo Buzalaf
• Associated scholarship(s): 19/04541-2 - Inclusion of a recombinant sugarcane cistatin in dental products for the protection of dental enamel, BP.PIPE

Abstract

Cystatins are reversible inhibitors of cysteine peptidases present in all forms of life. Human cystatin B has recently been identified in the acquired dental enamel film as an acid-resistant protein, which demonstrates its potential to be used in dental products for protection against caries and tooth erosion. However, the cost of this cystatin is expansive, which makes its inclusion in commercial products unfeasible. Alternatively, we studied the inhibitory potential and the interaction capacity with the dental enamel of a recombinant sugarcane cystatin, called CaneCPI-5, which was produced in E. coli. CaneCPI-5 demonstrated a strong interaction force with enamel and significantly reduced initial erosion. Due to the good results a patent was deposited and an article published in an important international journal of dentistry, we now intend to evaluate the production of CaneCPI-5 in Pichia pastoris with the advantage that protein is secreted into the induction medium, which makes the production process simpler and economically more favorable. However, it is necessary to verify if the protein produced in Pichia pastoris has the same protection against initial erosion. To do so, we will re-evaluate the protective capacity of the solution containing CaneCPI-5 against initial erosion and further evaluate the inclusion of this protein in another vehicle, a dental gel, in order to define the best vehicle for commercial use in preventing erosion. For this, in vitro experiments with bovine enamel blocks incubated with solution or gel containing CaneCPI-5 and then exposed to citric acid will be performed and then the surface microhardness tests will be performed to measure softening of the enamel. The data will be evaluated in relation to normality and homogeneity, to select the appropriate statistical test (p <0.05). In addition, the cytotoxicity of CaneCPI-5 in fibroblasts culture will be evaluated in order to prove their safety, an essential step for inclusion in commercial dental products. (AU)