IADR Abstract Archives
Sheep Wool Keratin Based Biocomposite Scaffold for Potential Use in Regenerative Endodontics
Objectives: Objective of the present study is to develop a novel biodegradable, biocompatible and antimicrobial biocomposite comprising Keratin-Chitosan-Tricalcium Phosphate (KCTPs) for application as scaffold in Regenerative Endodontics. Extracted Keratin proteins from sheep wool have a favorable 3D matrix that allows for cellular attachment and proliferation while Chitosan has been introduced due to its broad spectrum antimicrobial activity. Tri-calcium phosphate (TCP) used in the matrix to maintain structural and mechanical performance of the composite.
Methods: Low molecular weight keratins were extracted from sheep wool using a novel cost effective chemical free method and characterised by gel electrophoresis(SDS-PAGE) for molecular weight distribution and Fourier transform infrared spectroscopy (FTIR) for protein structure. KCTPs consisiting of different ratios of Keratin, Chitosan and TCP were fabricated by solution casting and drying method. Micro structural analysis was done by Scanninig Electron Micrscopy (SEM) and Micro computed tomography(Micro CT). Cytocompatiblity was analysed with MDPC 23 (Odontoblast like cells) and OCCM 30 (Cementoblast like Cells) according to ISO 10993-Part 5 using direct testing method with Mineral Trioxide Aggregate (MTA) as control.The minimum bactericidal concentration (MBC) of KCTPs against Streptococcus mutans and Enterococcus faecalis were assessed visually by using MTT calorimetric assay .
Results: The electrophoresis patterns of the keratin powder showed bands of high protein density in the low fraction range (3.5-15 kDa) and the FTIR spectra inferred keratin adopts α-helical conformation with admixture of β-sheet.SEM of the scaffold revealed well-connected nanostructures 3D matrix of KCTPs with irregular shaped crystals of TCP dispersed among them. Micro-CT detected interconnected porous structure.The scaffold induced adherence and statistically significant proliferation rate of both MDPC 23 and OCCM 30 cells when compared to the control. The scaffold displayed bactericidal activity at a minimum concentration of 3600 μg/ml.
Conclusions: The present study reported fabrication of a novel KCTPs biocomposite by a relatively low cost method and established its cytocompatibility and antimicrobial efficacy, making it a promising scaffold material in Regenerative Endodontic Therapy.
Methods: Low molecular weight keratins were extracted from sheep wool using a novel cost effective chemical free method and characterised by gel electrophoresis(SDS-PAGE) for molecular weight distribution and Fourier transform infrared spectroscopy (FTIR) for protein structure. KCTPs consisiting of different ratios of Keratin, Chitosan and TCP were fabricated by solution casting and drying method. Micro structural analysis was done by Scanninig Electron Micrscopy (SEM) and Micro computed tomography(Micro CT). Cytocompatiblity was analysed with MDPC 23 (Odontoblast like cells) and OCCM 30 (Cementoblast like Cells) according to ISO 10993-Part 5 using direct testing method with Mineral Trioxide Aggregate (MTA) as control.The minimum bactericidal concentration (MBC) of KCTPs against Streptococcus mutans and Enterococcus faecalis were assessed visually by using MTT calorimetric assay .
Results: The electrophoresis patterns of the keratin powder showed bands of high protein density in the low fraction range (3.5-15 kDa) and the FTIR spectra inferred keratin adopts α-helical conformation with admixture of β-sheet.SEM of the scaffold revealed well-connected nanostructures 3D matrix of KCTPs with irregular shaped crystals of TCP dispersed among them. Micro-CT detected interconnected porous structure.The scaffold induced adherence and statistically significant proliferation rate of both MDPC 23 and OCCM 30 cells when compared to the control. The scaffold displayed bactericidal activity at a minimum concentration of 3600 μg/ml.
Conclusions: The present study reported fabrication of a novel KCTPs biocomposite by a relatively low cost method and established its cytocompatibility and antimicrobial efficacy, making it a promising scaffold material in Regenerative Endodontic Therapy.