Abstract:Background and Objectives: Bacterial collagenase from Closrtidium histolyticum is widely used as a clinical tool in the nonsurgical treatment of Dupuytren’s disease in eye’s disorders treatment, for enzymatic debridement, for accelerated resorption of catgut sutures. Collagenase main feature is its ability to digest key protein of the animal extracellular matrix – collagen. Dynamic Light Scattering (DLS) technique allows for investigation of collagen and collagenase solutions in conditions close to physiological. Varying the solution parameters (pH, temperature, solvent type) and adding activators or inhibitors of collagenase one can simulate the processes in living organisms. Materials and Methods: DLS method enables one to assess the translation diffusion coefficient of particles in solutions by analyzing the characteristic time of scattered light intensity fluctuations. Experiments were carried out using the photon-correlation spectrometer Photocor-Complex with diode laser (wavelength 647 nm, power 25 mW). Collagen type I from calf skin and Bacterial collagenase from Clostridium histolyticum type IA produced by Sigma-Aldrich were used. Results: Using the DLS method we have experimentally obtained: dependence of the translation diffusion coefficient dependence on pH in collagenase water solutions (pI 6.0), translation diffusion coefficient on time dependencies of scattering particles in “collagen+collagenase” Tris-HCl buffer solution without additives and with addition of CaCl$_2$ . Conclusion: DLS enables us to monitor the dynamics of collagen biodegradation in real time. Different phy siological states can be simulated for in-vitro investigation by varying temperature, solution formula and collagen-collagenase ratio.
Keywords:collagen type I, bacterial collagenase, calcium chloride, activator, biodegradation, fluorescence spectroscopy, dynamic light scattering.
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