Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2015, 159(1):067-076 | 10.5507/bp.2012.115
Aim: The purpose of this study was to prepare a coladerm-beta glucan membrane (CBGM) and to evaluate its biocompatibility, cytotoxicity, antimicrobial activity, genotoxicity and mutagenicity.
Methods: The biocompatibility of the membrane was studied on the base of cell adhesion and colonization of human fibroblasts on the biomaterial surface by light microscopy. The MTT test and LDH level determination in the culture medium removed from the control and cells treated on the membrane, were used for viability and cytotoxic evaluations. Flow cytometry and gel electrophoresis were used for analysis of cell cycle and death. The antimicrobial activity of CBGM was tested using the qualitative dilution method. Ames bacteria gene mutation test and Comet assay were used for mutagenic and genotoxic studies.
Results: MTT and LDH tests confirmed that CBGM is a non-toxic biomaterial. Flow cytometry and gel electrophoresis demonstrated that the membrane did not affect the cell cycle and did not induce either necrotic or apoptotic cell death. CBGM exhibited antibacterial activity against G- bacteria E. sakazakii, S. marcescens, E. coli and agains G+ sporogenic bacteria B. cereus. No antifungal activity was detected. The membrane did not induce mutagenicity in the bacterial reverse mutation test in Salmonella Typhimurium strains. Similarly, the comet assay showed that the tested fibroblast cells growing with/without the membrane did not show any statistically significant DNA damage.
Conclusions: The CBGM has good biocompatibility, no cytotoxicity/genotoxicity/mutagenicity and it can be included as a potential scaffold for tissue engineering.
Received: July 4, 2012; Accepted: December 19, 2012; Prepublished online: February 14, 2013; Published: March 9, 2015