Integration of cellulases into bacterial cellulose: Toward bioabsorbable cellulose composites

Cellulose biodegradation resulting from enzymolysis generally occurs in nature rather than in the human body because of the absence of cellulose degrading enzymes. In order to achieve in-vivo degradation in human body for in-vivo tissue regeneration applications, we developed a bioaborbable bacterial cellulose (BBC) material, which integrates one or more cellulose degrading enzymes (cellulases), and demonstrated its degradability in vitro using buffers with pH values relevant to wound environments. We introduced a double lyophilizing process to retain the microstructure of the bacterial cellulose as well as the activity of embedded enzymes allowing for long-term storage of the material, which only requires hydration before use. Enzymes and their combinations have been examined to optimize the in-vitro degradation of the BBC material. In-vitro studies revealed that acidic cellulases from Trichoderma viride showed reasonable activity for pH values ranging from 4.5 to 6.0. A commercial cellulase (cellulase-5000) did not show good activity at pH 7.4, but its degrading ability increased when used in conjunction with a β-glucosidase from Bacillus subtilis or a β-glucosidase from Trichoderma sp. Given the harmless glucose product of the enzymatic degradation of cellulose, the BBC material may be ideal for many wound care and tissue engineering applications for the bioabsorbable purpose.