Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering

Abstract

The aim of this study was to characterize the physicochemical properties of bacterial cellulose (BC) membranes functionalized with osteogenic growth peptide (OGP) and its C-terminal pentapeptide OGP[10-14], and to evaluate in vitro osteoinductive potential in early osteogenesis, besides, to evaluate cytotoxic, genotoxic and/or mutagenic effects. Peptide incorporation into the BC membranes did not change the morphology of BC nanofibers and BC crystallinity pattern. The characterization was complemented by Raman scattering, swelling ratio and mechanical tests. In vitro assays demonstrated no cytotoxic, genotoxic or mutagenic effects for any of the studied BC membranes. Culture with osteogenic cells revealed no difference in cell morphology among all the membranes tested. Cell viability/proliferation, total protein content, alkaline phosphatase activity and mineralization assays indicated that BC-OGP membranes enabled the highest development of the osteoblastic phenotype in vitro. In conclusion, the negative results of cytotoxicity, genotoxicity and mutagenicity indicated that all the membranes can be employed for medical supplies, mainly in bone tissue engineering/regeneration, due to their osteoinductive properties.

Bacterial Cellulose Membranes as a Potential Drug Delivery System for Photodynamic Therapy of Skin Cancer

“…The cytotoxicity potential of pure BC was already evaluated by Saska et al, 52 which demonstrated absence of in vitro cytotoxicity effects of BC membranes. In the present study, cytotoxicity assay demonstrated that BC-ClAlPc membrane was non-cytotoxic in CHO-K1 cells.…”

Bacterial Cellulose

“…BC has been proposed for application in tissue engineering as a scaffold for cartilage [66,67], bone repair [68][69][70][71], vascular grafts [64,[72][73][74][75] and neural repair [75][76][77][78], or as a barrier membrane for guided bone regeneration [57,58,60,70,71,79]. In addition several composite materials have been developed with the purpose of rendering BC bioactive such as hydroxyapatite (HA) [79][80][81][82][83][84][85], collagen [70,[86][87][88], gelatin [85,89], peptides [71,90] and some polymers such as chitosan [35], silk [90][91][92][93], polyhydroxybutyrate [94,95] and poly(2-hydroxyethyl methacrylate) [96].…”

“…In addition several composite materials have been developed with the purpose of rendering BC bioactive such as hydroxyapatite (HA) [79][80][81][82][83][84][85], collagen [70,[86][87][88], gelatin [85,89], peptides [71,90] and some polymers such as chitosan [35], silk [90][91][92][93], polyhydroxybutyrate [94,95] and poly(2-hydroxyethyl methacrylate) [96]. The incorporation in BC of antimicrobial agents such as propolis, chitosan, potassium sorbate and silver nanoparticles has been already mentioned.…”

“…Other peptides have been incorporated to BC as the osteogenic growth peptide (OGP: H 2 N-ALKRQGRTLYGFGG-OH) and its C-terminal pentapeptide H 2 N-YGFGG-OH [OGP ( [10][11][12][13][14])], which confer an osteoinductive property [71]. OGP is identical to the C-terminal amino acid sequence of histone H4.…”

“…In vitro studies have revealed that BC or BC functionalized with OGP peptides enable the highest development of the osteoblastic phenotype. Moreover they have shown negative results for cytotoxicity, genotoxicity and mutagenicity indicating that they can be employed for bone tissue engineering [71].…”