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The purpose of this study was to evaluate the bone regeneration efficacy of an 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)-cross-linked collagen membrane for guided bone regeneration (GBR). A non-cross-linked collagen membrane (Control group), and an EDC-cross-linked collagen membrane (Test group) were used in this study. In vitro, mechanical, and degradation testing and cell studies were performed. In the animal study, 36 artificial bone defects were formed in the mandibles of six beagles. Implants were inserted at the time of bone grafting, and membranes were assigned randomly. Eight weeks later, animals were sacrificed, micro-computed tomography was performed, and hematoxylin-eosin stained specimens were prepared. Physical properties (tensile strength and enzymatic degradation rate) were better in the Test group than in the Control group. No inflammation or membrane collapse was observed in either group, and bone volumes (%) in defects around implants were similar in the two groups (p > 0.05). The results of new bone areas (%) analysis also showed similar values in the two groups (p > 0.05). Therefore, it can be concluded that cross-linking the collagen membranes with EDC is the method of enhancing the physical properties (tensile strength and enzymatic degradation) of the collagen membranes without risk of toxicity.

This study aimed to compare two methods of crosslinking collagen type I on implanted titanium surfaces, that is, using glutaraldehyde (GA) or gamma-rays (GRs), in a beagle dog model. For in vivo experiments, implants were allocated to three groups and applied to mandibular bone defects in beagle dogs; Group SLA; non-treated Sandblasted, large grit, acid-etched (SLA) implants, Group GA; SLA implants coated with GA crosslinked collagen type I, Group GR; SLA surface implants coated with collagen type I and crosslinked using 25 kGy of 60Co gamma radiation. New bone μCT volumes were obtained, and histologic and histometric analyses were performed in regions of interest. The GR group had significantly better new bone areas (NBAs) and bone to implant contact (BIC) results than the SLA group (p < 0.05), but the GA and GR groups were similar in this respect. New bone volumes and inter-thread bone densities (ITBD) were non-significantly different in the three groups (p > 0.05). Within the limits of this study, gamma-ray collagen crosslinking on titanium implants can be considered a substitute for glutaraldehyde crosslinking.

The purpose of this study was to evaluate the bone-generating ability of a new bovine-derived xenograft (S1-XB) containing hydrogel. For control purposes, we used Bio-Oss and Bone-XB bovine-derived xenografts. S1-XB was produced by mixing Bone-XB and hydrogel. Cell proliferation and differentiation studies were performed to assess cytotoxicities and cell responses. For in vivo study, 8 mm-sized cranial defects were formed in 16 rabbits, and then the bone substitutes were transplanted into defect sites in the four study groups, that is, a Bio-Oss group, a Bone-XB group, an S1-XB group, and a control (all n = 4); in the control group defects were left empty. Eight weeks after surgery, new bone formation areas were measured histomorphometrically. In the cell study, extracts of Bio-Oss, Bone-XB, and S1-XB showed good results in terms of the osteogenic differentiation of human mesenchymal stem cells (hMSCs) and no cytotoxic reaction was evident. No significant difference was observed between mean new bone areas in the Bio-Oss (36.93 ± 4.27%), Bone-XB (35.07 ± 3.23%), and S1-XB (30.80 ± 6.41%) groups, but new bone area was significantly smaller in the control group (18.73 ± 5.59%) (p < 0.05). Bovine-derived bone graft material containing hydrogel (S1-XB) had a better cellular response and an osteogenic effect similar to Bio-Oss.