Physical, mechanical, and biological properties of collagen membranes for guided bone regeneration: a comparative in vitro study

Abstract

Background: To provide a reference for clinical selection of collagen membranes by analyzing the properties of three kinds of collagen membranes widely used in clinics: Bio-Gide membrane from porcine dermis (PD), Heal-All membrane from bovine dermis (BD), and Lyoplant membrane from bovine pericardium (BP).

Methods: The barrier function of three kinds of collagen membranes were evaluated by testing the surface morphology, mechanical properties, hydrophilicity, and degradation rate of collagen membranes in collagenase and artificial saliva. In addition, the bioactivity of each collagen membrane as well as the proliferation and osteogenesis of MC3T3-E1 cells were evaluated. Mass spectrometry was also used to analyze the degradation products.

Results: The BP membrane had the highest tensile strength and Young's modulus as well as the largest water contact angle. The PD membrane exhibited the highest elongation at break, the smallest water contact angle, and the lowest degradation weight loss. The BD membrane had the highest degradation weight loss, the highest number of proteins in its degradation product, the strongest effect on the proliferation of MC3T3-E1 cells, and the highest expression level of osteogenic genes.

Conclusions: The PD membrane is the best choice for shaping and maintenance time, while the BD membrane is good for osteogenesis, and the BP membrane is suitable for spatial maintenance. To meet the clinical requirements of guided bone regeneration, using two different kinds of collagen membranes concurrently to exert their respective advantages is an option worth considering.

Keywords: Barrier membrane; Bioactivity; Collagen membrane; Degradation products; Guided bone regeneration.

Fig. 1

Gross appearance (scale bar: 1 cm) and SEM (scale bar: 50 μm) images of collagen membranes

Fig. 2

Comparison of the physical characteristics of different collagen membranes. a Tensile strength, b elongation at break, and c Young’s modulus of collagen membranes in the dry and wet state. The error bars represent ± SD. Two-way analysis of variance analysis. ns: not significant; ****P < 0.0001 vs PD; #### P < 0.0001 vs BP. n = 3 for all samples. d Water contact angle of collagen membranes. Mean ± SD shown. n = 5 for all samples

Fig. 3

Comparison of the degradation of different collagen membranes. a Photographs of collagen membranes before and 21 days after degradation using collagenase. Degradation weight loss (%) of the collagen membranes incubated with artificial saliva (b) and collagenase (c) after 7, 14, and 21 days. The error bars represent ± SD. Two-way analysis of variance analysis. ns: not significant, *P < 0.05, ***P <0.001, ****P < 0.0001 vs PD. n = 3 for all samples. d SDS-PAGE-Tricine gel electrophoresis. M: low molecular weight marker. Lane 1 and 2: PD incubated with or without collagenase; Lane 3 and 4: BP incubated with or without collagenase; Lane 5 and 6: BD incubated with or without collagenase; Lane 7: Collagenase solution

Fig. 4

Effects of collagen membranes degradation products on the proliferation and differentiation of MC3T3-E1 cells. CCK-8 proliferation assay performed with and without collagen membranes degradation products after 1, 3, and 5 days. b Alkaline phosphatase (ALP) activity of MC3T3-E1 cells incubated with or without collagen membranes degradation products for 1, 7, and 14 days. ALP activity was normalized by total cellular protein amounts. qPCR analysis of osteoblastic marker (c) runt-related transcription factor 2 (Runx2), (d) osteocalcin (OC), and (e) collagen type I (Col-1) relative expression in MC3T3-E1 cells incubated with or without collagen membranes degradation products for 7 and 14 days. The error bars represent ± SD. Two-way analysis of variance analysis. ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.05, ****P < 0.0001. n = 3 for all samples

Fig. 5

Analysis of peptides identified by LC–MS after in vitro degradation of three collagen membranes. a Identified proteins and peptides. b Length distribution of peptides (amino acids). X-axis represents the length of the peptides, and y-axis represents count of peptides. c Protein origin distribution