Research Paper
Bone Regeneration
Sequential delivery of BMP-2 and BMP-7 for bone regeneration using a heparinized collagen membrane

https://doi.org/10.1016/j.ijom.2015.02.015Get rights and content

Abstract

To investigate the effect of the sequential delivery of bone morphogenetic proteins BMP-2 and BMP-7 on bone regeneration in rat calvarial defects (40 Sprague-Dawley rats, 8 mm defect size), all animals were treated with a hydroxyapatite (HA)/tricalcium phosphate (TCP) bone graft covered with a collagen membrane. The experimental groups were as follows: (1) control group: unmodified collagen (no treatment); (2) BMP-2 group: 5 μg of BMP-2; (3) hep-BMP-7 group: 5 μg BMP-7 chemically bound to heparinized collagen; and (4) BMP-2/hep-BMP-7 group: 2.5 μg BMP-7 bound to heparinized collagen and subsequently treated with 2.5 μg BMP-2. Defect healing was examined at 2 and 8 weeks after surgery. The BMP-2 group showed the largest new bone area at week 2 (29.3 ± 7.3%; P = 0.009); new bone areas in the hep-BMP-7 and BMP-2/hep-BMP-7 groups were similar (11.8 ± 3.4% and 12.9 ± 5.71%, respectively; P = 0.917). After 8 weeks, the BMP-2/hep-BMP-7 group showed the largest new bone area (43.3 ± 6.2%), followed by the BMP-2 and hep-BMP-7 groups (P = 0.013). Accordingly, in comparison with single deliveries of BMP-2 and BMP-7, sequential delivery of BMP-2 and BMP-7 using a heparinized collagen membrane significantly induced new bone formation with a smaller quantity of BMP-2 in rat calvarial defects.

Section snippets

Materials

Toluidine blue O (TBO), (3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), N-hydroxysuccinimide (NHS), and 2-morpholinoethane sulphonic acid buffer (MES) were purchased from Sigma Aldrich (St. Louis, MO, USA); heparin sulphate salt was obtained from Acros Organics (Morris Plains, NJ, USA). Collagen membranes were purchased from GENOSS (Suwon, South Korea), and ErhBMP-2 (referred to as BMP-2) and ErhBMP-7 (referred to as BMP-7) were obtained from PeproTech (Rocky Hill, NJ, USA). All other

Heparinization of collagen membranes

A schematic diagram of the designed sequential BMP delivery carrier is illustrated in Fig. 1. The morphologies of collagen membranes before and after heparin conjugation are presented in Fig. 3a. Nanofibrous collagen bundles (fibre diameter 243 ± 56 nm) were observed on the surfaces of membranes. Heparinized membranes had similar nanofibre diameters, and although some rupture of collagen fibres was observed on membrane surfaces, the fibrous structure of the collagen was maintained. The conjugation

Discussion

Controlled or sustained growth factor release is considered essential for proper bone regeneration, and many trials have been performed on the use of BMPs to enhance bone regeneration.14 BMPs are multifunctional growth factors that belong to the TGF-β family, and BMP-2 and BMP-7, in particular, play key roles in osteoblast differentiation. The conventional strategy for delivering BMPs to bone defects involves their adsorption onto the surfaces of scaffolds, but when delivered in this manner

Funding

This study was supported by a Dental Research Grant (#PNUDH-DER 2013-01) from the Pusan National University Dental Hospital.

Competing interests

None declared.

Ethical approval

The Ethics Committee on Animal Experimentation of the Korean Atomic Energy Research Institute (KAERI-IACUC-2013-004).

Patient consent

Not required.

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    These authors contributed equally in this study.

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