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Functionalization of multilayered DNA-coatings with bone morphogenetic protein 2
J J J P van den Beucken1, X F Walboomers, O C Boerman
1Department of Periodontology and Biomaterials, Radboud University Nijmegen Medical Center 309, PO Box 9101, 6500 HB, Nijmegen, the Netherlands.
Summary
This study demonstrates functionalizing DNA coatings with bone morphogenetic protein 2 (BMP-2) via different loading methods. BMP-2 loaded coatings show sustained release and impact osteoblast-like cell mineralization, with superficial and double-layer loading accelerating calcium deposition.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Multilayered DNA coatings offer a versatile platform for biomolecule immobilization.
- Bone morphogenetic protein 2 (BMP-2) is a key osteoinductive factor crucial for bone regeneration.
- Controlled delivery of BMP-2 is essential for effective bone tissue engineering strategies.
Purpose of the Study:
- To functionalize multilayered DNA coatings with BMP-2 using distinct loading modalities.
- To investigate the release kinetics of BMP-2 from these functionalized coatings.
- To evaluate the in vitro biological effects of BMP-2 loaded DNA coatings on osteoblast-like cells.
Main Methods:
- Fabrication of multilayered DNA coatings using poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) and DNA via electrostatic self-assembly (ESA).
- Loading of radiolabeled BMP-2 into coatings using superficial (s), deep (d), and double-layer (dl) modalities.
- Quantification of BMP-2 loading and release using radiolabeling.
- In vitro assessment of osteoblast-like cell behavior (proliferation, differentiation, mineralization, morphology).
Main Results:
- Successful functionalization of DNA coatings with BMP-2 via s, d, and dl loading modalities, with incorporation ratios observed as s+(4*d)=dl.
- All loading methods resulted in an initial burst release of BMP-2, followed by sustained release.
- Superficial (s) and double-layer (dl) BMP-2 loaded coatings accelerated calcium deposition in osteoblast-like cells without affecting proliferation.
- Deep (d) loaded coatings decreased calcium deposition in osteoblast-like cells.
Conclusions:
- Multilayered DNA coatings can be effectively functionalized with BMP-2 using different loading strategies.
- The loading modality significantly influences BMP-2 release kinetics and its biological impact on osteoblast-like cells.
- Superficial and double-layer BMP-2 loading show promise for enhancing bone mineralization in vitro.