Biointerface control of electrospun fiber scaffolds for bone regeneration: engineered protein link to mineralized surface.
Control over the interface of biomaterials that favors the initial adhesion and subsequent differentiation of stem cells is one of the key strategies in bone tissue engineering. Here we engineer the interface of biopolymer electrospun fiber matrices with a fusion protein of fibronectin 9-10 domain (FNIII9-10) and osteocalcin (OCN), aiming to stimulate mesenchymal stem cell (MSC) functions, including initial adhesion, growth and osteogenic differentiation. In particular, a specific tethering of FNIII9-10-OCN protein was facilitated by the hydroxyapatite (HA) mineralization of the biopolymer surface through a molecular recognition of OCN to the HA crystal lattice. The FNIII9-10-OCN anchorage to the HA-mineralized fiber was observed to be highly specific and tightly bound to preserve stability over a long period. Initial cell adhesion levels, as well as the spreading shape and process, of MSCs within 24h were strikingly different between the fibers linked with and without fusion protein. Significant up-regulations in the mRNA expression of adhesion signaling molecules occurred with the fusion protein link, as analyzed by the reverse transcriptase polymerase chain reaction. The expression of a series of osteogenic-related genes at later stages, over 2-3weeks, was significantly improved in the fusion protein-tailored fiber, and the osteogenic protein levels were highly stimulated, as confirmed by immunofluorescence imaging and fluorescence-activated cell sorting analyses. In vivo study in a rat calvarium model confirmed a higher quantity of new bone formation in the fiber linked with fusion protein, and a further increase was noticed when the MSCs were tissue-engineered with the fusion protein-linked fiber. Collectively, these results indicate that FN-OCN fusion protein links via HA mineralization is a facile tool to generate a biointerface with cell-attractive and osteogenic potential, and that the engineered fibrous matrix is a potential bone regenerative scaffold.
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea.
Jae Ho
JH
Lee
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea.
Jeong-Hui
JH
Park
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea.
Ahmed
A
El-Fiqi
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea.
Joong-Hyun
JH
Kim
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea.
Ye-Rang
YR
Yun
Department of Biochemistry, Medical College, Inha University, Republic of Korea.
Jun-Hyeog
JH
Jang
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Biomaterials Science, College of Dentistry, Dankook University, Republic of Korea.
Cheol-Min
CM
Han
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea.
Eun-Jung
EJ
Lee
Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Republic of Korea; Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University Graduate School, Republic of Korea; Department of Biochemistry, Medical College, Inha University, Republic of Korea. Electronic address: kimhw@dku.edu.
Hae-Won
HW
Kim
D000818:Animals; D001483:Base Sequence; D001861:Bone Regeneration; D002113:Calcification, Physiologic; D002448:Cell Adhesion; D017931:DNA Primers; D008297:Male; D008855:Microscopy, Electron, Scanning; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D054457:Tissue Scaffolds
D017931:DNA Primers
Cell adhesion; Electrospun fiber scaffolds; Mineralized surface; Osteogenic differentiation; Protein modification