Efficacy of rhBMP-2 Loaded PCL/β-TCP/bdECM Scaffold Fabricated by 3D Printing Technology on Bone Regeneration

This study was undertaken to evaluate the effect of 3D printed polycaprolactone (PCL)/β-tricalcium phosphate (β-TCP) scaffold containing bone demineralized and decellularized extracellular matrix (bdECM) and human recombinant bone morphogenetic protein-2 (rhBMP-2) on bone regeneration. Scaffolds were divided into PCL/β-TCP, PCL/β-TCP/bdECM, and PCL/β-TCP/bdECM/BMP groups. In vitro release kinetics of rhBMP-2 were determined with respect to cell proliferation and osteogenic differentiation. These three reconstructive materials were implanted into 8 mm diameter calvarial bone defect in male Sprague-Dawley rats. Animals were sacrificed four weeks after implantation for micro-CT, histologic, and histomorphometric analyses. The findings obtained were used to calculate new bone volumes (mm3) and new bone areas (%). Excellent cell bioactivity was observed in the PCL/β-TCP/bdECM and PCL/β-TCP/bdECM/BMP groups, and new bone volume and area were significantly higher in the PCL/β-TCP/bdECM/BMP group than in the other groups (p < .05). Within the limitations of this study, bdECM printed PCL/β-TCP scaffolds can reproduce microenvironment for cells and promote adhering and proliferating the cells onto scaffolds. Furthermore, in the rat calvarial defect model, the scaffold which printed rhBMP-2 loaded bdECM stably carries rhBMP-2 and enhances bone regeneration confirming the possibility of bdECM as rhBMP-2 carrier.

[1]  K. Kusumoto,et al.  Experimental studies on bone inducing activity of composites of atelopeptide type I collagen as a carrier for ectopic osteoinduction by rhBMP-2. , 1995, Biochemical and biophysical research communications.

[2]  Yuanhua Lin,et al.  Calcium ion release and osteoblastic behavior of gelatin/beta-tricalcium phosphate composite nanofibers fabricated by electrospinning , 2012 .

[3]  I. Noh,et al.  Characterization of low-molecular-weight hyaluronic acid-based hydrogel and differential stem cell responses in the hydrogel microenvironments. , 2009, Journal of biomedical materials research. Part A.

[4]  W. Bowen,et al.  Hydrogels derived from demineralized and decellularized bone extracellular matrix , 2013, Acta biomaterialia.

[5]  J. Wozney,et al.  Augmentation of Alveolar Bone and Dental Implant Osseointegration: Clinical Implications of Studies with rhBMP-2 A Comprehensive Review , 2001, The Journal of bone and joint surgery. American volume.

[6]  Dong-Woo Cho,et al.  Fabrication of blended polycaprolactone/poly(lactic-co-glycolic acid)/β-tricalcium phosphate thin membrane using solid freeform fabrication technology for guided bone regeneration. , 2013, Tissue engineering. Part A.

[7]  Myron Nevins,et al.  De novo bone induction by recombinant human bone morphogenetic protein-2 (rhBMP-2) in maxillary sinus floor augmentation. , 2005, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[8]  A. Burstein,et al.  The healing of segmental bone defects induced by demineralized bone matrix. A radiographic and biomechanical study. , 1984, The Journal of bone and joint surgery. American volume.

[9]  D W Hutmacher,et al.  The stimulation of healing within a rat calvarial defect by mPCL-TCP/collagen scaffolds loaded with rhBMP-2. , 2009, Biomaterials.

[10]  J. Wozney,et al.  Periodontal repair in dogs: evaluation of rhBMP-2 carriers. , 1996, The International journal of periodontics & restorative dentistry.

[11]  R L Reis,et al.  Bone morphogenetic proteins in tissue engineering: the road from the laboratory to the clinic, part I (basic concepts) , 2008, Journal of tissue engineering and regenerative medicine.

[12]  Karoly Jakab,et al.  Tissue engineering by self-assembly and bio-printing of living cells , 2010, Biofabrication.

[13]  Yunn-Shiuan Liao,et al.  3D Printing Bioceramic Porous Scaffolds with Good Mechanical Property and Cell Affinity , 2015, PloS one.

[14]  Sung‐Wook Choi,et al.  Effects of different rhBMP-2 release profiles in defect areas around dental implants on bone regeneration , 2015, Biomedical materials.

[15]  Dong-Woo Cho,et al.  Stimulation of healing within a rabbit calvarial defect by a PCL/PLGA scaffold blended with TCP using solid freeform fabrication technology , 2012, Journal of Materials Science: Materials in Medicine.

[16]  J. Shim,et al.  Development and Assessment of a 3D-Printed Scaffold with rhBMP-2 for an Implant Surgical Guide Stent and Bone Graft Material: A Pilot Animal Study , 2017, Materials.

[17]  Yun Lu,et al.  Segmental bone regeneration using an rhBMP-2-loaded gelatin/nanohydroxyapatite/fibrin scaffold in a rabbit model. , 2009, Biomaterials.

[18]  G. Balian,et al.  The use of demineralized bone matrix in the repair of segmental defects. Augmentation with extracted matrix proteins and a comparison with autologous grafts. , 1986, The Journal of bone and joint surgery. American volume.

[19]  J. Hollinger,et al.  Demineralized bone matrix in bone repair: History and use☆ , 2012, Advanced Drug Delivery Reviews.

[20]  Dong-Woo Cho,et al.  Effects of 3D-Printed Polycaprolactone/β-Tricalcium Phosphate Membranes on Guided Bone Regeneration , 2017, International journal of molecular sciences.

[21]  B. Guillaume,et al.  Clinical Performance of a Highly Porous Beta-TCP as the Grafting Material for Maxillary Sinus Augmentation , 2014, Implant dentistry.

[22]  Sophia P Pilipchuk,et al.  3D-printed Bioresorbable Scaffold for Periodontal Repair , 2015, Journal of dental research.

[23]  Min Soo Bae,et al.  The effect of immobilization of heparin and bone morphogenic protein-2 (BMP-2) to titanium surfaces on inflammation and osteoblast function. , 2011, Biomaterials.

[24]  Hollinger,et al.  Sustained release emphasizing recombinant human bone morphogenetic protein-2. , 1998, Advanced drug delivery reviews.

[25]  K. Char,et al.  Delivery of bone morphogenetic protein-2 and substance P using graphene oxide for bone regeneration , 2014, International journal of nanomedicine.

[26]  D. Cho,et al.  Systemically replicated organic and inorganic bony microenvironment for new bone formation generated by a 3D printing technology , 2016 .

[27]  M. Casal,et al.  Bone morphogenetic proteins in tissue engineering: the road from laboratory to clinic, part II (BMP delivery) , 2008, Journal of tissue engineering and regenerative medicine.

[28]  George P McCabe,et al.  Maintenance of human hepatocyte function in vitro by liver-derived extracellular matrix gels. , 2010, Tissue engineering. Part A.

[29]  E. Brunner,et al.  Nonparametric Analysis of Ordered Categorical Data in Designs with Longitudinal Observations and Small Sample Sizes , 2000 .

[30]  Dong-Woo Cho,et al.  Surface modification with fibrin/hyaluronic acid hydrogel on solid-free form-based scaffolds followed by BMP-2 loading to enhance bone regeneration. , 2011, Bone.

[31]  Jung-Bo Huh,et al.  Effect of immobilization of the recombinant human bone morphogenetic protein 2 (rhBMP-2) on anodized implants coated with heparin for improving alveolar ridge augmentation in beagle dogs: Radiographic observations , 2013 .

[32]  E. Sachlos,et al.  Making tissue engineering scaffolds work. Review: the application of solid freeform fabrication technology to the production of tissue engineering scaffolds. , 2003, European cells & materials.

[33]  M J Yaszemski,et al.  Three-dimensional culture of rat calvarial osteoblasts in porous biodegradable polymers. , 1998, Biomaterials.

[34]  T. Adachi,et al.  Framework for optimal design of porous scaffold microstructure by computational simulation of bone regeneration. , 2006, Biomaterials.

[35]  Dong-Woo Cho,et al.  Cell adhesion and proliferation evaluation of SFF-based biodegradable scaffolds fabricated using a multi-head deposition system , 2009, Biofabrication.

[36]  D. Markel,et al.  Characterization of the inflammatory response to four commercial bone graft substitutes using a murine biocompatibility model , 2012, Journal of inflammation research.

[37]  F. O'Brien Biomaterials & scaffolds for tissue engineering , 2011 .

[38]  J. Shim,et al.  Comparative Efficacies of Collagen-Based 3D Printed PCL/PLGA/β-TCP Composite Block Bone Grafts and Biphasic Calcium Phosphate Bone Substitute for Bone Regeneration , 2017, Materials.

[39]  Deok‐Ho Kim,et al.  Printing three-dimensional tissue analogues with decellularized extracellular matrix bioink , 2014, Nature Communications.

[40]  H. Jennissen,et al.  Molecular Modelling of Bone Morphogenetic Protein-2 (BMP-2) by 3D-Rapid Prototyping Molekulares Modellieren des knochenmorphogenetischen Proteins-2 (BMP-2) mit Hilfe des 3D-Rapid Prototyping , 2001 .