Three dimensional printed macroporous polylactic acid/hydroxyapatite composite scaffolds for promoting bone formation in a critical-size rat calvarial defect model

Abstract We have explored the applicability of printed scaffold by comparing osteogenic ability and biodegradation property of three resorbable biomaterials. A polylactic acid/hydroxyapatite (PLA/HA) composite with a pore size of 500 μm and 60% porosity was fabricated by three-dimensional printing. Three-dimensional printed PLA/HA, β-tricalcium phosphate (β-TCP) and partially demineralized bone matrix (DBM) seeded with bone marrow stromal cells (BMSCs) were evaluated by cell adhesion, proliferation, alkaline phosphatase activity and osteogenic gene expression of osteopontin (OPN) and collagen type I (COL-1). Moreover, the biocompatibility, bone repairing capacity and degradation in three different bone substitute materials were estimated using a critical-size rat calvarial defect model in vivo. The defects were evaluated by micro-computed tomography and histological analysis at four and eight weeks after surgery, respectively. The results showed that each of the studied scaffolds had its own specific merits and drawbacks. Three-dimensional printed PLA/HA scaffolds possessed good biocompatibility and stimulated BMSC cell proliferation and differentiation to osteogenic cells. The outcomes in vivo revealed that 3D printed PLA/HA scaffolds had good osteogenic capability and biodegradation activity with no difference in inflammation reaction. Therefore, 3D printed PLA/HA scaffolds have potential applications in bone tissue engineering and may be used as graft substitutes in reconstructive surgery.

[1]  J O Hollinger,et al.  The critical size defect as an experimental model to test bone repair materials. , 1990, The Journal of craniofacial surgery.

[2]  Harold Alexander,et al.  Biological Response of Intramedullary Bone to Poly-L-Lactic Acid , 1991 .

[3]  A. Mikos,et al.  Osteoblast function on synthetic biodegradable polymers. , 1994, Journal of biomedical materials research.

[4]  C. Friedman,et al.  Synthetic bone graft substitutes. , 1994, Otolaryngologic clinics of North America.

[5]  A. Mikos,et al.  Degradation of polydispersed poly(L-lactic acid) to modulate lactic acid release. , 1995, Biomaterials.

[6]  C. M. Agrawal,et al.  Sterilization, toxicity, biocompatibility and clinical applications of polylactic acid/polyglycolic acid copolymers. , 1996, Biomaterials.

[7]  F. Gage,et al.  Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Zhang,et al.  Effect(s) of the demineralization process on the osteoinductivity of demineralized bone matrix. , 1997, Journal of periodontology.

[9]  K. Schomacker,et al.  Improved osteoinduction of cortical bone allografts: A study of the effects of laser perforation and partial demineralization , 1997, Journal of Orthopaedic Research.

[10]  C. Vacanti,et al.  An overview of tissue engineered bone. , 1999, Clinical orthopaedics and related research.

[11]  R Langer,et al.  In vitro generation of osteochondral composites. , 2000, Biomaterials.

[12]  Osteoconductivity of an injectable and bioresorbable poly(propylene glycol-co-fumaric acid) bone cement. , 2000, Biomaterials.

[13]  P. Gruber,et al.  Polylactic Acid Technology , 2000 .

[14]  M. Bostrom,et al.  An Unexpected Outcome During Testing of Commercially Available Demineralized Bone Graft Materials: How Safe Are the Nonallograft Components? , 2001, Spine.

[15]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[16]  H A Merten,et al.  Degradation characteristics of alpha and beta tri-calcium-phosphate (TCP) in minipigs. , 2002, Journal of biomedical materials research.

[17]  R. Legeros,et al.  Properties of osteoconductive biomaterials: calcium phosphates. , 2002, Clinical orthopaedics and related research.

[18]  F. Kloss,et al.  Degradation characteristics of α and β tri-calcium-phosphate (TCP) in minipigs , 2002 .

[19]  C Krettek,et al.  Comparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatite, beta-tricalcium phosphate and demineralized bone matrix. , 2003, Biomaterials.

[20]  Hans-Dieter John,et al.  Histomorphometric analysis of natural bone mineral for maxillary sinus augmentation. , 2004, The International journal of oral & maxillofacial implants.

[21]  G. Vunjak‐Novakovic,et al.  Osteogenic differentiation of human bone marrow stromal cells on partially demineralized bone scaffolds in vitro. , 2004, Tissue engineering.

[22]  D. Kaplan,et al.  In vitro and in vivo evaluation of differentially demineralized cancellous bone scaffolds combined with human bone marrow stromal cells for tissue engineering. , 2005, Biomaterials.

[23]  M. Shaw,et al.  Biodegradable HA-PLA 3-D porous scaffolds: effect of nano-sized filler content on scaffold properties. , 2005, Acta biomaterialia.

[24]  P. Coulthard,et al.  Interventions for replacing missing teeth: bone augmentation techniques for dental implant treatment. , 2006, The Cochrane database of systematic reviews.

[25]  J. Tamura,et al.  A 5-7 year in vivo study of high-strength hydroxyapatite/poly(L-lactide) composite rods for the internal fixation of bone fractures. , 2006, Biomaterials.

[26]  A. Boccaccini,et al.  Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. , 2006, Biomaterials.

[27]  Cato T Laurencin,et al.  Polymers as biomaterials for tissue engineering and controlled drug delivery. , 2006, Advances in biochemical engineering/biotechnology.

[28]  Ralph Müller,et al.  Nondestructive micro-computed tomography for biological imaging and quantification of scaffold-bone interaction in vivo. , 2007, Biomaterials.

[29]  T. Bauer,et al.  Bone graft substitutes , 2007, Skeletal Radiology.

[30]  H. De Bruyn,et al.  A literature review on biomaterials in sinus augmentation procedures. , 2007, Clinical implant dentistry and related research.

[31]  B. McAllister,et al.  Bone augmentation techniques. , 2007, Journal of periodontology.

[32]  Pierre-Yves Zambelli,et al.  Repair of critical size defects in the rat cranium using ceramic-reinforced PLA scaffolds obtained by supercritical gas foaming. , 2007, Journal of biomedical materials research. Part A.

[33]  P. Moy,et al.  Which hard tissue augmentation techniques are the most successful in furnishing bony support for implant placement? , 2007, The International journal of oral & maxillofacial implants.

[34]  Cato T Laurencin,et al.  Tissue engineering of bone: material and matrix considerations. , 2008, The Journal of bone and joint surgery. American volume.

[35]  Yilin Cao,et al.  Evaluation of Partially Demineralized Osteoporotic Cancellous Bone Matrix Combined with Human Bone Marrow Stromal Cells for Tissue Engineering: An In Vitro and In Vivo Study , 2008, Calcified Tissue International.

[36]  Heungsoo Shin,et al.  Nanofibrous poly(lactic acid)/hydroxyapatite composite scaffolds for guided tissue regeneration. , 2008, Macromolecular bioscience.

[37]  M Navarro,et al.  Biomaterials in orthopaedics , 2008, Journal of The Royal Society Interface.

[38]  C. Ohtsuki,et al.  Review Paper: Behavior of Ceramic Biomaterials Derived from Tricalcium Phosphate in Physiological Condition , 2008, Journal of biomaterials applications.

[39]  Dong Han,et al.  Ectopic Osteogenesis by Ex Vivo Gene Therapy Using Beta Tricalcium Phosphate as a Carrier , 2008, Connective tissue research.

[40]  T. Nagy,et al.  Micro-computed tomographic analysis of bone healing subsequent to graft placement. , 2009, Journal of biomedical materials research. Part B, Applied biomaterials.

[41]  David Dean,et al.  Effect of initial cell seeding density on early osteogenic signal expression of rat bone marrow stromal cells cultured on cross-linked poly(propylene fumarate) disks. , 2009, Biomacromolecules.

[42]  Huipin Yuan,et al.  Heterotopic bone formation by nano-apatite containing poly(D,L-lactide) composites. , 2010, European cells & materials.

[43]  A. Janorkar,et al.  Poly(lactic acid) modifications , 2010 .

[44]  S. H. Keshel,et al.  Clinical, Cosmetic and Investigational Dentistry Dovepress the Comparative Effectiveness of Demineralized Bone Matrix, Beta-tricalcium Phosphate, and Bovine-derived Anorganic Bone Matrix on Inflammation and Bone Formation Using a Paired Calvarial Defect Model in Rats , 2022 .

[45]  Yufeng Zheng,et al.  Cell responses and hemocompatibility of g-HA/PLA composites , 2011, Science China Life Sciences.

[46]  Wenjie Zhang,et al.  LvBMP-2 gene-modified BMSCs combined with calcium phosphate cement scaffolds for the repair of calvarial defects in rats , 2011, Journal of materials science. Materials in medicine.

[47]  Amy J Wagoner Johnson,et al.  A review of the mechanical behavior of CaP and CaP/polymer composites for applications in bone replacement and repair. , 2011, Acta biomaterialia.

[48]  T. Sawase,et al.  Lateral bone augmentation with newly developed β-tricalcium phosphate block: an experimental study in the rabbit mandible. , 2011, Clinical oral implants research.

[49]  Rozalia Dimitriou,et al.  Bone regeneration: current concepts and future directions , 2011, BMC medicine.

[50]  J. Schrooten,et al.  A calcium-induced signaling cascade leading to osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells. , 2012, Biomaterials.

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

[52]  Wenbo Jiang,et al.  Morphology, wettability, and mechanical properties of polycaprolactone/hydroxyapatite composite scaffolds with interconnected pore structures fabricated by a mini‐deposition system , 2012 .

[53]  D. Grijpma,et al.  Influence of polymer molecular weight in osteoinductive composites for bone tissue regeneration. , 2013, Acta biomaterialia.

[54]  A. Azim,et al.  Evaluation of horizontal ridge augmentation using beta tricalcium phosphate and demineralized bone matrix: A comparative study , 2013, Journal of clinical and experimental dentistry.

[55]  Wenbo Jiang,et al.  Three dimensional melt-deposition of polycaprolactone/bio-derived hydroxyapatite composite into scaffold for bone repair , 2013, Journal of biomaterials science. Polymer edition.

[56]  F. A. Sheikh,et al.  Air jet spinning of hydroxyapatite/poly(lactic acid) hybrid nanocomposite membrane mats for bone tissue engineering. , 2013, Colloids and surfaces. B, Biointerfaces.

[57]  D. Grijpma,et al.  Controlling dynamic mechanical properties and degradation of composites for bone regeneration by means of filler content. , 2013, Journal of the mechanical behavior of biomedical materials.

[58]  Marcus Abboud,et al.  Comparison of three hydroxyapatite/β-tricalcium phosphate/collagen ceramic scaffolds: an in vivo study. , 2014, Journal of biomedical materials research. Part A.

[59]  S. Dorozhkin Bioceramics from calcium orthophosphates , 2015 .

[60]  Robert Liska,et al.  3D printing of biomaterials , 2015 .