In vitro cellular responses to scaffolds containing two microencapulated growth factors.

Growth factors play an important role in the complex cascade of tissue events in periodontal regeneration, although optimal methods of delivery remain to be identified. We hypothesize that multiple delivery of growth factors, particularly via a microparticle-containing scaffold, will enhance cellular events leading to periodontal regeneration. In this study, cellular responses of periodontal ligament fibroblasts (PDLFs) in scaffolds containing microparticles (MPs) loaded with either bone morphogenetic protein (BMP)-2, insulin-like growth factor (IGF)-1, or a mixture of both MPs were evaluated, and the dual-MP-containing scaffold exhibited the release of different proteins in a sustained and independent fashion. When PDLF-seeded scaffolds were cultured in a flow perfusion bioreactor, cell metabolism and proliferation of PDLFs were significantly increased within 3 days in all IGF-1-containing scaffolds compared with those in groups lacking IGF-1 and particulate delivery enhanced these effects between 3 and 7 days. The dual-MP-containing group showed the most positive results. Both the BMP-2-in-MP and IGF-1-in-MP groups showed greater effects of alkaline phosphatase activity, more osteocalcin and osteopontin production, and more calcium deposition compared with matched GF-adsorbed groups. All osteoblastic markers were at their highest in the dual-MP-containing group at all detected time points. The combined results suggest that our dual-MP-containing scaffold can be used as a cell vehicle to positively affect cell behavior, thus exhibiting the potential to be a candidate scaffold for future periodontal tissue engineering.

[1]  F. Schwarz,et al.  Regeneration of periodontal tissues: combinations of barrier membranes and grafting materials - biological foundation and preclinical evidence: a systematic review. , 2008, Journal of clinical periodontology.

[2]  J. Jansen,et al.  Effect of dual growth factor delivery on chondrogenic differentiation of rabbit marrow mesenchymal stem cells encapsulated in injectable hydrogel composites. , 2009, Journal of biomedical materials research. Part A.

[3]  Alain Meunier,et al.  Bovine BMP osteoinductive potential enhanced by functionalized dextran-derived hydrogels. , 2005, Biomaterials.

[4]  Yan Jin,et al.  Enhancement of periodontal tissue regeneration by locally controlled delivery of insulin-like growth factor-I from dextran-co-gelatin microspheres. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[5]  R. Genco,et al.  Periodontal regeneration. , 2014, Journal of periodontology.

[6]  Dhirendra S Katti,et al.  Growth factor-delivery systems for tissue engineering: a materials perspective , 2006, Expert review of medical devices.

[7]  Tatsuo Nakamura,et al.  Novel approach to regeneration of periodontal tissues based on in situ tissue engineering: effects of controlled release of basic fibroblast growth factor from a sandwich membrane. , 2003, Tissue engineering.

[8]  Antonios G. Mikos,et al.  Flow Perfusion Enhances the Calcified Matrix Deposition of Marrow Stromal Cells in Biodegradable Nonwoven Fiber Mesh Scaffolds , 2005, Annals of Biomedical Engineering.

[9]  Q. Jin,et al.  Current concepts in periodontal bioengineering. , 2005, Orthodontics & craniofacial research.

[10]  S. Froum,et al.  Current concepts of periodontal regeneration. A review of the literature. , 2002, The New York State Dental Journal.

[11]  Antonios G Mikos,et al.  Effect of flow perfusion on the osteogenic differentiation of bone marrow stromal cells cultured on starch-based three-dimensional scaffolds. , 2003, Journal of biomedical materials research. Part A.

[12]  Yan Jin,et al.  Novel glycidyl methacrylated dextran (Dex-GMA)/gelatin hydrogel scaffolds containing microspheres loaded with bone morphogenetic proteins: formulation and characteristics. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[13]  Giuseppe Polimeni,et al.  Biology and principles of periodontal wound healing/regeneration. , 2006, Periodontology 2000.

[14]  Yan Jin,et al.  Periodontal regeneration using novel glycidyl methacrylated dextran (Dex-GMA)/gelatin scaffolds containing microspheres loaded with bone morphogenetic proteins. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[15]  Sha Huang,et al.  Release of bioactive BMP from dextran-derived microspheres: a novel delivery concept. , 2006, International journal of pharmaceutics.

[16]  G. Xiao,et al.  Bone Morphogenetic Protein 2 Induces Dental Follicle Cells to Differentiate Toward a Cementoblast/Osteoblast Phenotype , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[17]  Eben Alsberg,et al.  Dual growth factor delivery and controlled scaffold degradation enhance in vivo bone formation by transplanted bone marrow stromal cells. , 2004, Bone.

[18]  Fabiana Quaglia,et al.  Bioinspired tissue engineering: the great promise of protein delivery technologies. , 2008, International journal of pharmaceutics.

[19]  G N King,et al.  Bone morphogenetic protein-2 stimulates cell recruitment and cementogenesis during early wound healing. , 2001, Journal of clinical periodontology.

[20]  A. E. Elçin,et al.  Differentiation of human embryonic stem cells on periodontal ligament fibroblasts in vitro. , 2008, Artificial organs.

[21]  D. Puleo,et al.  In vitro effects of combined and sequential delivery of two bone growth factors. , 2004, Biomaterials.

[22]  M. Somerman,et al.  Periodontal ligament cells and gingival fibroblasts respond differently to attachment factors in vitro. , 1989, Journal of periodontology.

[23]  J. Tessmar,et al.  Matrices and scaffolds for protein delivery in tissue engineering. , 2007, Advanced drug delivery reviews.

[24]  David L Kaplan,et al.  Growth factor gradients via microsphere delivery in biopolymer scaffolds for osteochondral tissue engineering. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[25]  Yan Jin,et al.  Localized delivery of growth factors for periodontal tissue regeneration: Role, strategies, and perspectives , 2009, Medicinal research reviews.

[26]  J. Jansen,et al.  Degradable hydrogel scaffolds for in vivo delivery of single and dual growth factors in cartilage repair. , 2007, Osteoarthritis and cartilage.

[27]  Michael J Yaszemski,et al.  Retention of in vitro and in vivo BMP-2 bioactivities in sustained delivery vehicles for bone tissue engineering. , 2008, Biomaterials.

[28]  Jeremy J Mao,et al.  Matrices and scaffolds for drug delivery in dental, oral and craniofacial tissue engineering. , 2007, Advanced drug delivery reviews.

[29]  S. Lynch,et al.  A new era in periodontal and periimplant regeneration: use of growth-factor enhanced matrices incorporating rhPDGF. , 2006, Compendium of continuing education in dentistry.

[30]  Francesca Ungaro,et al.  Controlled drug delivery in tissue engineering. , 2008, Advanced drug delivery reviews.

[31]  J. Fiorellini,et al.  A phase I/II clinical trial to evaluate a combination of recombinant human platelet-derived growth factor-BB and recombinant human insulin-like growth factor-I in patients with periodontal disease. , 1997, Journal of periodontology.

[32]  R. Coletta,et al.  Effects of enamel matrix derivative and transforming growth factor-beta1 on human periodontal ligament fibroblasts. , 2007, Journal of clinical periodontology.

[33]  U. Ripamonti Recapitulating development: a template for periodontal tissue engineering. , 2007, Tissue engineering.

[34]  Yan Jin,et al.  Current approaches and challenges in making a bio-tooth. , 2008, Tissue engineering. Part B, Reviews.

[35]  E L Scheller,et al.  Tissue engineering: state of the art in oral rehabilitation. , 2009, Journal of oral rehabilitation.

[36]  Wei Zhang,et al.  The synergetic bone-forming effects of combinations of growth factors expressed by adenovirus vectors on chitosan/collagen scaffolds. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[37]  Vasif Hasirci,et al.  Sequential growth factor delivery from complexed microspheres for bone tissue engineering. , 2008, Biomaterials.

[38]  T. Sugaya,et al.  Effect of recombinant human platelet-derived growth factor-BB and bone morphogenetic protein-2 application to demineralized dentin on early periodontal ligament cell response. , 1999, Journal of periodontal research.

[39]  J C Petit,et al.  Periodontal tissue regeneration by combined applications of recombinant human osteogenic protein-1 and bone morphogenetic protein-2. A pilot study in Chacma baboons (Papio ursinus). , 2001, European journal of oral sciences.

[40]  Antonios G Mikos,et al.  Dual growth factor delivery from degradable oligo(poly(ethylene glycol) fumarate) hydrogel scaffolds for cartilage tissue engineering. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[41]  R. Genco,et al.  Informational Paper: Oral Reconstructive and Corrective Considerations in Periodontal Therapy. , 2005, Journal of periodontology.

[42]  Antonios G Mikos,et al.  Gelatin as a delivery vehicle for the controlled release of bioactive molecules. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[43]  A. Mikos,et al.  Spatial and temporal localization of transforming growth factor-beta1, bone morphogenetic protein-2, and platelet-derived growth factor-A in healing tooth extraction sockets in a rabbit model. , 2003, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[44]  W. Otto,et al.  Fluorimetric DNA assay for cell growth estimation. , 1992, Analytical biochemistry.

[45]  William V Giannobile,et al.  Growth factor delivery for oral and periodontal tissue engineering , 2006, Expert opinion on drug delivery.