Tri‐Layered Nanocomposite Hydrogel Scaffold for the Concurrent Regeneration of Cementum, Periodontal Ligament, and Alveolar Bone

A tri-layered scaffolding approach is adopted for the complete and concurrent regeneration of hard tissues-cementum and alveolar bone-and soft tissue-the periodontal ligament (PDL)-at a periodontal defect site. The porous tri-layered nanocomposite hydrogel scaffold is composed of chitin-poly(lactic-co-glycolic acid) (PLGA)/nanobioactive glass ceramic (nBGC)/cementum protein 1 as the cementum layer, chitin-PLGA/fibroblast growth factor 2 as the PDL layer, and chitin-PLGA/nBGC/platelet-rich plasma derived growth factors as the alveolar bone layer. The tri-layered nanocomposite hydrogel scaffold is cytocompatible and favored cementogenic, fibrogenic, and osteogenic differentiation of human dental follicle stem cells. In vivo, tri-layered nanocomposite hydrogel scaffold with/without growth factors is implanted into rabbit maxillary periodontal defects and compared with the controls at 1 and 3 months postoperatively. The tri-layered nanocomposite hydrogel scaffold with growth factors demonstrates complete defect closure and healing with new cancellous-like tissue formation on microcomputed tomography analysis. Histological and immunohistochemical analyses further confirm the formation of new cementum, fibrous PDL, and alveolar bone with well-defined bony trabeculae in comparison to the other three groups. In conclusion, the tri-layered nanocomposite hydrogel scaffold with growth factors can serve as an alternative regenerative approach to achieve simultaneous and complete periodontal regeneration.

[1]  T. Matsuyama,et al.  Regenerative effect of basic fibroblast growth factor on periodontal healing in two-wall intrabony defects in dogs. , 2010, Journal of clinical periodontology.

[2]  Dietmar W Hutmacher,et al.  Multiphasic construct studied in an ectopic osteochondral defect model , 2014, Journal of The Royal Society Interface.

[3]  M. Leite,et al.  Characterization and induction of cementoblast cell proliferation by bioactive glass nanoparticles , 2012, Journal of tissue engineering and regenerative medicine.

[4]  D. Hutmacher,et al.  Multiphasic Scaffolds for Periodontal Tissue Engineering , 2014, Journal of dental research.

[5]  K. Chennazhi,et al.  Periodontal Specific Differentiation of Dental Follicle Stem Cells into Osteoblast, Fibroblast, and Cementoblast. , 2015, Tissue engineering. Part C, Methods.

[6]  H. Arzate,et al.  Bone Regeneration in Rat Cranium Critical-Size Defects Induced by Cementum Protein 1 (CEMP1) , 2013, PloS one.

[7]  J C Middleton,et al.  Synthetic biodegradable polymers as orthopedic devices. , 2000, Biomaterials.

[8]  S. Deepthi,et al.  Bilayered construct for simultaneous regeneration of alveolar bone and periodontal ligament. , 2016, Journal of biomedical materials research. Part B, Applied biomaterials.

[9]  B. Kramer,et al.  The Composition of Bone. IV. Primary Calcification. , 1928 .

[10]  R. Schüle,et al.  FHL2 mediates dexamethasone‐induced mesenchymal cell differentiation into osteoblasts by activating Wnt/β‐catenin signaling‐dependent Runx2 expression , 2008, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[11]  Y. Yatomi,et al.  Optimized preparation method of platelet-concentrated plasma and noncoagulating platelet-derived factor concentrates: maximization of platelet concentration and removal of fibrinogen. , 2012, Tissue engineering. Part C, Methods.

[12]  S. Li,et al.  The effect of different platelet‐rich plasma concentrations on proliferation and differentiation of human periodontal ligament cells in vitro , 2007, Cell proliferation.

[13]  M. Miloro,et al.  Bone healing in a rabbit mandibular defect using platelet-rich plasma. , 2010, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[14]  R. J. Alves,et al.  Synthesis and Characterization of Poly(D,L-Lactide-co-Glycolide) Copolymer , 2012 .

[15]  K. Hiller,et al.  Influence of autogenous platelet concentrate on combined GTR/graft therapy in intrabony defects: a 7-year follow-up of a randomized prospective clinical split-mouth study. , 2012, Journal of clinical periodontology.

[16]  H. Birkedal‐Hansen,et al.  Proteins of the periodontium , 1977, Calcified Tissue Research.

[17]  G. Gheysen,et al.  Bioglass composites: a potential material for dental application. , 1983, Biomaterials.

[18]  M. Rinaudo,et al.  Chitin and chitosan: Properties and applications , 2006 .

[19]  Y. Shimabukuro,et al.  Periodontal Regeneration by Fgf-2 (bfgf) in Primate Models on Behalf Of: International and American Associations for Dental Research Periodontal Regeneration by Fgf-2 (bfgf) in Primate Models , 2001 .

[20]  Wei Fan,et al.  A biphasic scaffold design combined with cell sheet technology for simultaneous regeneration of alveolar bone/periodontal ligament complex. , 2012, Biomaterials.

[21]  L. Díaz de León,et al.  Human cementum protein extract promotes chondrogenesis and mineralization in mesenchymal cells. , 1996, Journal of periodontal research.

[22]  Remo Guidieri Res , 1995, RES: Anthropology and Aesthetics.

[23]  Saso Ivanovski,et al.  Advanced tissue engineering scaffold design for regeneration of the complex hierarchical periodontal structure. , 2014, Journal of clinical periodontology.

[24]  Rohit T Rao,et al.  An overview of recent patents on musculoskeletal interface tissue engineering , 2016, Connective tissue research.

[25]  Hector F Rios,et al.  Biomimetic hybrid scaffolds for engineering human tooth-ligament interfaces. , 2010, Biomaterials.

[26]  Roach Hi Why Does Bone Matrix Contain Non-Collagenous Proteins? The Possible Roles of Osteocalcin, Osteonectin, Osteopontin and Bone Sialoprotein in Bone Mineralisation and Resorption , 1994 .

[27]  Hermann Ehrlich,et al.  Chitin and chitosan in selected biomedical applications , 2014 .

[28]  Gibson Wa,et al.  Histochemistry of the periodontal ligament. II. The phosphatases. , 1967, Periodontics.

[29]  R. Reis,et al.  Preparation and in vitro characterization of novel bioactive glass ceramic nanoparticles. , 2009, Journal of biomedical materials research. Part A.

[30]  H. Arzate,et al.  Cementum proteins: role in cementogenesis, biomineralization, periodontium formation and regeneration. , 2015, Periodontology 2000.

[31]  Rui L Reis,et al.  A tissue engineering approach for periodontal regeneration based on a biodegradable double-layer scaffold and adipose-derived stem cells. , 2014, Tissue engineering. Part A.

[32]  Y. Izumi,et al.  Periodontal Tissue Regeneration Using Fibroblast Growth Factor -2: Randomized Controlled Phase II Clinical Trial , 2008, PloS one.

[33]  H. Arzate,et al.  Cementum protein 1 (CEMP1) induces differentiation by human periodontal ligament cells under three‐dimensional culture conditions , 2012, Cell biology international.

[34]  K. Chennazhi,et al.  Chitin Scaffolds in Tissue Engineering , 2011, International journal of molecular sciences.

[35]  In-Yong Kim,et al.  Chitosan and its derivatives for tissue engineering applications. , 2008, Biotechnology advances.

[36]  L. Guan,et al.  Periodontal regeneration using a bilayered PLGA/calcium phosphate construct. , 2011, Biomaterials.

[37]  Peng Shang,et al.  Three-dimensional printed multiphase scaffolds for regeneration of periodontium complex. , 2014, Tissue engineering. Part A.