Dental pulp tissue engineering with bFGF-incorporated silk fibroin scaffolds

Abstract

The clinical translation of regenerative endodontics demands further development of suitable scaffolds. Here, we assessed the possibility of using silk fibroin scaffold for pulp regeneration with dental pulp stem cells (DPSCs) and basic fibroblast growth factor (bFGF) in ectopic root canal transplantation model. Porous silk fibroin scaffolds were fabricated using freeze-drying technique (with or without bFGF incorporation), and characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy. DPSCs were isolated, characterized, seeded onto scaffolds, and inserted into the tooth root fragments. Cell viability and morphology were tested in the 3D model in vitro using CCK8 assay and SEM. Furthermore, the ectopic transplantation model was used to verify the generation of pulp-like tissue in DPSCs seeded silk fibroin scaffold with bFGF, as examined by histological analysis. DPSCs seeded in silk fibroin scaffold survived, exhibited cytoplasmic elongation in scaffolds at least 4 weeks in culture. bFGF promoted DPSCs viability in tooth fragments/scaffolds (TSS) between 7 and 28 days. Pulp-like tissue was generated in the bFGF-incorporated TSS with DPSCs. Histologically, the generated tissue was shown to be with well vascularity, have new matrix deposition and dentin-like tissue formation, and consist of both the transplanted and host-derived cells. Collectively, these data support the use of bFGF-incorporated silk fibroin scaffold as a highly promising scaffold candidate for future treatment concepts in regenerative endodontics to save teeth.

Disclosing the physiology of pulp tissue for vital pulp therapy

“…Some carriers that have previously been proposed were collagen (Hu et al 1998, Kikuchi et al 2007, Bezerra Da Silva et al 2008, gelatin Ishimatsu et al 2009), agarose (Chaussain et al 2009), sodium alginate (Oliva-Rodriguez et al 2011), chitosan . Current in vitro evidence suggests other potential drug delivery systems for bioactive molecules, such as biodegradable polymer matrix of lactide and glycolide (Mathieu et al 2013), porous silk fibroin scaffolds (Yang et al 2015), poly-2-hydroxyethyl methacrylate (polyHEMA)-based hydrogel (Takeda et al 2015), tricalcium phosphate microsphere-hydrogel composite (Lee et al 2014). In addition, biomaterials that can be used as scaffolds for VPT have recently been suggested and could enhance proliferation and differentiation of human dental pulp cells, such as polycaprolactone/submicron bioactive glass hybrid composites (Wang et al 2016a), a chitosan-based scaffold (Bellamy et al 2016) and keratin hydrogel (Sharma et al 2016).…”

Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering

“…seeded 2×10 6  DPSCs/per silk fiber/collagen/hexafluoro-2-propanol (HFIP) scaffold and observed that they formed soft dental pulp. 65 However, the disadvantages associated with these cells is their heterogeneity which can affect its osteogenic potential. Moreover, several investigators have also reported the limited potential of DMSCs to regenerate bone in vivo due to its inability to promote de novo bone formation in the form of cortical bone and not vascularized bone as noted in a human mandibular defect.…”

Hyaluronic acid hydrogels incorporating platelet lysate enhance human pulp cell proliferation and differentiation

“…These therapies explore the endogenous regenerative potential of the endodontic tissues and procure to enhance it by combining biopolymers with adequate biochemical stimuli [1]. Among the most reported biopolymers for endodontic therapy are hyaluronic acid (HA) [2][3][4], chitosan [5], alginate [6], gelatin [7][8][9], collagen [10,11], fibrin [12], and silk fibroin [13]. These natural-origin molecules resemble the extracellular matrix (ECM) components, providing unique properties to the specific regenerative approach, thereby enhancing their biocompatibility and biological recognition.…”