Jun-Hyeog Jang

Electrospun materials as potential platforms for bone tissue engineering

Nanofibrous materials produced by electrospinning processes have attracted considerable interest in tissue regeneration, including bone reconstruction. A range of novel materials and processing tools have been developed to mimic the native bone extracellular matrix for potential applications as tissue engineering scaffolds and ultimately to restore degenerated functions of the bone. Degradable polymers, bioactive inorganics and their nanocomposites/hybrids nanofibers with suitable mechanical properties and bone bioactivity for osteoblasts and progenitor/stem cells have been produced. The surface functionalization with apatite minerals and proteins/peptides as well as drug encapsulation within the nanofibers is a promising strategy for achieving therapeutic functions with nanofibrous materials. Recent attempts to endow a 3D scaffolding technique to the electrospinning regime have shown some promise for engineering 3D tissue constructs. With the improvement in knowledge and techniques of bone-targeted nanofibrous matrices, bone tissue engineering is expected to be realized in the near future.

Membrane of hybrid chitosan-silica xerogel for guided bone regeneration

Chitosan-silica xerogel hybrid membranes were fabricated using a sol-gel process and their potential applications in guided bone regeneration (GBR) were investigated in terms of their in vitro cellular activity and in vivo bone regeneration ability. TEM observation revealed that the silica xerogel was dispersed in the chitosan matrix on the nanoscale. The hybrid membrane showed superior mechanical properties to chitosan in the wet state and the rapid induction of calcium phosphate minerals in simulated body fluid, reflecting its excellent in vitro bone bioactivity. Osteoblastic cells were observed to adhere well and grow actively on the hybrid membrane to a level higher than that observed on the chitosan membrane. The alkaline phosphatase activity of the cells was also much higher on the hybrid than on the chitosan membrane. The in vivo study in a rat calvarial model demonstrated significantly enhanced bone regeneration using the hybrid membrane compared to that observed using the pure chitosan one. Histomorphometric analysis performed 3 weeks after implantation revealed a fully closed defect in the hybrid membrane, whereas there was only 57% defect closure in the chitosan membrane.

Apatite-mineralized polycaprolactone nanofibrous web as a bone tissue regeneration substrate

Degradable synthetic polymers with a nanofibrous structure have shown great promise in populating and recruiting cells for the reconstruction of damaged tissues. However, poor cell affinity and lack of bioactivity have limited their potential usefulness in bone regeneration. We produced polymeric nanofiber poly(epsilon-caprolactone) (PCL) with its surface mineralized with bone-like apatite for use as bone regenerative and tissue engineering matrices. PCL was first electrospun into a nanofibrous web, and the surface was further mineralized with apatite following a series of solution treatments. The surface of the mineralized PCL nanofiber was observed to be almost fully covered with nanocrystalline apatites. Through mineralization, the wettability of the nanofiber matrix was greatly improved. Moreover, the murine-derived osteoblastic cells were shown to attach and grow actively on the apatite-mineralized nanofibrous substrate. In particular, the mineralized PCL nanofibrous substrate significantly stimulated the expression of bone-associated genes, including Runx2, collagen type I, alkaline phosphatase, and osteocalcin, when compared with the pure PCL nanofiber substrate without mineralization. The currently developed polymer nanofibrous web with the bioactive mineralized surface is considered to be potentially useful as bone regenerative and tissue engineering matrices.

Bioactivity improvement of poly(ε-caprolactone) membrane with the addition of nanofibrous bioactive glass

Nanofibrous glass with a bioactive composition was added to a degradable polymer poly(epsilon-caprolactone) (PCL) to produce a nanocomposite in thin membrane form ( approximately 260 microm). The bioactivity and osteoblastic responses of the nanocomposite membrane were examined and compared with those of a pure PCL membrane. Glass nanofibers with diameters in the range of hundreds of nanometers were added to a PCL solution at 20 wt.%, and the mixture was stirred vigorously and air dried. The obtained nanocomposite membrane showed that many chopped glass nanofibers formed by the mixing step were embedded uniformly into the PCL matrix. The nanocomposite membrane induced the rapid formation of apatite-like minerals on the surface when immersed in a simulated body fluid. Murine-derived osteoblastic cells (MC3T3-E1) grew actively over the nanocomposite membrane with cell viability significantly improved compared with those on the pure PCL membrane. Moreover, the osteoblastic activity, as assessed by the expression of alkaline phosphatase, was significantly higher on the nanocomposite membrane than on the pure PCL membrane. The currently developed nanocomposite of the bioactive glass-added PCL might find applications in the bone regeneration areas such as the guided bone regeneration (GBR) membrane.

Therapeutic-designed electrospun bone scaffolds: Mesoporous bioactive nanocarriers in hollow fiber composites to sequentially deliver dual growth factors.

A novel therapeutic design of nanofibrous scaffolds, holding a capacity to load and deliver dual growth factors, that targets bone regeneration is proposed. Mesoporous bioactive glass nanospheres (MBNs) were used as bioactive nanocarriers for long-term delivery of the osteogenic enhancer fibroblast growth factor 18 (FGF18). Furthermore, a core-shell structure of a biopolymer fiber made of polyethylene oxide/polycaprolactone was introduced to load FGF2, another type of cell proliferative and angiogenic growth factor, safely within the core while releasing it more rapidly than FGF18. The prepared MBNs showed enlarged mesopores of about 7 nm, with a large surface area and pore volume. The protein-loading capacity of MBNs was as high as 13% when tested using cytochrome C, a model protein. The protein-loaded MBNs were smoothly incorporated within the core of the fiber by electrospinning, while preserving a fibrous morphology. The incorporation of MBNs significantly increased the apatite-forming ability and mechanical properties of the core-shell fibers. The possibility of sequential delivery of two experimental growth factors, FGF2 and FGF18, incorporated either within the core-shell fiber (FGF2) or within MBNs (FGF18), was demonstrated by the use of cytochrome C. In vitro studies using rat mesenchymal stem cells demonstrated the effects of the FGF2-FGF18 loadings: significant stimulation of cell proliferation as well as the induction of alkaline phosphate activity and cellular mineralization. An in vivo study performed on rat calvarium defects for 6 weeks demonstrated that FGF2-FGF18-loaded fiber scaffolds had significantly higher bone-forming ability, in terms of bone volume and density. The current design utilizing novel MBN nanocarriers with a core-shell structure aims to release two types of growth factors, FGF2 and FGF18, in a sequential manner, and is considered to provide a promising therapeutic scaffold platform that is effective for bone regeneration.

Effects of Fibroblast Growth Factor-2 on the Expression and Regulation of Chemokines in Human Dental Pulp Cells

BACKGROUND Fibroblast growth factor-2 (FGF-2) participates in both hematopoiesis and osteogenesis; however, the effects of FGF-2 on chemokines during odontoblastic differentiation have not been reported. This study investigated whether human dental pulp cells (HDPCs) treated with FGF-2 could express chemokines during differentiation into odontoblastic cells and sought to identify its underlying mechanism of action. METHODS To analyze differentiation, we measured alkaline phosphatase (ALP) activity, calcified nodule formation by alizarin red staining, and marker RNA (mRNA) expression by reverse-transcriptase polymerase chain reaction (RT-PCR). Expression of chemokines, such as interleukin-6 (IL-6), IL-8, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), and MIP-3α, were evaluated by RT-PCR. RESULTS ALP activity, the mineralization, and mRNA expression for odontoblastic markers were enhanced by FGF-2 in HDPCs. FGF-2 also up-regulated the expression of IL-6, IL-8, MCP-1, MIP-1α, and MIP-3α mRNAs, which were attenuated by inhibitors of p38, ERK1/2 and p38 MAP kinases, protein kinase C, phosphoinositide-3 kinase, and NF-κB. CONCLUSION Taken together, these data suggest that FGF-2 plays a role not only as a differentiation inducing factor in the injury repair processes of pulpal tissue but also as a positive regulator of chemokine expression, which may help in tissue engineering and pulp regeneration using HDPCs. However, the fate of odontoblastic or osteoblastic differentiation, effective local delivery for FGF-2, interaction of chemotatic and odontogenic factors, and other limitations will need to be overcome before a major modality for the treatment of pulp disease.

Effect of Calcium Phosphate Cements on Growth and Odontoblastic Differentiation in Human Dental Pulp Cells

OBJECTIVE Calcium phosphate cements (CPCs) are an interesting class of bone substitute materials. However, the biological effects of CPCs have not been well studied in human dental pulp cells (HDPCs). The purpose of this study was to investigate the effects of CPCs on the mechanical properties, growth, and odontoblastic differentiation in HDPCs compared with Portland cement (PC) and mineral trioxide aggregate (MTA). METHODS Experimental CPCs either containing chitosan (Ch-CPC) or without chitosan (CPC) were composed from the alpha-tricalcium phosphate powder. Setting time, compressive strength measurements, cell growth, alkaline phosphatase (ALP) activity, the levels of messenger RNA for differentiation-related genes, and mineralization of the HDPCs on various cements were assessed. RESULTS The setting time for CPC-Ch was 7.5 minutes, which was significantly less than the 8.6 minutes for the CPC. On the seventh day of immersion, the compressive strength of CPC-CH reached 13.1 MPa, which was higher than 10.8 MPa of CPC. CPC and Ch-CPC-treated cells showed decreased cell proliferation but increased the levels of ALP activity, enhanced mineralized nodule formation, and upregulated odontoblastic markers messenger RNA including osteonectin, osteopontin, bone sialoprotein, dentin matrix protein-1, matrix extracellular phosphoglycoprotein, and dentin sialophosphoprotein (DSPP), compared with untreated control. The response of CPC and CP-CPC were similar to that of PC and MTA. However, the adhesion, growth, and differentiation in Ch-CPC-treated cells were similar to that in the CPC. CONCLUSION CPC may be useful for pulp-capping applications based on its abilities to promote HDPC differentiation.

Biomimetic approach to dental implants

Titanium, as an implant material, is regarded to be durable and biocompatible, which allows functional replacement of missing teeth. Successful dental implantation depends on an osseointegration phenomenon, a direct structural and functional binding reaction between bone and implant. It is well known that physicochemical characteristics of the dental implant surface, such as roughness, topography, chemistry, and electrical charge affect the biological reactions occurring at the interface of tissue and implant. Therefore, considerable efforts have been made to modify the surface of titanium implants which are based on mechanical, physical and chemical treatments. Recently, biological molecules were introduced onto the surface of implants to stimulate osteogenic cells in the early stage of implantation and consequently accelerate bone formation around implant and subsequent rapid implant stabilization. A range of extracellular matrix components, designed peptides, and growth factors have been proposed as the biological moiety. In this review, we address several issues related to the biology of dental implants and discuss biomimetic modification of the implant surface as a novel approach to obtain successful osseointegration.

Effects of surface modification on the resin-transfer moulding (RTM) of glass-fibre/unsaturated-polyester composites

Abstract The effects of glass fibre surface modification on the flow characteristics of unsaturated polyester (UPE) resin were investigated in the resin-transfer moulding (RTM) process. γ-Methacryloxypropyl trimethoxy silane (γ-MPS) was used as a glass fibre surface modifier. It was found that surface energy of glass fibre was decreased by γ-MPS treatment by advancing contact-angle measurement. Unsteady state permeability of glass fabric preforms was measured according to Darcys law. The apparent permeability of γ-MPS-treated glass fabric preforms was slightly lower than that of untreated fabric performs because of the macro/micro flow induced by capillary action. The void contents and the flexural properties of the cured glass-fibre/UPE composites were estimated and morphological study of the glass-fibre/UPE composites was also performed by SEM. When the fibre surface was treated with γ-MPS, the void content and the flexural properties of the glass-fibre/UPE composites were different in different regions of the mould cavity.

Silica xerogel-chitosan nano-hybrids for use as drug eluting bone replacement.

Silica xerogel-chitosan hybrids containing vancomycin were fabricated by the sol–gel process at room temperature and their potential as a drug eluting bone replacement was evaluated in terms of their mechanical properties and drug release behaviors. Regardless of the content of chitosan, all of the prepared hybrids had a uniform mesoporous structure, which would allow the effectual loading of vancomycin. As the content of chitosan was increased, the strength, strain to failure, and work of fracture of the hybrids were significantly enhanced, while the elastic modulus was decreased. These changes in the mechanical properties were mainly attributed to the mitigation of the brittleness of the silica xerogel through its hybridization with the flexible chitosan phase. In addition, the initial burst-effect was remarkably reduced by increasing the content of chitosan. The hybrids with more than 30% chitosan could release the vancomycin for an extended period of time in a controlled manner.