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Inflammatory responses are frequently associated with the expression of inflammatory cytokines and severe osteoclastogenesis, which significantly affect the efficacy of biomaterials. Recent findings have suggested that interferon (IFN)-γ and zoledronate (Zol) are effective inhibitors of osteoclastogenesis. However, little is known regarding the utility of IFN-γ and Zol in bone tissue engineering. In this study, we generated rat models by generating critically sized defects in calvarias implanted with an alpha-tricalcium phosphate/collagen sponge (α-TCP/CS). At four weeks post-implantation, the rats were divided into IFN-γ, Zol, and control (no treatment) groups. Compared with the control group, the IFN-γ and Zol groups showed remarkable attenuation of severe osteoclastogenesis, leading to a significant enhancement in bone mass. Histomorphometric data and mRNA expression patterns in IFN-γ and Zol-injected rats reflected high bone-turnover with increased bone formation, a reduction in osteoclast numbers, and tumor necrosis factor-α expression. Our results demonstrated that the administration of IFN-γ and Zol enhanced bone regeneration of α-TCP/CS implants by enhancing bone formation, while hampering excess bone resorption.

Cell sheet technology is a scaffold-free method for tissue reconstruction. A sheet-shaped scaffold would be suitable for the regeneration of periodontal membrane. We designed a stem cell sheet combining human mesenchymal stromal cells (hMSCs) and a 10-µm thick biological apatite (BAp) membrane fabricated with an ArF pulsed laser ablation for periodontal regeneration. X-ray diffraction showed that crystalline hydroxyapatite (HAp) was present in BAp and HAp membranes after post-annealing. Energy dispersive analysis of the BAp membrane revealed peaks of Na and Mg in addition to Ca and P. Approximately 3×10(4) hMSCs were cultured on BAp and HAP membranes for 7 and 14 days. From in vitro assays, hMSCs grew faster and had higher osteoblast differentiation when cultured on the BAp membrane than did the cell culture on the HAp membrane. Stem cell sheets combined with a BAp membrane may have potential applications in guided bone regeneration and osteoconductive scaffolds.

This study aimed to establish an in vitro human periodontal ligament-like tissue (HPdLLT) by three-dimensional culturing of human periodontal ligament fibroblasts (HPdLFs) in a porous poly-L-lactide (PLLA) matrix modified hydrophilically with ammonia solution. After ammonia modification, the surface roughness and culture-medium-soaking-up ability of the PLLA matrix increased, whereas the contact angle of water drops decreased. The thickness, porosity, and pore size of the PLLA matrix were 400±50 μm, 83.3%, and 75-150 μm, respectively. HPdLFs (1×10(5) cells) were seeded on the modified PLLA matrix and centrifuged to facilitate seeding into its interior and cultured for 14 days. Scanning electron microscope (SEM) observation, proliferation assay, picrosirius-red staining, and real-time polymerase chain reaction (RT-PCR) for type-1 collagen (COL1), periodontal ligament associated protein-1 (PLAP-1), fibroblast growth factor-2 (FGF-2), and alkaline phosphatase (ALP) mRNA were conducted on days 1, 3, 7, and 14. HPdLFs were observed entirely from the surface to the rear side of the matrix. Cell proliferation analysis, SEM observation, and picrosirius-red staining showed both progressive growth of 3D-cultured HPdLFs and extracellular matrix maturation by the secretion of COL1 and type 3 collagen (COL3) from days 1 to 14. Expressions of COL1, PLAP-1, and FGF-2 mRNA suggested the formation of cellular components and supplementation of extracellular components. Expressions of ALP, COL1, and PLAP-1 mRNA suggested the osteogenic potential of the HPdLLT. The results indicated in vitro HPdLLT formation, and it could be used in future periodontal ligament tissue engineering to achieve optimal periodontal regeneration., (Copyright © 2013. Published by Elsevier B.V.)

A novel, injectable bone tissue engineering material was developed that consisted of beta-tricalcium phosphate (beta-TCP) beads as the solid phase and alginate as the gel phase. To prepare the instantaneously formed composite scaffold, an aqueous calcium chloride solution was dried on the surface of beta-TCP beads and crosslinked with an alginic acid sodium solution, thereby forming stable beta-TCP beads and alginate gel which were injectable via a syringe. This biodegradable composite was a three-dimensional (3D) material that could be used as an injectable scaffold for bone tissue engineering. In particular, the composite with 2.0 wt% alginate concentration exhibited a compressive strength of 69 kPa in dry conditions, which was significantly higher than that exhibited by 1.0 wt%. Furthermore, mesenchymal stem cells (MSC) were 3D-cultured within the composite and then investigated for osteogenic markers. MSC-loaded composite was subjected to scanning electron microscope (SEM) examination and implanted subcutaneously for in vivo experiment. Results showed that the scaffold provided support for osteogenic differentiation. In light of the encouraging results obtained, this novel injectable composite material may be useful for bone tissue engineering.

Objectives: The purpose of this study was to evaluate and compare the osteoblastic differentiation ability of dedifferentiated fat (DFAT) cells and adipose stem cells (ASCs) from the buccal fat pad (BFP). Materials and methods: We isolated human DFAT cells and ASCs from the BFP of a patient who underwent oral and maxillofacial surgery and then analyzed their cell surface antigens by flow cytometry. Then, the cells were cultured in osteogenic medium for 14 days. Measurement of bone-specific alkaline phosphatase (BAP), osteocalcin (OCN), and calcium deposition and alizarin red staining were performed to evaluate the osteoblastic differentiation ability of both cell types. Results: ASCs and DFAT cells were positive for CD90 and CD105 and negative for CD11b, CD34, and CD45. BAP (days 3 and 7), OCN (day 14), and calcium deposition (days 7 and 14) within DFAT cell cultures were significantly higher than those in ASC cultures. The alizarin red-stained area in DFAT cell cultures, which indicates mineralized matrix deposition, was stained more strongly than that in ASC cultures. Conclusions: The cell surface antigens of ASCs and DFAT cells tend to be similar. Furthermore, the osteoblastic differentiation ability of human DFAT cells is higher than that of ASCs from the BFP. Clinical relevance: Isolation of DFAT cells from the BFP has an esthetic advantage because the BFP can be obtained via the oral cavity without injury to the external body surface. Therefore, we consider that DFAT cells from the BFP are an ideal cell source for bone tissue engineering. [ABSTRACT FROM AUTHOR]

To date, there has been no study on the development of novel regimens based on the following tissue engineering principles: seeding and culturing mesenchymal stem cells (MSCs) on a scaffold before surgery or injecting cultured MSCs into a scaffold during surgery. The purpose of this study was to assess the in viuo osteogenic ability of scaffold/MSCs implanted beneath the periosteum of the cranial bone of rats in three different sample groups: one in which MSCs were pre-seeded and cultured on a scaffold to produce the 3-D woven fabric scaffold/MSC composite using osteo-lineage induction medium, one in which cultured MSCs produced by osteo- lineage induction in cell cultivation flasks were injected into a scaffold during surgery and a control group, in which only the 3-D woven fabric scaffold was implanted. The results indicate that pre-seeding MSCs on a scaffold leads to a higher osteogenic ability than injecting cultured MSCs into a scaffold during surgery. [ABSTRACT FROM AUTHOR]

Summary: For many years, bone graft substitutes have been used to reconstruct bone defects in orthopedic and dental fields. However, synthetic bone substitutes such as hydroxyapatite or β-tricalcium phosphate have no osteoinductive or osteogenic abilities. Bone tissue engineering has also been promoted as an alternative approach to regenerating bone tissue. To succeed in bone tissue engineering, osteoconductive scaffolding biomaterials should provide a suitable environment for osteogenic cells and provide local controlled release of osteogenic growth factors. In addition, the scaffold for the bone graft substitute should biodegrade to replace the newly formed bone. Recent advances in bone tissue engineering have allowed the creation of composite scaffolds with tailored functional properties. This review focuses on composite scaffolds that consist of synthetic ceramics and natural polymers as drug delivery carriers for alveolar bone tissue engineering. [Copyright &y& Elsevier]

Synthetic biomaterials have been developed and used for bone grafting. Here, we developed a biodegradable sponge composite for bone tissue engineering by combining β-tricalcium phosphate (β-TCP) and collagen. In addition, we sought to determine the optimal β-TCP granules/collagen ratio by evaluating and bone formation in vivo. Porous β-TCP granules were mixed with atelocollagen hydrochloride solution at various ratios -- 0.02, 0.05, 0.1, and 0.2 g/mL. The resultant mixtures were freeze-dried and subjected to dehydrothermal treatment in vacuo. The final composites obtained were designated β-TCP/collagen sponge composites (β-TCP/CS). Through compression testing, it was found that the stress values for β-TCP/CS (0.2 g/mL) were higher than those of the other three composites over the whole strain range. Histological evaluation at four weeks after implantation revealed that the collagen sponge had degraded and newly formed bone was present on the surface of the β-TCP granules. At 12 weeks, the β-TCP granules were completely degraded and remodeling of the lamellar bone was observed. [ABSTRACT FROM AUTHOR]