Dentin-Pulp Complex Tissue Regeneration via Three-Dimensional Cell Sheet Layering

Hu, Yan; Oshima, Masamitsu; Raju, Resmi; Raman, Swarnalakshmi; Sekine, Keitaro; Waskitho, Arief; Inoue, Miho; Inoue, Masahisa; Baba, Otto; Morita, Tsuyoshi; Miyagi, Mayu; Matsuka, Yoshizo

doi:10.1089/ten.tec.2021.0171

Cited by 8 publications

(4 citation statements)

References 53 publications

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“…Our findings further support the use of Alg and CMC bioinks for tissue regeneration as previously shown in other tissues including bone tissue engineering [86,87]. Future studies, similar to dentin regeneration reports published by our group and others [88][89][90], using in vitro culture of cellladen scaffolds in differentiation media accompanied by in vivo studies using pre-clinical enamel regeneration models could further validate the suitability of the Alg-CMC bioinks for dental enamel tissue regeneration. Taken together, our findings suggest that Alg-CMC is a promising candidate to be used as bioink for enamel tissue engineering.…”

Section: Discussionsupporting

confidence: 89%

Bioprinting of alginate-carboxymethyl chitosan scaffolds for enamel tissue engineering in vitro

et al. 2022

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Three dimensional (3D) bioprinting has been emerging enabling the incorporation of living cells into the biomaterials (such a mixture is referred as a bioink) to create scaffolds. However, the bioinks available for scaffold bioprinting are limited, particularly for dental tissue engineering, due to the complicated, yet compromised, printability, mechanical and biological properties simultaneously imposed on the bioinks. This paper shows the development of a novel bioink from carboxymethyl chitosan and alginate for bioprinting scaffolds for enamel tissue regeneration. Carboxymethyl chitosan is used because to its antibacterial ability and superior cell interaction properties, while alginate was added to enhance printability and mechanical properties as well as to regulate the degradation rate. The bioinks with three mixture ratios of alginate and carboxymethyl chitosan (2-4, 3-3 and 4-2) are prepared, and then printed into the calcium chloride crosslinker solution (100 mM) to form a 3D structure of scaffolds. The printed scaffolds are characterized in terms of structural, swelling, degradation, and mechanical properties, followed by their in vitro characterization for enamel tissue regeneration. The results show that the bioinks with higher concentrations of alginate are more viscous and need higher pressure for printing; while the printed scaffolds are highly porous and show a high degree of printability and structural integrity. The hydrogels with higher carboxymethyl chitosan ratios have higher swelling ratios, faster degradation rates, and lower compressive modulus. Dental epithelial cell line, HAT-7, could maintain high viability in the printed constructs after 1, 7 and 14 days of culture. HAT-7 cells are also able to maintain their morphology and secrete alkaline phosphatase after 14 days of culture in the 3D printed scaffolds, suggesting the capacity of these cells for mineral deposition and enamel-like tissue formation. Taken together, alginate-carboxymethyl chitosan may be suitable to print scaffolds with dental epithelial cells for enamel tissue regeneration.

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Section: Discussionsupporting

confidence: 89%

Bioprinting of alginate-carboxymethyl chitosan scaffolds for enamel tissue engineering in vitro

et al. 2022

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“…Previous studies have already examined the role of hDPSCs in the dental pulp tissue regeneration in tooth root fragments through a scaffold-free strategy using SHED-derived cells. The major difficulty was developing the complex histological dental pulp structure that possesses highly organized physiologic patterns [ 32 ].…”

Section: Discussionmentioning

confidence: 99%

Decellularized Dental Pulp, Extracellular Vesicles, and 5-Azacytidine: A New Tool for Endodontic Regeneration

et al. 2022

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Dental pulp is a major component of the dental body that serves to maintain the tooth life and function. The aim of the present work was to develop a system that functions as a growth-permissive microenvironment for dental pulp regeneration using a decellularized dental pulp (DDP) matrix, 5-Aza-2′-deoxycytidine (5-Aza), and Extracellular Vesicles (EVs) derived from human Dental Pulp Stem Cells (hDPSCs). Human dental pulps extracted from healthy teeth, scheduled to be removed for orthodontic purpose, were decellularized and then recellularized with hDPSCs. The hDPSCs were seeded on DDP and maintained under different culture conditions: basal medium (CTRL), EVs, 5-Aza, and EVs+-5-Aza. Immunofluorescence staining and Western blot analyses were performed to evaluate the proteins’ expression related to dentinogenesis, such as ALP, RUNX2, COL1A1, Vinculin, DMP1, and DSPP. Protein contents found in the DDP recellularized with hDPSCs were highly expressed in samples co-treated with EVs and 5-Aza compared to other culture conditions. This study developed a DDP matrix loaded by hDPSCs in co-treatment with EVs, which might enhance the dentinogenic differentiation with a high potentiality for endodontic regeneration.

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“…In a separate report, researchers implanted layered cell sheets prepared from rat dental pulp (DP) cells into the subrenal capsule of nude mice, and observed new bone development at 8 weeks post-implantation. 114 Ueyama et al 115 further performed the transplantation of osteogenic bone marrow stem cell (BMSC) sheets into maxillofacial bone defects in rats, thereby promoting extensive new bone development in the implanted region at 8 weeks post-surgery. A number of studies have also highlighted the beneficial effects of osteogenic cell sheet application in the context of delayed bone union or nonunion, which is a complex process that can be shaped by a range of mechanical and/or biological factors.…”

Section: Preparation Of Cell Sheetsmentioning

confidence: 99%

Cell Sheet Technology as an Engineering-Based Approach to Bone Regeneration

You¹,

Lu²,

Li³

et al. 2022

IJN

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Bone defects that are congenital or the result of infection, malignancy, or trauma represent a challenge to the global healthcare system. To address this issue, multiple research groups have been developing novel cell sheet technology (CST)-based approaches to promote bone regeneration. These methods hold promise for use in regenerative medicine because they preserve cellcell contacts, cell-extracellular matrix interactions, and the protein makeup of cell membranes. This review introduces the concept and preparation system of the cell sheet (CS), explores the application of CST in bone regeneration, highlights the current states of the bone regeneration via CST, and offers perspectives on the challenges and future research direction of translating current knowledge from the lab to the clinic.

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Dentin-Pulp Complex Tissue Regeneration via Three-Dimensional Cell Sheet Layering

Cited by 8 publications

References 53 publications

Bioprinting of alginate-carboxymethyl chitosan scaffolds for enamel tissue engineering in vitro

Bioprinting of alginate-carboxymethyl chitosan scaffolds for enamel tissue engineering in vitro

Decellularized Dental Pulp, Extracellular Vesicles, and 5-Azacytidine: A New Tool for Endodontic Regeneration

Cell Sheet Technology as an Engineering-Based Approach to Bone Regeneration

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