Three-dimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame

Regenerative medicine aims to restore damaged tissues and mainly takes advantage of human mesenchymal stromal cells (hMSCs), either alone or combined with three-dimensional scaffolds. The scaffold is generally considered a support, and its contribution to hMSC proliferation and differentiation is unknown or poorly investigated. The aim of this study was to evaluate the capability of an innovative three-dimensional gelatin–chitosan hybrid hydrogel scaffold (HC) to activate the osteogenic differentiation process in hMSCs. We seeded hMSCs from adipose tissue (AT-hMSCs) and bone marrow (BM-hMSCs) in highly performing HC of varying chitosan content in the presence of growing medium (GM) or osteogenic medium (OM) combined with Fetal Bovine Serum (FBS) or human platelet lysate (hPL). We primarily evaluated the viability and the proliferation of AT-hMSCs and BM-hMSCs under different conditions. Then, in order to analyse the activation of osteogenic differentiation, the osteopontin (OPN) transcript was absolutely quantified at day 21 by digital PCR. OPN was expressed under all conditions, in both BM-hMSCs and AT-hMSCs. Cells seeded in HC cultured with OM+hPL presented the highest OPN transcript levels, as expected. Interestingly, both BM-hMSCs and AT-hMSCs cultured with GM+FBS expressed OPN. In particular, BM-hMSCs cultured with GM+FBS expressed more OPN than those cultured with GM+hPL and OM+FBS; AT-hMSCs cultured with GM+FBS presented a lower expression of OPN when compared with those cultured with GM+hPL, but no significant difference was detected when compared with AT-hMSCs cultured with OM+FBS. No OPN expression was detected in negative controls. These results show the capability of HC to primarily and independently activate osteogenic differentiation pathways in hMCSs. Therefore, these scaffolds may be considered no more as a simple support, rather than active players in the differentiative and regenerative process.

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Mineralization of 3D Osteogenic Model Based on Gelatin-Dextran Hybrid Hydrogel Scaffold Bioengineered with Mesenchymal Stromal Cells: A Multiparametric Evaluation

Materials ◽

10.3390/ma14143852 ◽

2021 ◽

Vol 14 (14) ◽

pp. 3852

Author(s):

Federica Re ◽

Luciana Sartore ◽

Elisa Borsani ◽

Matteo Ferroni ◽

Camilla Baratto ◽

...

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Bone Repair ◽

Active Role ◽

Ionization Mass ◽

Hydrogel Scaffold ◽

Hybrid Hydrogel ◽

Hydrogel Scaffolds ◽

Human Platelet Lysate ◽

Identical Composition

Gelatin–dextran hydrogel scaffolds (G-PEG-Dx) were evaluated for their ability to activate the bone marrow human mesenchymal stromal cells (BM-hMSCs) towards mineralization. G-PEG-Dx1 and G-PEG-Dx2, with identical composition but different architecture, were seeded with BM-hMSCs in presence of fetal bovine serum or human platelet lysate (hPL) with or without osteogenic medium. G-PEG-Dx1, characterized by a lower degree of crosslinking and larger pores, was able to induce a better cell colonization than G-PEG-Dx2. At day 28, G-PEG-Dx2, with hPL and osteogenic factors, was more efficient than G-PEG-Dx1 in inducing mineralization. Scanning electron microscopy (SEM) and Raman spectroscopy showed that extracellular matrix produced by BM-hMSCs and calcium-positive mineralization were present along the backbone of the G-PEG-Dx2, even though it was colonized to a lesser degree by hMSCs than G-PEG-Dx1. These findings were confirmed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), detecting distinct lipidomic signatures that were associated with the different degree of scaffold mineralization. Our data show that the architecture and morphology of G-PEG-Dx2 is determinant and better than that of G-PEG-Dx1 in promoting a faster mineralization, suggesting a more favorable and active role for improving bone repair.

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Long-Term Cryostorage of Mesenchymal Stem Cell-Containing Hybrid Hydrogel Scaffolds Based on Fibrin and Collagen

Gels ◽

10.3390/gels6040044 ◽

2020 ◽

Vol 6 (4) ◽

pp. 44

Author(s):

Marfa N. Egorikhina ◽

Yulia P. Rubtsova ◽

Diana Ya. Aleynik

Keyword(s):

Tissue Engineering ◽

Proliferative Activity ◽

Experimental Protocol ◽

Hydrogel Scaffold ◽

Hybrid Hydrogel ◽

Hydrogel Scaffolds ◽

High Viability ◽

Scaffold Structure ◽

Difficult Issue

The most difficult issue when using tissue engineering products is enabling the ability to store them without losing their restorative capacity. The numbers and viability of mesenchymal stem cells encapsulated in a hydrogel scaffold after cryostorage at −80 °C (by using, individually, two kinds of cryoprotectors—Bambanker and 10% DMSO (Dimethyl sulfoxide) solution) for 3, 6, 9, and 12 months were determined, with subsequent assessment of cell proliferation after 96 h. The analysis of the cellular component was performed using fluorescence microscopy and the two fluorochromes—Hoechst 3334 and NucGreenTM Dead 488. The experimental protocol ensured the preservation of cells in the scaffold structure, retaining both high viability and proliferative activity during storage for 3 months. Longer storage of scaffolds led to their significant changes. Therefore, after 6 months, the proliferative activity of cells decreased. Cryostorage of scaffolds for 9 months led to a decrease in cells’ viability and proliferative activity. As a result of cryostorage of scaffolds for 12 months, a decrease in viability and proliferative activity of cells was observed, as well as pronounced changes in the structure of the hydrogel. The described scaffold cryostorage protocol could become the basis for the development of storage protocols for such tissue engineering products, and for helping to extend the possibilities of their clinical use while accelerating their commercialization.

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Self-assembled three-dimensional hierarchical graphene hybrid hydrogels with ultrathin β-MnO2 nanobelts for high performance supercapacitors

Journal of Materials Chemistry A ◽

10.1039/c4ta04921g ◽

2015 ◽

Vol 3 (4) ◽

pp. 1540-1548 ◽

Cited By ~ 69

Author(s):

Sheng Zhu ◽

Hui Zhang ◽

Ping Chen ◽

Lin-Hui Nie ◽

Chuan-Hao Li ◽

...

Keyword(s):

Self Assembly ◽

High Performance ◽

Three Dimensional ◽

The Self ◽

Mass Loading ◽

Hybrid Hydrogel ◽

Precise Control ◽

Ultrafine Structure ◽

Hybrid Hydrogels ◽

Self Assembled

A facile protocol for the self-assembly of the rGO/β-MnO2 hybrid hydrogel with ultrafine structure and precise control of mass-loading for high performance supercapacitors is reported.

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Tissue-Engineered Cartilage in Three-Dimensional Cultured Chondrocytes by Ultrasound Stimulation and Collagen Sponge as Scaffold

Volume 2: Biomedical and Biotechnology Engineering ◽

10.1115/imece2009-11107 ◽

2009 ◽

Cited By ~ 1

Author(s):

Toshihiko Shiraishi ◽

Ietomo Matsunaga ◽

Shin Morishita ◽

Ryohei Takeuchi ◽

Tomoyuki Saito ◽

...

Keyword(s):

Mechanical Properties ◽

Three Dimensional ◽

Collagen Sponge ◽

Cartilage Tissue ◽

Mechanical Environment ◽

Ultrasound Stimulation ◽

Scaffold Structure ◽

Matrix Production ◽

Engineered Cartilage ◽

Cultured Chondrocytes

This paper describes the effects of ultrasound stimulation on chondrocytes in three-dimensional culture in relation to the production of regenerative cartilage tissue, using collagen sponges as a carrier and supplementation with hyaluronic acid (used in the conservative treatment of osteoarthritis). It has been shown that cell proliferation and matrix production can be facilitated by considering the mechanical environment of the cultured chondrocytes and the mechanical properties of the scaffold structure used in the culture and of the stimulation used.

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Collagen Expression in Tissue Engineered Cartilage of Aged Human Articular Chondrocytes in a Rotating Bioreactor

The International Journal of Artificial Organs ◽

10.1177/039139880302600407 ◽

2003 ◽

Vol 26 (4) ◽

pp. 319-330 ◽

Cited By ~ 34

Author(s):

S. Marlovits ◽

B. Tichy ◽

M. Truppe ◽

D. Gruber ◽

W. Schlegel

Keyword(s):

Electron Microscopy ◽

Collagen Type ◽

Articular Chondrocytes ◽

Three Dimensional ◽

Collagen Type I ◽

Human Articular Chondrocytes ◽

Type I ◽

Transmission Electron ◽

Low Shear Stress ◽

Engineered Cartilage

This study describes the culture and three-dimensional assembly of aged human articular chondrocytes under controlled oxygenation and low shear stress in a rotating-wall vessel. Chondrocytes cultured in monolayer were released and placed without any scaffold as a single cell suspension in a rotating bioreactor for 12 weeks. Samples were analyzed with immunohistochemistry, molecular biology and electron microscopy. During serial monolayer cultures chondrocytes dedifferentiated to a “fibroblast-like” structure and produced predominantly collagen type I. When these dedifferentiated cells were transferred to the rotating bioreactor, the cells showed a spontaneous aggregation and formation of solid tissue during the culture time. Expression of collagen type II and other components critical for the extracellular cartilage matrix could be detected. Transmission electron microscopy revealed a fine network of randomly distributed collagen fibrils. This rotating bioreactor proves to be a useful tool for providing an environment that enables dedifferentiated chondrocytes to redifferentiate and produce a cartilage-specific extracellular matrix.

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Three-dimensional changes of noses after transplantation of implant-type tissue-engineered cartilage for secondary correction of cleft lip–nose patients

Regenerative Therapy ◽

10.1016/j.reth.2017.09.001 ◽

2017 ◽

Vol 7 ◽

pp. 72-79 ◽

Cited By ~ 9

Author(s):

Kazuto Hoshi ◽

Yuko Fujihara ◽

Hideto Saijo ◽

Kumiko Kurabayashi ◽

Hideyuki Suenaga ◽

...

Keyword(s):

Cleft Lip ◽

Three Dimensional ◽

Dimensional Changes ◽

Engineered Cartilage ◽

Cleft Lip Nose

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Seismic and Robustness Design of Steel Frame Buildings

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.763.116 ◽

2018 ◽

Vol 763 ◽

pp. 116-123 ◽

Cited By ~ 4

Author(s):

Massimiliano Ferraioli ◽

Angelo Lavino ◽

Alberto Mandara ◽

Marianna Donciglio ◽

Antonio Formisano

Keyword(s):

Progressive Collapse ◽

Three Dimensional ◽

Design Procedure ◽

Steel Frame ◽

Fundamental Parameters ◽

Frame Buildings ◽

Steel Frame Buildings ◽

European Standards ◽

Three Dimensional Models ◽

Column Removal Scenario

In this paper, a design procedure that combines both progressive collapse design under column removal scenario and capacity design to produce a hierarchy of design strengths is presented. The procedure develops in the context of the European Standards, using the classification of European steel sections and considering the seismic design features. Three-dimensional models of typical multi-storey steel frame buildings are employed in numerical analysis. The design for progressive collapse is carried out with three types of analysis, namely linear static, nonlinear static and nonlinear dynamic. Since the behaviour following sudden column loss is likely to be inelastic and possibly implicate catenary effects, both geometric and material nonlinearities are considered. The influence of the fundamental parameters involved in seismic and robustness design is finally investigated.

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