Encapsulation of chondrocytes in injectable alkali-treated collagen gels prepared using poly(ethylene glycol)-based 4-armed star polymer

Taguchi, Tetsushi; Xu, Liming; Kobayashi, Haruo; Taniguchi, Akiyoshi; Kataoka, Kazunori; Tanaka, Junzo

doi:10.1016/j.biomaterials.2004.04.029

Cited by 84 publications

(57 citation statements)

References 21 publications

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“…RNA Extraction and Real Time PCR Analysis-The mRNA levels of differentiation-related marker genes were determined by quantitative real time PCR as described previously (27). Briefly, total RNA was extracted at various points with ISOGEN (Nippon Gene, Japan).…”

Section: Expression and Purification Of Recombinant Mousementioning

confidence: 99%

Reuptake of Extracellular Amelogenin by Dental Epithelial Cells Results in Increased Levels of Amelogenin mRNA through Enhanced mRNA Stabilization

Harada

Yokohama-Tamaki

et al. 2006

Journal of Biological Chemistry

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Amelogenin is an extracellular matrix protein secreted by ameloblasts and is a major component of enamel matrix. Recently, in addition to their role in enamel formation, the biological activity of enamel proteins in the process of cell differentiation has recently become widely appreciated. In this study, we examined the biological activity of amelogenin on ameloblast differentiation. Recombinant mouse amelogenin (rm-amelogenin) enhanced the expression of endogenous amelogenin mRNA in a cultured dental epithelial cell line (HAT-7), despite a lack of increased amelogenin promoter activity. To solve this discrepancy, we analyzed the effects of rmamelogenin on the stability of amelogenin mRNA. The half-life of amelogenin mRNA is extremely short, but in the presence of rmamelogenin its half-life was extended three times longer than the control. Furthermore, we showed the entry of exogenous fluorescein isothiocyanate-conjugated rm-amelogenin into the cytoplasm of HAT-7 cells. It follows from our results that exogenous amelogenin increases amelogenin mRNA levels through stabilization of mRNA in the cytoplasm of HAT-7 cells. Here we speculated that during differentiation, dental epithelial cells utilize a unique mechanism for increasing the production of amelogenin, the reuptake of secreted amelogenin.

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Section: Expression and Purification Of Recombinant Mousementioning

confidence: 99%

Reuptake of Extracellular Amelogenin by Dental Epithelial Cells Results in Increased Levels of Amelogenin mRNA through Enhanced mRNA Stabilization

Harada

Yokohama-Tamaki

et al. 2006

Journal of Biological Chemistry

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“…Multifunctional PEG cross-linking systems are either linear or exhibit branched chains with terminal active ester groups that react with the primary amine groups of the building blocks [13,18]. Poly (ethylene glycol) ether tetrasuccinimidyl glutarate (4S-StarPEG), a multi-armed functional PEG molecule, has been reported as an effective cross-linker of hydrogels [18,19] and implantable fibres [13]. The resulting scaffolds demonstrated improved stability and mechanical properties in physiological environments while exhibiting good biocompatibility [18,19].…”

Section: U N C O R R E C T E D P R O O Fmentioning

confidence: 99%

“…Poly (ethylene glycol) ether tetrasuccinimidyl glutarate (4S-StarPEG), a multi-armed functional PEG molecule, has been reported as an effective cross-linker of hydrogels [18,19] and implantable fibres [13]. The resulting scaffolds demonstrated improved stability and mechanical properties in physiological environments while exhibiting good biocompatibility [18,19]. Similarly, succinimidyl carboxymethyl ester (SCM-PEG-SCM), a linear functionalised PEG cross-linker, has been shown to improve stability and mechanical properties of collagen hydrogels with good biocompatibility [17,20].…”

Section: U N C O R R E C T E D P R O O Fmentioning

confidence: 99%

Cross-linking of a biopolymer-peptide co-assembling system

et al. 2017

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The ability to guide molecular self-assembly at the nanoscale into complex macroscopic structures could enable the development of functional synthetic materials that exhibit properties of natural tissues such as hierarchy, adaptability, and self-healing. However, the stability and structural integrity of these kinds of materials remains a challenge for many practical applications. We have recently developed a dynamic biopolymer-peptide co-assembly system with the capacity to grow and undergo morphogenesis into complex shapes. Here we explored the potential of different synthetic (succinimidyl carboxymethyl ester [SCM-PEG-SCM], poly (ethylene glycol) ether tetrasuccinimidyl glutarate [4S-StarPEG] and glutaraldehyde) and natural (genipin) cross-linking agents to stabilize membranes made from these biopolymer-peptide co-assemblies. We investigated the cross-linking efficiency, resistance to enzymatic degradation, and mechanical properties of the different cross-linked membranes. We also compared their biocompatibility by assessing the metabolic activity and morphology of adipose-derived stem cells (ADSC) cultured on the different membranes. While all cross-linkers successfully stabilized the system under physiological conditions, membranes cross-linked with genipin exhibited better resistance in physiological environments, improved stability under enzymatic degradation, and a higher degree of in vitro cytocompatibility compared to the other cross-linking agents. The results demonstrated that genipin is an attractive candidate to provide functional structural stability to complex self-assembling structures for potential tissue engineering or in vitro model applications. Statement of SignificanceMolecular self-assembly is widely used for the fabrication of complex functional biomaterials to replace and/or repair any tissue or organ in the body. However, maintaining the stability and corresponding functionality of these kinds of materials in physiological conditions remains a challenge. Chemical cross-linking strategies (natural or synthetic) have been used in an effort to improve their structural integrity. Here we investigate key performance parameters of different cross-linking strategies for stabilising self-assembled materials with potential biomedical applications using a novel protein-peptide co-assembling membrane as proof-of-concept. From the different cross-linkers tested, the natural cross-linker genipin exhibited the best performance. This cross-linker successfully enhanced the mechanical properties of the system enabling the maintenance of the structure in physiological conditions without compromising its bioactivity and biocompatibility. Altogether, we provide a systematic characterization of cross-linking alternatives for self-assembling materials focused on biocompatibility and stability and demonstrate that genipin is a promising alternative for the cross-linking of such materials with a wide variety of potential applications such as in tissue engineering and drug delivery.

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“…Chondrocytes were isolated under sterile conditions from the femoral-patellar groove of a calf [14]. Cartilage fragments were sharply curetted off the articular surface of the joint and digested using type II collagenase (Sigma Chemical, Missouri, USA).…”

Section: Chondrocyte Seeding and Spheroid Culturing On Micropatternedmentioning

confidence: 99%

Chondrocyte spheroids on microfabricated PEG hydrogel surface and their noninvasive functional monitoring

Otsuka

Nagamura

Kaneko

et al. 2012

Science and Technology of Advanced Materials

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A two-dimensional microarray of 10 000 (100 × 100) chondrocyte spheroids was constructed with a 100 µm spacing on a micropatterned gold electrode that was coated with poly(ethylene glycol) (PEG) hydrogels. The PEGylated surface as a cytophobic region was regulated by controlling the gel structure through photolithography. In this way, a PEG hydrogel was modulated enough to inhibit outgrowth of chondrocytes from a cell adhering region in the horizontal direction, which is critical for inducing formation of three-dimensional chondrocyte aggregations (spheroids) within 24 h. We further report noninvasive monitoring of the cellular functional change at the cell membrane using a chondrocyte-based field effect transistor. This measurement is based on detection of extracellular potential change induced as a result of the interaction between extracellular matrix protein secreted from spheroid and substrate at the cell membrane. The interface potential change at the cell membrane/gate interface can be monitored during the differentiation of spheroids without any labeling materials. Our measurements of the time evolution of the interface potential provide important information for understanding the uptake kinetics for cellular differentiation.

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Encapsulation of chondrocytes in injectable alkali-treated collagen gels prepared using poly(ethylene glycol)-based 4-armed star polymer

Cited by 84 publications

References 21 publications

Reuptake of Extracellular Amelogenin by Dental Epithelial Cells Results in Increased Levels of Amelogenin mRNA through Enhanced mRNA Stabilization

Reuptake of Extracellular Amelogenin by Dental Epithelial Cells Results in Increased Levels of Amelogenin mRNA through Enhanced mRNA Stabilization

Cross-linking of a biopolymer-peptide co-assembling system

Chondrocyte spheroids on microfabricated PEG hydrogel surface and their noninvasive functional monitoring

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