scholarly journals Establishment of Collagen: Hydroxyapatite/BMP-2 Mimetic Peptide Composites

Materials ◽  
2020 ◽  
Vol 13 (5) ◽  
pp. 1203
Author(s):  
Liane Schuster ◽  
Nina Ardjomandi ◽  
Marita Munz ◽  
Felix Umrath ◽  
Christian Klein ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cost Effective ◽  
Quartz Crystals ◽  
Mimetic Peptide ◽  
Glutamic Acid Residue ◽  
Hydroxyapatite Composite ◽  
Mimetic Peptides ◽  
Periosteal Cells

Extensive efforts were undertaken to develop suitable biomaterials for tissue engineering (TE) applications. To facilitate clinical approval processes and ensure the success of TE applications, bioinspired concepts are currently focused on. Working on bone tissue engineering, we describe in the present study a method for biofunctionalization of collagen/hydroxyapatite composites with BMP-2 mimetic peptides. This approach is expected to be fundamentally transferable to other tissue engineering fields. A modified BMP-2 mimetic peptide containing a negatively charged poly-glutamic acid residue (E7 BMP-2 peptide) was used to bind positively charged hydroxyapatite (HA) particles by electrostatic attraction. Binding efficiency was biochemically detected to be on average 85% compared to 30% of BMP-2 peptide without E7 residue. By quartz crystal microbalance (QCM) analysis, we could demonstrate the time-dependent dissociation of the BMP-2 mimetic peptides and the stable binding of the E7 BMP-2 peptides on HA-coated quartz crystals. As shown by immunofluorescence staining, alkaline phosphatase expression is similar to that detected in jaw periosteal cells (JPCs) stimulated with the whole BMP-2 protein. Mineralization potential of JPCs in the presence of BMP-2 mimetic peptides was also shown to be at least similar or significantly higher when low peptide concentrations were used, as compared to JPCs cultured in the presence of recombinant BMP-2 controls. In the following, collagen/hydroxyapatite composite materials were prepared. By proliferation analysis, we detected a decrease in cell viability with increasing HA ratios. Therefore, we chose a collagen/hydroxyapatite ratio of 1:2, similar to the natural composition of bone. The following inclusion of E7 BMP-2 peptides within the composite material resulted in significantly elevated long-term JPC proliferation under osteogenic conditions. We conclude that our advanced approach for fast and cost-effective scaffold preparation and biofunctionalization is suitable for improved and prolonged JPC proliferation. Further studies should prove the functionality of composite scaffolds in vivo.

scholarly journals Neural plate targeting by in utero nanoinjection (NEPTUNE) reveals a role for Sptbn2 in neurulation and abdominal wall closure

2020 ◽  
Author(s):  
Katrin Mangold ◽  
Jan Mašek ◽  
Jingyan He ◽  
Urban Lendahl ◽  
Elaine Fuchs ◽  
...  
Keyword(s):  
Cost Effective ◽  
Cell Types ◽  
Neural Plate ◽  
Spinocerebellar Ataxia Type ◽  
Loss Of Function ◽  
Conditional Expression ◽  
Cost Effective Technique ◽  
The Brain

ABSTRACTGene variants associated with disease are efficiently identified with whole genome sequencing or GWAS, but validation in vivo lags behind. We developed NEPTUNE (neural plate targeting by in utero nanoinjection), to rapidly and flexibly introduce gene expression-modifying viruses to the embryonic murine neural plate prior to neurulation, to target the future adult nervous system. Stable integration in >95% of cells in the brain enabled long-term gain- or loss-of-function, and conditional expression was achieved using mini-promotors for cell types of interest. Using NEPTUNE, we silenced Sptbn2, a gene associated with Spinocerebellar ataxia type 5 (SCA5) in humans. Silencing of Sptbn2 induced severe neural tube defects and embryo resorption, suggesting that SPTBN2 in-frame and missense deletions in SCA5 reflect hypomorphic or neomorphic functions, not loss of function. In conclusion, NEPTUNE offers a novel, rapid and cost-effective technique to test gene function in brain development, and can reveal loss of function phenotypes incompatible with life.

Long-term in vivo response to citric acid-based nanocomposites for orthopaedic tissue engineering

2011 ◽  
Vol 22 (9) ◽  
pp. 2131-2138 ◽  
Author(s):  
Eun Ji Chung ◽  
Pradeep Kodali ◽  
William Laskin ◽  
Jason L. Koh ◽  
Guillermo A. Ameer
Keyword(s):  
Tissue Engineering ◽  
Citric Acid

Novel Electrospun Nanocomposites With Controllable Compositional Gradient and Degradation Kinetics for Vascular Tissue Engineering

2009 ◽  
Author(s):  
Walter Bonani ◽  
Antonella Motta ◽  
Claudio Migliaresi ◽  
Wei Tan
Keyword(s):  
Tissue Engineering ◽  
Vascular Tissue ◽  
Degradation Kinetics ◽  
Interpenetrating Networks ◽  
Compositional Gradient ◽  
Small Vessels ◽  
Synthetic Materials ◽  
Fiber Materials

Vascular graft materials currently used in the medical field are often made from bioinert synthetic materials such as polytetrafluoroethylene (PTFE). The high long-term failure rate of these materials in the replacement of small vessels is known to be associated with the lack of proper signaling events by PTFE to vascular cells causing adverse hemodynamic, inflammatory or coagulatory conditions. Tissue engineering approaches emerge as a promising method to obtain replacement vessels. These approaches are often based on homogeneous constructs or multilayer of homogeneous constructs are yet to demonstrate capability of controlling the integration of tissue engineering construct in vivo better for long-term patency. Therefore, constant and pressing is the demand for scaffold constructs which can provide not only proper mechanical support, but also precise molecular cues and degradation kinetics to facilitate the proper remodeling and integration process in vivo over the time for long-term patency. To this end, we have developed and demonstrated a novel double-electrospinning apparatus to obtain interpenetrating networks of nanofibers made from different polymers in a tailored proportion with heterogeneous gradient patterns of fiber materials and functional biomolecules.

scholarly journals Tissue engineering of cartilage using poly-ɛ-caprolactone nanofiber scaffolds seeded in vivo with periosteal cells

2010 ◽  
Vol 18 (7) ◽  
pp. 981-991 ◽  
Author(s):  
M.E. Casper ◽  
J.S. Fitzsimmons ◽  
J.J. Stone ◽  
A.O. Meza ◽  
Y. Huang ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Nanofiber Scaffolds ◽  
Periosteal Cells

scholarly journals Excellency of Hydroxyapatite Composite Scaffolds for Bone Tissue Engineering

Biomaterials ◽  
2020 ◽  
Author(s):  
Mohammad Shariful Islam ◽  
Mohammad Abdulla-Al-Mamun ◽  
Alam Khan ◽  
Mitsugu Todo
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Surface Reactivity ◽  
Growth Factor Delivery ◽  
Composite Scaffolds ◽  
In Vivo Experiments ◽  
Hydroxyapatite Composite

The hydroxyapatite [HAp, Ca10(PO4)6(OH)2] has a variety of applications in bone fillers and replacements due to its excellent bioactivity and osteoconductivity. It comprises the main inorganic component of hard tissues. Among the various approaches, a composite approach using several components like biopolymer, gelatin, collagen, and chitosan in the functionalization of scaffolds with HAp has the prospective to be an engineered biomaterial for bone tissue engineering. HAp composite scaffolds have been developed to obtain a material with different functionalities such as surface reactivity, bioactivity, mechanical strength, and capability of drug or growth factor delivery. Several techniques and processes for the synthesis and fabrication of biocompatible HAp composite scaffolds suitable for bone regeneration are addressed here. Further, this chapter described the excellences of various HAp composite scaffolds used in in vitro and in vivo experiments in bone tissue engineering.

scholarly journals Potential Research Tool of Stem Cells from Human Exfoliated Deciduous Teeth: Lentiviral Bmi-1 Immortalization with EGFP Marker

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Siqi Yao ◽  
Lingping Tan ◽  
Huan Chen ◽  
Xiaojun Huang ◽  
Wei Zhao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Stem Cell ◽  
Cell Line ◽  
Fluorescent Protein ◽  
Deciduous Teeth ◽  
Early Passage ◽  
Human Exfoliated Deciduous Teeth

Stem cells from human exfoliated deciduous teeth (SHED) are a favourable source for tissue engineering, for its great proliferative capacity and the ease of collection. However, the transplantation of stem cells and the study of stem cell-based tissue engineering require massive stem cells. After long-term expansion, stem cells face many challenges, including limited lifespan, senescence, and loss of stemness. Therefore, a cell line capable of overcoming those problems should be built. In this study, we generated a Bmi-1-immortalized SHED cell line with an enhanced green fluorescent protein (EGFP) marker (SHED-Bmi1-EGFP) using lentiviral transduction. We compared this cell line with the original SHED for cell morphology under a microscope. The expression of Bmi-1 was detected with Western blot. Replicative lifespan determination and colony-forming efficiency assessment were using to assay proliferation capability. Senescence-associated β-galactosidase assay was performed to assay the senescence level of cells. Moreover, multipotency, karyotype, and tumour formation in nude mice of SHED and SHED-Bmi1-EGFP were also tested. Our results confirmed that Bmi-1 immortalization did not affect the main features of SHED. SHED-Bmi1-EGFP could be passaged for a long time and stably expressed EGFP. SHED-Bmi1-EGFP at a late passage showed low activity of β-galactosidase and similar multilineage differentiation as SHED at an early passage. The immortalized cells had no potential tumourigenicity ability in vivo. Moreover, we provided some suggestions for potential applications of the immortalized SHED cell line with the EGFP marker. Thus, the immortalized cell line we built can be used as a functional tool in the lab for long-term studies of SHED and stem cell-based regeneration.

Screening of mimetic peptides for CD14 binding site with LBP and antiendotoxin activity of mimetic peptide in vivo and in vitro

2009 ◽  
Vol 58 (1) ◽  
pp. 45-53 ◽  
Author(s):  
Z. Xu ◽  
G. S. Qian ◽  
Q. Li ◽  
Q. J. Feng ◽  
G. M. Wu ◽  
...  
Keyword(s):  
Binding Site ◽  
Mimetic Peptide ◽  
Mimetic Peptides
Sign in / Sign up

Export Citation Format

Share Document