scholarly journals Methods of Cryoprotectant Preservation: Allogeneic Cellular Bone Grafts and Potential Effects

2019 ◽  
Vol 2019 ◽  
pp. 1-7
Author(s):  
W. Blake Martin ◽  
Renaud Sicard ◽  
Shabnam M. Namin ◽  
Timothy Ganey
Keyword(s):  
Bone Grafting ◽  
Bone Cells ◽  
Cell Types ◽  
Osteogenic Potential ◽  
Fusion Procedure ◽  
Long Term Storage ◽  
Local Bone ◽  
Surgical Fusion ◽  
Cellular Bone ◽  
Autologous Grafts

Debridement of the bone surface during a surgical fusion procedure initiates an injury response promoting a healing cascade of molecular mediators released over time. Autologous grafts offer natural scaffolding to fill the bone void and to provide local bone cells. Commercial bone grafting products such as allografts, synthetic bone mineral products, etc., are used to supplement or to replace autologous grafts by supporting osteoinductivity, osteoconductivity, and osteogenesis at the surgical site. To assure osteogenic potential, preservation of allogeneic cells with cryoprotectants has been developed to allow for long-term storage and thus delivery of viable bone cells to the surgical site. Dimethyl sulfoxide (DMSO) is an intracellular cryoprotectant commonly used because it provides good viability of the cells post-thaw. However, there is known cytotoxicity reported for DMSO when cells are stored above cryogenic temperatures. For most cellular bone graft products, the cryoprotectant is incorporated with the cells into the other mineralized bone and demineralized bone components. During thawing, the DMSO may not be sufficiently removed from allograft products compared to its use in a cell suspension where removal by washing and centrifugation is available. Therefore, both the allogeneic cell types in the bone grafting product and the local cell types at the bone grafting site could be affected as cytotoxicity varies by cell type and by DMSO content according to reported studies. Overcoming cytotoxicity may be an additional challenge in the formation of bone at a wound or surgical site. Other extracellular cryoprotectants have been explored as alternatives to DMSO which preserve without entering the cell membrane, thereby providing good cellular viability post-thaw and might abrogate the cytotoxicity concerns.

scholarly journals Bergamot Polyphenol Fraction Exerts Effects on Bone Biology by Activating ERK 1/2 and Wnt/β-Catenin Pathway and Regulating Bone Biomarkers in Bone Cell Cultures

Nutrients ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 1305 ◽  
Author(s):  
Arturo Pujia ◽  
Cristina Russo ◽  
Samantha Maurotti ◽  
Roberta Pujia ◽  
Vincenzo Mollace ◽  
...  
Keyword(s):  
Bone Cells ◽  
Citrus Fruit ◽  
Cell Types ◽  
Bone Cell ◽  
In Vivo Studies ◽  
Bone Biology ◽  
High Concentration ◽  
Mineral Density ◽  
Differentiation Markers

Epidemiological studies show that fruit consumption may modulate bone mineral density. However, data regarding the effect of the Citrus bergamia Risso (Bergamot orange), a citrus fruit containing a high concentration of flavonoids, on bone health are still lacking. In this study, we investigated the effects of Bergamot polyphenols on the Wnt/β-catenin pathway in two distinct bone cell types (Saos-2 and MG63). Findings showed that exposure to 0.01 and 0.1 mg/mL doses upregulate β-catenin expression (p = 0.001), osteoblast differentiation markers (e.g., RUNX2 and COL1A), and downregulate RANKL (p = 0.028), as compared to the control. Our results highlight, for the first time, that Bergamot polyphenols act on bone cells through the β-catenin pathway. In vivo studies are necessary to fully understand Bergamot’s role against bone resorption.

scholarly journals High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction

Micromachines ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 387
Author(s):  
Jianxiong Zhang ◽  
Yawei Hu ◽  
Xiaoqing Wang ◽  
Peng Liu ◽  
Xiaofang Chen
Keyword(s):  
Gene Delivery ◽  
High Throughput ◽  
Large Scale ◽  
Cell Types ◽  
Long Term Storage ◽  
Efficient Gene ◽  
Genetic Modifications ◽  
Media Change ◽  
Virus Packaging ◽  
Nih 3T3

Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types.

Uni- and Bilateral Instrumented Posterolateral Fusion of the Lumbar Spine With Local Bone Grafting

Spine ◽  
2011 ◽  
Vol 36 (26) ◽  
pp. E1744-E1748 ◽  
Author(s):  
Seiji Ohtori ◽  
Takana Koshi ◽  
Munetaka Suzuki ◽  
Masashi Takaso ◽  
Masaomi Yamashita ◽  
...  
Keyword(s):  
Lumbar Spine ◽  
Bone Grafting ◽  
Posterolateral Fusion ◽  
Local Bone

scholarly journals Composite Membranes of Poly(ε-caprolactone) with Bisphosphonate-Loaded Bioactive Glasses for Potential Bone Tissue Engineering Applications

Molecules ◽  
2019 ◽  
Vol 24 (17) ◽  
pp. 3067 ◽  
Author(s):  
Zoi Terzopoulou ◽  
Diana Baciu ◽  
Eleni Gounari ◽  
Theodore Steriotis ◽  
Georgia Charalambopoulou ◽  
...  
Keyword(s):  
Bone Cells ◽  
Cell Attachment ◽  
Composite Membranes ◽  
Osteogenic Potential ◽  
Bioactive Glasses ◽  
Noticeable Increase ◽  
Different Types ◽  
Thin Membranes ◽  
Osteogenic Effect ◽  
Effect Of The Composition

Poly(ε-caprolactone) (PCL) is a bioresorbable synthetic polyester with numerous biomedical applications. PCL membranes show great potential in guided tissue regeneration because they are biocompatible, occlusive and space maintaining, but lack osteoconductivity. Therefore, two different types of mesoporous bioactive glasses (SiO2-CaO-P2O5 and SiO2-SrO-P2O5) were synthesized and incorporated in PCL thin membranes by spin coating. To enhance the osteogenic effect of resulting membranes, the bioglasses were loaded with the bisphosphonate drug ibandronate prior to their incorporation in the polymeric matrix. The effect of the composition of the bioglasses as well as the presence of absorbed ibandronate on the physicochemical, cell attachment and differentiation properties of the PCL membranes was evaluated. Both fillers led to a decrease of the crystallinity of PCL, along with an increase in its hydrophilicity and a noticeable increase in its bioactivity. Bioactivity was further increased in the presence of a Sr substituted bioglass loaded with ibandronate. The membranes exhibited excellent biocompatibility upon estimation of their cytotoxicity on Wharton’s Jelly Mesenchymal Stromal Cells (WJ-SCs), while they presented higher osteogenic potential in comparison with neat PCL after WJ-SCs induced differentiation towards bone cells, which was enhanced by a possible synergistic effect of Sr and ibandronate.

scholarly journals Cells Derived from Human Long Bone Appear More Differentiated and More Actively Stimulate Osteoclastogenesis Compared to Alveolar Bone-Derived Cells

2020 ◽  
Vol 21 (14) ◽  
pp. 5072
Author(s):  
Cindy Kelder ◽  
Cornelis J. Kleverlaan ◽  
Marjolijn Gilijamse ◽  
Astrid D. Bakker ◽  
Teun J. de Vries
Keyword(s):  
Tissue Engineering ◽  
Mononuclear Cells ◽  
Alveolar Bone ◽  
Bone Cells ◽  
Transmembrane Protein ◽  
Cell Types ◽  
Long Bone ◽  
Long Bones ◽  
Peripheral Blood Mononuclear ◽  
Alveolar Bone Cells

Osteoblasts derived from mouse skulls have increased osteoclastogenic potential compared to long bone osteoblasts when stimulated with 1,25(OH)2 vitamin D3 (vitD3). This indicates that bone cells from specific sites can react differently to biochemical signals, e.g., during inflammation or as emitted by bioactive bone tissue-engineering constructs. Given the high turn-over of alveolar bone, we hypothesized that human alveolar bone-derived osteoblasts have an increased osteogenic and osteoclastogenic potential compared to the osteoblasts derived from long bone. The osteogenic and osteoclastogenic capacity of alveolar bone cells and long bone cells were assessed in the presence and absence of osteotropic agent vitD3. Both cell types were studied in osteogenesis experiments, using an osteogenic medium, and in osteoclastogenesis experiments by co-culturing osteoblasts with peripheral blood mononuclear cells (PBMCs). Both osteogenic and osteoclastic markers were measured. At day 0, long bones seem to have a more late-osteoblastic/preosteocyte-like phenotype compared to the alveolar bone cells as shown by slower proliferation, the higher expression of the matrix molecule Osteopontin (OPN) and the osteocyte-enriched cytoskeletal component Actin alpha 1 (ACTA1). This phenotype was maintained during the osteogenesis assays, where long bone-derived cells still expressed more OPN and ACTA1. Under co-culture conditions with PBMCs, long bone cells also had a higher Tumor necrose factor-alfa (TNF-α) expression and induced the formation of osteoclasts more than alveolar bone cells. Correspondingly, the expression of osteoclast genes dendritic cell specific transmembrane protein (DC-STAMP) and Receptor activator of nuclear factor kappa-Β ligand (RankL) was higher in long bone co-cultures. Together, our results indicate that long bone-derived osteoblasts are more active in bone-remodeling processes, especially in osteoclastogenesis, than alveolar bone-derived cells. This indicates that tissue-engineering solutions need to be specifically designed for the site of application, such as defects in long bones vs. the regeneration of alveolar bone after severe periodontitis.

Chemokines in Oral Inflammatory Diseases: Apical Periodontitis and Periodontal Disease

2007 ◽  
Vol 86 (4) ◽  
pp. 306-319 ◽  
Author(s):  
T.A. Silva ◽  
G.P. Garlet ◽  
S.Y. Fukada ◽  
J.S. Silva ◽  
F.Q. Cunha
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Bone Cells ◽  
Inflammatory Diseases ◽  
Cell Types ◽  
Cellular Responses ◽  
Experimental Models ◽  
Periodontal Tissue ◽  
Oral Diseases ◽  
Specific Receptors

The inflammatory oral diseases are characterized by the persistent migration of polymorphonuclear leukocytes, monocytes, lymphocytes, plasma and mast cells, and osteoblasts and osteoclasts. In the last decade, there has been a great interest in the mediators responsible for the selective recruitment and activation of these cell types at inflammatory sites. Of these mediators, the chemokines have received particular attention in recent years. Chemokine messages are decoded by specific receptors that initiate signal transduction events, leading to a multitude of cellular responses, including chemotaxis and activation of inflammatory and bone cells. However, little is known about their role in the pathogenesis of inflammatory oral diseases. The purpose of this review is to summarize the findings regarding the role of chemokines in periapical and periodontal tissue inflammation, and the integration, into experimental models, of the information about the role of chemokines in human diseases.

scholarly journals Extracellular Vesicles in Tumors: A Potential Mediator of Bone Metastasis

2021 ◽  
Vol 9 ◽  
Author(s):  
Shenglong Li ◽  
Wei Wang
Keyword(s):  
Bone Metastasis ◽  
Extracellular Vesicles ◽  
Lipid Membrane ◽  
Bone Cells ◽  
Membrane Vesicles ◽  
Metastatic Tumor ◽  
Cell Types ◽  
Biological Phenomena ◽  
Breast And Prostate Cancer ◽  
Metastatic Sites

As one of the most common metastatic sites, bone has a unique microenvironment for the growth and prosperity of metastatic tumor cells. Bone metastasis is a common complication for tumor patients and accounts for 15–20% of systemic metastasis, which is only secondary to lung and liver metastasis. Cancers prone to bone metastasis include lung, breast, and prostate cancer. Extracellular vesicles (EVs) are lipid membrane vesicles released from different cell types. It is clear that EVs are associated with multiple biological phenomena and are crucial for intracellular communication by transporting intracellular substances. Recent studies have implicated EVs in the development of cancer. However, the potential roles of EVs in the pathological exchange of bone cells between tumors and the bone microenvironment remain an emerging area. This review is focused on the role of tumor-derived EVs in bone metastasis and possible regulatory mechanisms.

scholarly journals A Dedifferentiation Strategy to Enhance the Osteogenic Potential of Dental Derived Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Francesco Paduano ◽  
Elisabetta Aiello ◽  
Paul Roy Cooper ◽  
Benedetta Marrelli ◽  
Irina Makeeva ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Cell Fate ◽  
Therapeutic Potential ◽  
Cell Types ◽  
Osteogenic Potential ◽  
Alternative Source ◽  
Dental Tissues ◽  
Numerous Cell ◽  
First Time

Dental stem cells (DSCs) holds the ability to differentiate into numerous cell types. This property makes these cells particularly appropriate for therapeutic use in regenerative medicine. We report evidence that when DSCs undergo osteogenic differentiation, the osteoblast-like cells can be reverted back to a stem-like state and then further differentiated toward the osteogenic phenotype again, without gene manipulation. We have investigated two different MSCs types, both from dental tissues: dental follicle progenitor stem cells (DFPCs) and dental pulp stem cells (DPSCs). After osteogenic differentiation, both DFPCs and DPSCs can be reverted to a naïve stem cell-like status; importantly, dedifferentiated DSCs showed a greater potential to further differentiate toward the osteogenic phenotype. Our report aims to demonstrate for the first time that it is possible, under physiological conditions, to control the dedifferentiation of DSCs and that the rerouting of cell fate could potentially be used to enhance their osteogenic therapeutic potential. Significantly, this study first validates the use of dedifferentiated DSCs as an alternative source for bone tissue engineering.

scholarly journals Evolutionary insights into primate skeletal gene regulation using a comparative cell culture model

2021 ◽  
Author(s):  
Genevieve Housman ◽  
Emilie Briscoe ◽  
Yoav Gilad
Keyword(s):  
Gene Expression ◽  
Gene Regulation ◽  
Bone Cells ◽  
Cell Types ◽  
Differentially Expressed ◽  
Functional Genomic ◽  
Cell Culture Model ◽  
Culture Model ◽  
Study Gene Regulation

AbstractThe evolution of complex skeletal traits in primates was likely influenced by both genetic and environmental factors. Because skeletal tissues are notoriously challenging to study using functional genomic approaches, they remain poorly characterized even in humans, let alone across multiple species. The challenges involved in obtaining functional genomic data from the skeleton, combined with the difficulty of obtaining such tissues from nonhuman apes, motivated us to consider an alternative in vitro system with which to comparatively study gene regulation in skeletal cell types. Specifically, we differentiated six human and six chimpanzee induced pluripotent stem cell lines (iPSCs) into mesenchymal stem cells (MSCs) and subsequently into osteogenic cells (bone cells). We validated differentiation using standard methods and collected single-cell RNA sequencing data from over 100,000 cells across multiple samples and replicates at each stage of differentiation. While most genes that we examined display conserved patterns of expression across species, hundreds of genes are differentially expressed (DE) between humans and chimpanzees within and across stages of osteogenic differentiation. Some of these interspecific DE genes show functional enrichments relevant in skeletal tissue trait development. Moreover, topic modeling indicates that interspecific gene programs become more pronounced as cells mature. Overall, we propose that this in vitro model can be used to identify interspecific regulatory differences that may have contributed to skeletal trait differences between species.Author SummaryPrimates display a range of skeletal morphologies and susceptibilities to skeletal diseases, but the molecular basis of these phenotypic differences is unclear. Studies of gene expression variation in primate skeletal tissues are extremely restricted due to the ethical and practical challenges associated with collecting samples. Nevertheless, the ability to study gene regulation in primate skeletal tissues is crucial for understanding how the primate skeleton has evolved. We therefore developed a comparative primate skeletal cell culture model that allows us to access a spectrum of human and chimpanzee cell types as they differentiate from stem cells into bone cells. While most gene expression patterns are conserved across species, we also identified hundreds of differentially expressed genes between humans and chimpanzees within and across stages of differentiation. We also classified cells by osteogenic stage and identified additional interspecific differentially expressed genes which may contribute to skeletal trait differences. We anticipate that this model will be extremely useful for exploring questions related to gene regulation variation in primate bone biology and development.

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