scholarly journals Regulation of Decellularized Tissue Remodeling via Scaffold-Mediated Lentiviral Delivery in Anatomically-Shaped Osteochondral Constructs

2018 ◽  
Author(s):  
Christopher R. Rowland ◽  
Katherine A. Glass ◽  
Adarsh R. Ettyreddy ◽  
Catherine C. Gloss ◽  
Jared Matthews ◽  
...  
Keyword(s):  
Transforming Growth Factor Beta ◽  
Protein Delivery ◽  
Spatial Organization ◽  
Transforming Growth Factor ◽  
Interleukin 1 ◽  
Bone Morphogenetic Protein 2 ◽  
Cartilage Tissue ◽  
Site Specific ◽  
Engineer Cartilage

AbstractCartilage-derived matrix (CDM) has emerged as a promising scaffold material for tissue engineering of cartilage and bone due to its native chondroinductive capacity and its ability to support endochondral ossification. Because it consists of native tissue, CDM can undergo cellular remodeling, which can promote integration with host tissue and enables it to be degraded and replaced by neotissue over time. However, enzymatic degradation of decellularized tissues can occur unpredictably and may not allow sufficient time for mechanically competent tissue to form, especially in the harsh inflammatory environment of a diseased joint. The goal of the current study was to engineer cartilage and bone constructs with the ability to inhibit aberrant inflammatory processes caused by the cytokine interleukin-1 (IL-1), through scaffold-mediated delivery of lentiviral particles containing a doxycycline-inducible IL-1 receptor antagonist (IL-1Ra) transgene on anatomically-shaped CDM constructs. Additionally, scaffold-mediated lentiviral gene delivery was used to facilitate spatial organization of simultaneous chondrogenic and osteogenic differentiation via site-specific transduction of a single mesenchymal stem cell (MSC) population to overexpress either chondrogenic, transforming growth factor-beta 3 (TGF-β3), or osteogenic, bone morphogenetic protein-2 (BMP-2), transgenes. Controlled induction of IL-1Ra expression protected CDM hemispheres from inflammation-mediated degradation, and supported robust bone and cartilage tissue formation even in the presence of IL-1. In the absence of inflammatory stimuli, controlled cellular remodeling was exploited as a mechanism for fusing concentric CDM hemispheres overexpressing BMP-2 and TGF-β3 into a single bi-layered osteochondral construct. Our findings demonstrate that site-specific delivery of inducible and tunable transgenes confers spatial and temporal control over both CDM scaffold remodeling and neotissue composition. Furthermore, these constructs provide a microphysiological, in vitro, joint, organoid model with site-specific, tunable, and inducible protein delivery systems for examining the spatiotemporal response to pro-anabolic and/or inflammatory signaling across the osteochondral interface.

scholarly journals Cytokine production by primary bone marrow megakaryocytes

Blood ◽  
1994 ◽  
Vol 84 (12) ◽  
pp. 4151-4156 ◽  
Author(s):  
S Jiang ◽  
JD Levine ◽  
Y Fu ◽  
B Deng ◽  
R London ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Interleukin 1 ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Tnf Alpha ◽  
Necrosis Factor Alpha ◽  
Gm Csf

Primary human bone marrow megakaryocytes were studied for their ability to express and release cytokines potentially relevant to their proliferation and/or differentiation. The purity of the bone marrow megakaryocytes was assessed by morphologic and immunocytochemical criteria. Unstimulated marrow megakaryocytes constitutively expressed genes for interleukin-1 beta (IL-1 beta), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha), by the polymerase chain reaction (PCR) and Northern blot analysis. At the protein level, megakaryocytes secreted significant amounts of IL-1 beta (53.6 +/- 3.6 pg/mL), IL-6 (57.6 +/- 15.6 pg/mL), and GM-CSF (24 +/- 4 pg/mL) but not TNF-alpha. Exposure of human marrow megakaryocytes to IL-1 beta increased the levels of IL-6 (87.3 +/- 2.3 pg/mL) detected in the culture supernatants. Transforming growth factor- beta was also able to stimulate IL-6, IL-1 beta, and GM-CSF secretion, but was less potent than stimulation with phorbol-12-myristate-13- acetate (PMA). The secreted cytokines acted additively to maintain and increase the number of colony-forming unit-megakaryocytes colonies (approximately 35%). These studies demonstrate the production of multiple cytokines by isolated human bone marrow megakaryocytes constitutively or stimulated in vitro. The capacity of human megakaryocytes to synthesize several cytokines known to modulate hematopoietic cells supports the concept that there may be an autocrine mechanism operative in the regulation of megakaryocytopoiesis.

scholarly journals Interacting cytokines regulate in vitro human megakaryocytopoiesis

Blood ◽  
1989 ◽  
Vol 73 (3) ◽  
pp. 671-677 ◽  
Author(s):  
E Bruno ◽  
ME Miller ◽  
R Hoffman
Keyword(s):  
Transforming Growth Factor Beta ◽  
Colony Formation ◽  
Transforming Growth Factor ◽  
Interleukin 1 ◽  
Interleukin 4 ◽  
Recombinant Erythropoietin ◽  
Hematopoietic Growth Factors ◽  
Macrophage Colony ◽  
Stimulating Factor

The effects of hematopoietic growth factors on in vitro human megakaryocytopoiesis were studied using a serum-depleted culture system. Both recombinant interleukin-3 (r-IL-3) and recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) increased megakaryocyte (MK) colony formation (P less than .01) above that observed in baseline cultures. Recombinant interleukin-4 (rIL-4) and interleukin 1 alpha (rIL-1 alpha) failed either to promote MK colony formation alone or to increase rIL-3 or rGM-CSF promoted colony formation. Recombinant erythropoietin (rEpo) and purified thrombocytopoiesis-stimulating factor (TSF) did not increase (P greater than .05) MK colony formation when added alone but synergized with rIL- 1 alpha, leading to a twofold increase in MK colony formation. Such a synergistic relationship was not observed between rIL-4 and rEpo. In addition, TSF enhanced the ability of rIL-3 but not rGM-CSF to promote MK colony formation. Addition of rEpo to optimal or suboptimal concentrations of rGM-CSF or suboptimal concentrations of rIL-3 resulted in a significant increase (P less than .05) in the total number of MK-containing colonies, due to the appearance of multilineage colonies containing MKs. The addition of rEpo to optimal concentrations of rIL-3 resulted in increased numbers of multilineage colonies containing MKs; however, the number of total MK-containing colonies was not significantly increased when compared to assays containing rIL-3 alone. By contrast, transforming growth factor-beta (TGF-beta) inhibited both rIL-3, and rGM-CSF promoted MK colony formation, with optimal inhibition resulting in a 35%-45% reduction of MK colony formation.

Cytokines modulate phagocytosis and intracellular digestion of collagen fibrils by fibroblasts in rabbit periosteal explants. Inverse effects on procollagenase production and collagen phagocytosis

1995 ◽  
Vol 108 (10) ◽  
pp. 3307-3315 ◽  
Author(s):  
E. van der Zee ◽  
V. Everts ◽  
K. Hoeben ◽  
W. Beertsen
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Interleukin 1 ◽  
Collagen Fibrils ◽  
Fibrillar Collagen ◽  
Tgf Beta ◽  
Intracellular Digestion ◽  
Collagen Phagocytosis

Degradation of fibrillar collagen may occur in the extracellular space by enzymes, such as the metalloproteinase collagenase, or in the lysosomal apparatus of fibroblasts following phagocytosis. As the mechanisms involved in the regulation of the latter process are unknown, we investigated possible modulating effects of the cytokines epidermal growth factor (EGF), platelet-derived growth factor (PDGF), interleukin-1 alpha (IL-1 alpha) and transforming growth factor-beta (TGF-beta) on both collagen phagocytosis and the release of collagenase in an in vitro model employing periosteal tissue explants. The data demonstrated that the level of intracellular collagen digestion could be influenced by cytokines: IL-1 alpha inhibited and TGF-beta enhanced phagocytosis of fibrillar collagen by periosteal fibroblasts, whereas the cytokines had an opposite effect on the release of procollagenase. In combination, IL-1 alpha and TGF-beta proved to have an antagonizing effect on either parameter. PDGF and EGF had no effect on phagocytosis or collagenase release. The level of phagocytosed collagen correlated positively with the actual breakdown of collagen as assessed by the release of hydroxyproline but negatively with the level of released procollagenase. Our findings demonstrated that cytokines are able to modulate both the phagocytosis of collagen fibrils by fibroblasts and their subsequent intracellular breakdown, as well as the release of procollagenase, an enzyme considered crucial for extracellular collagenolysis. Moreover, our data show a negative correlation between these two parameters. It is concluded that IL-1 alpha, EGF and TGF-beta may be important in modulating the contribution of the intracellular and extracellular route of collagen breakdown.

scholarly journals Interacting cytokines regulate in vitro human megakaryocytopoiesis

Blood ◽  
1989 ◽  
Vol 73 (3) ◽  
pp. 671-677 ◽  
Author(s):  
E Bruno ◽  
ME Miller ◽  
R Hoffman
Keyword(s):  
Transforming Growth Factor Beta ◽  
Colony Formation ◽  
Transforming Growth Factor ◽  
Interleukin 1 ◽  
Interleukin 4 ◽  
Recombinant Erythropoietin ◽  
Hematopoietic Growth Factors ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract The effects of hematopoietic growth factors on in vitro human megakaryocytopoiesis were studied using a serum-depleted culture system. Both recombinant interleukin-3 (r-IL-3) and recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) increased megakaryocyte (MK) colony formation (P less than .01) above that observed in baseline cultures. Recombinant interleukin-4 (rIL-4) and interleukin 1 alpha (rIL-1 alpha) failed either to promote MK colony formation alone or to increase rIL-3 or rGM-CSF promoted colony formation. Recombinant erythropoietin (rEpo) and purified thrombocytopoiesis-stimulating factor (TSF) did not increase (P greater than .05) MK colony formation when added alone but synergized with rIL- 1 alpha, leading to a twofold increase in MK colony formation. Such a synergistic relationship was not observed between rIL-4 and rEpo. In addition, TSF enhanced the ability of rIL-3 but not rGM-CSF to promote MK colony formation. Addition of rEpo to optimal or suboptimal concentrations of rGM-CSF or suboptimal concentrations of rIL-3 resulted in a significant increase (P less than .05) in the total number of MK-containing colonies, due to the appearance of multilineage colonies containing MKs. The addition of rEpo to optimal concentrations of rIL-3 resulted in increased numbers of multilineage colonies containing MKs; however, the number of total MK-containing colonies was not significantly increased when compared to assays containing rIL-3 alone. By contrast, transforming growth factor-beta (TGF-beta) inhibited both rIL-3, and rGM-CSF promoted MK colony formation, with optimal inhibition resulting in a 35%-45% reduction of MK colony formation.

scholarly journals In Vitro Hair Dermal Papilla Cells Induction by Fagraea berteroana, a Tree of the Marquesan Cosmetopoeia (French Polynesia)

Cosmetics ◽  
2021 ◽  
Vol 8 (1) ◽  
pp. 13
Author(s):  
Kristelle Hughes ◽  
Raimana Ho ◽  
Claire Chazaud ◽  
Stéphanie Hermitte ◽  
Stéphane Greff ◽  
...  
Keyword(s):  
Transforming Growth Factor Beta ◽  
Hair Growth ◽  
Transforming Growth Factor ◽  
Dermal Papilla ◽  
Fold Increase ◽  
French Polynesia ◽  
Bone Morphogenetic Protein 2 ◽  
Dermal Papilla Cells ◽  
Growth State

Fagraea berteroana is a tree used in traditional medicine in various islands of the South Pacific. Here, we studied its hair growth-inducing properties as suggested by one of its Marquesan ethno-uses in haircare. The ethyl acetate extract of the fruits of F. berteroana (FEAE) and four resulting fractions (FEAE-F0, FEAE-F1, FEAE-F2, and FEAE-F3) were tested on hair follicle dermal papilla cells to determine their cell proliferative activity. Furthermore, RT-qPCR analysis enabled gene modulation analysis, while immunostaining of the β-catenin protein was used to follow protein regulation. We found that the plant extracts induced a controlled, dose-dependent cell proliferation. FEAE-F0 simultaneously down-regulated Bone Morphogenetic Protein 2 (BMP2) mRNA expression and upregulated Cyclin-D1 (CCND1) gene expression, which suggests an involvement in the regulation of the Wnt and Transforming Growth Factor beta (TGFβ) pathways that control the hair cycle. FEAE-F0 exhibited a 1.34-fold increase of nuclear β-catenin protein. This is indicative of an active hair growth state. Thus, we conclude that FEAE-F0 could be an innovative candidate in hair care, which opens interesting leads to promote the Marquesan cosmetopoeia.

scholarly journals The in vitro production of endochondral bone from bovine periosteal explants

2021 ◽  
Author(s):  
Kristina Collavino
Keyword(s):  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Bone Morphogenetic Protein 2 ◽  
Cartilage Tissue ◽  
Osseous Tissue ◽  
In Vitro Production ◽  
Endochondral Bone ◽  
Media Types

In the present study, periosteal tissue explants were explored as a substratum for the production of endochondral bone tissue in vitro. Endochondral bone is formed when mesenchymal stem cells (MSCs) proceed through the chondrogenic lineage to produce a transitory cartilage template which eventually is ossified. The periosteum is of interest as this tissue is found enveloping long bones and has been shown to contain a resident cellular population capable of generating endochondral bone. Endochondral ossification was induced in periosteal explants through the successive application of chondrogenic and hypertrophic/osteogenic media simulating the in vivo progression of the process. Different chondrogenic and osteogenic media types were utilized in order to assess the best method for producing osseous tissue. The results indicated that endochondral bone could be produced from periosteum tissue in vitro. It was determined that chondrogenic culture with transforming growth factor β1 (TGFβ1) led to the development of immature (resting or proliferative) cartilage tissue while chondrogenic culture with bone morphogenetic protein 2 (BMP2) produced mature (hypertrophic or calcified) cartilage and osseous tissue. Osteogenic media generally failed to improve ossification in cartilaginous explants but did affect their progression through the endochondral process. Cartilaginous periosteum explants responded differently to osteogenic media types based on the method of chondrogenic pre-induction. Immature cartilage formed under TGFβ1 induction underwent maturation in osteogenic media with triiodothyronine (T3). Mature cartilage formed under BMP2 continued to undergo maturation in the presence of osteogenic media with BMP2 or T3. Overall, these findings suggested that BMP2 is crucial in the development of endochondral bone from periosteal explants in vitro and that the osteogenic media is unnecessary in promoting this process.

scholarly journals The in vitro production of endochondral bone from bovine periosteal explants

2021 ◽  
Author(s):  
Kristina Collavino
Keyword(s):  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Bone Morphogenetic Protein 2 ◽  
Cartilage Tissue ◽  
Osseous Tissue ◽  
In Vitro Production ◽  
Endochondral Bone ◽  
Media Types

In the present study, periosteal tissue explants were explored as a substratum for the production of endochondral bone tissue in vitro. Endochondral bone is formed when mesenchymal stem cells (MSCs) proceed through the chondrogenic lineage to produce a transitory cartilage template which eventually is ossified. The periosteum is of interest as this tissue is found enveloping long bones and has been shown to contain a resident cellular population capable of generating endochondral bone. Endochondral ossification was induced in periosteal explants through the successive application of chondrogenic and hypertrophic/osteogenic media simulating the in vivo progression of the process. Different chondrogenic and osteogenic media types were utilized in order to assess the best method for producing osseous tissue. The results indicated that endochondral bone could be produced from periosteum tissue in vitro. It was determined that chondrogenic culture with transforming growth factor β1 (TGFβ1) led to the development of immature (resting or proliferative) cartilage tissue while chondrogenic culture with bone morphogenetic protein 2 (BMP2) produced mature (hypertrophic or calcified) cartilage and osseous tissue. Osteogenic media generally failed to improve ossification in cartilaginous explants but did affect their progression through the endochondral process. Cartilaginous periosteum explants responded differently to osteogenic media types based on the method of chondrogenic pre-induction. Immature cartilage formed under TGFβ1 induction underwent maturation in osteogenic media with triiodothyronine (T3). Mature cartilage formed under BMP2 continued to undergo maturation in the presence of osteogenic media with BMP2 or T3. Overall, these findings suggested that BMP2 is crucial in the development of endochondral bone from periosteal explants in vitro and that the osteogenic media is unnecessary in promoting this process.

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