TGF-β1/CD105 signaling controls vascular network formation within growth factor sequestering hyaluronic acid hydrogels

ABSTRACT Mouse capillary endothelial cells (1G11 cell line) embedded in type I collagen gels undergo in vitro angiogenesis. Cells rapidly reorganize and form capillary-like structures when stimulated with serum. Transforming growth factor β1 (TGF-β1) alone can substitute for serum and induce cell survival and tubular network formation. This TGF-β1-mediated angiogenic activity depends on phosphatidylinositol 3-kinase (PI3K) and p42/p44 mitogen-activated protein kinase (MAPK) signaling. We showed that specific inhibitors of either pathway (wortmannin, LY-294002, and PD-98059) all suppressed TGF-β1-induced angiogenesis mainly by compromising cell survival. We established that TGF-β1 stimulated the expression of TGF-α mRNA and protein, the tyrosine phosphorylation of a 170-kDa membrane protein representing the epidermal growth factor (EGF) receptor, and the delayed activation of PI3K/Akt and p42/p44 MAPK. Moreover, we showed that all these TGF-β1-mediated signaling events, including tubular network formation, were suppressed by incubating TGF-β1-stimulated endothelial cells with a soluble form of an EGF receptor (ErbB-1) or tyrphostin AG1478, a specific blocker of EGF receptor tyrosine kinase. Finally, addition of TGF-α alone poorly stimulated angiogenesis; however, by reducing cell death, it strongly potentiated the action of TGF-β1. We therefore propose that TGF-β1 promotes angiogenesis at least in part via the autocrine secretion of TGF-α, a cell survival growth factor, activating PI3K/Akt and p42/p44 MAPK.

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Chitosan-Polylactide/Hyaluronic Acid Complex Microspheres as Carriers for Controlled Release of Bioactive Transforming Growth Factor-β1

Pharmaceutics ◽

10.3390/pharmaceutics10040239 ◽

2018 ◽

Vol 10 (4) ◽

pp. 239 ◽

Cited By ~ 4

Author(s):

Qing Min ◽

Jiaoyan Liu ◽

Jing Li ◽

Ying Wan ◽

Jiliang Wu

Keyword(s):

Hyaluronic Acid ◽

Growth Factor ◽

Transforming Growth Factor ◽

Sodium Tripolyphosphate ◽

Early Stage ◽

Transforming Growth Factor Β1 ◽

Acid Complex ◽

Solubility In Water ◽

Protection Method ◽

Tgf Β1

Chitosan(CH)-polylactide(PLA) copolymers containing varied PLA percentages were synthesized using a group-protection method and one of them with solubility in water-based solvents was used to prepare CH-PLA/hyaluronic acid (HA) complex microspheres for the delivery of transforming growth factor-β1 (TGF-β1). An emulsification processing method was developed for producing TGF-β1-loaded CH-PLA/HA microspheres using sodium tripolyphosphate (TPP) as ionic crosslinker and the size of the microspheres was devised to the micron level in order to achieve high encapsulating efficiency. The encapsulating efficiency, swelling property and release administration of the microspheres could be synergistically regulated by PLA component, the applied TPP dose and the incorporated HA amount. In comparison to CH/HA microspheres, the CH-PLA/HA microspheres had greatly reduced TGF-β1 release rates and were able to administrate the TGF-β1 release at controlled rates over a significant longer period of time. The released TGF-β1 was detected to be bioactive when compared to the free TGF-β1. These results suggest that the presently developed CH-PLA/HA complex microspheres have promising potential in delivering TGF-β1 for cartilage repair applications where the applied TGF-β1 amount in the early stage needs to be low whilst the sustained TGF-β1 release at an appropriate dose in the later stage has to be maintained

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Modulatory effects of TGF-β1 and BMP6 on thecal angiogenesis and steroidogenesis in the bovine ovary

Reproduction ◽

10.1530/rep-19-0311 ◽

2020 ◽

Vol 159 (4) ◽

pp. 397-408 ◽

Cited By ~ 3

Author(s):

D Mattar ◽

M Samir ◽

M Laird ◽

P G Knight

Keyword(s):

Endothelial Cell ◽

Growth Factor ◽

Network Formation ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Luteal Function ◽

Cell Network ◽

Progesterone Secretion ◽

Alk5 Inhibitor ◽

Tgf Β1

Angiogenesis plays an integral role in follicular and luteal development and is positively regulated by several intra-ovarian factors including vascular endothelial growth factor A (VEGFA) and fibroblast growth factor 2 (FGF2). Various transforming growth factor-β (TGF-β) superfamily members function as intra-ovarian regulators of follicle and luteal function, but their potential roles in modulating ovarian angiogenesis have received little attention. In this study, we used a bovine theca interna culture model (exhibiting characteristics of luteinization) to examine the effects of TGF-β1 and bone morphogenetic protein 6 (BMP6) on angiogenesis and steroidogenesis. VEGFA/FGF2 treatment promoted endothelial cell network formation but had little or no effect on progesterone and androstenedione secretion or expression of key steroidogenesis-related genes. TGF-β1 suppressed basal and VEGFA/FGF2-induced endothelial cell network formation and progesterone secretion, effects that were reversed by an activin receptor-like kinase 5 (ALK5) inhibitor (SB-431542). The ALK5 inhibitor alone raised androstenedione secretion and expression of several transcripts including CYP17A1. BMP6 also suppressed endothelial cell network formation under VEGFA/FGF2-stimulated conditions and inhibited progesterone secretion and expression of several steroidogenesis-related genes under basal and VEGFA/FGF2-stimulated conditions. These effects were reversed by an ALK1/2 inhibitor (K02288). Moreover, the ALK1/2 inhibitor alone augmented endothelial network formation, progesterone secretion, androstenedione secretion and expression of several steroidogenesis-related genes. The results indicate dual suppressive actions of both TGF-β1 and BMP6 on follicular angiogenesis and steroidogenesis. Further experiments are needed to unravel the complex interactions between TGF-β superfamily signalling and other regulatory factors controlling ovarian angiogenesis and steroidogenesis.

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Tethered TGF-β1 in a Hyaluronic Acid-Based Bioink for Bioprinting Cartilaginous Tissues

International Journal of Molecular Sciences ◽

10.3390/ijms23020924 ◽

2022 ◽

Vol 23 (2) ◽

pp. 924

Author(s):

Julia Hauptstein ◽

Leonard Forster ◽

Ali Nadernezhad ◽

Jürgen Groll ◽

Jörg Teßmar ◽

...

Keyword(s):

Hyaluronic Acid ◽

Growth Factors ◽

Growth Factor ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Cartilage Regeneration ◽

Cartilaginous Tissues ◽

Dual Stage ◽

Tgf Β1

In 3D bioprinting for cartilage regeneration, bioinks that support chondrogenic development are of key importance. Growth factors covalently bound in non-printable hydrogels have been shown to effectively promote chondrogenesis. However, studies that investigate the functionality of tethered growth factors within 3D printable bioinks are still lacking. Therefore, in this study, we established a dual-stage crosslinked hyaluronic acid-based bioink that enabled covalent tethering of transforming growth factor-beta 1 (TGF‑β1). Bone marrow-derived mesenchymal stromal cells (MSCs) were cultured over three weeks in vitro, and chondrogenic differentiation of MSCs within bioink constructs with tethered TGF‑β1 was markedly enhanced, as compared to constructs with non-covalently incorporated TGF‑β1. This was substantiated with regard to early TGF‑β1 signaling, chondrogenic gene expression, qualitative and quantitative ECM deposition and distribution, and resulting construct stiffness. Furthermore, it was successfully demonstrated, in a comparative analysis of cast and printed bioinks, that covalently tethered TGF‑β1 maintained its functionality after 3D printing. Taken together, the presented ink composition enabled the generation of high-quality cartilaginous tissues without the need for continuous exogenous growth factor supply and, thus, bears great potential for future investigation towards cartilage regeneration. Furthermore, growth factor tethering within bioinks, potentially leading to superior tissue development, may also be explored for other biofabrication applications.

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Low Degree Hyaluronic Acid Crosslinking Inducing the Release of TGF-Β1 in Conditioned Medium of Wharton’s Jelly-Derived Stem Cells

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2019.307 ◽

2019 ◽

Vol 7 (10) ◽

pp. 1572-1575 ◽

Cited By ~ 1

Author(s):

Nora Ariyati ◽

Kusworini Kusworini ◽

Nurdiana Nurdiana ◽

Yohanes Widodo Wirohadidjojo

Keyword(s):

Stem Cells ◽

Hyaluronic Acid ◽

Growth Factor ◽

Conditioned Medium ◽

Sandwich Elisa ◽

Wharton’S Jelly ◽

Crosslinking Degree ◽

Wharton's Jelly ◽

Low Degree ◽

Tgf Β1

BACKGROUND: Presently, the application of stem cells and their paracrine effect for anti-ageing therapy has commenced. Wharton's jelly-derived stem cell conditioned medium (WJSCs-CM) is renowned for increasing proliferation, migrate ageing skin fibroblasts and increase consumption of extracellular transforming growth factor-β (TGF-β). With more than 85% of frequently used dermal filler procedures are hyaluronic acid fillers (HA), a mixture of both with optimal HA crosslinking degree has not yet been identified. AIM: This study aimed to determine the discrepancies in the results of various HA crosslinking degree in WJSCs-CM concerning various levels of growth factors (GF). METHODS: Conditioned medium was obtained from mesenchymal stem cells Wharton’s jelly of the newborn umbilical cord with caesarean section procedure, fabricated with hypoxia method (HCM). HA was obtained from preparations on the market with crosslinking degrees of 3%, 4%, and 10%. GF levels were measured using sandwich ELISA method based on the protocol provided by anti-TGF-β1, platelet-derived growth factor (PDGF), and basic fibroblast growth factor (bFGF) antibody producers (Cloud-Clone Corp®, Texas, USA). RESULTS: Low degree HA crosslinking (3% and 4%) elevated TGF-β1 release in WJSCs-CM. HA crosslinking did not provoke increased levels of PDGF and bFGF in WJSCs-CM, both at low and higher degrees. CONCLUSION: Low degree HA crosslinking induced the increase of TGF-β1 release in WJSCs-CM.

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Biocompatibility and Tolerability of a Purely Bicarbonate-Buffered Peritoneal Dialysis Solution

Peritoneal Dialysis International ◽

10.1177/089686080902900610 ◽

2009 ◽

Vol 29 (6) ◽

pp. 647-655 ◽

Cited By ~ 32

Author(s):

Lars Weiss ◽

Bernd Stegmayr ◽

Gudrun Malmsten ◽

Mattias Tejde ◽

Henrik Hadimeri ◽

...

Keyword(s):

Renal Function ◽

Hyaluronic Acid ◽

Peritoneal Dialysis ◽

Growth Factor ◽

Membrane Integrity ◽

Residual Renal Function ◽

Fluid Exchange ◽

Tnf Α ◽

Peritoneal Dialysis Fluid ◽

Tgf Β1

Background Novel peritoneal dialysis solutions are characterized by a minimal content of glucose degradation products and a neutral pH. Many studies have shown the biocompatibility of neutral lactate-buffered solutions; however, until now, the effect of purely bicarbonate-buffered solutions has not been intensively studied in vivo. Methods This study was an open label, prospective, crossover multicenter trial to investigate the biocompatibility of a purely bicarbonate-buffered solution (bicPDF) by measuring biocompatibility parameters such as cancer antigen 125 (CA125) in peritoneal effluent. 55 patients were enrolled in the study. After a 2-week run-in phase, 53 patients could be randomized into 2 groups, starting with either standard lactate-buffered peritoneal dialysis fluid (SPDF) for 12 weeks (phase 1) and then switching to bicPDF for 12 weeks (phase 2), or vice versa. Overnight peritoneal effluents were collected at baseline and at the end of phases 1 and 2 and were tested for CA125, hyaluronic acid, vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interferon gamma (IFNγ), and transforming growth factor-beta1 (TGF-β1). Total ultrafiltration and residual renal function were also assessed. At the end of the study, pain during fluid exchange and dwell was evaluated using special questionnaires. Results 34 patients completed the study; 27 of them provided data for analysis of the biocompatibility parameters. CA125 levels in overnight effluent were significantly higher with bicPDF (61.9 ± 33.2 U/L) than with SPDF (18.6 ± 18.2 U/L, p < 0.001). Hyaluronic acid levels were significantly lower after the use of bicPDF (185.0 ± 119.6 ng/mL) than after SPDF (257.4 ± 174.0 ng/mL, p = 0.013). Both TNF-α and TGF-β1 showed higher levels with the use of bicPDF than with SPDF. No differences were observed for IL-6, VEGF, or IFNγ levels. We observed an improvement in the glomerular filtration rate with the use of bicPDF but no differences were observed for total fluid loss. Pain scores could be analyzed in 23 patients: there was no difference between the solutions. Conclusions The use of a purely bicarbonate-buffered low-glucose degradation product solution significantly changes most of the peritoneal effluent markers measured, suggesting an improvement in peritoneal membrane integrity. Additionally, it seems to have a positive effect on residual renal function.

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