Mechanical Stimulation Increases Collagen Type I and Collagen Type III Gene Expression of Stem Cell: Collagen Sponge Constructs for Patellar Tendon Repair

2007 ◽  
Author(s):  
Natalia Juncosa-Melvin ◽  
Karl S. Matlin ◽  
Robert W. Holdcraft ◽  
Victor S. Nirmalanandhan ◽  
David L. Butler
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Soft Tissues ◽  
Collagen Type ◽  
Tendon Repair ◽  
Collagen Type I ◽  
Collagen Sponge ◽  
Type I ◽  
Collagen Type Iii ◽  
Initial Stiffness

Tendons (rotator cuff, Achilles and patellar tendons) are among the most commonly injured soft tissues [1]. Many repairs/reconstructions have been attempted using sutures, resorbable biomaterials, autografts, and allografts, but with varying success. A tissue engineered repair using mesenchymal stem cells (MSCs) is attractive [2–4] but often lacks initial stiffness and strength [5].

Mechanical Stimulation Increases Collagen Type I and Collagen Type III Gene Expression of Stem Cell–Collagen Sponge Constructs for Patellar Tendon Repair

2007 ◽  
Vol 13 (6) ◽  
pp. 1219-1226 ◽  
Author(s):  
Natalia Juncosa-Melvin ◽  
Karl S. Matlin ◽  
Robert W. Holdcraft ◽  
Victor S. Nirmalanandhan ◽  
David L. Butler
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Patellar Tendon ◽  
Mechanical Stimulation ◽  
Collagen Type ◽  
Tendon Repair ◽  
Collagen Type I ◽  
Collagen Sponge ◽  
Type I ◽  
Collagen Type Iii

scholarly journals The Effects of Bone Marrow-Derived Mesenchymal Stem Cells and Fascia Wrap Application to Anterior Cruciate Ligament Tissue Engineering

2005 ◽  
Vol 14 (10) ◽  
pp. 763-773 ◽  
Author(s):  
Zigang Ge ◽  
James Cho Hong Goh ◽  
Eng Hin Lee
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Collagen Type ◽  
Cruciate Ligament ◽  
Collagen Type I ◽  
Fascia Lata ◽  
Type I ◽  
Collagen Type Iii ◽  
Group I ◽  
Anterior Cruciate

After an anterior cruciate ligament (ACL) injury, surgical reconstructions are necessary in most cases, either with autografts, allografts, or artificial ligaments. Potential tissue-engineered ligaments would circumvent the disadvantages apparent in these methods. While seeding of mesenchymal stem cells (MSCs) and fascia wrap could potentially improve tissue regeneration and mechanical properties, their exact roles were evaluated in the current study. Knitted biodegradable scaffolds of poly-L-lactic acid (PLLA) and poly-glycolic-lactic acid (PGLA) yarns were used to reconstruct ACL in 48 rabbits. These were divided into four equal groups: only knitted scaffolds were used in group I; knitted scaffolds and mesenchymal stem cells were used in group II; knitted scaffolds, MSCs, and fascia lata were used in group III; knitted scaffolds and fascia lata were used in group IV. Carboxyfluorescein diacetate (CFDA)-labeled MSCs were used to trace the fate of seeded cells in groups II and III. Histology, Western blot analysis, and mechanical properties of reconstructed ACL were analyzed after 20 weeks. Fibroblast ingrowths were seen in all four groups while CFDA-labeled MSCs could be found after 8 weeks of implantation in groups II and III. Both the amount of collagen type I and collagen type III in groups III and IV were significantly higher than in group II, which was much higher than in group I. Both maximal tensile loads and stiffness of the reconstructed ACLs in groups I, II, III, and IV were significantly lower than normal controls after 20 weeks of implantation. It is concluded that MSCs could promote synthesis of collagen type I and collagen type III in tissue-engineered ligaments, while fascia wraps have stronger effects. Both MSC seeding and fascia wrap could not enhance ultimate tensile load and stiffness.

scholarly journals Tenogenically differentiated adipose-derived stem cells are effective in Achilles tendon repair in vivo

2018 ◽  
Vol 9 ◽  
pp. 204173141881118 ◽  
Author(s):  
Jolanta B Norelli ◽  
Dawid P Plaza ◽  
Drew N Stal ◽  
Anish M Varghese ◽  
Haixiang Liang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Collagen Type ◽  
Tendon Repair ◽  
Collagen Type I ◽  
Platelet Derived Growth Factor ◽  
Type I ◽  
Adipose Derived Stem Cells ◽  
Adipose Derived Stem Cell

The purpose of this study was to characterize rat adipose-derived stem cells, induce adipose-derived stem cell tenogenesis, and analyze adipose-derived stem cell effects on tendon repair in vivo. Adipose-derived stem cells demonstrated an immunomodulatory, pro-angiogenic, and pro-proliferatory profile in vitro. Tenogenesis was induced for 1, 7, 14, and 21 days with 24 combinations of growth differentiation factor-5, 6, and 7 and platelet-derived growth factor–BB. Adipose-derived stem cells expression of scleraxis and collagen type I increased the most after 14 days of induction with growth differentiation factor-6 and platelet-derived growth factor–BB. Achilles excision defects injected with hydrogel alone (Gp2), with undifferentiated (Gp3) adipose-derived stem cells, or tenogenically differentiated (Gp4) adipose-derived stem cells exhibited improved tissue repair compared with untreated tendons (Gp1). Addition of adipose-derived stem cells improved tissue cytoarchitecture and increased expression of collagen type I and III, scleraxis, and tenomodulin. Adipose-derived stem cells significantly improved biomechanical properties (ultimate load and elastic toughness) over time more than hydrogel alone, while tenogenically differentiated adipose-derived stem cells improved the mean histological score and collagen fiber dispersion range closest to normal tendon. In addition, tendon sections treated with GFP-adipose-derived stem cells exhibited green fluorescence and positive GFP immunostaining on microscopy confirming the in vivo survival of adipose-derived stem cells that were injected into tendon defects to support the effects of adipose-derived stem cells on tissue up to 4.5 weeks post injury.

Long-Term Effects of Collagen-Binding Mesenchymal Stem Cells Affinity Peptide on Skin Regeneration and Scar Formation Running Title: MSC Capturing Collagen and Skin Regeneration

2020 ◽  
Author(s):  
Huili Wang ◽  
Xin Yan ◽  
Yue Lin ◽  
Shuqin Wang ◽  
Yanan Jiang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Collagen Type ◽  
Collagen Scaffold ◽  
Collagen Type I ◽  
Skin Regeneration ◽  
Type I ◽  
Skin Defect ◽  
Collagen Type Iii ◽  
Collagen Binding

Abstract The medical significance of mesenchymal stem cells (MSCs) is not only providing a promising strategy for cutaneous wound healing, but also bringing alternative for the patients whose wounds or burns were difficult to treat and heal. In our previous researches, we produced a fusion protein (Collagen/CBD-E7 peptide) which had high affinity to MSCs and then bound it to collagen scaffold with a CBD (collagen-binding domain). When applied to full-thickness skin defect porcine models, MSCs could be specifically raised to target area which helps accelerate coalescence at early stage. However, scar is an indispensable part of the tissue repair, arising after almost every dermal injury with abnormal appearance and functional disturbance. It usually takes one to three years for the scars to pale and mature. Thus, in this study, we focused on the structure repair of skin defect more than one year after the wounds healing. We found that at the 16w after operation, the proportion of collagen type III was more than collagen type I in Collagen/CBD-E7 peptide group, which indicates a higher healing quality comparatively. While at the 64w, the ratio between collagen type I and collagen type III reached a balance of natural skin tissues. In addition, cutaneous appendages were observed in Collagen/CBD-E7 peptide scaffold group at the 16w and 64w after surgery. From this point of view, the functional collagen scaffold treatments may be able to improve the cosmetic outcome of injury, burns or trauma, and scars are no longer an inevitable consequence of skin healing. This will also imply that humans would no longer be suffering from the scars caused by skin trauma, which is of significant orthopaedic meaning.

scholarly journals Tensile Stimulation of Murine Stem Cell–Collagen Sponge Constructs Increases Collagen Type I Gene Expression and Linear Stiffness

2009 ◽  
Vol 15 (9) ◽  
pp. 2561-2570 ◽  
Author(s):  
Kumar Chokalingam ◽  
Natalia Juncosa-Melvin ◽  
Shawn A. Hunter ◽  
Cynthia Gooch ◽  
Chris Frede ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Collagen Type ◽  
Collagen Type I ◽  
Collagen Sponge ◽  
Type I ◽  
I Gene ◽  
Stimulation Of

scholarly journals SAT0298 IS INTERLEUKIN 6 A FACTOR OF FIBROGENESIS IN DERMAL FIBROBLASTS?

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1094.1-1094
Author(s):  
A. S. Siebuhr ◽  
P. Juhl ◽  
M. Karsdal ◽  
A. C. Bay-Jensen
Keyword(s):  
Gene Expression ◽  
Type I Collagen ◽  
Collagen Type ◽  
Collagen Type I ◽  
Dermal Fibroblasts ◽  
Type I ◽  
Dependent Manner ◽  
Full Time ◽  
Collagen Formation ◽  
Type Iii

Background:Interleukin 6 (IL-6) is known to have both pro- and anti-inflammatory properties, depending on the receptor activation. The classical IL-6 signaling via the membrane bound receptor is mainly anti-inflammatory, whereas signaling through the soluble receptor (sIL-6R) is pro-inflammatory/pro-fibrotic. However, the direct fibrotic effect of IL-6 stimulation on dermal fibroblasts is unknown.Objectives:We investigated the fibrotic effect of IL-6 + sIL-6R in a dermal fibroblast model and assessed fibrosis by neo-epitope biomarkers of extracellular matrix proteins.Methods:Primary healthy human dermal fibroblasts were grown for up to 17 days in DMEM medium with 0.4% fetal calf serum, ficoll (to produce a crowded environment) and ascorbic acid. IL-6 [1-90 nM]+sIL-6R [0.1-9 nM] alone or in combination with TGFβ [1 nM] were tested in three different donors. TGFβ [1 nM], PDGF-AB [3 nM] and non-stimulated cells (w/o) were used as controls. Tocilizumab (TCZ) with TGFβ + IL-6 + sIL-6R stimulation was tested in one donor. Collagen type I, III and VI formation (PRO-C1, PRO-C3 and PRO-C6) and fibronectin (FBN-C) were evaluated by validated ELISAs (Nordic Bioscience). Western blot analysis investigated signal cascades. Gene expression of selected ECM proteins was analyzed. Statistical analyses included One-way and 2-way ANOVA and area under the curve analysis.Results:formation by the end of the culture period. The fibronectin and collagen type VI signal were consistent between the three tested donors, whereas the formation of type III collagen was only increased in one donor, but in several trials. Type I collagen formation was unchanged by IL-6 + sIL-6R stimulation. The gene expression of type I collagen was induced by IL-6 + sIL-6R. Western blot analysis validated trans-signaling by the IL-6+sIL-6R stimulation as expected.IL-6 + sIL-6R stimulation in combination with TGFβ decreased fibronectin levels compared to TGFβ alone but did not reach the level of unstimulated fibroblasts. The formation of collagen type IV was generally unchanged with IL-6 + sIL-6R + TGFβ compared to TGFβ alone. Collagen type I and III formation was more scattered in the signals when IL-6 + sIL-6R was in combination with TGFβ, as the biomarker level could be either decreased or increased compared to TGFβ alone. In two studies the type I collagen level was synergistic increased by IL-6 + sIL-6R + TGFβ, whereas another study found the level to be decreased compared to TGFβ alone. The gene expression of fibronectin and type I collagen was increased with TGFβ +IL-6+sIL-6R compared to TGFβ alone.Inhibition of IL-6R by TCZ in combination with IL-6 + sIL-6R did only decrease the fibronectin level with the lowest TCZ concentration (p=0.03). TCZ alone decreased the fibronectin level in a dose-dependent manner (One-way ANOVA p=0.0002).Conclusion:We investigated the fibrotic response of dermal fibroblasts to IL-6 + sIL-6R stimulation. IL-6 modulated the fibronectin level and modulated the collagen type III formation level in a somewhat dose-dependent manner. In combination with TGFβ, IL-6 decreased collagen type I and IV formation and fibronectin. However, in this study inhibition of IL-6R by TCZ did not change the fibrotic response of the dermal fibroblasts. This study indicated that IL-6 did not induce collagen formation in dermal fibroblasts, except type III collagen formation with high IL-6 concentration.Figure:Disclosure of Interests:Anne Sofie Siebuhr Employee of: Nordic Bioscience, Pernille Juhl Employee of: Nordic Bioscience, Morten Karsdal Shareholder of: Nordic Bioscience A/S., Employee of: Full time employee at Nordic Bioscience A/S., Anne-Christine Bay-Jensen Shareholder of: Nordic Bioscience A/S, Employee of: Full time employee at Nordic Bioscience A/S.

scholarly journals Endothelial cell growth factor and heparin regulate collagen gene expression in keloid fibroblasts

1991 ◽  
Vol 278 (3) ◽  
pp. 863-869 ◽  
Author(s):  
E M L Tan ◽  
J Peltonen
Keyword(s):  
Gene Expression ◽  
Endothelial Cell ◽  
Collagen Synthesis ◽  
Type I Collagen ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Gene ◽  
Endothelial Cell Growth Factor ◽  
Endothelial Cell Growth

Keloids are benign cutaneous tumours characterized by excess deposition of collagen, specifically type I collagen. We report here that collagen biosynthesis, as measured by hydroxyproline synthesis, was markedly inhibited by 65-80% by the combination of endothelial cell growth factor (ECGF) supplement and heparin in keloid fibroblast cultures. Fibroblast cultures that were incubated with ECGF alone also demonstrated a measurable decrease of approx. 50% in collagen synthesis compared with control cultures. The inhibition of collagen synthesis was related to the down-regulation of collagen gene expression. Quantitative measurements of mRNA-cDNA hybrids revealed that the gene expression of collagen type I was decreased by more than 80% by heparin and ECGF. Markedly diminished levels of mRNA encoding collagen type I were also observed in cultures incubated with ECGF alone. The results show that ECGF and heparin elicit a negative regulatory effect on collagen production, and that this inhibition is due largely to the down-regulation of the pro-alpha 1(I) of type I collagen gene. Furthermore, ECGF has a potent suppressive effect, and heparin provides an additive effect to this inhibitory phenomenon.

scholarly journals TOB1 Deficiency Enhances the Effect of Bone Marrow-Derived Mesenchymal Stem Cells on Tendon-Bone Healing in a Rat Rotator Cuff Repair Model

2016 ◽  
Vol 38 (1) ◽  
pp. 319-329 ◽  
Author(s):  
Yulei Gao ◽  
Yinquan Zhang ◽  
Yanghu Lu ◽  
Yi Wang ◽  
Xingrui Kou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Rotator Cuff ◽  
Bone Healing ◽  
Rotator Cuff Repair ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Cuff Repair

Background/Aims: This study investigated the effect of silencing TOB1 (Transducer of ERBB2, 1) expression in bone marrow-derived mesenchymal stem cells (MSCs) on MSC-facilitated tendon-bone healing in a rat supraspinatus repair model. Methods: Rat MSCs were transduced with a recombinant lentivirus encoding short hairpin RNA (shRNA) against TOB1. MSC cell proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. The effect of MSCs with TOB1 deficiency on tendon-bone healing in a rat rotator cuff repair model was evaluated by biomechanical testing, histological analysis and collagen type I and II gene expression. An upstream regulator (miR-218) of TOB1 was determined in MSCs. Results: We found that knockdown of TOB1 significantly increased the proliferative activity of rat MSCs in vitro. When MSCs with TOB1 deficiency were injected into injured rat supraspinatus tendon-bone junctions, the effect on tendon-bone healing was enhanced compared to treatment with control MSCs with normal TOB1 expression, as evidenced by elevated levels of ultimate load to failure and stiffness, increased amount of fibrocartilage and augmented expression of collagen type I and type II genes. In addition, we found that the TOB1 3′ untranslated region is a direct target of miR-218. Similar to the effect of TOB1 deficiency, overexpression of miR-218 effectively promoted tendon-bone healing in rat. Conclusion: These results suggest that TOB1 may play a negative role in the effect of MSCs on tendon-bone healing, and imply that expression of TOB1 may be regulated by miR-218.

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