Implantation of adipose-derived stem cells for tailored therapy using collagen Type I scaffolds on liver fibrosis

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.

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Adipose-Derived Stem Cells Ameliorate Ischemia-Reperfusion Injury in a Rat Skin Free Flap Model

Journal of Reconstructive Microsurgery ◽

10.1055/s-0038-1648246 ◽

2018 ◽

Vol 34 (08) ◽

pp. 601-609 ◽

Cited By ~ 2

Author(s):

Javier Casado ◽

Elena Abellán ◽

Francisco Vela ◽

Verónica Álvarez ◽

Alejandra Usón ◽

...

Keyword(s):

Stem Cells ◽

Collagen Type ◽

Ischemia Reperfusion ◽

Collagen Type I ◽

Control Group ◽

Type I ◽

Adipose Derived Stem Cells ◽

Skin Flaps ◽

Flap Survival ◽

Scaffold Group

Background Ischemia-reperfusion (I/R) injury is inevitable during free tissue transfers. When the period of ischemia exceeds the tissue tolerance, it causes necrosis and flap failure. The aim of this study was to investigate the effects of adipose-derived stem cells (ASCs) embedded in a collagen type I scaffold on the survival of free skin flaps to counteract I/R injury. Methods Left superficial caudal epigastric skin flaps (3 × 6 cm) were performed in 28 Wistar rats that were divided into four groups. The flaps elevated in the animals of the control group did not suffer any ischemic insult, and the vascular pedicle was not cut. All other flaps were subjected to 8 hours of ischemia prior to revascularization: I/R control group (8 hours of ischemia), I/R scaffold group (8 hours of ischemia + collagen type I scaffold), and I/R scaffold–ASCs group (8 hours of ischemia + collagen type I scaffold with rat ASCs embedded). Transit-time ultrasound blood flow measurements were performed. After 7 days, the areas of flap survival were measured and tissues were stained with hematoxylin/eosin and Masson's trichrome stain for histological analysis. Results The mean percentage flap survival area was significantly higher in the ASCs-treated flaps (I/R scaffold–ASCs group) compared with the ischemic controls (I/R control group and I/R scaffold group). Higher vascular proliferation and lower severity of necrosis and inflammatory changes were seen histologically in the samples of the ASCs-treated group. No significant difference in blood flow was detected between groups. Conclusion Subcutaneous administration of ASCs embedded on a collagen type I scaffold reduces tissue damage after I/R injury in microvascular free flaps.

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Collagen Type I and Type II Expression Evaluation on Cartilage Defect Regeneration Treated with Dwikora–Ferdiansyah–Lesmono–Purwati (DFLP) Scaffold Supplemented with Adipose–Derived Stem Cells (ASCs) or Secretome: An In-Vivo Study

Qanun Medika - Medical Journal Faculty of Medicine Muhammadiyah Surabaya ◽

10.30651/jqm.v4i2.4377 ◽

2020 ◽

Vol 4 (2) ◽

pp. 225

Author(s):

Adrianto Prasetyo Perbowo ◽

Dwikora Novembri Utomo ◽

Lukas Widhiyanto ◽

Primadenny Ariesa Airlangga ◽

Purwati Purwati

Keyword(s):

Stem Cells ◽

Collagen Type ◽

Collagen Type I ◽

Cartilage Defects ◽

Type I ◽

Adipose Derived Stem Cells ◽

Type Ii ◽

Collagen Type Ii ◽

Significant Difference

Abstract Cell-based therapies such as Scaffold, stem cells, and secretome, are one of the alternatives to enhance the regeneration of hyaline-like cartilage in cases of cartilage defects. This study is an in-vivo experiment using animal models, in which we apply a composite of DFLP (Dwikora-Ferdiansyah-Lesmono-Purwati) Scaffold and Adipose-Derived Stem Cells (ASCs) or Secretome to an injury model on the distal femoral trochlea of New Zealand White Rabbits. The animals were divided into four groups: (1) control (K); (2) Scaffold only (S); (3) Scaffold + ASCs (SA); (4) Scaffold + Secretome (SS). Animals were terminated in the 12th week, and an immunohistochemistry (IHC) evaluation for Collagen type I and II were done. Statistical analysis shows that collagen type I IHC between groups shows no significant difference (p = 0.546). Collagen type II IHC shows significant difference between groups (p = 0,016). The findings in this study showed that Scaffold + ASCs group and Scaffold + Secretome have better collagen type II expression compared to the control group. DFLP Scaffold composite with ASCs or Secretome shows potential for cartilage regeneration therapy by increasing type II collagen expression as in hyaline-like cartilage which may be used for regenerative therapy for cartilage defects. Keywords : DFLP Scaffold; Adipose-Derived Stem Cells (ASCs); Secretome; Collagen Type I; Collagen Type IICorrespondence : [email protected]

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Electromagnetic fields enhance chondrogenesis of human adipose-derived stem cells in a chondrogenic microenvironment in vitro

Journal of Applied Physiology ◽

10.1152/japplphysiol.01216.2012 ◽

2013 ◽

Vol 114 (5) ◽

pp. 647-655 ◽

Cited By ~ 33

Author(s):

Chung-Hwan Chen ◽

Yi-Shan Lin ◽

Yin-Chih Fu ◽

Chih-Kuang Wang ◽

Shun-Cheng Wu ◽

...

Keyword(s):

Stem Cells ◽

Electromagnetic Field ◽

Cell Viability ◽

Chondrogenic Differentiation ◽

Collagen Type ◽

Cartilage Tissue ◽

Pulse Electromagnetic Field ◽

Type I ◽

Adipose Derived Stem Cells ◽

Pemf Treatment

We tested the hypothesis that electromagnetic field (EMF) stimulation enhances chondrogenesis in human adipose-derived stem cells (ADSCs) in a chondrogenic microenvironment. A two-dimensional hyaluronan (HA)-coated well (2D-HA) and a three-dimensional pellet culture system (3D-pellet) were used as chondrogenic microenvironments. The ADSCs were cultured in 2D-HA or 3D-pellet, and then treated with clinical-use pulse electromagnetic field (PEMF) or the innovative single-pulse electromagnetic field (SPEMF) stimulation. The cytotoxicity, cell viability, and chondrogenic and osteogenic differentiations were analyzed after PEMF or SPEMF treatment. The modules of PEMF and SPEMF stimulations used in this study did not cause cytotoxicity or alter cell viability in ADSCs. Both PEMF and SPEMF enhanced the chondrogenic gene expression (SOX-9, collagen type II, and aggrecan) of ADSCs cultured in 2D-HA and 3D-pellet. The expressions of bone matrix genes (osteocalcin and collagen type I) of ADSCs were not changed after SPEMF treatment in 2D-HA and 3D-pellet; however, they were enhanced by PEMF treatment. Both PEMF and SPEMF increased the cartilaginous matrix (sulfated glycosaminoglycan) deposition of ADSCs. However, PEMF treatment also increased mineralization of ADSCs, but SPEMF treatment did not. Both PEMF and SPEMF enhanced chondrogenic differentiation of ADSCs cultured in a chondrogenic microenvironment. SPEMF treatment enhanced ADSC chondrogenesis, but not osteogenesis, when the cells were cultured in a chondrogenic microenvironment. However, PEMF enhanced both osteogenesis and chondrogenesis under the same conditions. Thus the combination of a chondrogenic microenvironment with SPEMF stimulation can promote chondrogenic differentiation of ADSCs and may be applicable to articular cartilage tissue engineering.

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WNT-5A regulates TGF-β-related activities in liver fibrosis

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00160.2016 ◽

2017 ◽

Vol 312 (3) ◽

pp. G219-G227 ◽

Cited By ~ 21

Author(s):

Leonie Beljaars ◽

Sara Daliri ◽

Christa Dijkhuizen ◽

Klaas Poelstra ◽

Reinoud Gosens

Keyword(s):

Liver Fibrosis ◽

Functional Role ◽

Collagen Type ◽

Collagen Type I ◽

Matrix Proteins ◽

Type I ◽

Human Livers

WNT-5A is a secreted growth factor that belongs to the noncanonical members of the Wingless-related MMTV-integration family. Previous studies pointed to a connection between WNT-5A and the fibrogenic factor TGF-β warranting further studies into the functional role of WNT-5A in liver fibrosis. Therefore, we studied WNT-5A expressions in mouse and human fibrotic livers and examined the relation between WNT-5A and various fibrosis-associated growth factors, cytokines, and extracellular matrix proteins. WNT-5A gene and protein expressions were significantly increased in fibrotic mouse and human livers compared with healthy livers. Regression or therapeutic intervention in mice resulted in decreased hepatic WNT-5A levels paralleled by lower collagen levels. Immunohistochemical analysis showed WNT-5A staining in fibrotic septa colocalizing with desmin staining indicating WNT-5A expression in myofibroblasts. In vitro studies confirmed WNT-5A expression in this cell type and showed that TGF-β significantly enhanced WNT-5A expression in contrast to PDGF-BB and proinflammatory cytokines IL-1β and TNF-α. Additionally, TGF-β induces the expression of the WNT receptors FZD2 and FZD8. After silencing of WNT-5A, reduced levels of collagen type I, vimentin, and fibronectin in TGF-β-stimulated myofibroblasts were measured compared with nonsilencing siRNA-treated controls. Interestingly, the antifibrotic cytokine IFNγ suppressed WNT-5A in vitro and in vivo. IFNγ-treated fibrotic mice showed significantly less WNT-5A expression compared with untreated fibrotic mice. In conclusion, WNT-5A paralleled collagen I levels in fibrotic mouse and human livers. WNT-5A expression in myofibroblasts is induced by the profibrotic factor TGF-β and plays an important role in TGF-β-induced regulation of fibrotic matrix proteins, whereas its expression can be reversed upon treatment, both in vitro and in vivo. NEW & NOTEWORTHY This study describes the localization and functional role of WNT-5A in human and mouse fibrotic livers. Hepatic WNT-5A expression parallels collagen type I expression. In vivo and in vitro, the myofibroblasts were identified as the key hepatic cells producing WNT-5A. WNT-5A is under control of TGF-β and its activities are primarily profibrotic.

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Effect of Allogeneic Bone Marrow-mesenchymal Stem Cells (BM-MSCs) to Accelerate Burn Healing of Rat on the Expression of Collagen Type I and Integrin α2β1

Pakistan Journal of Biological Sciences ◽

10.3923/pjbs.2016.345.351 ◽

2016 ◽

Vol 19 (8-9) ◽

pp. 345-351 ◽

Cited By ~ 3

Author(s):

Gusti Revilla ◽

Eryati Darwin ◽

Yanwirasti . ◽

Fedik A. Rantam

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Collagen Type ◽

Collagen Type I ◽

Allogeneic Bone Marrow ◽

Type I ◽

Allogeneic Bone ◽

Marrow Mesenchymal Stem Cells

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

Cellular Physiology and Biochemistry ◽

10.1159/000438632 ◽

2016 ◽

Vol 38 (1) ◽

pp. 319-329 ◽

Cited By ~ 16

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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