Aloe Vera/Collagen Mixture Induces Integrin α1β1 and PECAM-1Genes Expression in Human Adipose-Derived Stem Cells

Purpose: Natural biomaterials are a key base in tissue engineering, and collagen, as the maincontent of the extracellular matrix (ECM), is frequently used in tissue engineering. Aloe verahas some therapeutic effects on ulcers, therefore, the use of this natural resource has alwaysbeen considered for improving collagen function. We aimed to evaluate the effect of Aloe vera/Collagen blended on cell viability, cell attachment, and angiogenic potential by determining ofintegrin α1β1 and platelet endothelial cell adhesion molecule (PECAM-1) genes expression inhuman adipose-derived stem cells (hASCs).Methods: In this study, hASCs after harvesting of adipose tissues from abdominal subcutaneousadipose tissue and isolation, were cultured in four groups of control, collagen gel, Aloe veragel, and Aloe vera/collagen blended in vitro environment at 24h and then cell viability wasassessed by MTT (3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium) assay. Integrin α1β1 andPECAM-1 genes expression were evaluated by real-time RT-PCR.Results: The results of MTT showed that the combination of Aloe vera/collagen was retained thecell viability at the normal range and improved it. In real-time RT-PCR results, integrin α1β1 andPECAM-1 gene expression were increased in the Aloe vera/collagen blended group comparedto the control group.Conclusion: For tissue engineering purposes, Aloe vera improves collagen properties in theculture of hASCs by increasing the expression of the integrin α1β1 and PECAM-1 genes.<br />

Download Full-text

Hypoxia-preconditioned adipose-derived stem cells combined with scaffold promote urethral reconstruction by up-regulation of angiogenesis and glycolysis

10.21203/rs.3.rs-70182/v2 ◽

2020 ◽

Author(s):

Xiang Wan ◽

Min-kai Xie ◽

Huan Xu ◽

Zi-wei Wei ◽

Hai-jun Yao ◽

...

Keyword(s):

Stem Cells ◽

Umbilical Vein ◽

Urethral Reconstruction ◽

Therapeutic Effects ◽

Adipose Derived Stem Cells ◽

Promising Alternative ◽

Hypoxia Preconditioning ◽

Novel Scaffold

Abstract Rationale: Tissue engineering is a promising alternative for urethral reconstruction, and adipose-derived stem cells (ADSCs) are widely used as seeding cells. Hypoxia preconditioning can significantly enhance the therapeutic effects of ADSCs. The low oxygen tension of postoperative wound healing is inevitable and may facilitate the nutritional function of ADSCs. This study aimed to investigate if hypoxia preconditioned ADSCs, compared to normxia preconditioned ADSCs, combined with scaffold could better promote urethral reconstruction and exploring the underlying mechanism.Methods: In vitro, paracrine cytokines and secretomes that were secreted by hypoxia- or normoxia-preconditioned ADSCs were added to cultures of human umbilical vein endothelial cells (HUVECs) to measure their functions. In vivo, hypoxia- or normoxia-preconditioned ADSCs were seeded on a porous nanofibrous scaffold for urethral repair on a defect model in rabbits.Results: The in vitro results showed that hypoxia could enhance the secretion of VEGFA by ADSCs, and hypoxia-preconditioned ADSCs could enhance the viability, proliferation, migration, angiogenesis and glycolysis of HUVECs (p < 0.05). After silencing VEGFA, angiogenesis and glycolysis were significantly inhibited (p < 0.05). The in vivo results showed that compared to normoxia-preconditioned ADSCs, hypoxia-preconditioned ADSCs combined with scaffolds led to a larger urethral lumen diameter, preserved urethral morphology and enhanced angiogenesis (p < 0.05). Conclusions: Hypoxia preconditioning of ADSCs combined with scaffold could better promote urethral reconstruction by upregulating angiogenesis and glycolysis. Hypoxia-preconditioned ADSCs combined with novel scaffold may provide a promising alternative treatment for urethral reconstruction.

Download Full-text

A Preliminary Study of Constructing Tissue Engineered Oral Mucosa

10.21203/rs.3.rs-299660/v1 ◽

2021 ◽

Author(s):

Qin Zhang ◽

Yu-Jing Tian ◽

Tao Ding ◽

De-Hong Femg

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Small Intestine ◽

Oral Mucosa ◽

Porous Membrane ◽

Adipose Derived Stem Cells ◽

Oral Keratinocytes ◽

Cell Scaffold ◽

Compound Graft

Abstract Background: Adipose derived stem cells (ADSCs) have a great potential for tissue-engineering purposes, and they may be introduced in oral mucosa tissue engineering for urethroplasty. This study was aim to develop a tissue-engineered oral mucosa through seeding oral keratinocytes (OKs) and adipose derived stem cells (ADSCs) on small intestine submucosa (SIS). Methods: SIS was obtained from porcine small intestine, and OKs and ADSCs were obtained from canine sources and were cultured and expanded in vitro. The two cell lines were seeded on the two surfaces of the SIS, and the cell-scaffold compound graft was cultured in an air fluid level for 1 week.Results: The SIS exhibited a porous membrane, and no cells were found through HE staining. The model cultured with OK-SIS only formed a thin and loose epithelium. Whereas the model cultured with OK-SIS-ADSC was much thicker and denser. Conclusion: The co-cultured of ADSC and OK grew well on the SIS in which the OKs formed a multilayer of epithelium. So it is feasible to construct a tissue-engineered oral mucosa graft with ADSCs, OKs and SIS. The ADSCs contribute to a thicker epithelium formation.

Download Full-text

Comparison of PLGA/Fibrin and PLGA/Hyaluronic Acid Scaffolds for Chondrogenesis of Human Adipose-Derived Stem Cells

International Journal of Medical Laboratory ◽

10.18502/ijml.v7i2.2920 ◽

2020 ◽

Author(s):

Zeinolabedin Sharifian ◽

Batool Hashemibeni ◽

Majid Pourentezari ◽

Ali Valiani ◽

Mohammad Mardani ◽

...

Keyword(s):

Gene Expression ◽

Tissue Engineering ◽

Stem Cells ◽

Hyaluronic Acid ◽

Real Time ◽

Cartilage Regeneration ◽

Type Ii Collagen ◽

Control Group ◽

Adipose Derived Stem Cells ◽

X Gene

Background and Aims: Tissue engineering is a relatively novel field that has been intensely developing during recent years and has shown to be excessively promising when used for cartilage regeneration. Scaffolds represent important components for tissue engineering. Materials and Methods: The Poly Lactic-Co-Glycolic Acid (PLGA) impregnated with fibrin and hyaluronic acid (HA) produce hybrid scaffolds. human adipose-derived stem cells (hADSCs) were seeded in scaffolds and cultured in chondrogenic media. The viability of cells in different groups was assessed by MTT. The expression of chondrogenic related genes [Sox9, type II collagen (Col II), Aggrecan(AGG)] and type X collagen (Col X) was quantified by real-time polymerase chain reaction. Results: The results of the real-time PCR showed SOX9, AGG and Col X gene expression in the control groups being significantly lower than the other groups (p≤0.05). It also demonstrated Col II gene expression in the control group being significantly lower than the PLGA/Fibrin and PLGA/Fibrin/HA groups (p≤0.05). The Col X gene expression of cells in PLGA/HA and PLGA/Fibrin/HA groups significantly decreased in comparison with the PLGA/Fibrin group (p≤0.05). Conclusions: These conclusions indicate that administration of PLGA/ Fibrin and PLGA/HA scaffolds, particularly PLGA/Fibrin/ HA, motivates chondrogenesis in hADSCs. This can be diminished by decreasing hypertrophic markers and increasing characteristic markers of hyaline cartilage.

Download Full-text

The Therapeutic Effects of Adipose-Derived Stem Cells and Recombinant Peptide Pieces on Mouse Model of DSS Colitis

Cell Transplantation ◽

10.1177/0963689718782442 ◽

2018 ◽

Vol 27 (9) ◽

pp. 1390-1400 ◽

Cited By ~ 6

Author(s):

Reiko Iwazawa ◽

Sayako Kozakai ◽

Tsukasa Kitahashi ◽

Kentaro Nakamura ◽

Ken-ichiro Hata

Keyword(s):

Stem Cells ◽

Cell Therapy ◽

Mouse Models ◽

The Body ◽

Toxicity Tests ◽

Therapeutic Effects ◽

Adipose Derived Stem Cells ◽

Recombinant Peptide ◽

General Toxicity

Cell therapies using adipose-derived stem cells (ADSCs) have been used to treat inflammatory bowel disease (IBD) in human and dog. We previously reported the CellSaic technique, which uses a recombinant scaffold to enhance the efficacy of cell therapy. To examine whether this technique can be applied to cell therapy for colitis, we evaluated the efficacy of CellSaic in colitis mouse models. Colitis mouse models were developed by administering dextran sulfate sodium (DSS) to C57BL/6 mice for 7 days. Then CellSaic comprising human/canine ADSCs (1.2 × 106 cells) or human/canine ADSCs only (1.2 × 106 cells) were administered to the mice. The body weights were measured, and the colon length measurements and histological evaluations were conducted at 7 days after administration. After in vitro culture of human ADSC (hADSC) CellSaic and hADSC spheroids in medium containing TNFα, the levels of the anti-inflammatory protein TSG-6 in each supernatant were measured. Furthermore, we conducted tumorigenicity and general toxicity tests of canine ADSC (cADSC) CellSaic in NOG mice for 8 weeks. In the colitis mouse models, the ADSC CellSaic group presented recovery of body weight and colon length compared with the ADSC-only group. Histological analysis showed that ADSC CellSaic decreased the number of inflammatory cells and repaired ulceration. In vitro, hADSC CellSaic secreted 3.1-fold more TSG-6 than the hADSCs. In addition, tumorigenicity and general toxicity of cADSC CellSaic were not observed. This study suggests that human and canine ADSC CellSaic has a therapeutic effect of colitis in human and dogs.

Download Full-text

A Preliminary Study of Constructing Tissue Engineered Oral Mucosa

International Journal of Dentistry and Oral Health ◽

10.16966/2378-7090.377 ◽

2021 ◽

Vol 7 (6) ◽

Author(s):

Zhang Q ◽

Tian YJ ◽

Ding T ◽

Feng DH

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Small Intestine ◽

Oral Mucosa ◽

Porous Membrane ◽

Adipose Derived Stem Cells ◽

Oral Keratinocytes ◽

Cell Scaffold ◽

Compound Graft

Adipose derived stem cells (ADSCs) have a great potential for tissue-engineering purposes, and they may be introduced in oral mucosa tissue engineering for urethroplasty. This study was aimed to develop a tissue-engineered oral mucosa through seeding oral keratinocytes (OKs) and adipose derived stem cells (ADSCs) on small intestine submucosa (SIS). From August 2018 to October 2019, an observational study was conducted in the laboratory of our hospital, to develop a tissue-engineered oral mucosa.SIS was obtained from porcine small intestine, and OKs and ADSCs were obtained from canine sources and were cultured and expanded in vitro. The two cell lines were seeded on the two surfaces of the SIS, and the cell-scaffold compound graft was cultured in an air fluid level for 1 week. The SIS exhibited a porous membrane, and no cells were found through HE staining. The model cultured with OK-SIS only formed a thin and loose epithelium. Whereas the model cultured with OK-SIS-ADSC was much thicker and denser. The co-cultured of ADSC and OK grew well on the SIS in which the OKs formed a multilayer of epithelium. So it is feasible to construct a tissue-engineered oral mucosa graft with ADSCs, OKs and SIS. The ADSCs contribute to a thicker epithelium formation.

Download Full-text

Hybrid Sponge-Like Scaffolds Based on Ulvan and Gelatin: Design, Characterization and Evaluation of Their Potential Use in Bone Tissue Engineering

Materials ◽

10.3390/ma13071763 ◽

2020 ◽

Vol 13 (7) ◽

pp. 1763

Author(s):

Leto-Aikaterini Tziveleka ◽

Andreas Sapalidis ◽

Stefanos Kikionis ◽

Eleni Aggelidou ◽

Efterpi Demiri ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Mechanical Performance ◽

Cell Attachment ◽

Sulfated Polysaccharide ◽

Hybrid Scaffolds

Ulvan, a bioactive natural sulfated polysaccharide, and gelatin, a collagen-derived biopolymer, have attracted interest for the preparation of biomaterials for different biomedical applications, due to their demonstrated compatibility for cell attachment and proliferation. Both ulvan and gelatin have exhibited osteoinductive potential, either alone or in combination with other materials. In the current work, a series of novel hybrid scaffolds based on crosslinked ulvan and gelatin was designed, prepared and characterized. Their mechanical performance, thermal stability, porosity, water-uptake and in vitro degradation ability were assessed, while their morphology was analyzed through scanning electron microscopy. The prepared hybrid ulvan/gelatin scaffolds were characterized by a highly porous and interconnected structure. Human adipose-derived mesenchymal stem cells (hADMSCs) were seeded in selected ulvan/gelatin hybrid scaffolds and their adhesion, survival, proliferation, and osteogenic differentiation efficiency was evaluated. Overall, it was found that the prepared hybrid sponge-like scaffolds could efficiently support mesenchymal stem cells’ adhesion and proliferation, suggesting that such scaffolds could have potential uses in bone tissue engineering.

Download Full-text

Hypoxia-preconditioned adipose-derived stem cells combined with scaffold promote urethral reconstruction by upregulation of angiogenesis and glycolysis

Stem Cell Research & Therapy ◽

10.1186/s13287-020-02052-4 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Xiang Wan ◽

Min-kai Xie ◽

Huan Xu ◽

Zi-wei Wei ◽

Hai-jun Yao ◽

...

Keyword(s):

Stem Cells ◽

Umbilical Vein ◽

Urethral Reconstruction ◽

Therapeutic Effects ◽

Adipose Derived Stem Cells ◽

Promising Alternative ◽

Hypoxia Preconditioning ◽

Novel Scaffold

Abstract Rationale Tissue engineering is a promising alternative for urethral reconstruction, and adipose-derived stem cells (ADSCs) are widely used as seeding cells. Hypoxia preconditioning can significantly enhance the therapeutic effects of ADSCs. The low oxygen tension of postoperative wound healing is inevitable and may facilitate the nutritional function of ADSCs. This study aimed to investigate if hypoxia-preconditioned ADSCs, compared to normoxia-preconditioned ADSCs, combined with scaffold could better promote urethral reconstruction and exploring the underlying mechanism. Methods In vitro, paracrine cytokines and secretomes that were secreted by hypoxia- or normoxia-preconditioned ADSCs were added to cultures of human umbilical vein endothelial cells (HUVECs) to measure their functions. In vivo, hypoxia- or normoxia-preconditioned ADSCs were seeded on a porous nanofibrous scaffold for urethral repair on a defect model in rabbits. Results The in vitro results showed that hypoxia could enhance the secretion of VEGFA by ADSCs, and hypoxia-preconditioned ADSCs could enhance the viability, proliferation, migration, angiogenesis, and glycolysis of HUVECs (p < 0.05). After silencing VEGFA, angiogenesis and glycolysis were significantly inhibited (p < 0.05). The in vivo results showed that compared to normoxia-preconditioned ADSCs, hypoxia-preconditioned ADSCs combined with scaffolds led to a larger urethral lumen diameter, preserved urethral morphology, and enhanced angiogenesis (p < 0.05). Conclusions Hypoxia preconditioning of ADSCs combined with scaffold could better promote urethral reconstruction by upregulating angiogenesis and glycolysis. Hypoxia-preconditioned ADSCs combined with novel scaffold may provide a promising alternative treatment for urethral reconstruction.

Download Full-text

In Vitro Cell Adhesion, Proliferation and Differentiation of Adipose Derived Stem Cells on Tulbaghia violacea Loaded Polycaprolactone (PCL) Nanofibers

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2019.2184 ◽

2019 ◽

Vol 9 (11) ◽

pp. 1485-1498 ◽

Cited By ~ 1

Author(s):

Lerato N. Madike ◽

M. Pillay ◽

Ketul C. Popat

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Cell Adhesion ◽

The Body ◽

Osteogenic Potential ◽

Adipose Derived Stem Cells ◽

Cell Proliferation And Differentiation ◽

Proliferation And Differentiation ◽

Osteocalcin Production

Tissue engineering has been used for decades to restructure, replace and repair damaged tissue in the body. However, there are a number of challenges that have been identified, with the biggest one currently being the development of scaffolds with the ideal properties that can promote cell-scaffold interactions to enhance cell proliferation and differentiation. There is currently very little research on the incorporation of extracts of medicinal plants in scaffold fabrication with the aim of enhancing the surface properties of the scaffold. For this study, Tulbaghia violacea-based PCL scaffolds were fabricated and evaluated for their osteogenic potential on adipose derived stem cells (ADSCs) in osteogenic media. The short-term studies illustrated enhanced cell adhesion and proliferation with low levels of toxicity as well as the formation of elongated cells in the T. violacea-based scaffolds when compared to the control PCL scaffold. The long term studies indicated increased alkaline phosphate activity (ALP) in the T. violacea scaffolds when compared to PCL and overall higher levels of osteocalcin production over a period of 3 weeks. Immunofluorescence imaging of marker proteins also illustrated that the T. violacea incorporated scaffolds supported better osteocalcin production which is a specific extracellular matrix (ECM) marker for cartilaginous tissue. These results support the incorporation of T. violacea plant extracts for the enhancement of nanofiber scaffolds with the potential for tissue engineering applications.

Download Full-text

Adipose-Derived Stem Cells in Novel Approaches to Breast Reconstruction: Their Suitability for Tissue Engineering and Oncological Safety

Breast Cancer Basic and Clinical Research ◽

10.1177/1178223417726777 ◽

2017 ◽

Vol 11 ◽

pp. 117822341772677 ◽

Cited By ~ 13

Author(s):

Niamh O’Halloran ◽

Donald Courtney ◽

Michael J Kerin ◽

Aoife J Lowery

Keyword(s):

Breast Cancer ◽

Tissue Engineering ◽

Stem Cells ◽

Breast Reconstruction ◽

Disease Recurrence ◽

Adipose Derived Stem Cells ◽

Oncological Safety ◽

Patients With Cancer

Adipose-derived stem cells (ADSCs) are rapidly becoming the gold standard cell source for tissue engineering strategies and hold great potential for novel breast reconstruction strategies. However, their use in patients with breast cancer is controversial and their oncological safety, particularly in relation to local disease recurrence, has been questioned. In vitro, in vivo, and clinical studies using ADSCs report conflicting data on their suitability for adipose tissue regeneration in patients with cancer. This review aims to provide an overview of the potential role for ADSCs in breast reconstruction and to examine the evidence relating to the oncologic safety of their use in patients with breast cancer.

Download Full-text