Increasing number of fibroblast, capillary and collagen amount in rat’s achilles tendon with diabetic mellitus after application of stromal vascular fraction derived from adipose tissue

2019 ◽  
Author(s):  
Panji Sananta ◽  
Langga Sintong ◽  
Sri Andarini ◽  
Respati Suryanto Dradjat
2020 ◽  
Vol 5 (3) ◽  
pp. 46-50
Author(s):  
Natal'ya Slesarenko ◽  
Aleksei Zharikov

The aim of the study is identification of features of Achilles tendon healing in the conditions of auto-transplantation of the cellular element, reflecting the regenerative activity of stromal-vascular fraction. Modern regenerative treat-ment methods for animal with tendon injuries involve stromal-vascular cells application of adipose tissue. They represent cell pool with high plasticity, proliferative activity, and ability to secrete multiple angiogenic factors. How-ever, there is no data on morphological and morphometric regenerative equivalents of their use in case of tendon ligaments injury. Micro morphometrical indicators are presented that substantiate the effectiveness of using autolo-gous cellular elements in case of Achilles tendon injury. The influence of stromal-vascular fraction from rat adipose tissue on regenerative process during healing of an induced tendon tissue defect was evaluated which lead to an increase in the thickness of bundles of the first and the second orders in the experimental group, significant differ-ences in comparison with the control of the fibroblastic cell pool, and the number of blood capillaries of these ani-mals. Based on the positive data of the autologous cell pool on the regenerative ability of the Achilles tendon, its practicability for veterinary medicine was scientifically justified. The experimental model of the study is a fancy rat (n=20). Achilles tendon was the research material. A complex methodological approach was used, including ex-perimental modeling, micro morphological studies, micro morphometry, and statistical data processing. It was re-vealed that experimental animals injected with stromal-vascular fraction, micro morphometric transformations of connective tissue were detected, aimed at thickening of bundles of the first and second orders, the transition of fi-broblasts to structured and ordered fibrocytes (tenocytes), acceleration of collagenogenesis and vascularization enhancement.


2014 ◽  
Vol 222 (2) ◽  
pp. 201-215 ◽  
Author(s):  
Jillian L Rourke ◽  
Shanmugam Muruganandan ◽  
Helen J Dranse ◽  
Nichole M McMullen ◽  
Christopher J Sinal

Chemerin is an adipose-derived signaling protein (adipokine) that regulates adipocyte differentiation and function, immune function, metabolism, and glucose homeostasis through activation of chemokine-like receptor 1 (CMKLR1). A second chemerin receptor, G protein-coupled receptor 1 (GPR1) in mammals, binds chemerin with an affinity similar to CMKLR1; however, the function of GPR1 in mammals is essentially unknown. Herein, we report that expression of murineGpr1mRNA is high in brown adipose tissue and white adipose tissue (WAT) and skeletal muscle. In contrast to chemerin (Rarres2) andCmklr1,Gpr1expression predominates in the non-adipocyte stromal vascular fraction of WAT. Heterozygous and homozygousGpr1-knockout mice fed on a high-fat diet developed more severe glucose intolerance than WT mice despite having no difference in body weight, adiposity, or energy expenditure. Moreover, mice lackingGpr1exhibited reduced glucose-stimulated insulin levels and elevated glucose levels in a pyruvate tolerance test. This study is the first, to our knowledge, to report the effects ofGpr1deficiency on adiposity, energy balance, and glucose homeostasisin vivo. Moreover, these novel results demonstrate that GPR1 is an active chemerin receptor that contributes to the regulation of glucose homeostasis during obesity.


2021 ◽  
Vol 22 (15) ◽  
pp. 7920
Author(s):  
Myroslava Mytsyk ◽  
Giulia Cerino ◽  
Gregory Reid ◽  
Laia Gili Sole ◽  
Friedrich S. Eckstein ◽  
...  

The therapeutic potential of mesenchymal stromal/stem cells (MSC) for treating cardiac ischemia strongly depends on their paracrine-mediated effects and their engraftment capacity in a hostile environment such as the infarcted myocardium. Adipose tissue-derived stromal vascular fraction (SVF) cells are a mixed population composed mainly of MSC and vascular cells, well known for their high angiogenic potential. A previous study showed that the angiogenic potential of SVF cells was further increased following their in vitro organization in an engineered tissue (patch) after perfusion-based bioreactor culture. This study aimed to investigate the possible changes in the cellular SVF composition, in vivo angiogenic potential, as well as engraftment capability upon in vitro culture in harsh hypoxia conditions. This mimics the possible delayed vascularization of the patch upon implantation in a low perfused myocardium. To this purpose, human SVF cells were seeded on a collagen sponge, cultured for 5 days in a perfusion-based bioreactor under normoxia or hypoxia (21% and <1% of oxygen tension, respectively) and subcutaneously implanted in nude rats for 3 and 28 days. Compared to ambient condition culture, hypoxic tension did not alter the SVF composition in vitro, showing similar numbers of MSC as well as endothelial and mural cells. Nevertheless, in vitro hypoxic culture significantly increased the release of vascular endothelial growth factor (p < 0.001) and the number of proliferating cells (p < 0.00001). Moreover, compared to ambient oxygen culture, exposure to hypoxia significantly enhanced the vessel length density in the engineered tissues following 28 days of implantation. The number of human cells and human proliferating cells in hypoxia-cultured constructs was also significantly increased after 3 and 28 days in vivo, compared to normoxia. These findings show that a possible in vivo delay in oxygen supply might not impair the vascularization potential of SVF- patches, which qualifies them for evaluation in a myocardial ischemia model.


2021 ◽  
Vol 82 (1) ◽  
Author(s):  
Anirban Mandal ◽  
Ajeet Kumar Jha ◽  
Dew Biswas ◽  
Shyamal Kanti Guha

Abstract Background The study was conducted to assess the characterization, differentiation, and in vitro cell regeneration potential of canine mesenteric white adipose tissue-derived mesenchymal stem cells (AD-MSCs). The tissue was harvested through surgical incision and digested with collagenase to obtain a stromal vascular fraction. Mesenchymal stem cells isolated from the stromal vascular fraction were characterized through flow cytometry and reverse transcription-polymerase chain reaction. Assessment of cell viability, in vitro cell regeneration, and cell senescence were carried out through MTT assay, wound healing assay, and β-galactosidase assay, respectively. To ascertain the trilineage differentiation potential, MSCs were stained with alizarin red for osteocytes, alcian blue for chondrocytes, and oil o red for adipocytes. In addition, differentiated cells were characterized through a reverse transcription-polymerase chain reaction. Results We observed the elongated, spindle-shaped, and fibroblast-like appearance of cells after 72 h of initial culture. Flow cytometry results showed positive expression for CD44, CD90, and negative expression for CD45 surface markers. Population doubling time was found 18–24 h for up to the fourth passage and 30±0.5 h for the fifth passage. A wound-healing assay was used to determine cell migration rate which was found 136.9 ± 4.7 μm/h. We observed long-term in vitro cell proliferation resulted in MSC senescence. Furthermore, we also found that the isolated cells were capable of differentiating into osteogenic, chondrogenic, and adipogenic lineages. Conclusions Mesenteric white adipose tissue was found to be a potential source for isolation, characterization, and differentiation of MSCs. This study might be helpful for resolving the problems regarding the paucity of information concerning the basic biology of stem cells. The large-scale use of AD-MSCs might be a remedial measure in regenerative medicine.


2020 ◽  
Vol 52 (06) ◽  
pp. 521-532
Author(s):  
Constanze Kuhlmann ◽  
Thilo Ludwig Schenck ◽  
Elisabeth Maria Haas ◽  
Riccardo Giunta ◽  
Paul Severin Wiggenhauser

Zusammenfassung Hintergrund Nicht nur regenerative Therapie wie zellassistierter Lipotransfer (cell assisted lipotransfer) sondern auch präklinische experimentelle Studien verwenden in der Plastischen Chirurgie Stammzellen aus Fettgewebe, sogenannte Adipose tissue-derived Stem Cells (ASCs). Hierbei haben allerdings vom jeweiligen Stammzellspender abhängige Faktoren einen entscheidenden Einfluss auf die Zellausbeute und das regenerative Potential von ASCs und der Stromal vascular Fraction (SVF). Ziel dieser Übersichtsarbeit war es daher, diese Einflussfaktoren des Stammzellspenders darzustellen und anhand des aktuellen Wissenstands zu beurteilen. Methoden Es erfolgte eine intensive Literaturrecherche in der der National Library of Medicine, mit Fokus auf Einflussfaktoren der Stammzellspender, die eine Beeinflussung der Zellausbeute und des regenerativen Potentials von humanen ASCs und SVF in vorherigen Studien gezeigt haben. Ergebnisse Aktuell gibt es eine Vielzahl von Studien, welche sich mit den Einflussfaktoren des Stammzellspenders auseinandersetzen. Allerdings sind diese Faktoren sehr inhomogen und teilweise sogar widersprüchlich, so dass hier noch weiterer Forschungsbedarf besteht. Dennoch gibt es einige Faktoren, die gemäß der aktuellen Literatur gehäuft untersucht wurden: Alter, Geschlecht, Gewicht, Nebenerkrankungen (z. B. Diabetes, Lipödem) sowie spezielle Medikamente (Antidepressiva, Antihormontherapie) und Chemotherapie. Schlussfolgerung Wir empfehlen, bei experimentellen und klinischen Arbeiten mit ASCs/SVF eine Charakterisierung des Patientenkollektivs zu veröffentlichen, so dass mögliche Beeinflussungen durch oben genannte Faktoren kommuniziert werden und eine bessere Vergleichbarkeit von Studien ermöglicht wird. Darüber hinaus kann aber auch mit einer präzisen Anamnese und körperlichen Untersuchung vorab ein möglichst homogenes Patientenkollektiv für die Sammlung von Proben für wissenschaftliche Arbeiten konstruiert werden. Auch könnten die Ergebnisse dazu beitragen, den Erfolg zukünftiger ASC-basierter Therapien einzuschätzen.


2000 ◽  
Vol 279 (6) ◽  
pp. R2329-R2335 ◽  
Author(s):  
Paul Trayhurn ◽  
Jacqueline S. Duncan ◽  
Anne M. Wood ◽  
John H. Beattie

White adipose tissue (WAT) has been examined to determine whether the gene encoding metallothionein (MT), a low-molecular-weight stress response protein, is expressed in the tissue and whether MT may be a secretory product of adipocytes. The MT-1 gene was expressed in epididymal WAT, with MT-1 mRNA levels being similar in lean and obese ( ob/ ob) mice. MT-1 mRNA was found in each of the main adipose tissue sites (epididymal, perirenal, omental, subcutaneous), and there was no major difference between depots. Separation of adipocytes from the stromal-vascular fraction of WAT indicated that the MT gene (MT-1 and MT-2) was expressed in adipocytes themselves. Treatment of mice with zinc had no effect on MT-1 mRNA levels in WAT, despite strong induction of MT-1 expression in the liver. MT-1 gene expression in WAT was also unaltered by fasting or norepinephrine. However, administration of a β3-adrenoceptor agonist, BRL-35153A, led to a significant increase in MT-1 mRNA. On differentiation of fibroblastic preadipocytes to adipocytes in primary culture, MT was detected in the medium, suggesting that the protein may be secreted from WAT. It is concluded that WAT may be a significant site of MT production; within adipocytes, MT could play an antioxidant role in protecting fatty acids from damage.


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