scholarly journals Mechanical micronization of lipoaspirates for treatment of bleomycin-induced scleroderma in nude mice

2020 ◽  
Author(s):  
Wenqing Jiang ◽  
Jing Wang ◽  
Jiayan Lin ◽  
Shenglu Jiang ◽  
Yuping Quan ◽  
...  

Abstract Background Scleroderma is a chronic autoimmune disease that causes hardening of the skin. Adipose tissue is used as a regenerative treatment for scleroderma, and mechanical micronization of adipose tissue can concentrate stem cells, improving its therapeutic efficacy. Stromal vascular fraction gel (SVF gel) is produced by subjecting lipoaspirates to a series of mechanical processes. The present study aimed to assess the therapeutic effect of SVF gel on dermal scleroderma. Methods Scleroderma was induced in nude mice by daily subcutaneous injections of bleomycin (BLM) for 4 weeks. SVF gel, cell-enriched lipoaspirates (CAL), or Coleman fat (CF) (0.1 mL) was injected subcutaneously into local sclerosis lesions. Histological analyses, TUNEL assay, and qRT-PCR were conducted 4 weeks post-transplantation. Results SVF gel, CAL, and CV decreased dermal thickness in the scleroderma model (P < 0.05), and sclerosis was significantly decreased in the SVF gel and CAL groups. SVF gel and CAL decreased macrophage infiltration, pro-inflammatory cytokine levels, and fibrosis in transferred fat and/or skin lesions (P < 0.05). This revealed anti-inflammatory and anti-fibrotic effects of adipose-derived stem cells (ASCs) therapy in scleroderma (P < 0.05). SVF gel exhibited the most potent anti-inflammatory and anti-fibrotic effects among treatment groups (P < 0.05). Conclusion SVF gel and CAL significantly reversed dermal sclerosis, and were more effective than CF. The SVF gel group exhibited the lowest inflammation both in transferred fat and in skin lesions, and also the lowest fibrosis among the three groups. This study suggested that SVF gel was a more suitable adipose-based therapy for topical treatment of scleroderma patients.

2021 ◽  
Vol 22 (3) ◽  
pp. 1375
Author(s):  
María Carmen Carceller ◽  
María Isabel Guillén ◽  
María Luisa Gil ◽  
María José Alcaraz

Adipose tissue represents an abundant source of mesenchymal stem cells (MSC) for therapeutic purposes. Previous studies have demonstrated the anti-inflammatory potential of adipose tissue-derived MSC (ASC). Extracellular vesicles (EV) present in the conditioned medium (CM) have been shown to mediate the cytoprotective effects of human ASC secretome. Nevertheless, the role of EV in the anti-inflammatory effects of mouse-derived ASC is not known. The current study has investigated the influence of mouse-derived ASC CM and its fractions on the response of mouse-derived peritoneal macrophages against lipopolysaccharide (LPS). CM and its soluble fraction reduced the release of pro-inflammatory cytokines, adenosine triphosphate and nitric oxide in stimulated cells. They also enhanced the migration of neutrophils or monocytes, in the absence or presence of LPS, respectively, which is likely related to the presence of chemokines, and reduced the phagocytic response. The anti-inflammatory effect of CM may be dependent on the regulation of toll-like receptor 4 expression and nuclear factor-κB activation. Our results demonstrate the anti-inflammatory effects of mouse-derived ASC secretome in mouse-derived peritoneal macrophages stimulated with LPS and show that they are not mediated by EV.


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.


2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Ilaria Roato ◽  
Daniela Alotto ◽  
Dimas Carolina Belisario ◽  
Stefania Casarin ◽  
Mara Fumagalli ◽  
...  

Osteoarthritis is characterized by loss of articular cartilage also due to reduced chondrogenic activity of mesenchymal stem cells (MSCs) from patients. Adipose tissue is an attractive source of MSCs (ATD-MSCs), representing an effective tool for reparative medicine, particularly for treatment of osteoarthritis, due to their chondrogenic and osteogenic differentiation capability. The treatment of symptomatic knee arthritis with ATD-MSCs proved effective with a single infusion, but multiple infusions could be also more efficacious. Here we studied some crucial aspects of adipose tissue banking procedures, evaluating ATD-MSCs viability, and differentiation capability after cryopreservation, to guarantee the quality of the tissue for multiple infusions. We reported that the presence of local anesthetic during lipoaspiration negatively affects cell viability of cryopreserved adipose tissue and cell growth of ATD-MSCs in culture. We observed that DMSO guarantees a faster growth of ATD-MSCs in culture than trehalose. At last, ATD-MSCs derived from fresh and cryopreserved samples at −80°C and −196°C showed viability and differentiation ability comparable to fresh samples. These data indicate that cryopreservation of adipose tissue at −80°C and −196°C is equivalent and preserves the content of ATD-MSCs in Stromal Vascular Fraction (SVF), guaranteeing the differentiation ability of ATD-MSCs.


2019 ◽  
Vol 47 (11) ◽  
pp. 2577-2583 ◽  
Author(s):  
Naomasa Yokota ◽  
Mari Hattori ◽  
Tadahiko Ohtsuru ◽  
Masaki Otsuji ◽  
Stephen Lyman ◽  
...  

Background: Intra-articular injection of adipose-derived stem cells (ASCs) has shown promise for improving symptoms and cartilage quality in the treatment of osteoarthritis (OA). However, while most preclinical studies have been performed with plastic-adherent ASCs, most clinical trials are being conducted with the stromal vascular fraction (SVF), prepared from adipose tissue without prior culture. Purpose: To directly compare clinical outcomes of intra-articular injection with ASCs or SVF in patients with knee OA. Study Design: Cohort study; Level of evidence, 3. Methods: The authors retrospectively compared 6-month outcomes in 42 patients (59 knees) receiving intra-articular injection with 12.75 million ASCs and 38 patients (69 knees) receiving a 5-mL preparation of SVF. All patients had Kellgren-Lawrence grade 2, 3, or 4 knee OA and had failed standard medical therapy. The visual analog scale (VAS) pain score and Knee injury and Osteoarthritis Outcome Score (KOOS) at baseline and 1, 3, and 6 months after injection were considered as outcomes. Outcome Measures in Rheumatology–Osteoarthritis Research Society International (OMERACT-OARSI) criteria were also used to assess positive response. A repeated measures analysis of variance was used for comparison between the treatment groups. Results: No major complications occurred in either group. The SVF group had a higher frequency of knee effusion (SVF 8%, ASC 2%) and minor complications related to the fat harvest site (SVF 34%, ASC 5%). Both groups reported improvements in pain VAS and KOOS domains. Specifically, in the ASC group, symptoms improved earlier (by 3 months; P < .05) and pain VAS decreased to a greater degree (55%; P < .05) compared with the SVF group (44%). The proportion of OMERACT-OARSI responders in the ASC group was slightly higher (ASCs, 61%; SVF, 55%; P = .25). Conclusion: It was observed that both ASCs and SVF resulted in clinical improvement in patients with knee OA, but that ASCs outperform SVF in the early reduction of symptoms and pain with less comorbidity.


2017 ◽  
Vol 31 (07) ◽  
pp. 686-697 ◽  
Author(s):  
Aaron Stoker ◽  
Chantelle Bozynski ◽  
Keiichi Kuroki ◽  
Kevin Clarke ◽  
Jed Johnson ◽  
...  

AbstractStromal vascular fraction (SVF) contains a small number of mesenchymal stem cells and has been used as a treatment for osteoarthritis and cartilage injury. Due to limited evidence of successful cartilage regeneration with injected stem cell therapies, there is interest in combining cellular therapies with injectable scaffolding materials to increase intra-articular residence times of stem cells and improve tissue regeneration. However, the safety of intra-articular injection of SVF combined with injectable scaffolds is unestablished. Also, it is unclear if SVF therapy is superior to more easily prepared biologics, such as platelet-rich plasma (PRP). The purpose of this study was to assess the safety of SVF when combined with an injectable poly(L-lactide-co-glycolide) nanofiber scaffold and to provide a comparison of SVF therapy to PRP. A total of 12 Beagles had osteochondral defects created in both medial femoral condyles and 4 dogs each were allocated to treatment groups of SVF (n = 4), SVF plus PLGA scaffolding (n = 4), or leukoreduced PRP (n = 4). One knee in each dog received treatment, and the contralateral knee was sham treated with saline. Dogs were assessed over a 6-month period, and outcome measures included functional, radiographic, biochemical, and histological assessments. PRP treatment resulted in improvements in lameness scores and objective kinetic assessments of function. There were no statistically significant improvements in function, cartilage biochemical composition, or histology for SVF-treated knees. The combination of SVF and the injectable PLGA scaffold had worse outcomes than other groups including sham treatment based upon functional, biochemical, and histological assessments, raising concerns over the safety of this scaffold for intra-articular injection.


2012 ◽  
Vol 8 (10) ◽  
pp. 757-762 ◽  
Author(s):  
Pedro Pires de Carvalho ◽  
Katie M. Hamel ◽  
Robert Duarte ◽  
Andrew G. S. King ◽  
Masudul Haque ◽  
...  

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Jaewoo Pak ◽  
Jung Hun Lee ◽  
Wiwi Andralia Kartolo ◽  
Sang Hee Lee

Osteoarthritis (OA) is one of the most common debilitating disorders among the elderly population. At present, there is no definite cure for the underlying causes of OA. However, adipose tissue-derived stem cells (ADSCs) in the form of stromal vascular fraction (SVF) may offer an alternative at this time. ADSCs are one type of mesenchymal stem cells that have been utilized and have demonstrated an ability to regenerate cartilage. ADSCs have been shown to regenerate cartilage in a variety of animal models also. Non-culture-expanded ADSCs, in the form of SVF along with platelet rich plasma (PRP), have recently been used in humans to treat OA and other cartilage abnormalities. These ADSCs have demonstrated effectiveness without any serious side effects. However, due to regulatory issues, only ADSCs in the form of SVF are currently allowed for clinical uses in humans. Culture-expanded ADSCs, although more convenient, require clinical trials for a regulatory approval prior to uses in clinical settings. Here we present a systematic review of currently available clinical studies involving ADSCs in the form of SVF and in the culture-expanded form, with or without PRP, highlighting the clinical effectiveness and safety in treating OA.


2007 ◽  
Vol 293 (5) ◽  
pp. E1153-E1158 ◽  
Author(s):  
Hui Ren Zhou ◽  
Eun-Kyoung Kim ◽  
Hyojung Kim ◽  
Kate J. Claycombe

Studies showed that monocyte chemotactic protein-1 (MCP-1) concentrations are increased in obesity. In our current study, we demonstrate that plasma MCP-1 level in leptin-deficient ob/ob mice is significantly higher than in lean mice. Furthermore, we determined that basal adipose tissue MCP-1 mRNA levels are significantly higher in ob/ob mice compared with lean mice. To determine the mechanisms underlying obesity-associated increases in plasma and adipose tissue MCP-1 levels, we determined adipose tissue cell type sources of MCP-1 production. Our data show that adipose tissue stem cells (CD34+), macrophages (F4/80+), and stromal vascular fraction (SVF) cells express significantly higher levels of MCP-1 compared with adipocytes under both basal and lipopolysaccharide (LPS)-stimulated conditions. Furthermore, basal and LPS-induced MCP-1 secretion levels were the same for both adipose F4/80+ and CD34+ cells, whereas adipose CD34+ cells have twofold higher cell numbers (30% of total SVF cells) compared with F4/80+ macrophages (15%). Our data also show that CD34+ cells from visceral adipose tissue depots secrete significantly higher levels of MCP-1 ex vivo when compared with CD34+ cells from subcutaneous adipose tissue depots. Taken together, our data suggest that adipose CD34+ stem cells may play an important role in obesity-associated increases in plasma MCP-1 levels.


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