Stromal Vascular Fraction Group Research Articles

Beneficial effects of adipose-derived stromal vascular fraction on testicular injury caused by busulfan

The use of stem cells can attenuate testicular injury and promote sperm production. The adipose-derived stromal vascular fraction (SVF) has become an attractive cell source for cell-based therapies. In this study, we aimed to investigate the therapeutic efficacy of SVF on busulfan-induced testicular damage in rats. Twenty-four male rats were randomly divided into control, busulfan, SVF, and busulfan + SVF groups. Testicular damage was induced by intraperitoneal administration of busulfan (35 mg/kg). SVF obtained from human adipose tissue using Lipocube SVF™ was injected into rats 5 weeks after busulfan administration. At the end of the 8th week, rats were sacrificed, and histopathological, biochemical, and western blotting analyses were performed. No harmful effects of SVF on healthy testis tissue and sperm parameters were detected. SVF improved busulfan-induced oxidative stress in both testis tissue and serum. SVF injection to damaged testicular tissue resulted in increases in the healthy spermatozoon numbers and decreases in the abnormal tail numbers. Additionally, SVF increased bax/Bcl, DAZL, and TGF-β1 levels whereas decreased ATG5 and NF-kB levels. According to the results we obtained in this study, we suggest that SVF is beneficial in restoring damaged tissue by primarily being a multipotent cell source, by inhibiting oxidative stress and converting necrotic cell death to apoptotic cell death. In the future, clinical applications should bring higher benefits. Since SVF is the patient’s own tissue, being harmless, it will offer an advantageous supportive treatment option for patients already weakened by cancer and anticancer therapy.

Efficacy of the immediate adipose-derived stromal vascular fraction autograft on functional sensorimotor recovery after spinal cord contusion in rats.

Spinal cord injuries (SCI) lead to functional alteration with important consequences such as motor and sensory disorders. The repair strategies developed to date remain ineffective. The adipose tissue-derived stromal vascular fraction (SVF) is composed of a cocktail of cells with trophic, pro-angiogenic and immunomodulatory effects. Numerous therapeutic benefits were shown for tissue reconstitution, peripheral neuropathy and for the improvement of neurodegenerative diseases. Here, the therapeutic efficacy of SVF on sensorimotor recovery after an acute thoracic spinal cord contusion in adult rats was determined. Male Sprague Dawley rats (n = 45) were divided into 3 groups: SHAM (without SCI and treatment), NaCl (animals with a spinal lesion and receiving a saline injection through the dura mater) and SVF (animals with a spinal lesion and receiving a fraction of fat removed from adipocytes through the dura mater). Some animals were sacrificed 14days after the start of the experiment to determine the inflammatory reaction by measuring the interleukin-1β, interleukin-6 and Tumor Necrosis Factor-α in the lesion area. Other animals were followed once a week for 12weeks to assess functional recovery (postural and locomotor activities, sensorimotor coordination). At the end of this period, spinal reflexivity (rate-dependent depression of the H-reflex) and physiological adjustments (ventilatory response to metabosensitive muscle activation following muscle fatigue) were measured with electrophysiological tools. Compared to non-treated animals, results indicated that the SVF reduced the endogenous inflammation and increased the behavioral recovery in treated animals. Moreover, H-reflex depression and ventilatory adjustments to muscle fatigue were found to be comparable between SHAM and SVF groups. Our results highlight the effectiveness of SVF and its high therapeutic potential to improve sensorimotor functions and to restore the segmental sensorimotor loop and the communication between supra- and sub-lesional spinal cord regions after traumatic contusion.

A Novel Intervention That Prevents Vocal Fold Scarring

We evaluated the preventive efficacy of stromal vascular fraction (SVF) for vocal fold scar in a rabbit model. Animal model. The study included 40 male New Zealand white rabbits: 20 received vocal fold scar surgery served as normal controls (control group). The other 20 received the same vocal fold scar surgery with SVF injection (SVF group) Histological and high-speed video analyses of vocal fold vibration were performed 4 weeks after scar surgery and SVF injection. The maximum amplitude of vocal fold vibration was used to assess vocal fold vibration. A real-time PCR study was also performed to evaluate the scar regeneration and remodeling including TGF-ß1, IL-6, procollagen-1, MMP-2, 9, and HAS-2, 3. Vocal fold vibration analyses indicated that the maximum amplitude differences in the vibration of the SVF group were significantly higher than the control group. The histological findings showed that the collagen density ratio were significantly lower in the SVF group compared to the control group. Real-time Polymerase Chain Reaction (PCR) study showed significant increases of MMP-2, 9 and HAS-2, 3, and a decrease of TGF-ß1, IL-6, procollagen-1 in the SVF group compared to the control group. Based on the vocal fold vibration study, histological findings, and real-time PCR study, SVF injection showed preventive activity and improvement of vocal fold vibration for vocal fold scar in a rabbit model.

Clinical Outcome of Minimally Manipulated, Mechanically Isolated Autologous Adipose Tissue-Derived Stromal Vascular Fraction (Sahaj Therapy®) in Knee Osteoarthritis-Randomized Controlled Trial.

Regenerative therapy has shown promising results in the treatment of osteoarthritis (OA) knee with Kellgren-Lawrence (KL) Grades I-III. We compared the safety, efficacy, functional, and clinical outcomes of intra-articular implantation of autologous adipose tissue-derived stromal vascular fraction (SVF) isolated using direct ultrasonic cavitation (Sahaj therapy-Cell Innovation Patented Technology) and saline injection in knee osteoarthritis. The present prospective observational study was conducted over 3years. We enrolled 120 patients in our study, where four patients got excluded as they did not meet the inclusion criteria. The remaining 116 patients were randomized into two groups, one with autologous adipose tissue-derived SVF and the other group with saline injection. A comparison of mean KOOS and VAS scores at different follow-ups was done using Paired 't' test. A p value of < 0.05 was considered significant. The results show that the SVF group had significantly higher KOOS scores (78.49 ± 6.54 in the SVF group vs 59.19 ± 5.14 in the saline group), respectively (p < 0.001). Similarly, the SVF group had significantly lesser VAS scores (3.17 ± 0.94 in the SVF group vs 3.89 ± 1.04 in the saline group), respectively (p < 0.001). Autologous adipose tissue-derived SVF is a better choice for treating knee osteoarthritis. For individuals with degenerative osteoarthritis, autologous SVF grafting in the same surgical procedure is an innovative and promising treatment modality. Even after 3years of follow-up, the study participants with OA knee have shown a good clinical and functional outcome.

Heterogeneous Cells as well as Adipose-Derived Stromal Cells in Stromal Vascular Fraction Contribute to Enhance Anabolic and Inhibit Catabolic Factors in Osteoarthritis.

The stromal-vascular fraction (SVF), comprising heterogeneous cell populations and adipose-derived stromal cells (ADSCs), has therapeutic potential against osteoarthritis (OA); however, the underlying mechanism remains elusive. This study aimed to investigate the therapeutic effects of heterogeneous cells in rabbit SVF on rabbit chondrocytes. Rabbit SVF and ADSCs were autografted into knees at OA onset. The SVF (1 × 105) and low-dose ADSCs (lADSC; 1 × 104) groups adjusted for their stromal cell content were compared. Animals were euthanized 8 and 12 weeks after OA onset for macroscopic and histological analyses of OA progression and synovitis. Immunohistochemical and real-time polymerase chain reaction assessments were conducted. In vitro, immune-fluorescent double staining was performed for SVF to stain macrophages with F4/80, CD86(M1), and CD163(M2). OA progression was markedly suppressed, and synovitis was reduced in the SVF groups (OARSI histological score 8W: 6.8 ± 0.75 vs. 3.8 ± 0.75, p = 0.001; 12W: 8.8 ± 0.4 vs. 5.4 ± 0.49, p = 0.0002). The SVF groups had higher expression of collagen II and SOX9 in cartilage and TGF-β and IL-10 in the synovium, lower expression of MMP-13, and lower macrophage M1/M2 ratio than the lADSC groups. Immunofluorescent double staining revealed a markedly higher number of M2 than that of M1 macrophages in the SVF. The therapeutic effects of SVF on chondrocytes were superior than those of lADSCs, with enhanced anabolic and inhibited catabolic factors. Heterogeneous cells, mainly M2 macrophages in the SVF, enhanced growth factor secretion and chondrocyte-protective cytokines, thus benefiting chondrocytes and knee joint homeostasis. Overall, the SVF is a safe, relatively simple, and a useful treatment option for OA.

Effect of Adipose Derived Stromal Vascular Fraction on The Early Inflammatory Phase of Burn Wound Healing in Male Rat Model

Objectives: Severe burn treatment remains challenging. Research has investigated the efficacy of stem cells and stromal vascular fraction (SVF) in many types of injuries. SVF has advantages for its heterogenous population of cells and avoidance of culturing and ethical issues. SVF can be isolated either enzymatically or mechanically. Enzymatically isolated SVF reduced inflammation and enhanced neovascularization and re-epithelization in the treatment of burn injury. This study highlighted the efficacy of applying mechanically isolated SVF on the early inflammatory phase of deep partial-thickness burn by investigating toll-like receptor 4 (TLR4) signaling pathway.&#x0D; Methods: 30 male Wistar rats with deep partial-thickness burns were assigned into: the control group, silver sulfadiazine (SSD) cream group, and mechanically isolated SVF group. Morphological, histopathological evaluation of inflammation, and immunohistochemical analysis for (TLR4, TNF-α, IL‐1β, and IL-6) were evaluated for the first eight days post-treatment.&#x0D; Results: Morphologically, the SVF group significantly reduced edema and increased wound bed dryness on day one compared to the control group (P=0.0001) and to the SSD group (P=0.001). Histopathologically, the SVF group significantly reduced inflammation four days post-treatment compared with the control group (P=0.045). The SVF group significantly reduced TLR4 protein expression on days four and eight post-treatment compared with the control group (P= 0.001, P= 0.042, respectively). The TNF-α protein expression in the SVF group was significantly lower on days four and eight post-treatment compared with the control group (P= 0.046, P= 0.046, respectively) and with the SSD group (P= 0.008, P= 0.001, respectively). The IL‐1β expression was significantly reduced in the SVF group compared to the control group on day four post-treatment (P= 0.017). There were no significant differences in IL-6 expressions between all groups on both days.&#x0D; Conclusion: The mechanical isolation of SVF has an early anti-inflammatory impact on deep partial-thickness burn injury. This effect could be through inhibiting TLR4, TNF-α, and IL‐1β pathways. This could partially explain the mechanism behind SVF efficacy in the healing process of burn injury.

Adipose-Derived Stromal Vascular Fractions Are Comparable With Allogenic Human Umbilical Cord Blood–Derived Mesenchymal Stem Cells as a Supplementary Strategy of High Tibial Osteotomy for Varus Knee Osteoarthritis

To compare the clinical, radiologic, and second-look arthroscopic outcomes of high tibial osteotomy (HTO) with stromal vascular fraction (SVF) implantation versus human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) transplantation and identify the association between cartilage regeneration and HTO outcomes. Patients treated with HTO for varus knee osteoarthritis between March 2018 and September 2020 were retrospectively identified. In this retrospective study, among 183 patients treated with HTO for varus knee osteoarthritis between March 2018 and September 2020, patients treated with HTO with SVF implantation (SVF group; n= 25) were pair-matched based on sex, age, and lesion size with those who underwent HTO with hUCB-MSC transplantation (hUCB-MSC group; n= 25). Clinical outcomes were evaluated using the International Knee Documentation Committee score and Knee Injury and Osteoarthritis Outcome Score. Radiological outcomes evaluated were the femorotibial angle and posterior tibial slope. All patients were evaluated clinically and radiologically before surgery and during follow-up. The mean final follow-up periods were 27.8 ± 3.6 (range 24-36) in the SVF group and 28.2 ± 4.1 (range, 24-36) in the hUCB-MSC group (P= 0.690). At second-look arthroscopic surgery, cartilage regeneration was evaluated using the International Cartilage Repair Society (ICRS) grade. A total of 17 male and 33 female patients with a mean age of 56.2 years (range, 49-67 years) were included. At the time of second-look arthroscopic surgery (mean, 12.6 months; range, 11-15 months in the SVF group and 12.7 months; range, 11-14 months in the hUCB-MSC group, P= .625), the mean International Knee Documentation Committee score and Knee Injury and Osteoarthritis Outcome Score in each group significantly improved (P < .001 for all), and clinical outcomes at final follow-up further improved in both groups when compared with the values at second-look arthroscopic surgery (P < .05 for all). Overall ICRS grades, which significantly correlated with clinical outcomes, were similar between groups with no significant differences (P= .170 for femoral condyle and P= .442 for tibial plateau). Radiologic outcomes at final follow-up showed improved knee joint alignment relative to preoperative conditions but showed no significant correlation with clinical outcomes or ICRS grade in either group (P > .05 for all). Improved clinical and radiological outcomes and favorable cartilage regeneration were seen after surgery for varus Knee OA in both SVF and hUCB-MSC groups. Level III, retrospective comparative study.

Arthroscopic Implantation of Adipose-Derived Stromal Vascular Fraction Improves Cartilage Regeneration and Pain Relief in Patients With Knee Osteoarthritis

PurposeTo compare the pain relief and cartilage repair status of patients with knee osteoarthritis who received arthroscopic treatment with or without stromal vascular fraction (SVF) implantation.MethodsWe retrospectively evaluated the patients who were examined with 12-month follow-up magnetic resonance imaging (MRI) after arthroscopic treatment for knee osteoarthritis from September 2019 to April 2021. Patients were included in this study if they had grade 3 or 4 knee osteoarthritis according to the Outerbridge classification in MRI. The visual analog scale (VAS) was used for pain assessment over the follow-up period (baseline and at 1-, 3-, 6-, and 12-month follow-ups). Cartilage repair was evaluated using follow-up MRIs based on Outerbridge grades and the Magnetic Resonance Observation of Cartilage Repair Tissue scoring system.ResultsAmong 97 patients who received arthroscopic treatment, 54 patients received arthroscopic treatment alone (conventional group) and 43 received arthroscopic treatment along with SVF implantation (SVF group). In the conventional group, the mean VAS score decreased significantly at 1-month post-treatment compared with baseline (P < .05), and gradually increased from 3 to 12 months’ post-treatment (all P < .05). In the SVF group, the mean VAS score decreased until 12 months post-treatment compared with baseline (all P < .05 except P = .780 in 1-month vs 3-month follow-ups). Significantly greater pain relief was reported in the SVF group than in the conventional group at 6 and 12 months’ post-treatment (all P < .05). Overall, Outerbridge grades were significantly greater in the SVF group than in the conventional group (P < .001). Similarly, mean Magnetic Resonance Observation of Cartilage Repair Tissue scores were significantly greater (P < .001) in the SVF group (70.5 ± 11.1) than in the conventional group (39.7 ± 8.2).ConclusionsThe results regarding pain improvement and cartilage regeneration and the significant correlation between pain and MRI outcomes at 12-months follow-up indicate that the arthroscopic SVF implantation technique may be useful for repairing cartilage lesions in knee osteoarthritis.Level of EvidenceLevel III, retrospective comparative study. To compare the pain relief and cartilage repair status of patients with knee osteoarthritis who received arthroscopic treatment with or without stromal vascular fraction (SVF) implantation. We retrospectively evaluated the patients who were examined with 12-month follow-up magnetic resonance imaging (MRI) after arthroscopic treatment for knee osteoarthritis from September 2019 to April 2021. Patients were included in this study if they had grade 3 or 4 knee osteoarthritis according to the Outerbridge classification in MRI. The visual analog scale (VAS) was used for pain assessment over the follow-up period (baseline and at 1-, 3-, 6-, and 12-month follow-ups). Cartilage repair was evaluated using follow-up MRIs based on Outerbridge grades and the Magnetic Resonance Observation of Cartilage Repair Tissue scoring system. Among 97 patients who received arthroscopic treatment, 54 patients received arthroscopic treatment alone (conventional group) and 43 received arthroscopic treatment along with SVF implantation (SVF group). In the conventional group, the mean VAS score decreased significantly at 1-month post-treatment compared with baseline (P < .05), and gradually increased from 3 to 12 months’ post-treatment (all P < .05). In the SVF group, the mean VAS score decreased until 12 months post-treatment compared with baseline (all P < .05 except P = .780 in 1-month vs 3-month follow-ups). Significantly greater pain relief was reported in the SVF group than in the conventional group at 6 and 12 months’ post-treatment (all P < .05). Overall, Outerbridge grades were significantly greater in the SVF group than in the conventional group (P < .001). Similarly, mean Magnetic Resonance Observation of Cartilage Repair Tissue scores were significantly greater (P < .001) in the SVF group (70.5 ± 11.1) than in the conventional group (39.7 ± 8.2). The results regarding pain improvement and cartilage regeneration and the significant correlation between pain and MRI outcomes at 12-months follow-up indicate that the arthroscopic SVF implantation technique may be useful for repairing cartilage lesions in knee osteoarthritis.

The Efficacy and Safety of a Human Perirenal Adipose Tissue-Derived Stromal Vascular Fraction in an Interstitial Cystitis Rat Model

Background:Interstitial cystitis (IC) is a chronic and intractable disease that can severely deteriorate patients’ quality of life. Recently, stem cell therapy has been introduced as a promising alternative treatment for IC in animal models. We aimed to verify the efficacy and safety of the human perirenal adipose tissue-derived stromal vascular fraction (SVF) in an IC rat model.Methods:From eight-week-old female rats, an IC rat model was established by subcutaneous injection of 200 μg of uroplakin3A. The SVF was injected into the bladder submucosal layer of IC rats, and pain scale analysis, awakening cytometry, and histological and gene analyses of the bladder were performed. For the in vivo safety analysis, genomic DNA purification and histological analysis were also performed to check tumorigenicity and thrombus formation.Results:The mean pain scores in the SVF 20 μl group were significantly lower on days 7 and 14 than those in the control group, and bladder intercontraction intervals were significantly improved in the SVF groups in a dose-dependent manner. Regeneration of the bladder epithelium, basement membrane, and lamina propria was observed in the SVF group. In the SVF groups, however, bladder fibrosis and the expression of inflammatory markers were not significantly improved compared to those in the control group.Conclusion:This study demonstrated that a perirenal adipose tissue-derived SVF is a promising alternative for the management of IC in terms of improving bladder pain and overactivity.

Micronized Cellular Adipose Matrix Promotes the Therapeutic Effect of an Artificial Nerve Conduit in Peripheral Nerve Gap Injury.

Stromal vascular fraction (SVF) isolated from adipose tissue has been shown to be beneficial for treating peripheral nerve injuries. Micronized cellular adipose matrix (MCAM) is an SVF-rich micronized fat tissue obtained by a series of simple mechanical processes. This study assessed the therapeutic effect of MCAM for peripheral nerve injury. Microscopic evaluation of the cell phenotype and functions was performed to determine the adipose-derived stem cell content of the MCAM. An artificial nerve conduit (ANC) filled with MCAM was implanted into a sciatic nerve defect in immunodeficient mice. Comparisons of this treatment with an autograft, an ANC filled with SVF cells, and an ANC alone were made based on electrophysiologic characteristics, Sciatic Functional Index, and histologic analyses of regenerated nerve fiber and myelination using electron microscopy, and the preventive effect on innervated muscle atrophy. MCAM contained many cells with a phenotype and differentiation potency similar to those of ADSCs. The implantation experiment indicated that MCAM enhanced the efficiency of functional and structural recovery and prevented atrophy of the innervated muscle. These effects were significantly improved compared with the control group (ANC only) and comparable to those in the SVF group, whereas the improvement did not reach the same level of the autograft group. Injection of MCAM into an ANC accelerated nerve regeneration compared with use of an ANC alone, which indicates that MCAM is a promising transplant material for treatment of peripheral nerve injury and an alternative to use of SVF cells. Micronized cellular adipose matrix, which can be harvested and isolated from adipose tissue with a simple device, has been shown for the first time to be highly useful as an implantable material for new peripheral nerve regeneration.

Лечение остеоартроза коленных суставов на основе применения клеточных технологий

Osteoarthritis (OA) is the most common chronic disease of joint and a major cause of joint pain and disability. Traditional therapies for knee osteoarthritis show little clinical benefit, and the terminal treatment option is joint replacement. Currently, cell therapy is prospective for the treatment of knee osteoarthritis and has shown significant results in animal studies and human clinical trials. The aim: to investigate the treatment results in patients with knee osteoarthritis using adipose tissue stromal-vascular fraction and synovial mesenchymal stem cells.Methods. 60 patients with knee osteoarthritis were included in the study. The stage of knee osteoarthritis was assessed using the Kellgren-Lawrence classification, and patients with stage II-III of osteoarthritis were included in this study. After arthroscopic debridement, patients were divided into 2 groups. The first group was injected with the stromal-vascular fraction (SVF) of synovial membrane, and the second group was injected with adipose tissue mesenchymal stem cells (SVF). The minimum clinical and radiologic follow-up period was 12 months. Results. The assessment of the joints condition after 12 months using different scales showed that in both experimental groups there was a significant improvement of the functional status of the knee of the patients after treatment with MSC and SVF. Functional indicators of knee joints on the WOMAC scale improved by 14.7 and 13.4, on the KOOS scale by 23.6 and 26.4 in the SVF and MSC groups, respectively. Pain syndrome regressed by 1.8 and 2.1 on the VAS scale in the SVF and MSC groups, respectively.Conclusion. The results of both study groups showed that there is a significant difference between WOMAC, VAS and KOOS values before and after surgery (8, 12 weeks and 12 months) in the compared groups towards improvement

Mechanically isolated stromal vascular fraction improves healing of deep-partial thickness burn in rat model

Background: Despite the advances in treating burn injuries, burn severity remains one of the most challenging cases to be treated. As technologies advances, regenerative therapies using stem cells have been established and found to be effective and friendly as a regenerative tool. Stromal vascular fraction (SVF), extracted from adipose tissue, has a great potential for multipotent mesenchymal stem cells differentiation. Recent research has determined the therapeutic effects of SVF on burn injury. Previous finding has proved the efficacy of using SVF that is isolated enzymatically. This research aimed to examine the efficacy of allogenic mechanically isolated (MI) SVF in treating deep partial-thickness burns in a Wistar rat. Materials and Methods: After burn induction, 45 rats were divided into three groups of 15 as follows: group one (control, treated with phosphate-buffered saline), group two (treated with silver sulfadiazine (SSD), and group three (injected with SVF). SVF was harvested from the inguinal fat pad of six rats and mechanically processed. All injections were administered intradermally at the four edges of the burn to cover the entire wound bed. Morphological and histopathological analyses were performed for all groups at three different time points (4, 8, and 32 d post-treatment). Results: Treatment with MI SVF significantly reduced edema formation and dryness of the wound bed on day one compared to the control (P = 0.001). Histopathological results showed that SVF significantly reduced inflammation compared to the control (P = 0.045) on day one and increased neovascularization on day 8 (P = 0.016). Epithelial thickness was significantly greater in the SVF group compared to the SSD group (P = 0.034). Conclusions: The results of this study indicated the therapeutic potential of MI SVF on deep-partial-thickness burns by increasing neovascularization and epithelial thickness and reducing inflammation.

Intraoperative Administration of Adipose Stromal Vascular Fraction Does Not Improve Functional Outcomes in Young Patients with Anterior Cruciate Ligament Reconstruction.

Adipose stromal vascular fraction (SVF) has a versatile cellular system for biologically augmented therapies. However, there have been no clinical studies investigating the benefits of the augmentation of anterior cruciate ligament reconstruction (ACLR) with SVF. We conducted a retrospective study in assessing the effects of intraoperative SVF administration on the functional outcomes in young patients with ACLR. The enrolled patients were divided into the control group (ACLR only) and the SVF group (ACLR with SVF). The functional outcomes in both groups were assessed by the Lysholm knee scoring system, the Tegner activity scale, and the International Knee Documentation Committee (IKDC) subjective evaluation form, and compared at several time points during a 12-month follow-up. We found that the sex distribution and pre-surgery scores were similar in the two groups, whereas the mean age of the SVF group was higher than that of the control group (p = 0.046). The between-group analysis and generalized estimating equation model analysis revealed that, while patients in the SVF group significantly improved all their functional outcomes at 12 months after surgery, this improvement was not significantly different from the results of patients in the control group (Lysholm, p = 0.553; Tegner, p = 0.197; IKDC, p = 0.486). No side effects were observed in either group. We concluded that the intraoperative administration of SVF does not improve or accelerate functional recovery after ACLR in young patients.

Stromal Vascular Fraction Promotes Viability of Co-grafted Axial Skin Flaps in Rats Model.

Stromal vascular fraction (SVF) has been proved in promoting the vascularization of fascial flap through cell differentiation and paracrine effect and can be autologous transplanted without culture after isolation in vitro. We intend to establish a novel co-grafted flap model of rats to investigate the efficacy and mechanism of SVF on flaps and skinsin facilitating angiogenesis and immune regulation. 60 female Sprague Dawley rats were divided into the SVF group and the control group. A pedicled fascial flap combined with a free skin model was established, and 4×106 CM-DIl labeled SVF cells were transplanted into the fascia flap; the rats were executed on days 1, 2, 3, 7, 10 postoperatively (n=6). Flow cytometry was carried out to determine the cell proportion and surface marker of SVFs. The therapeutic effects of SVF were evaluated via Doppler blood perfusion imager, flap survival rates, histology, immunohistochemistry and immunofluorescence. The bioinformatic mechanism analysis was achieved by high-throughput RNAseq of mRNA and LncRNA. Flow cytometry confirmed SVF contains heterogeneous cellular composition, especially hematopoietic cells. Doppler blood perfusion imager showed SVF significantly improved flap survival with higher blood perfusion and survival rates. Immunohistochemistry of CD31 displayed higher level of angiogenesis in SVF-treated group, and CM-DIL-labeled SVF cells could survive and participate in revascularization, and RNA sequencing results revealed SVF promoted wound healing by facilitating intercellular adhesion, cell migration and positive immune response. SVF could reduce skin flap necrosis and activated neovascularization in rats by facilitating intercellular adhesion, cell migration and regulate positive immune response. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Paracrine effect of the stromal vascular fraction containing M2 macrophages on human chondrocytes through the Smad2/3 signaling pathway.

The adipose-derived stromal vascular fraction (SVF) is composed of a heterogeneous mix of adipose-derived stem cells (ADSCs), macrophages, pericytes, fibroblasts, blood, and other cells. Previous studies have found that the paracrine effects of SVF cells may be therapeutic, but their role in osteoarthritis treatment remains unclear. This study aimed to investigate the therapeutic effect of SVF cells on chondrocytes. Chondrocytes were seeded on culture plates alone (control) or cocultured with SVF or ADSCs on cell culture inserts. After 48 h of coculture, chondrocyte collagen II, tissue inhibitors of metalloproteinases-3 (TIMP-3), and matrix metalloproteinases-13 (MMP-13) messenger RNA (mRNA) expression levels were evaluated using reverse-transcription polymerase chain reaction, and the transforming growth factor-β (TGF-β) levels in the supernatant were measured using ELISA. Immunohistochemical staining and flow cytometry were used to evaluate the macrophages in the SVF. These macrophages were characterized according to phenotype using the F4/80, CD86, and CD163 markers. To determine whether the Smad2/3 signaling pathways were involved, the chondrocytes were pre-treated with a Smad2/3 phosphorylation inhibitor and stimulated with the SVF, and then Smad2/3 phosphorylation levels were analyzed using western blot. The mRNA expression levels of various paracrine factors and chondrocyte pellet size were also assessed. Collagen II and TIMP-3 expression were higher in the SVF group than in the ADSC group and controls, while MMP-13 expression was the highest in the ADSC group and the lowest in the controls. TGF-β levels in the SVF group were also elevated. Immunohistochemical staining and flow cytometry revealed that the macrophages in the SVF were of the anti-inflammatory phenotype. Western blot analysis showed that the SVF increased Smad2/3 phosphorylation, while Smad2/3 inhibitors decreased phosphorylation. Smad2/3 inhibitors also reduced the expression of various other paracrine factors and decreased chondrocyte pellet size. These findings suggested that the paracrine effect of heterogeneous cells, such as anti-inflammatory macrophages, in the SVF partly supports chondrocyte regeneration through TGF-β-induced Smad2/3 phosphorylation.

Mid-term prognosis of the stromal vascular fraction for knee osteoarthritis: a minimum 5-year follow-up study

BackgroundThe short-term safety and efficacy of stromal vascular fraction (SVF) in treating knee osteoarthritis (KOA) have been extensively studied but the mid-term and long-term prognoses remain unknown.Methods126 KOA patients were recruited and randomly assigned to SVF group and hyaluronic acid (HA) group (control group). The scores of visual analogue scale (VAS) and the Western Ontario and McMaster University Osteoarthritis Index (WOMAC) were assessed and compared between the two groups 1, 2, 3, and 5 years after treatment. The endpoint was defined as surgeries related to KOA or clinical scores exceeding the patient acceptable symptom state (PASS).ResultsThe VAS and WOMAC scores in the SVF group were significantly better than those in the HA group during the 5-year follow-up after treatment. The average responsive time to SVF treatment (61.52 months) was significantly longer than HA treatment (30.37 months). The adjusted Cox proportional hazards model showed that bone marrow lesion (BML) severity, body mass index (BMI) and treatment were independent risk factors and that the use of SVF reduced the risk of clinical failure by 2.602 times. The cartilage volume was reduced in both the SVF and control groups at 5 years but reduced less in the SVF group.ConclusionsUp to 5 years after SVF treatment, acceptable clinical state was present for approximately 60% of patients. BML severity and BMI were independent predictors of the prognosis.Trial Registry: This study was retrospectively registered at Chinses Clinical Trial Registry with identifier ChiCTR2100052818 and was approved by ethics committee of the First Affiliated Hospital of Zhejiang Chinese Medical University, number 2013-X-063.

Stromal-vascular fraction and adipose-derived stem cell therapies improve cartilage regeneration in osteoarthritis-induced rats

This study aimed to evaluate the effects of the stromal vascular fraction (SVF) and adipose-derived stem cells (ADSCs) on cartilage injury in an osteoarthritis (OA) rat model. Sodium iodoacetate (3 mg/50 μL) was used to induce OA in the left knee joint of rats. On day 14 after OA induction, 50 μL of SVF (5 × 106cells), ADSCs (1 × 106 cells), or 0.9% normal saline (NS) was injected into the left knee-joint cavity of each group. The macroscopic view and histological sections revealed that the articular cartilage in the NS group was damaged, inflamed, uneven and thin, and had hyperchromatic cell infiltration. Notably, the cartilage surface had recovered to nearly normal and appeared smooth and bright on day 14 in the SVF and ADSC groups. Additionally, the white blood cell counts in the SVF and ADSC groups were higher than those in the NS group on day 14. Plasma IL-1β levels on days 7 and 14 were reduced in the SVF and ADSC groups. These results indicated that both SVF and ADSC treatments may assist in articular cartilage regeneration after cartilage injury. Cell therapy may benefit patients with OA. However, clinical trials with humans are required before the application of SVF and ADSC treatments in patients with OA.

THE INFLUENCE OF REGENERATIVE TECHNOLOGIES ON RECOVERY PROCESSES IN LEG AFTER TRAUMATIC ISCHEMIA (EXPERIMENTAL STUDY)

Post-traumatic muscle ischemia results from severe injury and can lead to muscle dysfunction. Therefore, patient management and treatment are very significant in all periods of injury. New methods are performed, especially using regenerative technologies to avoid complications and improve long-term outcomes. Objective. To determine histological changes in the muscles of the injured limb after traumatic ischemia after injection of platelet-rich plasma,Bone marrow stem cell concentrate (BMAC), and Stromal vascular fraction (SVF) prepared from adipose tissue on the 5, 15, and 30 days. Material and methods. Experiments were conducted on rabbits (Chinchilla breed). A tourniquet imposed on a lower limb, from the middle third of the thigh to the ankle joint. After 6 hours, the tourniquet was removed. The animals were divided into four groups: control, platelet-rich plasma, bone marrow stem cell concentrate, and stromal vascular fraction prepared from adipose tissue — histological muscle changes provided by Tescan Mira 3 LMU (Czech Republic) in scanning transmission electron microscopy. Results. On the 5th day after the experiment were no significant histological changes in muscles but in the contrary on the 15 days after experiment in BMAC and SVF groups detected new muscle fibers formation in necrotic areas and myonucleus organization. On the 30th day new angiogenesis was detected around muscle fibers. Platelet-rich plasma group characterized by massive connective tissue formation in necrotic areas. Conclusions. Necrosis and progressive muscle hypotrophy are unavoidably for this type of injury. It was shown that BMAC and SVF could stimulate regeneration and angiogenesis. 

TGF-1 biomarker level evaluation on fracture healing in a murine model with a bone defect after stromal vascular fraction application.

Aim Bone defect is a challenge even for experienced orthopaedic surgeons and it is a significant cause of morbidity in patients and a source of high economic burden in health care. A severe bone defect is a condition whereby the bone tissue cannot undergo natural healing despite surgical stabilization and requires further surgical intervention. Stromal vascular fraction (SVF) is a heterogeneous cell population derived from adipose tissue that results from minimal manipulation of the adipose tissue itself. TGF is essential in maintaining and expanding mesenchymal stem cells or progenitors of osteoblasts. Furthermore, TGF-β signalling also triggers osteoprogenitor cell proliferation, early differentiation, and maintenance of osteoblasts in the bone healing process. The aim of this study was to determine the effect of administering SVF on bone defects' healing process assessed based on the TGF- β1. Methods This was an animal study involving twelve Wistar strain Rattus norvegicus. They were divided into three groups: negative group (normal rats), positive group (rats with bone defect without SVF application), and SVF group (rats with bone defect with SVF application). After 30 days, the rats were sacrificed, the TGF- β1 biomarker was evaluated (quantified using ELISA). Results TGF- β1 biomarker expressions were higher in the group with SVF application than in the group without SVF application. All comparisons of the SVF group and positive control group showed significant differences (p=0,000), respectively. Conclusion Giving SVF application could aid the healing process in a murine model with bone defect, marked by an increased level of TGF- β1.

The Effects of Autologous Fat Transfer in an In Vitro Model of Basal Joint Osteoarthritis

Basal joint osteoarthritis (OA) is a highly prevalent and debilitating condition. Recent clinical evidence suggests that autologous fat transfer (AFT) may be a promising, minimally invasive treatment for this condition. However, the mechanism of action is not fully understood. It is theorized that AFT reduces inflammation in the joint, functions to regenerate cartilage, or acts as a mechanical buffer. The purpose of this study was to better understand the underlying mechanism of AFT using an invitro model. We hypothesize that the addition of stromal vascular fraction (SVF) cells will cause a reduction in markers of inflammation. Articular chondrocytes were expanded in culture. Liposuction samples were collected from human subjects and processed similarly to AFT protocols to isolate SVF rich in adipose-derived stem cells. A control group was treated with standard growth media, and a positive control group (OA group) was treated with inflammatory cytokines. To mimic AFT, experimental groups received inflammatory cytokines and either a low or high dose of SVF. Expression of relevant genes was measured, including interleukin (IL)-1ß, IL-1 receptor antagonist, and matrix metalloproteinases (MMP). Compared to the OA group, significant decreases in IL-1ß, MMP3, and MMP13 expression on treatment day 3 were found in the high-dose SVF group, while MMP13 expression was also significantly decreased in the low-dose SVF group on day3. In this study, we found that SVF treatment reduced expression of IL-1ß, MMP3, and MMP13 in an invitro model of OA. These results suggest that an anti-inflammatory mechanism may be responsible for the clinical effects seen with AFT in the treatment of basal joint OA. An anti-inflammatory mechanism may be responsible for the clinical benefits seen with AFT for basal joint arthritis.