scholarly journals Mechanical process prior to cryopreservation of lipoaspirates maintains extracellular matrix integrity and cell viability: evaluation of the retention and regenerative potential of cryopreserved fat-derived product after fat grafting

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Jingwei Feng ◽  
Wansheng Hu ◽  
Mimi Lalrimawii Fanai ◽  
Shengqian Zhu ◽  
Jing Wang ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Viability ◽  
Retention Rate ◽  
Stromal Vascular Fraction ◽  
Explant Culture ◽  
Fat Grafting ◽  
Tissue Integrity ◽  
Immunocompromised Mice ◽  
Adipose Derived Stem Cell

Abstract Background Cryopreservation of fat grafts facilitates reinjection for later use. However, low temperature and thawing can disrupt tissues and cause lipid leakage, which raises safety concerns. Here, we compared the cryopreservation potential of stromal vascular fraction (SVF) gel processed from lipoaspirate with that of fat. Methods Human SVF gel and fat were cryopreserved at − 20 °C without cryoprotectant for 1 month. Fresh SVF gel and fat were used as controls. Tissue viability, adipose-derived stem cell (ASC) function, and the extracellular content were evaluated. At 3 months after transplanting the specimens to immunocompromised mice subcutaneously, the grafts were examined for retention, tissue engraftment, and inflammatory levels. The regenerative effect of cryopreserved SVF gel was evaluated in a murine ischemic wound healing model. Results At 1 month, the cell death rate in the SVF gel group was 36 ± 2%. The survived ASCs not only could be isolated via explant culture but also preserved colony-forming and differentiation. However, prolonged cryopreservation exacerbated apoptosis. Assessment of recovered tissues showed that the morphology, cell viability, and extracellular protein enrichment were better in SVF gel-preserved tissues than in frozen fat. At 3 months after lipotransfer, the retention ability of 1-month cryopreserved fat was 41.1 ± 9% compared to that of 1-month cryopreserved SVF gel. Immunostaining results showed that adipose tissue regeneration and integrity in the 1-month cryopreserved SVF gel group were superior to those of the cryopreserved fat group. The cryopreserved SVF gel also accelerated healing of the ischemic wound, compared with cryopreserved fat. Conclusion Cryopreserved SVF gel maintained tissue integrity and cell viability and resulted in a better long-term retention rate than that of cryopreserved fat. Cryopreserved SVF gel also showed superior regenerative potential and improved ischemic wound healing.

scholarly journals Enzyme-Crosslinked Gelatin Hydrogel with Adipose-Derived Stem Cell Spheroid Facilitating Wound Repair in the Murine Burn Model

Polymers ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 2997
Author(s):  
Ting-Yu Lu ◽  
Kai-Fu Yu ◽  
Shuo-Hsiu Kuo ◽  
Nai-Chen Cheng ◽  
Er-Yuan Chuang ◽  
...  
Keyword(s):  
Wound Healing ◽  
Wound Repair ◽  
Three Dimensional ◽  
Stromal Vascular Fraction ◽  
Skin Wound ◽  
3D Scaffold ◽  
Skin Wound Healing ◽  
Cell Spheroid ◽  
Cell Spheroids ◽  
Adipose Derived Stem Cell

Engineered skin that can facilitate tissue repair has been a great advance in the field of wound healing. A well-designed dressing material together with active biological cues such as cells or growth factors can overcome the limitation of using auto-grafts from patients. Recently, many studies showed that human adipose-derived stem cells (hASCs) can be used to promote wound healing and skin tissue engineering. hASCs have already been widely applied for clinical trials. hASCs can be harvested abundantly because they can be easily isolated from fat tissue known as the stromal vascular fraction (SVF). On the other hand, increasing studies have proven that cells from spheroids can better simulate the biological microenvironment and can enhance the expression of stemness markers. However, a three-dimensional (3D) scaffold that can harbor implanted cells and can serve as a skin-repaired substitute still suffers from deficiency. In this study, we applied a gelatin/microbial transglutaminase (mTG) hydrogel to encapsulate hASC spheroids to evaluate the performance of 3D cells on skin wound healing. The results showed that the hydrogel is not toxic to the wound and that cell spheroids have significantly improved wound healing compared to cell suspension encapsulated in the hydrogel. Additionally, a hydrogel with cell spheroids was much more effective than other groups in angiogenesis since the cell spheroid has the possibility of cell–cell signaling to promote vascular generation.

scholarly journals Characterized the Adipogenic Capacity of Adipose-Derived Stem Cell, Extracellular Matrix, and Microenvironment With Fat Components Grafting

2021 ◽  
Vol 9 ◽  
Author(s):  
Wenqing Jiang ◽  
Junrong Cai ◽  
Jingyan Guan ◽  
Yunjun Liao ◽  
Feng Lu ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Fluorescent Protein ◽  
Macrophage Polarization ◽  
Lethal Dose ◽  
Stromal Vascular Fraction ◽  
Fat Grafting ◽  
Tissue Integrity ◽  
Green Fluorescent

Background: Autologous fat grafting has been a widely used technique; however, the role of adipose-derived stem cells (ASCs), extracellular matrix (ECM), and microenvironment in fat regeneration are not fully understood.Methods: Lipoaspirates were obtained and processed by inter-syringe shifting to remove adipocytes, yielding an adipocyte-free fat (Aff). Aff was then exposed to lethal dose of radiation to obtain decellularized fat (Df). To further remove microenvironment, Df was rinsed with phosphate-buffered saline (PBS) yielding rinsed decellularized fat (Rdf). Green fluorescent protein (GFP) lentivirus (LV-GFP)-transfected ASCs were added to Df to generate cell-recombinant decellularized fat (Crdf). Grafts were transplanted subcutaneously into nude mice and harvested over 3 months.Results: Removal of adipocytes (Aff) didn’t compromise the retention of fat grafts, while additional removal of stromal vascular fraction (SVF) cells (Df) and microenvironment (Rdf) resulted in poor retention by day 90 (Aff, 82 ± 7.1% vs. Df, 28 ± 6.3%; p < 0.05; vs. Rdf, 5 ± 1.2%; p < 0.05). Addition of ASCs to Df (Crdf) partially restored its regenerative potential. Aff and Crdf exhibited rapid angiogenesis and M2-polarized macrophages infiltration, in contrast to impaired angiogenesis and M1-polarized inflammatory pattern in Df. GFP + ASCs participated in angiogenesis and displayed a phenotype of endothelial cells in Crdf.Conclusion: Adipose ECM and microenvironment have the capacity to stimulate early adipogenesis while ECM alone cannot induce adipogenesis in vivo. By directly differentiating into endothelial cells and regulating macrophage polarization, ASCs coordinate early adipogenesis with angiogenesis and tissue remodeling, leading to better long-term retention and greater tissue integrity.

scholarly journals Development of a System and Method for Automated Isolation of Stromal Vascular Fraction from Adipose Tissue Lipoaspirate

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Swathi SundarRaj ◽  
Abhijeet Deshmukh ◽  
Nancy Priya ◽  
Vidya S. Krishnan ◽  
Murali Cherat ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Point Of Care ◽  
Stromal Vascular Fraction ◽  
Fat Grafting ◽  
Automated System ◽  
Soft Tissue Reconstruction ◽  
Manual Method ◽  
Size Dependent ◽  
Tissue Reconstruction

Autologous fat grafting for soft tissue reconstruction is challenged by unpredictable long-term graft survival. Fat derived stromal vascular fraction (SVF) is gaining popularity in tissue reconstruction as SVF-enriched fat grafts demonstrate improved engraftment. SVF also has potential in regenerative medicine for remodeling of ischemic tissues by promoting angiogenesis. Since SVF cells do not require culture expansion, attempts are being made to develop automated devices to isolate SVF at the point of care. We report development of a closed, automated system to process up to 500 mL lipoaspirate using cell size-dependent filtration technology. The yield of SVF obtained by automated tissue digestion and filtration (1.17 ± 0.5 × 105 cells/gram) was equivalent to that obtained by manual isolation (1.15 ± 0.3 × 105;p= 0.8), and the viability of the cells isolated by both methods was greater than 90%. Cell composition included CD34+CD31− adipose stromal cells, CD34+CD31+ endothelial progenitor cells, and CD34−CD31+ endothelial cells, and their relative percentages were equivalent to SVF isolated by the manual method. CFU-F capacity and expression of angiogenic factors were also comparable with the manual method, establishing proof-of-concept for fully automated SVF isolation, suitable for use in reconstructive surgeries and regenerative medicine applications.

scholarly journals Efektivitas Sediaan Salep Limbah Kulit Singkong Sebagai Penyembuh Luka

2019 ◽  
Vol 5 (2) ◽  
pp. 133
Author(s):  
Widia Nila Risnadewi ◽  
Nadya Arianita Turisia ◽  
Alsifah Nurhidayati ◽  
Candra Dwipayana Hamdin
Keyword(s):  
Wound Healing ◽  
Wound Care ◽  
Healing Process ◽  
Ethanol Extract ◽  
Good Effect ◽  
Wound Healing Process ◽  
Tissue Integrity ◽  
Herbal Plants ◽  
Physical And Chemical

The skin is an important organ that functions to receive stimuli such as touch, pain and other influences from the outside. Some problems that occur on the skin can becaused because the skin is very vulnerable to trauma and injury. Wound is a condition of the breakdown of tissue integrity caused by violence both mechanical, physical and chemical in nature. In general, wound care in the community is done with a dressing accompanied by compresses betadine and normal saline because these materials are easily available. However, long-term use of the dressing can cause slow wound healing and various infections can appear. This causes people to look for other alternative treatments, one of them with herbal plants, namely cassava plants have the potential to be used as a wound healing and is able to inhibit bacteria because they are suspected to contain phenolic compounds. This study was a true experimental study, the group of test animals was divided into 6 groups, after that the making of wounds and smeared with bacteria were given ointments with 3 concentrations namely 5%, 10%, and 15%. Ointment extract of cassava skin waste gives effect to the wound healing process seen from the length of the wound. The ethanol extract ointment which has a good effect on wound healing is an ointment with a concentration of 10% and 15%.

A Systematic Review on Extracellular Vesicles-Enriched Fat Grafting: A Shifting Paradigm

2020 ◽  
Author(s):  
Mohammad Ghiasloo ◽  
Laura De Wilde ◽  
Kashika Singh ◽  
Patrick Tonnard ◽  
Alexis Verpaele ◽  
...  
Keyword(s):  
Systematic Review ◽  
Graft Survival ◽  
Extracellular Vesicles ◽  
Retention Rate ◽  
Fat Grafting ◽  
Retention Rates ◽  
Fat Graft ◽  
Cochrane Central Register

Abstract Background Recent evidence confirms that mesenchymal stem cells (MSCs) facilitate angiogenesis mainly through paracrine function. Extracellular vesicles (EVs) are regarded as key components of the cell secretome, possessing functional properties of their source cells. Subsequently, MSC-EVs have emerged as a novel cell-free approach to improve fat graft retention rate. Objectives To provide a systematic review of all studies reporting the use of MSC-EVs to improve graft retention rate. Methods A systematic search was undertaken using the Embase, PubMed and the Cochrane Central Register of Controlled Trials databases. Outcome measures included donor/receptor organism of the fat graft, study model, intervention groups, evaluation intervals, EV research data, in vitro and in vivo results. Results Of the total 1717 articles, 62 full-texts were screened. Seven studies reporting on 294mice were included. Overall, EV treated groups showed higher graft retention rates compared to untreated groups. Notably, retention rate was similar following EV- and MSC-treatment. In addition to reduced inflammation, graft enrichment with EVs resulted in early revascularization and better graft integrity. Interestingly, hypoxic preconditioning of MSCs improved their beneficial paracrine effects and led to a more proangiogenic EV population, as observed by both in vitro and in vivo results. Conclusions MSC-EVs appear to offer an interesting cell-free alternative to improve fat graft survival. While their clinical relevance remains to be determined, it is clear that not the cells, but their secretome is essential for graft survival. Thus, a paradigm shift from cell-assisted lipotransfer towards ‘secretome-assisted lipotransfer’ is well on its way.

scholarly journals A Clinical Study of Platelet-Rich Fibrin Combined With Autologous High-Density Fat Transplantation in Augmentation Rhinoplasty

2021 ◽  
pp. 014556132110169
Author(s):  
Dan Yan ◽  
Shuai-Hua Li ◽  
An-Li Zhang ◽  
Yao Xiao ◽  
Ze-Chun Huang
Keyword(s):  
Retention Rate ◽  
Statistical Significance ◽  
High Density ◽  
Control Group ◽  
Long Term Effects ◽  
Fat Transplantation ◽  
Augmentation Rhinoplasty ◽  
Platelet Rich Fibrin ◽  
Autologous Fat

Objective: This study was designed to analyze the clinical effect of autologous fat-granule transplantation in augmentation rhinoplasty and explore methods to improve the fat retention rate. Methods: A total of 70 enrolled patients were randomly divided into 2 groups: the platelet-rich fibrin (PRF) combined with high-density fat transplantation group (combined group) and the conventional fat-granule transplantation group (control group; n = 35 in each group). In the combined group, an appropriate amount of autologous fat was extracted and centrifuged, and the lower layer of high-density fat was taken and mixed with PRF isolated from whole blood for autotransplantation. In the control group, only fat was extracted and centrifuged for transplantation. The patients were followed up with for more than one year to observe the short- and long-term effects, complications, safety, and patient satisfaction. Results: Six months after the operation, the nasal shape was stable, the contour was higher and more stereoscopic than before, the average increase of nasal height was 3.0 mm in the combined group and 2.0 mm in the control group. No complications, such as fat embolism, infection, or necrosis occurred during the 1-year follow-up. The satisfaction rate between the 2 groups has statistical significance ( P < .05). Conclusion: Overall, PRF combined with autologous high-density fat transplantation is simple to perform, has a significantly increased fat-retention rate than the control group, and has stable long-term effects without obvious adverse reactions. A sufficient amount of fat and PRF transplantation can achieve a good orthopedic effect. Thus, this method can be widely used in clinical augmentation rhinoplasty.

scholarly journals Facial Rejuvenation with Concentrated Lipograft—A 12 Month Follow-Up Study

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 594
Author(s):  
Lukas Prantl ◽  
Eva Brix ◽  
Sally Kempa ◽  
Oliver Felthaus ◽  
Andreas Eigenberger ◽  
...  
Keyword(s):  
Stromal Vascular Fraction ◽  
Facial Rejuvenation ◽  
Fat Graft ◽  
Volume Loss ◽  
Patient’S Satisfaction ◽  
Resazurin Assay ◽  
Long Term Follow Up ◽  
The Difference

Lipofilling is a popular technique to treat volume loss in aging patients. The isolated adipose tissue is composed of adipocytes and stromal vascular fraction cells, which include adipose-derived stem cells (ASC). We hypothesize that the patient’s wrinkle severity scale (WSS) and patient’s satisfaction on the global aesthetic improvement scale (GAIS) can be improved after using concentrated lipoaspirate. Fourteen patients (54 years ± 11.09 years) with volume loss in the midface area underwent waterjet-assisted liposuction (Human Med AG, Schwerin, Germany). Fat was centrifuged in an ACP Double Syringe (Arthrex GmbH, Munich, Germany) using Rotofix 32A centrifuge (Andreas Hettich, GmbH & Co.KG, Tuttlingen, Germany). Homogenization was performed using the double syringe and a 1.4 mm female–female luerlock connector. After a second centrifugation, patients received periorbital (PO) and nasolabial (NL) lipografting. ASC count was performed after enzymatical digestion. Vitality of cells was assessed using a resazurin assay. During long-term follow up (12 months, n = 10), we found a high patient’s satisfaction (GAIS 1+/−0.52) and a good improvement of the WSS during short- and long-term follow-up. The ASC count of processed lipoaspirate was 2.1-fold higher than of unprocessed lipoaspirate (p < 0.001). The difference of ASC in sedimented and simply centrifuged lipoaspirate was also significant (p < 0.05). Facial rejuvenation with concentrated fat graft offers good results concerning objective aesthetic outcome and patient’s satisfaction.

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