Influence of adipose tissue and its derivative on wound repair and vascularization / 中国组织工程研究

BACKGROUND:Vascularization is essential for wound healing and functional recovery during soft tissue repair.Adipose tissue is believed to be the body's largest source of stem cells,and a number of different fat complexes have been developed for research and treatment.Its ability to promote angiogenesis and soft tissue repair has been extensively studied. OBJECTIVE:To review the progress of vascularization in soft tissue repair,and to summarize the preparation methods of adipose tissue and its derivative and their applications in vascularization and soft tissue repair.It is proven that adipose tissue and its derivative have excellent research value and clinical application prospects in vascular and soft tissue engineering. METHODS:PubMed,Web of Science and CNKI databases were used to search the related articles published from January 2010 to February 2023.The search terms were"soft tissue repair,wound healing,vascularization,angiogenesis,adipose tissue,stromal vascular fraction,adipose tissue-derived microvascular fragment,nanofat,adipose extracellular matrix/stromal vascular fraction gel"in Chinese and English.A small number of old classic literature was also included.An initial screening was performed by reading the titles and abstracts to exclude literature that was not relevant to the topic of the article,and 69 papers were finally included for the analysis of the result. RESULTS AND CONCLUSION:(1)Wound healing is an important physiological process,which mainly occurs when tissue is damaged,such as injury,surgery,burn,tumor,infection and vascular disease caused by tissue damage and defects.(2)Adequate vascularization of the wound site is essential for tissue repair,reconstruction of local homeostasis and functional recovery.(3)Adipose tissue is believed to be the body's largest pool of stem cells and a number of different fat components have been used for research and treatment.(4)Due to its inherent composition and preparation advantages,adipose tissue will continue to play an important role in tissue engineering research and therapy.

The effect of gauze filtration on the content of adipose stem cells in fat gel / 中国医师杂志

Objective:To study the effect of gauze filtration on the content of adipose stem cells in fat gel.Methods:In March 2022, fat samples from 5 patients who underwent abdominal fat aspiration at the Hunan Provincial People′s Hospital were collected, prepared fat ged, the fat gel was filtered 1-5 times with 2 layers of gauze, and the amount of fat gel passing through a 1 ml threaded syringe with a 27 G needle was measured after each filtration. At the same time, the content of fat stem cells in the filtered fat gel was detected by flow cytometry.Results:As the number of filters increases, the amount of fat gel passing through gradually increased. The increasing trend was obvious after the first to fourth filters, and the increasing trend slowed down after the fifth filter. After statistical analysis, the differences between the first filter and the second filter, the second filter and the third filter, and the third filter and the fourth filter were statistically significant ( P<0.05). There was no statistically significant difference between the fourth filter and the fifth filter ( P>0.05). As the number of filters increased, the content of adipose stem cells in the fat gel gradually decreased. The decreasing trend was obvious after the first to third filters, and the decreasing trend slowed down after the fourth filter. After statistical analysis, the differences between the first filter and the second filter, as well as the second filter and the third filter, were statistically significant (all P<0.05). However, there was no statistically significant difference between the third filter and the fourth filter, and the fourth filter and the fifth filter (all P>0.05). Conclusions:The more times gauze is filtered, the better the permeability of fat gel. However, the content of adipose stem cells will significantly decrease in the first three filters. Therefore, the number of times gauze is filtered for fat gel should be minimized as much as possible.

Conversion of stromal vascular fraction in the microenvironment of radiation-induced skin injuries and its clinical implications / 中华放射医学与防护杂志

Objective:To investigate the conversion of stromal vascular fraction (SVF) in the microenvironment of radiation-induced skin injuries to provide guidance for clinical applications.Methods:Based on a random number table, C57BL/6N mice were categorized into four groups: the blank control, negative control, acute injury, and chronic injury groups, with each group containing 25 mice. The backs of mice in the blank control, acute injury, and chronic injury groups were exposed to 15 Gy X-ray irradiation. Then, the mice in the negative control, acute injury, and chronic injury groups were injected subcutaneously with the SVF derived from B6/G-R mice. The survival of these mice was observed 1, 3, 7, 14, and 21 d after the injection through fluorescence tracing and in vivo imaging. Accordingly, the clinical SVF injection regimens were optimized based on the experimental result of mice. Finally, local SVF injection was performed on different frequencies for patients in different wound conditions, with the efficacy being observed. Results:The fluorescence of SVF was observed from the tissue slices of the acute injury, chronic injury, and negative control groups 14 d post-injection. The result showed that the fluorescence intensity of SVF 1, 3, and 7 d post-injection was in the order of the negative control group > the acute injury group > the chronic injury group. The acute injury group ranked at the top and the chronic injury group remained at the bottom 14 d after the injection. The fluorescence of SVF in each group was barely detected 21 d after the injection. Compared to the negative control group, the acute injury group exhibited statistical differences only 14 d post-injection ( t = 4.11, P < 0.05), while the chronic injury group displayed statistical differences 1, 3, 7, and 14 d after the injection ( t = 3.88-5.74, P < 0.05). Furthermore, the acute injury group exhibited significantly higher fluorescence intensity of SVF than the chronic injury group ( t = 4.73-8.38, P < 0.05). The half-life of SVF for the negative control, acute injury, and chronic injury groups was 6.336, 6.014, and 2.163 d, respectively. As indicated by the application of SVF transplantation based on traditional surgical protocols in the clinical trial, SVF can significantly promote wound repair, with earlier SVF transplantation being more beneficial for wound healing. Conclusions:The conversion of SVF differs in the microenvironments of acute and chronic radiation-induced skin injuries. This can serve as an essential guide for the administration timing and injection frequency of SVF in clinical applications.

Role of active ADM microparticle in skin wound healing of diabetic mice / 中华医学美学美容杂志

Objective:To evaluate the effects of adipose-derived stem cells (ADSCs) and ADM microparticle on diabetic wound healing.Methods:ADSCs was co-cultured with ADM microparticle in vitro. The models of diabetic nude mice were established by intraperitoneal injection of STZ and the full-thickness skin defects were designed on the back. All 24 diabetic mice were randomly divided into 4 group: experimental groups were transplanted with ADSCs and ADM microparticle and the other groups were transplanted with ADSCs, ADM microparticle and blank control group was set up. On the 7th and 14 th days, the wound healing rate of 3 mice randomly selected from each group was calculated, and the thickness between dermis and epidermis was measured by hematoxylin and eosin staining. The density of neovascularization was measured by immunohistochemical staining. The differences were compared between the groups.Results:Compared to the ADSCs groups, the mice of the experimental groups showed higher cell survival rate. The wound healing rate in the experimental groups was (86.0±2.7)% (7 days) and (98.5±1.1)% (14 days), thicker dermis-epidermis distance was (99.1±1.8) μm (7 days) and (124.3±4.3) μm (14 days) ( P<0.05), and higher density of neovascularization was noted. Conclusions:The transplantation with active ADM microparticle can significantly promote neovascularization and wound healing of diabetic wound.

Clinical effect of mid-face lifting combined with autologous compound fat transplantation in facial rejuvenation / 中华医学美学美容杂志

Objective:To investigate the clinical effect and safety of mid-face lifting combined with compound fat transplantation in facial rejuvenation.Methods:From October 2016 to May 2020, 26 patients with mid-facial aging were enrolled, including 10 males and 16 females, with an average age of (47±6) years (38-65 years). The facial superficial musculoaponeurotic system was stripped, folded and suspended through the temporal hairline incision to lift the middle of the face. Liposuction was performed and compound fat was prepared, in which structural fat was injected into the area marked with loss of facial volume before operation, and extravascular matrix component gel was injected into the static wrinkle dermis. Six and 12 months after operation, the global aesthetic improvement scale of the observer and the global aesthetic improvement scale of the patient were scored, and the postoperative complications and patients＇ satisfaction were counted.Results:At the end of 6 months, improvements in mid face were reported in all patients by blinded reviewers. 12 months after operation, 53.8% (14 cases) of patients had significant improvement in facial aging, 30.8% (8 cases) had moderate improvement, and 15.4% (4 cases) had slight improvement; 25 patients (96%) were satisfied with the postoperative results. Postoperative hemorrhage occurred in one patient, scar hyperplasia occurred in one patient, and no obvious complications occurred in the other patients.Conclusions:The method of midface lifting combined with compound fat transplantation can improve the facial soft tissue and supplement the facial missing capacity, improve the static wrinkles of the face, and comprehensively solve the aging problem of midface. It has a definite clinical effect, fewer complications, and is safe and reliable. It has definite clinical effect with less complications.

Alcance de las células madre derivadas de tejido adiposo / Scope of stem cells derived from adipose tissue

Introducción: En el tejido adiposo se han identificado células madre mesenquimales con capacidad autorrenovadora y multipotencial. Mediante digestión enzimática y centrifugado del lipoaspirado se libera una población heterogénea de células denominada fracción vascular estromal, con innumerables potencialidades terapéuticas en el campo de la medicina regenerativa. Objetivo: Actualizar el alcance de las células madre derivadas de tejido adiposo en la terapia regenerativa. Método: Se revisaron 38 artículos entre los años 2000 y 2019 en las bases de datos Scielo, ScienceDirect, Medline y Pubmed. Desarrollo: Las células de la fracción vascular estromal se caracterizan por su capacidad de generar tejido adiposo y vasos sanguíneos, y por la producción de factores de crecimiento que ayudan en la supervivencia de los adipocitos y la formación de una red vascular. El principal mecanismo de acción de las células madre derivadas de tejido adiposo parece deberse a su acción paracrina y a la sinergia con células endoteliales. En el campo de la medicina regenerativa se han utilizado en el tratamiento de cicatrices patológicas y de fibrosis deformantes con impotencia funcional, en las reconstrucciones de secuelas por cáncer y en el cierre precoz de zonas cruentas. Conclusiones: La lipotransferencia es un procedimiento con un mínimo de complicaciones que constituye una de las opciones terapéuticas más empleadas para corregir defectos en los tejidos, debido a que no solo es un medio de relleno, sino que también permite la regeneración y restauración tisular. La presencia de células madre en el tejido adiposo, unido a su accesibilidad, disponibilidad e histocompatibilidad, ha motivado su aplicación cada vez más expandida en la medicina estética, reconstructiva y regenerativa(AU)

Introduction: Mesenchymal stem cells with self-renewing and multipotential capacity have been identified in adipose tissue. By means of enzymatic digestion and centrifugation of the lipoaspirate a heterogeneous population of cells called vascular stromal fraction is released. It has innumerable therapeutic potentialities in the field of regenerative medicine. Objective: To update the scope of stem cells derived from adipose tissue in regenerative therapy. Method: 38 articles published between 2000 and 2019 in the Scielo, ScienceDirect, Medline and Pubmed databases were reviewed. Development: The cells of the vascular stromal fraction are characterized by generating adipose tissue and blood vessels and by the production of growth factors that help in the survival of adipocytes and the formation of a vascular network. The main mechanism of action of stem cells derived from adipose tissue appears to be due to their paracrine action and synergy with endothelial cells. Stem cells derived from adipose tissue have been used in regenerative medicine for the treatment of pathological scars and deforming fibrosis with functional impotence, in the reconstruction of cancer sequelae and in the early closure of bloody areas. Conclusions: Lipotransfer is a procedure with a minimum of complications that constitutes one of the most widely used therapeutic options to correct tissue defects, since it is not only a filling medium, but also allows tissue regeneration and restoration. The presence of stem cells in adipose tissue, together with their accessibility, availability and histocompatibility, has motivated their increasingly widespread application in aesthetic, reconstructive and regenerative medicine(AU)

Clinical outcome of hair transplantation combined with SVF GEL assisted fat transplantation in cicatricial alopecia / 中华医学美学美容杂志

Objective:To explore the clinical effects of hair transplantation combined with SVF GEL assisted fat transplantation in treating cicatricial alopecia.Methods:Eleven patients (5 males and 6 females, ranging from 22 to 41 years old) with cicatricial alopecia caused by burn or tumor excision in Shanghai Mylike Cosmetic Hospital from October 2017 to August 2019. All patients were treated with hair transplantation combined with SVF GEL assisted fat transplantation according to their scar areas.Results:Patients were followed up for 11 months on average, their hair grew well, with significantly improved appearances and no obvious complications. Hair follicle detection system was used to analyze the hair density after transplantation. The average survival rate reached 90.26% in accompany with desirable patients＇ satisfactory rates.Conclusions:Excellent results can be achieved by conducting hair transplantation combined with SVF GEL assisted fat transplantation in treating cicatricial alopecia, which is worth clinical application.

Clinical effect of stromal vascular fraction gel injection on local rhinoplasty / 中华医学美学美容杂志

Objective:To observe the application and clinical effect of stromal vascular fraction gel (SVF-gel) in local rhinoplasty.Methods:A total of 84 patients with mild nasal root and low bridge were divided into observation group and control group. In the observation group, liposuction with SVF gel extraction plus injection rhinoplasty was carried out; in the control group, liposuction plus injection rhinoplasty was carried out according to the height change of nasal root and bridge, the change of nasal frontal angle, the occurrence of complications and the satisfaction of patients after operation.Results:The 84 patients were followed up for 2 weeks to 24 months. The height of nasal root and bridge increased significantly and the lines were more harmonious. The absorption of the observation group was much lower than that of control group. Except 3 patients in the observation group and 8 patients in the control group, there were different degrees of absorption within 6 months after operation. All patients did not have complications such as fat liquefaction, necrosis, induration and infection in the injection area. Three patients in the observation group were satisfied with the results after reinjection, and one patient in the control group was satisfied with the results after reinjection, three patients were not satisfied with the results, and four patients were satisfied with the prosthesis augmentation rhinoplasty. In the observation group, there were 37 cases of excellent satisfaction, 5 cases of good satisfaction, and the overall satisfaction was 100%; in the control group, there were 20 cases of excellent satisfaction, 15 cases of good satisfaction, 7 cases of bad satisfaction, and the satisfaction was 83.3%. The former was better than the latter.Conclusions:In view of superiorities such as lower complications with long-lasting effect, high fat survival rate and high satisfaction in local augmentation rhinoplasty, SVF-gel injection is especially suitable for patients who cannot accept autologous bone, prosthesis and allograft tissue filler for augmentation rhinoplasty.

Separation of adipose-derived stromal vascular fraction cells by a variety of physical methods: A comparative study / 中国组织工程研究

BACKGROUND: Increasing attention has been paid to vascular components of the adipose-derived matrix and adipose-derived stem cells in tissue engineering. Existing methods for separating the vascular components of the adipose-derived matrix mainly include enzymatic and bolus injection, both of which have fatal disadvantages. OBJECTIVE: To search for a method for preparing adipose-derived stromal vascular fractions with high efficiency, safety, and simplicity. METHODS: The group without any treatment was used as the negative control, and the enzymatic hydrolysis method served as the positive control. The enzymatic hydrolysis method, traditional bolus method, modified bolus method, glass beads method and built-in ultrasonic waves method were compared through cell volume, survival rate, cell fragments, cell viability, increment rate and detection of microbial infection. The enzymatic hydrolysis method and the common bolus injection method were commonly used in the separation of vascular component cells of the fat source matrix; the improved bolus method was a method obtained by improving on the basis of the ordinary bolus method; the glass bead method was to use the glass bead to oscillate. The shear force generated was obtained by adding glass beads to the fat granules and shaking at 2 500 r/min for 9 minutes to prepare stromal vascular fraction cells. Using the built-in ultrasonic method, adipose tissue was treated at 25 W for 36 seconds to obtain stromal vascular fraction cells through a cavitation effect. RESULTS AND CONCLUSION: (1) The size of stromal vascular fraction cells isolated by five methods showed no significant difference (P > 0.05). (2)The cell viability was lowest in the negative control group, and highest in the enzymatic hydrolysis group. The cell viability in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups was significantly higher than that in the modified and traditional bolus groups (P < 0.05). (3) The cell survival rate and cell proliferation rate in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups were significantly higher than those in the modified and traditional bolus groups (P < 0.05). (4) The cell fragmentation rate and cell apoptosis rate in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups were significantly lower than those in the modified and traditional bolus groups (P < 0.05). (5) These results indicate that the built-in ultrasonic method and the glass bead method are better in enriching vascular components of the adipose-derived matrix. But glass bead method adds exogenous products, so it increases the risk of pollution. Built-in ultrasonic method inserts the ultrasound probe into the adipose tissue, but as long as the ultrasound probe is thoroughly sterilized, the risk of contamination is minimized. In general, the built-in ultrasonic method and the glass bead method are superior to modified and traditional bolus methods.

Chondrocyte cocultures with stromal vascular fraction of adipose tissue promote cartilage regeneration in vivo / 华西口腔医学杂志

OBJECTIVE@#This study aimed to compare the cartilage regeneration of the stromal vascular fraction (SVF) cells and adipose-derived mesenchymal stem cells (ASCs) cocultured with chondrocytes seeded on the scaffolds.@*METHODS@#The cellular morphologies and proliferation capabilities on the scaffolds were evaluated. The scaffolds with the cocul-ture of ASCs/SVF and chondrocytes were implanted into the full thickness cartilage defective rabbit joints for 10 weeks.@*RESULTS@#The cells seeded into the scaffolds showed good adhesion and proliferation. Implantation with SVF and chondrocytes revealed desirable in vitro healing outcomes.@*CONCLUSIONS@#The SVF cells were better than ASCs in terms of the formation of cartilage matrix in a coimplantation model. Without in vitro expansion, the SVF cells are good cell sources for cartilage repair.

Effect of human stromal vascular fraction gel on the treatment of patients with skin depressed scar and its mechanism / 中华烧伤杂志

Objective@#To observe content of cytokine in human stromal vascular fraction gel (SVF-GEL) and effect of SVF-GEL on biological behaviors of epidermal and dermal cells in vitro and clinical efficacy of SVF-GEL.@*Methods@#(1) SVF-GEL was prepared using liposuction aspirates harvested from females who received abdomen liposuction in author′s unit. SVF-GEL (1 mL) and high-glucose Dulbecco′s modified eagle medium (DMEM, 1 mL) were respectively cultured for 24 h with high-glucose DMEM containing 10% fetal calf serum, 10 g/L penicillin, and 10 g/L streptomycin, denoted as SVF-GEL group and negative control group, with 6 samples in each group. Content of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) in the supernatant was determined by enzyme-linked immunosorbent assay. (2) A number of 5×105 human skin fibroblasts (HSF) and HaCaT cells in logarithmic phase were inoculated and cultured in Transwell chambers for 12 h. All Transwell chambers containing cells were divided into SVF-GEL group (0.5 mL SVF-GEL was added for co-culture) and control group (0.5 mL high-glucose DMEM was added for co-culture), with 9 samples in each group for HSF and HaCaT cells. Scratch assay was performed after culture for 24 h, and residual scratch width was observed at post scratch hour (PSH) 0 (immediately), 24, and 48. Cell migration distance was measured at PSH 24 and 48. After culture for 24, 48, and 72 h, the number of living cell was counted using cell counter. (3) From June 2018 to June 2019, SVF-GEL was applied clinically to treat 15 patients with depressed scars on face, including 2 males and 13 females, aged 19 to 42 years. Survival condition of SVF-GEL and whether complications or not were observed 6 months after surgery. Before surgery and 6 months after surgery, depressed degree, color, and pliability of scar were compared. Vancouver Scar Scale (VSS) was employed to access color, vascularity, and pliability before surgery and 6 months after surgery, and total score was calculated. The number of patients with complete satisfaction or satisfaction was counted six months after surgery. Data were processed with analysis of variance of factorial design, paired samples t test, and Wilcoxon rank sum test.@*Results@#(1) The content of EGF in SVF-GEL group and negative control group was (316.6±12.8) and (3.4±0.6) pg/mL, and the content of VEGF in SVF-GEL group and negative control group was (568.67±12.19) and (4.93±0.16) pg/mL, with statistically significant differences between the two groups (t=48.777, 92.485, P<0.01). (2) Residual scratch widths of HSF and HaCaT in SVF-GEL group and control group were decreased gradually along with time elapse, in which those in SVF-GEL group at PSH 24 and 48 were less than those in control group. At PSH 24 and 48, cell migration distances of HSF and HaCaT in SVF-GEL group were more than those in control group (tHSF=-20.304, -43.516, tHaCaT=-15.060, -8.684, P<0.01). After culture for 24, 48, and 72 h, the number of living cell of HSF and HaCaT in SVF-GEL group was significantly more than that in control group (tHSF=-3.374, -6.809, -18.036, tHaCaT=-4.793, -6.028, -8.141, P<0.05 or P<0.01). (3) Six months after surgery, SVF-GEL grafted into patients survived well without complications, and depressed degree of scar ameliorated obviously with lightened pigmentation and softer texture as compared with before surgery. Compared with those before surgery, VSS scores of color, vascularity, and pliability, and total score of 15 patients with depressed scars on face were obviously decreased 6 months after surgery (Z=-2.06, -2.07, -2.07, t=-15.811, P<0.05 or P<0.01). One patient was satisfied with the clinical outcome, and the rest 14 patients were completely satisfied with the clinical outcomes.@*Conclusions@#SVF-GEL contains cytokines EGF and VEGF, which can enhance cell migration ability and proliferation ability of HSF and HaCaT cells and have obvious effects on depressed scars for clinical application.

Mechanism of promoted neovascularization by SVF after fat grafting / 中华整形外科杂志

Stromal vascular fraction(SVF)are the remaining cells after removing mature fat cells in the adipose tissue. Containing a certain amount of adipose derived stem cells(ADSCs), SVF also includes many other cells, which may have the potential of promoting angiogenesis. In this review, the role of SVF in angiogenesis after fat transplantation was summarized by intensive reading relative literature in recent years. The result is that angiogenesis and fat graft revascularization are regulated by various factors: SVF promotes secretion of a diverse array of cytokines and growth which are capable of stabilizing endothelium vascular network. ADSCs have the potential of differentiating into smooth muscle cells and endothelial cells which can coroperate to form new blood vessels.

Possibility of Injecting Adipose-Derived Stromal Vascular Fraction Cells to Accelerate Microcirculation in Ischemic Diabetic Feet: A Pilot Study

BACKGROUND AND OBJECTIVES: Beneficial effects of human adipose-derived stromal vascular fraction (SVF) cell injection on microcirculation have been recently reported in in vitro and in vivo studies. However, no clinical studies have reported its effect in diabetic patients who commonly experience compromised tissue perfusion, regardless of the status of intravascular blood flow. The present piloting study was designed to clinically examine the possibility of SVF cell injection to accelerate microcirculation, particularly in ischemic diabetic feet. METHODS: Ten diabetic feet were included to receive subcutaneous injection of SVF cells around wounds. Transcutaneous partial oxygen pressure (TcPO2) and cutaneous microvascular blood flow were measured before and every four weeks after cell injection until the 12th week visit. RESULTS: TcPO2 values increased from 31.3±7.4 before injection to 46.4±8.2 mmHg at 12 weeks after SVF injection (1.5-fold, p<0.05). Cutaneous microvascular blood flow levels increased from 34.0±21.1 before injection to 76.1±32.5 perfusion unit at 12 weeks after SVF injection (2.2-fold, p<0.05). There were no adverse events related to SVF cell injection. CONCLUSIONS: Results of this study demonstrate that adipose-derived SVF cell injection have the possibility to provide beneficial effects on microcirculation in ischemic diabetic feet.

Combination of stromal vascular fraction and Ad-COMP-Ang1 gene therapy improves long-term therapeutic efficacy for diabetes-induced erectile dysfunction / 亚洲男科学杂志(英文版)

Men with diabetic erectile dysfunction (ED) respond poorly to the currently available oral phosphodiesterase-5 inhibitors. Therefore, functional therapies for diabetic ED are needed. Stromal vascular fraction (SVF) and the adenovirus-mediated cartilage oligomeric matrix angiopoietin-1 (Ad-COMP-Ang1) gene are known to play critical roles in penile erection. We previously reported that SVF and Ad-COMP-Ang1 have only a short-term effect in restoring erectile function. Further improvements to ED therapy are needed for long-lasting effects. In the present study, we aimed to test if the combination of SVF and Ad-COMP-Ang1 could extend the erection effect in diabetic ED. We found that the combination therapy showed a long-term effect in restoring erectile function through enhanced penile endothelial and neural cell regeneration. Combination therapy with SVF and Ad-COMP-Ang1 notably restored cavernous endothelial cell numbers, pericyte numbers, endothelial cell-cell junctions, decreased cavernous endothelial cell permeability, and promoted neural regeneration for at least 4 weeks in diabetic mice. In summary, this is an initial description of the long-term effect of combination therapy with SVF and Ad-COMP-Ang1 in restoring erectile function through a dual effect on endothelial and neural cell regeneration. Such combination therapy may have therapeutic potential for the treatment of diabetic ED.

Effects of local transplantation of autologous adipose-derived stromal vascular fraction on the hyperplastic scar formation in rabbit ears and the mechanism / 中华烧伤杂志

Objective@#To explore the effects of local transplantation of autologous adipose-derived stromal vascular fraction (SVF) on the hyperplastic scar (HS) formation in rabbit ears and the mechanism.@*Methods@#Twenty-four New Zealand white rabbits were used to reproduce HSs by making four full-thickness skin defect wounds with a diameter of 1 cm on the ventral surface of left ear of each rabbit. Wound epithelization and local-tissue proliferation were observed, and wound healing (complete epithelization) time and formation time of HS were recorded. The 24 rabbits were divided into SVF group, pure DMEM group, and pure HS group according to the random number table, with 8 rabbits and 32 wounds in each group. On post injury day (PID) 25 (after the complete epithelization of wounds), 0.2 mL of low glucose DMEM medium containing CM-Dil labeled autologous SVF was injected into HSs of rabbits in SVF group, while the same amount of low glucose DMEM medium was injected into HSs of rabbits in pure DMEM group. The frequency of injection was once every 5 days, totally for 3 times. HSs of rabbits in pure HS group did not receive any treatment. On PID 40, HSs of rabbits′ ears in each group were harvested, then the histological form was observed by hematoxylin and eosin staining, the arrangement of collagen in HS was observed by Van Gieson staining, the distribution of CM-Dil-labeled SVF in the HS was observed with fluorescence microscope, and the mRNA expression and the protein expression of transforming growth factor β1 (TGF-β1), Smad3, and Smad7 in HS were determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction and Western blotting, respectively. Data were processed with one-way analysis of variance and Tukey test.@*Results@#(1) Complete epithelization time of wounds of rabbits′ ears was (20.0±2.0) d post injury, and HSs were formed on PID 25. On PID 40, HSs of rabbits′ ears in pure DMEM group and pure HS group were still in hyperplasia, while those in SVF group became smaller, flat, soft, and light colored. (2) On PID 40, compared with those in pure DMEM group and pure HS group, the number of epithelium foot like structures was more and the amount of inflammatory cells was less. The collagen of HSs of rabbits′ ears in SVF group was arranged more regularly with broader gap between collagens. (3) On PID 40, CM-Dil-labeled SVF could still be observed in the HSs of rabbits′ ears in SVF group. (4) On PID 40, compared with those in pure DMEM group and pure HS group, the mRNA expressions of TGF-β1 and Smad3 in the HSs of rabbits′ ears in SVF group were significantly down-regulated (P<0.05), while the mRNA expression of Smad7 was significantly up-regulated (P<0.05). There were no significant differences in the mRNA expressions of TGF-β1, Smad3, and Smad7 in the HSs of rabbits′ ears between pure DMEM group and pure HS group (P>0.05). (5) On PID 40, compared with those in pure DMEM group (0.74±0.03, 0.73±0.10, 0.54±0.09) and pure HS group (0.72±0.08, 0.71±0.12, 0.53±0.06), the protein expressions of TGF-β1 and Smad3 in the HSs of rabbits′ ears in SVF group (0.57±0.06, 0.42±0.09) were significantly down-regulated (P<0.05), while the protein expression of Smad7 (0.71±0.05) was significantly up-regulated (P<0.05). The protein expressions of TGF-β1, Smad3, and Smad7 in the HSs of rabbits′ ears in pure DMEM group and pure HS group were close (P>0.05).@*Conclusions@#Autologous SVF transplantation can inhibit the formation of HS in the early stage of scar formation of rabbit, the mechanism may be related to the TGF-β1/Smad signaling pathway.

Complex sternal and rib reconstruction with allogeneic material

Sternal malunion, or loss, developed after a median sternotomy cannot only be difficult to manage and treat, but also may diminish one’s quality-of-life drastically. The technique presented here represents a multispecialty approach in one stage for the reconstruction of an unstable thoracic cage. The procedure utilized a donated sternum and ribs. The sternum with ribs harvested from a single donor included adipose derived stromal vascular fraction (ADSVF) cells with marrow also from the same donor. Autologous muscle flaps, stabilized with acellular dermal matrix were utilized to provide a robust blood supply to the ADSVF cells and bone grafts. Acellular dermal matrix was used to construct the ribs and stabilize the plugs of stem cells and bone. These procedures, in the hands of multispecialty physicians, have led to several successful reconstructions involving complex chest wall deformities. This surgical intervention was performed in a one stage operation. This represents the first successful complete sternal transplant in a patient with return to normal activities and increased quality-of-life.

Efficacy and Safety of Autologous Stromal Vascular Fraction in the Treatment of Empty Nose Syndrome

OBJECTIVES: Regenerative treatment using stem cells may serve as treatment option for empty nose syndrome (ENS), which is caused by the lack of turbinate tissue and deranged nervous system in the nasal cavity. We aimed to assess the efficacy and safety of the autologous stromal vascular fraction (SVF) in the treatment of ENS. METHODS: In this prospective observational clinical study, we enrolled 10 ENS patients who volunteered to undergo treatment of ENS through the injection of autologous SVF. Data, including demographic data, pre- and postoperative Sino-Nasal Outcome Test-25 (SNOT-25) scores, overall patient satisfaction, and postoperative complications, were prospectively collected. Nasal secretion was assessed using the polyurethane foam absorption method, and the levels of biological markers were analyzed in both ENS group and control group using enzyme-linked immunosorbent assay. The SVF extracted from abdominal fat was diluted and injected into both inferior turbinates. RESULTS: Among the 10 initial patients, one was excluded from the study. Subjective satisfaction was rated as “much improved” in two and “no change” in seven. Among the improved patients, the mean preinjection SNOT-25 score was 55.0 and the score at 6 months after injection was 19.5. However, the average SNOT-25 score of nine participants at 6 months after injection (mean±standard deviation, 62.4±35.8) did not differ significantly from the baseline SNOT-25 score (70.1±24.7, P>0.05, respectively). Among the various inflammatory markers assessed, the levels of interleukin (IL)-1β, IL-8, and calcitonin gene-related peptide were significantly higher in ENS patients. Compared with preinjection secretion level, the nasal secretions from SVF-treated patients showed decreased expressions of IL-1β and IL-8 after injection. CONCLUSION: Although SVF treatment appears to decrease the inflammatory cytokine levels in the nasal mucosa, a single SVF injection was not effective in terms of symptom improvement and patient satisfaction. Further trials are needed to identify a more practical and useful regenerative treatment modality for patients with ENS.

Combination of stromal vascular fraction and Ad-COMP-Ang1 gene therapy improves long-term therapeutic efficacy for diabetes-induced erectile dysfunction / 亚洲男科学杂志(英文版)

Men with diabetic erectile dysfunction (ED) respond poorly to the currently available oral phosphodiesterase-5 inhibitors. Therefore, functional therapies for diabetic ED are needed. Stromal vascular fraction (SVF) and the adenovirus-mediated cartilage oligomeric matrix angiopoietin-1 (Ad-COMP-Ang1) gene are known to play critical roles in penile erection. We previously reported that SVF and Ad-COMP-Ang1 have only a short-term effect in restoring erectile function. Further improvements to ED therapy are needed for long-lasting effects. In the present study, we aimed to test if the combination of SVF and Ad-COMP-Ang1 could extend the erection effect in diabetic ED. We found that the combination therapy showed a long-term effect in restoring erectile function through enhanced penile endothelial and neural cell regeneration. Combination therapy with SVF and Ad-COMP-Ang1 notably restored cavernous endothelial cell numbers, pericyte numbers, endothelial cell-cell junctions, decreased cavernous endothelial cell permeability, and promoted neural regeneration for at least 4 weeks in diabetic mice. In summary, this is an initial description of the long-term effect of combination therapy with SVF and Ad-COMP-Ang1 in restoring erectile function through a dual effect on endothelial and neural cell regeneration. Such combination therapy may have therapeutic potential for the treatment of diabetic ED.

Construction of tissue engineering fat filler and survival of fat particles / 中华医学美学美容杂志

Objective To study the construction of tissue engineering fat filler and the survival of fat particles.Methods Stromal vascular fraction (SVF) was derived from inguinal subcutaneous fat of rabbits,mixed with autologus mature fat particles and with or without neuropeptide Y to construct PLGA composite tissue.The complex was subcutaneously transplantef into back sites of the rabbits.Based on the different combination five groups were divided:Group A:PLGA + mature fat particles without NPY;Group B:PLGA + mature fat particles + NPY;Group C:PLGA + complex of SVF with mature fat particles without NPY;Group D:PLGA + complex of SVF with mature fat particles + NPY;Group E:Complex of SVF with mature fat particles + small ball with NPY.Diffenrence of virous constructive ways and fat particle survival was evaluated by general observation,histological staining,fluorescence tracing at two weeks,one month and three month after operation.Results Group D was superior to groups A,B,C and E in survival volume,graft texture and vascularization at one mouth and three mouths.The fat average srvival rate was 57.5±2.5％.Fat cell grew well,and precursor cells proliferated and differentiated actively.Conclusions High quality tissue engineering materials are successfully established with SVF-mature fat particle complex,PLGA and NPY,which could obviously improve fat particle transplantation.

Effect of adipose tissue processing procedures in culture result: a study preliminary.

Background: There are various methods of processing adipose tissue before culture, depending on the adipose tissue samples. The aim of this study is to compare several modifications of culturing and sub-culturing procedures of adipose tissue to fit the condition in our laboratory. Method: This is a descriptive study that was done in the Immunology and Endocrinology Integrated Laboratory, University of Indonesia, from October 2009 to April 2010. Three adipose tissue processing procedures, various amount of seeding and two subculture methods were compared in term of cell yield and time needed. In the first procedure, collagenase-1 digestion was done in 30minutes, cell seeding were 24,000 and 36,000 per flask; in the second procedure, collagenase-1 digestion was done in 60minutes, cell seeding were 24,000, 48,000, and 72,000 per flask; and in the third procedure, the adipose tissue remnants from the first procedure were again digested for another 45 minutes, cell seeding were 74,000, and 148,000 per flask. Difference in subculture methods were the presence or absence of washing step. Result: Procedure 1 yielded the lowest amount of cell, and after culture, the cells grew very slow, and was contaminated before harvest of primary culture. Procedure-2 and -3 succeeded to yield primary cultures. Some of the cultures were contaminated, so that further subculture was not applicable, and only one tissue processing procedure (procedure 2: 60 minute collagenase-1 digestion, without lysis buffer, cell seeding 48,000 and 72,000) could complete the three subcultures. Though some of the procedures could not be completed, final result could be concluded. Conclusion: In this preliminary study, 60 minute colagenase-1 digestion with intermittent shaking every 5 minutes and cell seeding around 50,000 or more, followed by subculture method without washing step gave the best result.