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@#Abstract: Methods - （ ） To investigate the repairing effect of adipose derived stem cells ADSCs transplantation in - - Methods 1 bromopropane induced peripheral nerve injury in rats. A total of 45 specific pathogen free male SD rats were （ ） （ ） randomly assigned to the control group 15 rats and exposure group 30 rats . The rats in the exposure group were gavaged with - ， ， 1 bromopropane at a dose of 800 mg/kg body weight while the control group were given with equal volume of corn oil once a ， - day five days per week for eight weeks. The rat model of peripheral nerve injury induced by 1 bromopropane was established - ， - and was randomly assigned to the self recovery group and ADSCs group 12 rats in each group. ADSCs group and self recovery （ 6 ） ， group were injected with 0.5 mL contains 1×10 cells ADSCs or 0.5 mL phosphate buffer solution respectively by tail vein once a week for four weeks. The control group was not administered any treatment. The rats in each group were assessed using - - （ ） ， ， inclined plane test and Basso Beattie Bresnahan BBB score on the first day before treatment as well as 7 14 21 and 28 day ， after treatment. At the end of treatment the sciatic nerve was isolated for histopathological examination. The oxidative stress - indexes in the cerebral motor cortex were detected by colorimetric analysis. The levels of brain derived neurotrophic factor （ ） （ ） - Results BDNF and nerve growth factor NGF in the sciatic nerve were detected by enzyme linked immunosorbent assay. The - maximum tilt angle of the inclined plate and the BBB score were lower in self recovery group at the five time points of treatment （ P< ） all 0.05 compared with the control group. The maximum tilt angle of the inclined plate test was higher in ADSCs group at 21 （ P< ）， ， ， （ P< ）， and 28 days of treatment all 0.05 and the BBB score was higher at 7 14 21 and 28 days of treatment all 0.05 - （ ） when compared with the self recovery group. The level of malondialdehyde MDA in the cerebral motor cortex increased （P< ）， （ ） （ ） （ P< ）， 0.05 while the superoxide dismutase SOD activity and glutathione GSH level decreased all 0.05 and the levels （ P< ） - of BDNF and NGF in the sciatic nerve decreased all 0.05 in self recovery group compared with the control group. There was ， ， no significant difference in SOD activity and MDA GSH BDNF and NGF levels between ADSCs group and control group （ P> ） （P< ）， （ all 0.05 . The level of MDA in cerebral motor cortex decreased 0.05 the SOD activity and GSH level increased all P< ）， （P< ） - 0.05 and the level of BDNF in sciatic nerve increased 0.05 in ADSCs group compared with the self recovery group. Conclusion - - ADSCs transplantation can repair peripheral nerve injury induced by 1 bromopropane via anti oxidative stress and regulating the secretion of neurotrophic factors.
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Objective:To prepare human decellular adipose tissue matrix (DAT) as injectable homogenate by a specially developed high-speed soft tissue homogenizer with controllable temperature, and to investigate its cellular compatibility and filling effect.Methods:From March 2019 to March 2021, DAT was obtained in the 940th Hospital from normal adipose tissue after decellular treatment. The DAT was mixed with normal saline at the rate of 1: 12. The specially developed high-speed soft tissue homogenizer with controllable temperature was used to conduct homogenization at 803×g for 10 min. The produced DAT homogenizer could pass through the 27G needle smoothly. DAT homogenate was observed under scanning electron microscope and its cytocompatibility was detected. Finally, granular fat, DAT homogenate and DAT homogenate + ADSCs were injected into the back of rats respectively, and the filling effect, angiogenesis ability and inflammatory infiltration were compared.Results:After decellular treatment, adipose tissue changed into DAT without cellular residue. The particle size of DAT homogenate was about (749.91±136.79) nm, which was prepared by the specially developed temperature controlled high-speed soft tissue homogenater. The adhesion rate was (92.16±1.00) %, and the A value increased with time. The cell growth was good, and the homogenate had no toxicity to the cells. In vivo experiment, the filling effect of DAT homogenate and DAT homogenate + ADSCs was significantly better than that of granulated fat group ( P<0.01). In the granulated fat group, a large number of adipocytes were necrotic and fused to form oil sacs, while DAT homogenate, DAT homogenate + ADSCs showed no obvious degradation, and some adipocytes were generated. The results of CD31 staining indicated that the number of microvessels in DAT homogenate group and DAT homogenate + ADSCs group was higher than that in granulated fat group ( P<0.01). The results of CD68 staining indicated that the inflammatory infiltration in DAT homogenate group and DAT homogenate + ADSCs group was lower than that in granule fat group ( P<0.01). Conclusions:Using the self-developed temperature controlled high-speed soft tissue homogenate device, DAT could be prepared into DAT homogenate that can pass through 27G needle. It has good biocompatibility and filling advantages, and injective process is simple, and the trauma is small, and so it could be used as filling material.
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BACKGROUND: Adipose stem cell-free liquid extracts include adipose stem cell conditioned medium, adipose stem cell exosomes, because they do not contain cells and have the advantages of being easy to carry, store, and transport, which has gradually become one of the most promising therapies currently. OBJECTIVE: To review the research progress in the therapeutic application of adipose stem cell-free liquid extracts. METHODS: The first author searched the CNKI, Wanfang and PubMed databases for relevant articles published from January 2005 to March 2020. The key words were “adipose stem cell, stromal cell and conditioned medium”, “adipose stem cell, stromal cell and exosomes” in Chinese, and “adipose stem cell, adipose stromal cell and conditioned medium” and “adipose stem cell, adipose stromal cell and exosomes” in English. Repetitive articles and those lacking of originality were eliminated. Totally 123 articles were searched initially, and 62 articles were included in result analysis. RESULTS AND CONCLUSION: Adipose stem cell-free liquid extracts have broad therapeutic potential in a variety of disease areas, such as anti-aging, wound healing, scar recovery, and nerve regeneration. However, the specific mechanism of action is not very clear, and further research is needed.
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RESUMEN: En la enfermedad hepática crónica el trasplante ortotópico es la única alternativa terapéutica actual pero es limitada por falta de donantes. Ensayos con células madre adultas en daño hepático agudo evidencian promisorios resultados. El objetivo de este trabajo fue evaluar en ratas con daño hepático crónico la efectividad de la infusión de células madre adiposas humanas (CMAd-h). Ratas con fibrosis hepática inducida por tioacetamida fueron agrupadas en: grupo I control que no recibió tioacetamida ni células madre, grupo II recibió tioacetamida y suero fisiológico i.v., grupo III recibió tioacetamida y células madre adiposas 1 x 106/kg i.v. vía vena de la cola. La regeneración hepática histológica se evaluó por el index METAVIR, mientras las Macrophagocytus stellatus, células estrelladas a- SMA+ y células colágeno I+ por inmunohistoquímica; el daño funcional se evaluó por los niveles sanguíneos de los analitos Aspartato Aminotransferasa (AST), Alanina Aminotransferasa (ALT), Fosfatasa Alcalina (ALP), úrea y nitrógeno ureico (BUN) y hemograma. Los resultados muestran atenuación del daño estructural hepático evidenciado por disminución de los nódulos, del grado de lesión histológica en el score Metavir, y disminución de Macrophagocytus stellatus, células a-SMA+ y células colágeno tipo I+; funcionalmente hay reducción moderada de AST, ALT, urea, BUN y disminución moderada de células blancas pero efecto favorable sobre el volumen corpuscular media y la hemoglobina corpuscular media. Ocho semanas después de la infusión hay escasa población de CMAd-h en el hígado. En conclusión la infusión intravenosa de CMAd-h en ratas disminuye el daño funcional y estructural de la fibrosis hepática con escasa persistencia de CMAd-h en el parénquima hepático. A nuestro conocimiento este es el primer trabajo que evalúa el efecto de las CMAd-h en el modelo daño hepático crónico murino y la persistencia de las células trasplantadas.
SUMMARY: In chronic liver disease, orthotopic transplantation is the only current therapeutic alternative but it is limited due to lack of donors. Trials with adult stem cells in acute liver damage show promising results. The aim of this work was to evaluate the effectiveness of human adipose stem cell (h-ASC) infusion in rats with chronic liver damage. Rats with thioacetamide- induced liver fibrosis were grouped into: group I control that did not receive thioacetamide and h-ASC, group II received thioacetamide and saline i.v., group III received thioacetamide and h-ASC 1 x 106/ kg i.v. via tail vein. Histological liver regeneration was evaluated by METAVIR index, while Macrophagocytus stellatus (Kupffer cells), stellate cells a-SMA+ and collagen I+ cells by immunohistochemistry; functional damage was evaluated by blood levels of the analytes Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), Alkaline Phosphatase (ALP), Urea and Blood Urea Nitrogen (BUN) and hemogram. The results show attenuation of structural liver damage evidenced by decreased nodules, degree of histologic injury on Metavir score, and decreased Macrophagocytus stellatus, a-SMA+ cells and type I+ collagen cells; functionally there is moderate reduction of AST, ALT, urea, BUN and moderate decrease of white cells but favorable effect on mean corpuscular volume and mean corpuscular hemoglobin. Eight weeks after infusion there is a small population of h-ASC in the liver. In conclusion, intravenous infusion of h-ASC in rats reduces functional and structural damage of hepatic fibrosis with low persistence of h- ASC in the liver parenchyma. To our knowledge this is the first work that evaluates the effect of h-SC in the model of chronic murine liver damage and the persistence of transplanted cells.
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Animals , Female , Rats , Mesenchymal Stem Cell Transplantation/methods , Liver Cirrhosis, Experimental/therapy , Aspartate Aminotransferases/analysis , Immunohistochemistry , Treatment Outcome , Alanine Transaminase/analysis , Disease Models, Animal , Alkaline Phosphatase/analysis , Cell- and Tissue-Based Therapy/methods , Liver Cirrhosis, Experimental/pathologyABSTRACT
BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets.OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.
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BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets. OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.
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Objective: To prepare graphene oxide(GO)/polycaprolactone(PCL)composite matrix nanomaterials with different concentrations, and investigate effects of the nanomaterials on the myocardial differentiation of rat brown adipose stem cells (BASC)in vitro. Methods: The GO/PCL composite nanofiber materials were prepared by electrospinning technology. The biocompatibility of the nanomaterials was tested by the CCK-8 assay after cultivation of the BASC on the pure PCL and GO/PCL composite nanofibers, and the characterization of the nanofiber materials was performed by the scanning electron microscopy(SEM)and electrical conductivity measurement. The expression of cardiomyocyte-related proteins was examined by the cytological method and immunofluorescence staining. The GO/PCL composite nanofiber materials were systematically evaluated for their effect on the viability, proliferation and myocardial differentiation of BASC. Results: The GO/PCL composite nanofiber materials showed no obvious cytotoxicity at the 0.1 mg/ml GO concentration. The statistical results for the protein fluorescence intensities showed that the expression of the cardiomyocyte specific protein, α-actinin, and the intercalated disc-related protein, connexin-43(CX-43), was significantly increased in the GO/PCL group than in the PCL group(P<0.05). The cytoskeletal staining results showed that, compared with the PCL group, the cells in the GO/PCL group showed a long spindle-like stretch similar to the natural myocardial cell bundle, and the growth direction had a certain polarity. Conclusion: This study successfully prepared GO/PCL composite nanomaterials, which could promote the differentiation of BASCs into cardiomyocytes and the expression of intercalated disc-related proteins.
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OBJECTIVE@#To investigate the formation of gap junctions between Schwann cells derived from differentiated adipose stem cells implanted in a rat model of dyskinesia induced by brain injury and its positive effect in promoting functional recovery of the rats.@*METHODS@#In a rat model of hemiplegia induced by motor cortex injury, adipose stem cells or Schwann cells differentiated from adipose stem cells, either with or without RNAi-mediated silencing of Cx43, were transplanted orthotopically in the lesion. The recovery of the motor function of the rats was observed and scored after the transplantation. Rat brain tissues were sampled to detect the expressions of nerve growth factor (NGF) using Western blotting and RT-PCR.@*RESULTS@#All the 3 cell transplantation therapies obviously improved the motor function scores of the rats as compared with the control rats. The expression of NGF in the brain tissue was significantly lower in the control group than in the cell transplantation groups. NGF expression in the brain tissues of rats receiving transplantation of Schwann cells with Cx43 gene silencing was lower than that in rats receiving Schwann cells without Cx43 silencing, and was similar with that in rats transplanted with adipose stem cells. The results of RT-PCR showed that NGF mRNA level in the control group was significantly lower than that in the other 3 groups. NGF mRNA expression was the highest in Schwann cell group without Cx43 silencing, followed by adipose stem cell group, and then by Schwann cell group with Cx43 silencing.@*CONCLUSIONS@#In the rat model of dyskinesia induced by brain injury, transplantations of adipose stem cells and adipose stem cells-derived Schwann cells both promote the functional recovery of brain damage, in which gap junction protein Cx43 plays an important role to promote functional gap junction formation possibly by enhancing NGF expression.
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Animals , Rats , Brain Injuries , Dyskinesias , Gap Junctions , Rats, Sprague-Dawley , Schwann Cells , Stem CellsABSTRACT
Objective Comparison of induction time on the proliferation of induced adipose-derived stem cells (ADSC) to differentiate into Schwann-like cells (iSC).Methods From March,2017 to October,2018,ADSCs were isolated from inguinal adipose tissue of healthy adult female SD rats.Flow cytometry was performed to detect ADSC positive markers CD29,CD90 and negative marker CD45.iSC induction medium was used to culture ADSC.S-100 and GFAP were detected by immunofluorescence staining to confirm that ADSC had differentiated into iSC.Morphological changes of cells were observed by inverted microscope on day 1st,4th,7th,10th,13rd,16th and 19th after induction.MTS assay was used to evaluate cell proliferation ability.Tunel staining was applied to assess cell apoptosis.Results Both S100 and GFAP were expressed in iSC.On day 7th,the cell proliferation rate was significantly slower than that before induction (A value was 0.330±0.020 vs.0.400±0.004,P＜0.05).It was negatively correlated with induction time.On day 19th,the proliferation rate of iSC was lower than 50％ of the proliferation rate before induction (A value was 0.016±0.003 vs.0.400±0.004,P＜0.05).Apoptosis of iSC was more obvious than ADSC at the same time point.Conclusion The proliferation ability of ADSC-induced iSC is optimal within 7 days after induction.
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Objective The aim of this research was to produce apparatus of urinary diversion using silk-fibroin loading rab-bit adipose stem cells,and assess the effect of urinary diversion in a rabbit model.Methods Adipose stem cells were obtained and cultured in vitro,and flow cytometry analysis was performed to determine the adipose stem cells.These cultured adipose stem cells were used to seed on the silk-fibroin scaffolds,and after being incubated in the conditioned medium for 7 days,the a-bove compounds were made into apparatus of urinary diversion.This apparatus of urinary diversion was implanted into 20 rab-bits with radical cystectomy to develop urinary diversion.Five rabbits from each experimental group were euthanized at the spe-cific time points(1,2,3,4 months postoperatively),and the implants were harvested for histological and immunohistochemical a-nalysis.In the control group,silk-fibroins with unseeded cells(only silk-protein scaffolds)were also made into apparatus of uri-nary diversion and then used as urinary diversion on another 5 rabbits with the same process.Results Rabbits adipose stem cells were isolated and cultured successfully,and determined by flow cytometry.The silk-fibroin scaffolds were synthesized suc-cessfully.All rabbits were alive in the experimental group until the time of sacrifice.Histological and immunohistochemical anal-ysis showed multilayer uroepithelium coverage in the luminal surface of apparatus of urinary diversion,and as time went on,epi-thelial layers increased continuously.In the control group,all animals were dead within 3 weeks,and urine leakage,severe in-flammatory reaction and tissue destruction were found by autopsy.Conclusion The present experiment has successfully used silk-fibroin loading rabbit adipose stem cells to construct apparatus of urinary diversion,and demonstrates the feasibility of this kind of apparatus for urinary diversion in a rabbit model,which provides some experimental basis for clinical applications.
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Objective@#To compare the in vitro osteogenic ability of brown adipose stem cells (BADSC) and white adipose stem cells (WADSC), and to provide evidence for further research and clinical application of adipose-derived stem cells.@*Methods@#The brown fat under the scapula of SD rats and the white adipose tissue in the groin were isolated and obtained BADSC and WADSC. The morphology of the cells was observed by an inverted phase contrast microscope, and the cell count was used to detect the proliferative ability. After osteogenic induction, alkaline phosphatase (ALP) staining and alizarin red staining were performed. The expression of the osteogenic marker gene [Runt-related transcription factor-2 (RUNX2), osteocalcin] was detected by quantitative real-time PCR (qPCR).@*Results@#Both BADSC and WADSC were osteogenic. The ALP activity of BADSC was significantly greater than that of WADSC at each time point after osteogenic induction. After 5 weeks of osteogenic induction, BADSC formed a larger area of calcium nodules (accumulated optical density was 92 558±1 507), which was significantly greater than WADSC (accumulated optical density was 52 319±1 786) (t=29.81, P<0.05). The expression of BADSC osteogenic marker genes (RUNX2 and osteocalcin) was significantly higher than that of WADSC (P<0.05).@*Conclusions@#Both BADSC and WADSC have the potential for osteogenic differentiation, but BADSC has better osteogenic differentiation ability than WADSC.
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Adipose tissue-derived stem cell (ASCs) are an attractive source of stem cells with therapeutic applicability in various fields for regenerating damaged tissues because of their stemness characteristics. However, little has reported on evaluating adverse responses caused by human ASC therapy. Therefore, in the present study, a clinical assessment after human ASC transplantation into dogs was undertaken. A total of 12 healthy male dogs were selected and divided into four groups: saline infusion, saline bolus, ASC infusion, and ASC bolus groups. Physical assessment and blood analysis were performed following ASC transplantation, and the concentrations of angiogenic factors, and pro- and anti-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA). There were no adverse vital sign responses among the dogs. Blood analyses revealed no remarkable complete blood count or serum chemistry results. ELISA results for angiogenic and anti-inflammatory factors including matrix metalloproteinase 9 (MMP9), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and interleukin-10 (IL-10) were significantly higher in the two ASCs groups than in the controls. In conclusion, this study demonstrated that transplantation of human ASCs produced no adverse effects and could be used safely in dogs. In addition, human ASCs could be involved in modulating secretions of angiogenic factors including MMP9, VEGF, bFGF, and HGF and anti-inflammatory factor IL-10.
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Animals , Dogs , Humans , Male , Angiogenesis Inducing Agents , Blood Cell Count , Chemistry , Cytokines , Enzyme-Linked Immunosorbent Assay , Fibroblast Growth Factor 2 , Hepatocyte Growth Factor , Interleukin-10 , Matrix Metalloproteinase 9 , Stem Cell Transplantation , Stem Cells , Transplantation , Vascular Endothelial Growth Factor A , Vital SignsABSTRACT
Adipose stem cells (ASCs) are a type of adult stem cells that share common characteristics with typical mesenchymal stem cells. In the last decade, ASCs have been shown to be a useful cell resource for tissue regeneration. The major role of regenerative medicine in this century is based on cell therapy in which ASCs hold a key position. Active research on this new type of adult stem cell has been ongoing and these cells now have several clinical applications, including fat grafting, overcoming wound healing difficulties, recovery from local tissue ischemia, and scar remodeling. The application of cultured cells will increase the efficiency of cell therapy. However, the use of cultured stem cells is strictly controlled by government regulation to ensure patient safety. Government regulation is a factor that can limit more versatile clinical application of ASCs. In this review, current clinical applications of ASCs in plastic surgery are introduced. Future stem cell applications in clinical field including culturing and banking of ASCs are also discussed in this review.
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Humans , Adipose Tissue/cytology , Cicatrix/prevention & control , Ischemia/therapy , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Surgery, Plastic , Tissue Engineering , Wound HealingABSTRACT
PURPOSE: Adipose-derived stromal cells (ASCs) are readily harvested from lipoaspirated tissue or subcutaneous adipose tissue fragments. The stromal vascular fraction (SVF) is a heterogeneous set of cell populations that surround and support adipose tissue, which includes the stromal cells, ASCs, that have the ability to differentiate into cells of several lineages and contains cells from the microvasculature. The mechanisms that drive the ASCs into the osteoblast lineage are still not clear, but the process has been more extensively studied in bone marrow stromal cells. The purpose of this study was to investigate the osteogenic capacity of adipose derived SVF cells and evaluate bone formation following implantation of SVF cells into the bone defect of human phalanx. METHODS: Case 1 a 43-year-old male was wounded while using a press machine. After first operation, segmental bone defects of the left 3rd and 4th middle phalanx occurred. At first we injected the SVF cells combined with demineralized bone matrix (DBM) to defected 4th middle phalangeal bone lesion. We used P (L/DL)LA [Poly (70L-lactide-co-30DL-lactide) Co Polymer P (L/DL)LA] as a scaffold. Next, we implanted the SVF cells combined with DBM to repair left 3rd middle phalangeal bone defect in sequence. Case 2 was a 25-year-old man with crushing hand injury. Three months after the previous surgery, we implanted the SVF cells combined with DBM to restore right 3rd middle phalangeal bone defect by syringe injection. Radiographic images were taken at follow-up hospital visits and evaluated radiographically by means of computerized analysis of digital images. RESULTS: The phalangeal bone defect was treated with autologous SVF cells isolated and applied in a single operative procedure in combination with DBM. The SVF cells were supported in place with mechanical fixation with a resorbable macroporous sheets acting as a soft tissue barrier. The radiographic appearance of the defect revealed a restoration to average bone density and stable position of pharyngeal bone. Densitometric evaluations for digital X-ray revealed improved bone densities in two cases with pharyngeal bone defects, that is, 65.2% for 4th finger of the case 1, 60.5% for 3rd finger of the case 1 and 60.1% for the case 2. CONCLUSION: This study demonstrated that adipose derived stromal vascular fraction cells have osteogenic potential in two clinical case studies. Thus, these reports show that cells from the SVF cells have potential in many areas of clinical cell therapy and regenerative medicine, albeit a lot of work is yet to be done.
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Adult , Humans , Male , Adipose Tissue , Bone Density , Bone Matrix , Durapatite , Fingers , Follow-Up Studies , Hand Injuries , Hypogonadism , Mesenchymal Stem Cells , Microvessels , Mitochondrial Diseases , Ophthalmoplegia , Osteoblasts , Osteogenesis , Polymers , Regenerative Medicine , Stromal Cells , Subcutaneous Fat , Surgical Procedures, Operative , Syringes , Cell- and Tissue-Based TherapyABSTRACT
Cartilage is one of the earliest reconstructed tissues used in tissue engineering. Due to the lack of appropriate seeding cells, cartilage tissue engineering is, however, relatively lagged behind. With the emergence of stem cell research, adipose stem cells(ASCs) are introduced as seeding cells into tissue engineering for possessing many advantages such as wide spreading, large amount of cells available and easy to obtain. However, the outcome of tissue engineered cartilage construction by ASCs is not as ideal as that by bone marrow stem cells (BMSCs) yet. Low efficiency of ASC chondrogenesis is considered the major cause. This review summarizes the purification of adipose-derived cells, maintenance of sternness and optimization of ehondrogenie induction, which play vital roles in improving ASC s chondrogenesis.
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@#Objective To explore the changes of rabbit adipose stem cells(ASCs)and bone mesenchymal stem cells(BMSCs)cultured in vitro and the anabolism under basic fibroblast growth factor(bFGF).Methods BMSCs and ASCs were cultured with DMEM,DMEM/F12(2∶1)or α-MEM respectively.The 3rd generation ASCs and BMSCs were divided into 2 groups respectively:group A:ASCs cultured in chondrogenic medium(CM),group B:ASCs cultured in CM supplemented with bFGF 5 ng/ml,group C:BMSCs cultured in CM,group D:BMSCs cultured in CM supplemented with bFGF 5 ng/ml.Morphological changes were observed under inverted microscope.The 35SO42-incorporation and total hydroxyproline were measured.Results BMSCs and ASCs showed much higher growth rate when cultured in α-MEM medium comparison with that in DMEM or in DMEM/F12(2:1).Both stem cells attachment cultured in monolayer greatly increased and cell clones were abundant,while the cells attachment became rather difficult and cell clones were less after cutured in CM.All stem cells possessed a round-like morphology,and the cells in group B and D were more than that in the other 2 groups.The 35SO42-incorporation and total hydroxyproline synthesis of group B or D increased compared with that of group A or C,but there was no diference between group D and B.Conclusion The rabbit ASCs and BMSCs cultured in CM suppling with bFGF grow well and their metabolism increased.
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PURPOSE: Adipose tissue contains a population of pluripotent stem cells capable of differentiating along multiple mesenchymal cell lineages. It is well known that fat depots from different part of our body shows different nature not only in morphological aspect but also physiologic aspect. The authors compared the adipogenic potentials and osteogenic potentials of adipose stem cells from different anatomical sites of human. METHODS: After laparotomy by surgery team, the authors isolated these adipose stem cells successfully from 7 men with an average age of 58, and induced differentiation along adipogenic and osteogenic lineages in vitro. On the 14th day, cells cultured in adipogenic media differentiated into adipocytes in vitro, as evidenced by positive Oil Red O staining of lipid vacuoles. On the 21st day, cells cultured in osteogenic media differentiated into osteoblasts in vitro as demonstrated by Alizarin red staining of a calcified extracellular matrix. RESULTS: After exposure to adipogenic and osteogenic differentiation medium, subcutaneous adipose stem cells were found to possess greater adipogenic and osteogenic potentials than cells isolated from visceral adipose tissues. CONCLUSION: This study indicates that adipogenic and osteogenic potentials of adipose stem cells vary by their anatomical sites, with subcutaneous adipose stem cells exhibiting higher adipogenic and osteogenic potential than those isolated from visceral fat.