Adipose-derived Stem Cells Group Research Articles

Safety and therapeutic potential of allogeneic adipose-derived stem cell spray transplantation in ischemic cardiomyopathy: a phase I clinical trial

BackgroundIschemic cardiomyopathy, characterized by coronary artery atherosclerosis, impairs the myocardial tissue. Coronary artery bypass grafting (CABG) is commonly used to revascularize affected areas and improve patient survival rates; however, it can fail to enhance cardiac function. Impaired capillary blood flow may obstruct functional recovery, prompting interest in treatments, such as angiogenic factor administration. Adipose-derived stem cells (ADSCs), which are known for immune evasion, have shown the potential to construct capillary networks and improve myocardial function. This clinical trial aimed to evaluate the safety and efficacy of ADSC spray therapy combined with CABG.MethodsThis single-center, randomized, double-blind study involved patients with ischemic cardiomyopathy who were scheduled for CABG and who had a left ventricular ejection fraction ≤ 40%. The participants were randomized to receive CABG as well as ADSC spray therapy or placebo. The primary endpoints were safety, changes in late gadolinium-enhanced (LGE) magnetic resonance imaging (MRI) volumes, and feasibility. The secondary endpoints included left ventricular function, exercise tolerance, and heart failure symptoms.ResultsSeven patients were enrolled; of them, six were randomized to receive ADSC therapy (n = 3) or placebo (n = 3). The procedure was successfully completed with minimal adverse events. One patient in the ADSC group developed pleural effusion that was resolved with drainage. The LGE-MRI volumes decreased in the ADSC group but remained unchanged in the placebo group. Improvements in left ventricular function and exercise tolerance were noted in the ADSC group, with heart failure symptoms improving to New York Heart Association class I. In contrast, the placebo group showed no significant changes, with one patient experiencing worsening symptoms.ConclusionsADSC spray therapy combined with CABG demonstrated safety and efficacy at enhancing cardiac function. ADSC likely contributes to capillary network reconstruction, thereby augmenting the benefits of CABG. Future phase II and III trials are warranted to confirm its therapeutic efficacy and long-term outcomes. This novel approach represents a significant advancement in the treatment of ischemic cardiomyopathy and offers a viable strategy for improving myocardial function and patient prognosis.Trial registration This study was registered with the Japan Registry of Clinical Trials (jRCT2053190103) and ClinicalTrials.gov (NCT04695522)Graphical

Efficacy of adipose-derived stem cells in preventing peripheral nerve adhesion and promoting nerve regeneration: A laboratory investigation in a rat model

BackgroundNeurolysis alone or administration of anti-adhesion products after neurolysis is performed to treat peripheral nerve adhesion; however, the recovery of nerve function is poor. This study aimed to investigate the efficacy of adipose-derived stem cells (ADSCs) for peripheral nerve adhesion in a rat model. MethodsAs a nerve adhesion procedure, the neural bed was coagulated, and the epineurium of the sciatic nerve was sutured to the coagulated neural bed using nylon. Neurolysis was performed 6 weeks after the nerve adhesion procedure, and saline (control group) or ADSCs (ADSC group) were administered around the nerve where neurolysis was performed. Evaluations were performed 6 weeks after the administration. ResultsThe wet weight ratio of the tibialis anterior muscle and nerve conduction velocity, which are indicators of nerve regeneration, were significantly better, while tensile strength, which is an indicator of the severity of nerve adhesion, was significantly lower in the ADSC group than in the control group. In the nerve, the expression of interleukin-10 and transforming growth factor-β in the nerve was significantly higher and that of tumor necrosis factor-α was significantly lower in the ADSC group than in the control group. Furthermore, significantly fewer M1 macrophages and significantly more M2 macrophages were observed in the ADSC group than in the control group. In the perineural scar, significantly fewer perineural collagen fibers and significantly more vascularization were observed in the ADSC group than in the control group. ConclusionsADSCs prevented peripheral nerve adhesion by reducing perineural scarring and enhancing vascularization. Additionally, ADSCs promoted nerve regeneration by decreasing inflammatory cytokine levels and increasing anti-inflammatory cytokine levels, as ADSCs regulated macrophage polarization from M1 to M2 macrophages. These findings hold promise for using ADSCs to treat nerve adhesion.

The effect of adipose-derived stem cells (ADSC) treatment on kidney histopathological appearance on the Wistar rat models with grade five kidney trauma.

Kidney trauma is the most common urological trauma. Technological advances have made conservative management possible for almost all kidney trauma. However, grade five kidney trauma needs to be carefully examined dueto its various complications, especially late complications that often delayed in recognition thus forming irreversible morbidity, with the most common late complication is kidney damage due to ischemic and fibrotic process. This study aims to confirm the effect of Adipose-Derived Stem Cells (ADSC) onthe prevention of fibrosis in grade five kidney trauma using Wistar rat models, where the fibrosis process will be measured with histopathological examination which had features of glomerular sclerosis, tubular atrophy, and interstitial fibrosis in kidney tissue, then followed by histopathological scoring and total renal score. A total of 22 adult rats were divided into five groups: one healthy control group, two trauma groups without ADSC, and two others trauma groups with ADSC. Two different treatment times were set: two weeks and four weeks after treatment. The data were tested for normality (Shapiro-Wilk test), while differences between groups were assessed using one-way ANOVA or Kruskal-Wallis test if the distribution was not normal. For the result of total renal score, statistical analysis reveal a significant difference in the total renal score in the kidney trauma with ADSC group compared with kidney trauma without ADSC group in fourth week of observation (p=0.001). These findings highlighted ADSC capability toprevent fibrosis caused by grade five kidney trauma on the Wistar rat models, as proven by significantly reduced histopathological grading on fibrosis.

An ADSC-loaded dermal regeneration template promotes full-thickness wound healing

IntroductionFull-thickness wounds lead to delayed wound healing and scarring. Adipose-derived stem cell (ADSC) grafting promotes wound healing and minimizes scarring, but the low efficiency of grafting has been a challenge. We hypothesized that loading ADSCs onto a clinically widely used dermal regeneration template (DRT) would improve the efficacy of ADSC grafting and promote full-thickness wound healing. MethodsADSCs from human adipose tissue were isolated, expanded, and labeled with a cell tracker. Labeled ADSCs were loaded onto the DRT. The viability, the location of ADSCs on the DRT, and the abundance of ADSCs in the wound area were confirmed using CCK8 and fluorescence microscopy. Full-thickness wounds were created on Bama minipigs, which were applied with sham, ADSC, DRT, and ADSC-DRT. Wounds from the four groups were collected at the indicated time and histological analysis was performed. RNA-seq analysis was also conducted to identify transcriptional differences among the four groups. The identified genes by RNA-seq were verified by qPCR. Immunohistochemistry and western blotting were used to assess collagen deposition. In vitro, the supernatant of ADSCs was used to culture fibroblasts to investigate the effect of ADSCs on fibroblast transformation into myofibroblasts. ResultsADSCs were successfully isolated, marked, and loaded onto the DRT. The abundance of ADSCs in the wound area was significantly greater in the ADSC-DRT group than in the ADSC group. Moreover, the ADSC-DRT group exhibited better wound healing with improved re-epithelialization and denser collagen deposition than the other three groups. The RNA-seq results suggested that the application of the integrated ADSC-DRT system resulted in the differential expression of genes mainly associated with extracellular matrix remodeling. In vivo, wounds from the ADSC-DRT group exhibited an earlier increase in type III collagen deposition and alleviated scar formation. ADSCs inhibited the transformation of fibroblasts into myofibroblasts, along with increased levels of CTGF, FGF, and HGF in the supernatant of ADSCs. Wounds from the ADSC-DRT group had up-regulated expressions of CTGF, HGF, FGF, and MMP3. ConclusionThe integral of ADSC-DRT increased the efficacy of ADSC grafting, and promoted full-thickness wound healing with better extracellular matrix remodeling and alleviated scar formation.

One-step stromal vascular fraction therapy in osteoarthritis with tropoelastin-enhanced autologous stromal vascular fraction gel.

Background: Osteoarthritis (OA) is a debilitating degenerative joint disease, leading to significant pain and disability. Despite advancements, current regenerative therapies, such as mesenchymal stem cells (MSCs), face challenges in clinical efficacy and ethical considerations. This study aimed to evaluate the therapeutic potential of stromal vascular fraction gel (SVF-gel) in comparison to available treatments like hyaluronic acid (HA) and adipose-derived stem cells (ADSCs) and to assess the enhancement of this potential by incorporating tropoelastin (TE). Methods: We conducted a comparative laboratory study, establishing an indirect co-culture system using a Transwell assay to test the effects of HA, ADSCs, SVF-gel, and TE-SVF-gel on osteoarthritic articular chondrocytes (OACs). Chondrogenic and hypertrophic markers were assessed after a 72-hour co-culture. SVF-gel was harvested from rat subcutaneous abdominal adipose tissue, with its mechanical properties characterized. Cell viability was specifically analyzed for SVF-gel and TE-SVF-gel. The in vivo therapeutic effectiveness was further investigated in a rat model of OA, examining MSCs tracking, effects on cartilage matrix synthesis, osteophyte formation, and muscle weight changes. Results: Cell viability assays revealed that TE-SVF-gel maintained higher cell survival rates than SVF-gel. In comparison to the control, HA, and ADSCs groups, SVF-gel and TE-SVF-gel significantly upregulated the expression of chondrogenic markers COL 2, SOX-9, and ACAN and downregulated the hypertrophic marker COL 10 in OACs. The TE-SVF-gel showed further improved expression of chondrogenic markers and a greater decrease in COL 10 expression compared to SVF-gel alone. Notably, the TE-SVF-gel treated group in the in vivo OA model exhibited the most MSCs on the synovial surface, superior cartilage matrix synthesis, increased COL 2 expression, and better muscle weight recovery, despite the presence of fewer stem cells than other treatments. Discussion: The findings suggest that SVF-gel, particularly when combined with TE, provides a more effective regenerative treatment for OA by enhancing the therapeutic potential of MSCs. This combination could represent an innovative strategy that overcomes limitations of current therapies, offering a new avenue for patient treatment. Further research is warranted to explore the long-term benefits and potential clinical applications of this combined approach.

Neutrophil extracellular trap-related mechanisms in acne vulgaris inspire a novel treatment strategy with adipose-derived stem cells

Acne vulgaris is a type of chronic skin disorder caused by Propionibacterium acnes (P. acnes). Neutrophil extrinsic traps (NETs) play key role in many types of inflammatory skin diseases. Adipose-derived stem cells (ADSCs) was reported modulate immune responses and neutrophil activity. Here, we explored the potential role of ADSCs and the potential mechanism associated with neutrophil extracellular traps (NETs) in relieving acne vulgaris. In the P. acnes-infected ear skin model, histological staining was used to evaluate the inflammatory infiltration and NET formation in control, P. acnes, and P. acnes + ADSCs groups. Besides, western blot was used to detect the expression levels of cit-H3, MPO, and Nrf2 in ear tissue. In vitro, the immunofluorescence staining of MPO and cit-H3, and SYTOX green staining were performed to measure the NET formation. CCK-8 assay, EdU staining, and wound healing assay were used to detect the proliferation and migration abilities of keratinocytes. ELISA assay was utilized to detect the secretion of inflammatory cytokines. In P. acnes-infected ear skin, ADSC treatment significantly attenuated inflammation and NET formation via activating Nrf2 signaling pathway. In vitro, the conditioned medium of ADSCs reduced the formation of P. acne-induced NETs. Besides, ADSCs could inhibit that the NETs efficiently promoted the proliferation, migration, and inflammatory cytokine secretion of keratinocytes. Our study suggested that ADSCs could attenuate P. acne-related inflammation by inhibiting NET formation. This study provides a novel therapeutic perspective of ADSCs in combating acne vulgaris.

Three-dimensional bioengineered dermal derived matrix scaffold in combination with adipose-derived stem cells accelerate diabetic wound healing

Due to the multifactorial nature of diabetic wounds, the most effective treatments require combinatorial approach. Herein we investigated whether engraftment of a bioengineered three-dimensional dermal derived matrix scaffold (DDMS) in combination with adipose-derived stem cells (ADSs), could accelerate diabetic wound healing. Diabetic animals were randomly planned into the control group, DDMS group, ADS group, and DDMS+ADS group. On days 7, 14, and 21, tissue samples were obtained for stereological, molecular, and tensiometrical assessments. We found that the wound contraction rate, the total volumes of new epidermis and dermis, the numerical densities of fibroblasts and blood vessels, collagen density, and tensiometrical parameters were meaningfully greater in the treated groups than in the control group, and these changes were more obvious in the DDMS+ADS ones (p < 0.05). Moreover, the expression of TGF-β, bFGF, and VEGF genes were considerably upregulated in treated groups compared to the control group and were greater in the DDMS+ADS group (p < 0.05). This is while expression of TNF-α and IL-1β, as well as the numerical densities of neutrophils and macrophages decreased more considerably in the DDMS+ADS group than in the other groups (p < 0.05). Overall, it was found that using both DDMS engraftment and ADS transplantation has more impact on diabetic wound healing.

Therapeutic potential of ADSCs in diabetic wounds: a proteomics-based approach.

Diabetes mellitus (DM), a chronic metabolic disease characterized by elevated blood sugar, leads to delayed or non-healing wounds, increasing amputation risks, and placing a significant burden on patients and society. While extensive research has been conducted on adipose-derived stem cells (ADSCs) for promoting wound healing, there is a scarcity of studies focusing on diabetic wounds, particularly those employing proteomics and bioinformatics approaches. This study aimed to investigate the mechanisms by which ADSCs promote diabetic wound healing using proteomics and bioinformatics techniques. Healthy rat fat tissue was used to isolate ADSCs. A T2DM rat model with back wounds was established. The experimental group received ADSC injections around the wound, while the control group received PBS injections. Wound healing rates were documented and photographed on days 0, 3, 7, 10, and 14. On day 7, wound tissues were excised for HE and Masson's staining. Additionally, on day 7, tissues were analyzed for protein quantification using 4D-DIA, with subsequent GO and KEGG analyses for differentially expressed proteins (DEPs) and protein-protein interaction (PPI) network analysis using STRING database (String v11.5). Finally, Western blot experiments were performed on day 7 wounds to verify target proteins. In all measured days postoperatively, the wound healing rate was significantly higher in the ADSC group than in the PBS group (day 7: p < 0.001, day 10: p = 0.001, day 14: p < 0.01), except on day 3 (p > 0.05). Proteomic analysis identified 474 differentially expressed proteins, with 224 key proteins after PPI analysis (78 upregulated and 146 downregulated in the ADSC group). The main cellular locations of these proteins were "cellular anatomical entity" and "protein-containing complex", while the biological processes were "cellular processes" and "biological regulation". The primary molecular functions were "binding" and "catalytic activity", with GO enrichment focused on "Wnt-protein binding", "neural development", and "collagen-containing extracellular matrix". Further analysis of PPI network nodes using LASSO regression identified Thy1 and Wls proteins, significantly upregulated in the ADSC group, as potentially crucial targets for ADSC application in diabetic wound treatment.

Exosomal circRNA-0008302 from Adipose-derived Stem Cells Protects Against Myocardial Injury

Objective: This study was aimed at investigating the expression and therapeutic potential of circRNAs from adipose-derived stem cell (ADSC) exosomes (ADSC-Exos) for cardiomyocyte injury. Methods: We screened differentially expressed circRNAs between ADSCs and ADSC-Exos with a circRNA microarray. A differential circRNA-0008302 siRNA plasmid was constructed to obtain ADSC-Exo and ADSC Exommu-circ-0008302 siRNA, respectively; subsequently, M6200 cells were divided into a control group, hydrogen peroxide (H2O2) induced group, H2O2 + ADSC-Exo group, and H2O2 + ADSC-Exommu-circ-0008302 siRNA group, and cell viability was evaluated. Apoptosis and intracellular reactive oxygen species were measured. The expression levels of miR-466i-5p were evaluated, and western blotting was performed to detect the expression of methionine sulfoxide reductase A (MsrA) protein. Results: Expression of circ-0008302 was significantly more elevated in the ADSC-Exo group than the ADSCs group. The treatment protected cardiomyocytes against H2O2-induced oxidative injury. Mechanistically, circ-0008302 downregulated miR-466i-5p levels, thus promoting expression of the miR-466i-5p target gene MsrA in cardiomyocytes. Conclusions: ADSC-Exos play protective roles in mitigating myocardial injury by delivering circ-0008302 to cardiomyocytes; this circRNA targets miR-466i-5p and subsequently enhances the expression of MsrA.

Adipose-derived stem cells repair radiation-induced chronic lung injury via inhibiting TGF-β1/Smad 3 signaling pathway.

To investigate the effect of adipose-derived stem cells (ASCs) transplantation on radiation-induced lung injury (RILI), Sprague-Dawley rats were divided into phosphate-buffered saline (PBS) group, ASCs group, Radiation + PBS group, and Radiation + ASCs group. Radiation + PBS and Radiation + ASCs groups received single dose of 30 Gy X-ray radiation to the right chest. The Radiation + PBS group received 1 mL PBS suspension and Radiation + ASCs group received 1 mL PBS suspension containing 1 × 107 CM-Dil-labeled ASCs. The right lung tissue was collected on Days 30, 90, and 180 after radiation. Hematoxylin-eosin and Masson staining were performed to observe the pathological changes and collagen fiber content in the lung tissue. Immunohistochemistry (IHC) and western blot (WB) were used to detect levels of fibrotic markers collagen I (Collal), fibronectin (FN), as well as transforming growth factor-β1 (TGF-β1), p-Smad 3, and Smad 3. Compared with the non-radiation groups, the radiation groups showed lymphocyte infiltration on Day 30 after irradiation and thickened incomplete alveolar walls, collagen deposition, and fibroplasia on Days 90 and 180. ASCs relieved these changes on Day 180 (Masson staining, P = 0.0022). Compared with Radiation + PBS group, on Day 180 after irradiation, the Radiation + ASCs group showed that ASCs could significantly decrease the expressions of fibrosis markers Collal (IHC: P = 0.0022; WB: P = 0.0087) and FN (IHC: P = 0.0152; WB: P = 0.026) and inhibit the expressions of TGF-β1 (IHC: P = 0.026; WB: P = 0.0152) and p-Smad 3 (IHC: P = 0.0043; WB: P = 0.0087) in radiation-induced injured lung tissue. These indicated that ASCs could relieve RILI by inhibiting TGF-β1/Smad 3 signaling pathway.

Evaluating the Efficacy of Dedifferentiated Fat Cells (DFATs) vs Adipose-Derived Stem Cells (ASCs) in Enhancing the Viability of Fat Grafts.

Enhancing graft fat survival remains a paramount challenge in autologous fat transplantation surgeries. Dedifferentiated fat cells (DFATs) and adipose-derived stem cells (ASCs) represent 2 pivotal cells with potential to improve fat graft survival rates. In this study we aimed to compare the effectiveness of DFATs and ASCs in promoting fat graft survival, emphasizing their adipogenic and angiogenic capabilities. Both in vitro and in vivo experiments were conducted. In vitro assessments compared adipogenesis, angiogenesis, osteogenesis, chondrogenesis, cell migration abilities, and surface markers. For in vivo evaluation, a cell-assisted lipotransfer animal model was employed to gauge graft volume retention and histological morphology. Analysis techniques included hematoxylin and eosin staining, Western blotting, and real-time polymerase chain reaction. In vitro findings suggested a slight superiority of DFATs in adipogenesis and angiogenesis compared to ASCs. In vivo tests demonstrated both cell types surpassed the control in terms of graft volume retention, with the DFATs group marginally outperforming in retention rates and the ASC group presenting a slightly enhanced graft tissue structure. Our study underscores the distinct advantages of DFATs and ASCs in bolstering fat graft survival, offering potentially novel insights for plastic surgeons aiming to elevate fat graft survival rates.

Tumor Necrosis Factor-α-Induced Protein-8-like 2 Transfected Adipose-Derived Stem Cells Regulated the Dysfunction of Monocrotaline Pyrrole-Induced Pulmonary Arterial Smooth Muscle Cells and Pulmonary Arterial Endothelial Cells.

Pulmonary arterial hypertension (PAH) is characterized by pulmonary arterial endothelial cell (PAEC) dysfunction and pulmonary arterial smooth muscle cell (PASMC) activation. For decades, the therapies for PAH based on stem cells have been shown to be effective. Meanwhile, tumor necrosis factor-α-induced protein-8-like 2 (TIPE2) promote the viability of human amniotic mesenchymal stem cells. Therefore, we aimed to explore the role of TIPE2 in adipose-derived stem cells (ADSCs) and the function of TIPE2-transfected ADSCs in the regulation of PAH. We first explored the role and underlying molecular mechanism of TIPE2 in viability and migration of ADSCs. Moreover, the ADSCs transfected with TIPE2 were cocultured with monocrotaline pyrrole (MCTP)-stimulated PASMCs or PAECs. The effects and mechanisms of TIPE2-transfected ADSCs on MCTP-induced PASMCs and PAECs were further investigated. The results showed that TIPE2 overexpression promoted viability and migration of ADSCs by activating the TLR4-ERK1/2 pathway. In addition, TIPE2-transfected ADSCs inhibited the abnormal proliferation and the impaired apoptosis of PASMCs via NF-κB signaling and promoted the conversion of PASMCs from synthetic to contractile. Meanwhile, TIPE2-transfected ADSCs reduced the apoptosis, endothelial-to-mesenchymal transition, and migration of PAECs via PI3K/AKT signaling after MCTP treatment. MCTP-induced oxidative stress and inflammation of PAECs were significantly decreased by TIPE2-transfected ADSCs. In rat model, TIPE2-ADSCs administration further decreased the monocrotaline-induced increase in the right ventricular systolic pressure and ratio of right ventricle weight/left ventricle and septa weight (L + S) and right ventricle weight/body weight compared with the ADSCs group. In conclusion, TIPE2-transfected ADSCs dramatically attenuated the PAH via inhibiting the dysfunction of PASMCs and PAECs.

Comparative Regenerative Effects of Allogeneic Bone Marrow and Patellar Ligament Fat Pad Mesenchymal Stem Cells on Experimental Superficial Digital Flexor Tendonitis in New Zealand Rabbits

Background: Cell therapy is applied in tendonitis to speed the healing process of tendon tissue and restore its functional properties. Almost all types of stem cells can differentiate from the recipient cells after transplantation. Objectives: The main goal of this study is to compare the effects of two sources of mesenchymal stem cells on tendon regeneration. Methods: This study randomly divided 32 New Zealand rabbits into 4 groups. The bacterial collagenase was induced at the superficial digital flexor tendon (SDFT) of all rabbits, and the treatment was performed 48 hours after collagenase induction. Group 1 was treated with allogeneic bone marrow mesenchymal stem cells (BMMSCs). Group 2 was treated with adipose-derived stem cells (ADSCs) from the patellar ligament fat pad. Group 3 (sham group) was treated with 0.9% normal saline, and group 4 (control group) was left with no treatment. All rabbits were euthanized 2 and 4 weeks after surgery, and tendon samples were harvested. The histopathology was assessed by hematoxylin-eosin, Masson’s trichrome, and Vangieson’s dye, and tendon structure, fiber arrangement, cell nuclei, tissue inflammation, vascularity (angiogenesis), and density were surveyed. Results: The tendon healing process in the BMMSC and ADSC groups revealed better regeneration than the control and sham groups (P≤0.05). Significant changes (P≤0.05) in some microscopic parameters were seen by comparing the BMMSC and ADSC groups. Conclusion: According to the present study, the injection of mesenchymal stem cells (BMMSCs or ADSCs) showed beneficial results in tendon tissue healing. Furthermore, ADSCs showed better regeneration of the injured tendon tissue than BMMSCs.

Effect of Kartogenin combined with adipose-derived stem cells on tendon-bone healing after anterior cruciate ligament reconstruction

To investigate the effect of Kartogenin (KGN) combined with adipose-derived stem cells (ADSCs) on tendon-bone healing after anterior cruciate ligament (ACL) reconstruction in rabbits. After the primary ADSCs were cultured by passaging, the 3rd generation cells were cultured with 10 μmol/L KGN solution for 72 hours. The supernatant of KGN-ADSCs was harvested and mixed with fibrin glue at a ratio of 1∶1; the 3rd generation ADSCs were mixed with fibrin glue as a control. Eighty adult New Zealand white rabbits were taken and randomly divided into 4 groups: saline group (group A), ADSCs group (group B), KGN-ADSCs group (group C), and sham-operated group (group D). After the ACL reconstruction model was prepared in groups A-C, the saline, the mixture of ADSCs and fibrin glue, and the mixture of supernatant of KGN-ADSCs and fibrin glue were injected into the tendon-bone interface and tendon gap, respectively. ACL was only exposed without other treatment in group D. The general conditions of the animals were observed after operation. At 6 and 12 weeks, the tendon-bone interface tissues and ACL specimens were taken and the tendon-bone healing was observed by HE staining, c-Jun N-terminal kinase (JNK) immunohistochemical staining, and TUNEL apoptosis assay. The fibroblasts were counted, and the positive expression rate of JNK protein and apoptosis index (AI) were measured. At the same time point, the tensile strength test was performed to measure the maximum load and the maximum tensile distance to observe the biomechanical properties. Twenty-eight rabbits were excluded from the study due to incision infection or death, and finally 12, 12, 12, and 16 rabbits in groups A-D were included in the study, respectively. After operation, the tendon-bone interface of groups A and B healed poorly, while group C healed well. At 6 and 12 weeks, the number of fibroblasts and positive expression rate of JNK protein in group C were significantly higher than those of groups A, B, and D (P<0.05). Compared with 6 weeks, the number of fibroblasts gradually decreased and the positive expression rate of JNK protein and AI decreased in group C at 12 weeks after operation, with significant differences (P<0.05). Biomechanical tests showed that the maximum loads at 6 and 12 weeks after operation in group C were higher than in groups A and B, but lower than those in group D, while the maximum tensile distance results were opposite, but the differences between groups were significant (P<0.05). After ACL reconstruction, local injection of a mixture of KGN-ADSCs and fibrin glue can promote the tendon-bone healing and enhance the mechanical strength and tensile resistance of the tendon-bone interface.

Adipose-Derived Stem Cells Enhance Graft Incorporation and Mineralization in a Murine Model of Irradiated Mandibular Nonvascularized Bone Grafting.

Nonvascularized bone grafting represents a practical method of mandibular reconstruction. However, the destructive effects of radiotherapy on native bone preclude the use of nonvascularized bone grafts in head and neck cancer patients. Adipose-derived stem cells have been shown to enhance bone healing and regeneration in numerous experimental models. The purpose of this study was to determine the impact of adipose-derived stem cells on nonvascularized bone graft incorporation in a murine model of irradiated mandibular reconstruction. Thirty isogenic rats were randomly divided into 3 groups: nonvascularized bone graft (control), radiation with nonvascularized bone graft (XRT), and radiation with nonvascularized bone graft and adipose-derived stem cells (ASC). Excluding the control group, all rats received a human-equivalent dose of radiation. All groups underwent mandibular reconstruction of a critical-sized defect with a nonvascularized bone graft from the contralateral hemimandible. After a 60-day recovery period, graft incorporation and bone mineralization were compared between groups. Compared with the control group, the XRT group demonstrated significantly decreased graft incorporation (P = 0.011), bone mineral density (P = 0.005), and bone volume fraction (P = 0.001). Compared with the XRT group, the ASC group achieved a significantly increased graft incorporation (P = 0.006), bone mineral density (P = 0.005), and bone volume fraction (P = 0.013). No significant differences were identified between the control and ASC groups. Adipose-derived stem cells enhance nonvascularized bone graft incorporation in the setting of human-equivalent radiation.

Adipose delivered stem cells protect liver after ischemia-reperfusion injury by controlling autophagy

Objective: Ischemia-reperfusion(I/R) injury is an unavoidable side effect of liver surgery and transplantation. A potentially useful tool for cellular therapy and tissue engineering is adipose-derived stem cells (ADSCs).The process of autophagy is used by the cell to break down inappropriate molecules.The study's goal was to examine the impact of ADSCs on the autophagic pathway after rat hepatic ischemia-reperfusion injury. Material and Methods: Thirty male rats used in our study were divided into control, ADSC, ischemia, I/R, and I/R+ ADSC groups (n = 6). Liver tissues were stained with hematoxylin-eosin and histological changes were evaluated with Suzuki scoring. Immunoexpressions of transforming growth factor (TGF-β) and autophagy markers LC3B, p62 were analyzed using the immunohistochemical method. Results: As a result of histological evaluation the ischemia and I/R groups displayed sinusoid congestion, vacuolization, and necrosis in liver tissues. We showed that the immunostaining of LC3B and TGF- β were elevated, and p62 decreased in the rat liver from ischemia and I/R groups when compared to the control group. Conclusion: ADSCs reduced the excessive level of autophagy and structural damage to hepatocytes and the pathological alterations in the liver after ıschemia-reperfusion injury.

Role of Adipose Derived Stem Cells in Patients with Diabetic Foot Ulcers: Systematic Review and Meta-Analysis of Randomised Controlled Trials.

Few studies to date have investigated the role of adipose derived stem cells (ADSCs) in patients with diabetic foot ulcers (DFU). We aimed to conduct a systematic search of the literature to explore the available evidence behind ADSCs application in patients with DFU to establish if it has any added benefit regarding healing rate and healing time in this cohort of patients. The PubMed and Embase databases were searched for eligible studies. Only randomised controlled trials which investigated the impact of ADSCs alone on the healing of DFU were considered eligible and were included for the review. Reported healing rates, time to healing and procedure related complications were collected and analysed. The initial search resulted in 160 papers. Following duplicate removal, 131 papers were screened for eligibility. Only four trials met the study criteria and were included for the final review and analysis. 97 out of 189 patients who were included in the four studies received ADSCs for treatment of DFU whereas the remaining 92 patients received standard measures (control). The median participant age was 62, predominantly male (72.5%). Complete healing was achieved in 83.5% (n = 81) of patients in the ADSC group compared to 52% (n = 48) for patients in the control group at 12 months (OR = 4.8, 95%CI = 2.25 to 10.24, P < 0.0001). Mean healing time in the ADSC group ranged from 31 to 85 days whereas mean healing time in the control group ranged from 42 to 85 days (Pooled weighted mean difference = -10.832856, 95%CI = -22.44 to 0.77, P = 0.0673). No significant procedure related complications were reported in either group. The use of ADSCs in patients with DFU appears to demonstrate improved healing rates. The procedure of ADSC harvest and administration appears to be safe based on the initial reports. Large, randomised trials are needed to establish its role in patients with diabetic foot wounds.

Accelerated wound healing using three-dimensional amniotic membrane scaffold in combination with adipose-derived stem cells in a diabetic rat model

The most important factors in the non-optimal healing of diabetic wounds are the lack of a suitable scaffold in the wound site for the migration and replacement of cells, as well as the lack of blood supply and effective growth factors in the wound site. Herein we investigated whether a bioengineered micro-porous three-dimensional decellularized amniotic membrane-scaffold (DAMS) in combination with adipose-derived stem cells (ASCs) could promote healing in ischemic wounds in diabetic type 1 rat. The diabetic animals were randomly divided into non-treated (untreated group), engraftment by DAMS (DAMS group), transplanted by ASCs (ASC group), and DAMS in combination with ASCs (DAMS+ASC group). Stereological, immunohistochemical, molecular, and tensiometrical assessments were performed on post-surgical days 7, 14, and 21. We found that the rate of wound closure, the volumes of new epidermis and dermis, the numerical density of fibroblasts and blood vessels, the numbers of proliferating cells and collagen deposition as well as biomechanical properties of the healed wounds were significantly higher in the treatment groups in comparison to the untreated group, and were the highest in DAMS+ASC ones. The transcripts for TGF-β and VEGF genes were significantly upregulated in all treatment regimens compared to the untreated group and were the highest for DAMS+ASC group. This is while expression of TNF-α and IL-1β as well as cell density of neutrophils decreased more significantly in DAMS+ASC group as compared with other groups. Overall, it was found that using both DAMS engraftment and ASC transplantation has more impact on diabetic wound healing.

Adipose-Derived Stem Cells and Tacrolimus Improve Nerve Regeneration in a Rat Sciatic Nerve Defect Model.

This study compared the effect of undifferentiated adipose-derived stem cells (ADSCs) vs tacrolimus (FK506) in peripheral nerve regeneration in a rat sciatic nerve complete transection model. Forty Wistar rats were equally distributed in four groups. In the SHAM surgery group, the sciatic nerve was exposed and no further intervention was done. In the conduit-alone group (the SLN group), a 10-mm nerve gap was created and bridged with a fibrin conduit filled in with normal saline. In the FK506 group, the fibrin conduit was injected with soluble FK506. In the ADSC group, the conduit was impregnated with undifferentiated ADSCs. Nerve regeneration was assessed by means of walking track analysis, electromyography, and neurohistomorphometry. Clinically and microscopically, nerve regeneration was achieved in all groups at 12 weeks. Walking track analysis confirmed functional recovery in the FK506 and ADSC groups, but there was no difference between them. Recovery in function was also achieved in the SLN group, but it was inferior (P<.05). Electromyography demonstrated superior nerve regeneration in the FK506 and ADSC groups compared with the SLN group (P<.05), with no difference between the FK506 and ADSC groups. Similarly, histology showed no difference between the FK506 and ADSC groups, although both outperformed the SLN group (P<.05). No complications were observed. Successful peripheral nerve regeneration can be accomplished after a 10-mm nerve defect treated with nerve conduits. Superior nerve regeneration may be expected when the conduits are loaded with undifferentiated ADSCs or FK506, with similar outcomes for ADSCs and FK506. [Orthopedics. 2023;46(6):e353-e361.].

MP27-10 CRISPRA-ENGINEERED ADIPOSE-DERIVED STEM CELLS IMPROVE ERECTILE DYSFUNCTION IN DIABETIC RATS

You have accessJournal of UrologyCME1 Apr 2023MP27-10 CRISPRA-ENGINEERED ADIPOSE-DERIVED STEM CELLS IMPROVE ERECTILE DYSFUNCTION IN DIABETIC RATS Wenchao Xu, Hao Li, Taotao Sun, Jiaxin Wang, Tao Wang, and Jihong Liu Wenchao XuWenchao Xu More articles by this author , Hao LiHao Li More articles by this author , Taotao SunTaotao Sun More articles by this author , Jiaxin WangJiaxin Wang More articles by this author , Tao WangTao Wang More articles by this author , and Jihong LiuJihong Liu More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003255.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Due to the high incidence of diabetes mellitus (DM) and poor response to phosphodiesterase type 5 inhibitor in DM-induced erectile dysfunction (DMED) patients, new therapeutic strategies for DMED are needed. Adipose-derived stem cells (ADSCs) transplantation is considered a promising treatment for DMED but is limited by poor survival and efficacy after transplantation. This study explored the therapeutic effect of CRISPRa-engineered ADSCs overexpressing GPX4 (GPX4-ADSCs) on erectile function in diabetic rats. METHODS: Thirty-two SD rats were randomly divided into four groups: the control, DMED, ADSCs, GPX4-ADSCs groups. Among them, the control group rats were non-diabetic rats, and the latter four groups were all diabetic ED rats of the same age. CRISPRa system was used to overexpress GPX4 in ADSCs. The GPX4-ADSCs group was treated with intracavernous injections of GPX4-ADSCs, while the ADSCs group was treated with ADSCs. Four weeks after transplantation, intracavernous pressure (ICP) and mean arterial pressure (MAP) were measured by electrical stimulation of cavernous nerve, and penile tissue was collected for subsequent testing. RESULTS: ADSCs and GPX4-ADSCs transplantation did not affect the weight or the glucose levels of diabetic rats. Although both the ADSCs group and the GPX4-ADSCs group improved erectile function in diabetic rats, the GPX4-ADSCs group had better improvement than the ADSCs group. In the DMED group, oxidative levels in the corpus cavernosum increased, PI3K/AKT/eNOS signal expression was down-regulated, Caspase3 activity was enhanced, and TGFβ1-Smad2/3-Collagen IV pathway was upregulated, showing obvious oxidative stress, apoptosis and fibrosis. ADSCs and GPX4-ADSCs can reverse these changes caused by diabetes to some extent, and GPX4-ADSCs are more effective than ADSCs. CONCLUSIONS: The CRISPRa-engineered ADSCs overexpressing GPX4 had stronger efficacy in improving erectile function than ADSCs, and the mechanism may involve reducing oxidative stress, apoptosis, and fibrosis, which provides a new strategy for the treatment of DMED. Source of Funding: This research was funded by grants from the National Natural Science Foundation of China (No. 82001536 and No.81873831) and the Fundamental Research Funds for the Central Universities (HUST: YCJJ202201021) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e366 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Wenchao Xu More articles by this author Hao Li More articles by this author Taotao Sun More articles by this author Jiaxin Wang More articles by this author Tao Wang More articles by this author Jihong Liu More articles by this author Expand All Advertisement PDF downloadLoading ...