Adipose-derived Stem Cells Group Research Articles

Autologous fat or adipose-derived stem cell grafting in systemic sclerosis treatment: a systematic review and meta-analysis.

Systemic sclerosis (SSc) is characterised by vasculopathy and progressive fibrosis of the skin. The aim of this article is to analyse and summarise the efficacy and safety of autologous fat (AF), stromal vascular fraction (SVF) and adipose-derived stem cell (ADSC) grafting in the treatment of SSc, providing evidence for clinical application. The research involves the efficacy and safety of AF, SVF and ADSC grafting in the treatment of patients with SSc. The studies were screened and selected independently by two authors based on pre-specified criteria. The data extraction and quality assessment were also performed independently by two authors. Fifteen studies were eligible for inclusion. Skin thickness reduced following SVF or AF therapy, but there was no significant difference. All measures used to assess fingertip symptoms revealed a significant improvement. Notably, SVF and AF were found to have the most impact on Raynaud's phenomenon improvement. The ADSC group improved the most in terms of finger pain alleviation. SVF reported the highest proportion of adverse events, accounting for approximately half of the cases. AF, SVF, and ADSC all displayed therapeutic effects of improving SSc, but differences existed in the effects on different symptoms. Plastic surgeons should choose the most suitable treatment strategy after comprehensively evaluating the patient's clinical manifestations.

Exercise combined with administration of adipose-derived stem cells ameliorates neuropathic pain after spinal cord injury.

Experimental studies have shown that exercise and human adipose-derived stem cells (ADSCs) play positive roles in spinal cord injury (SCI). However, whether ADSCs and/or exercise have a positive effect on SCI-induced neuropathic pain is still unclear. Thus, there is a need to explore the effects of exercise combined with administration of ADSCs on neuropathic pain after SCI. In this study, a thoracic 11 (T11) SCI contusion model was established in adult C57BL/6 mice. Exercise was initiated from 7 days post-injury and continued to 28 days post-injury, and approximately 1 × 105 ADSCs were transplanted into the T11 spinal cord lesion site immediately after SCI. Motor function and neuropathic pain-related behaviors were assessed weekly using the Basso Mouse Scale, von Frey filament test, Hargreaves method, and cold plate test. Histological studies (Eriochrome cyanine staining and immunohistochemistry) were performed at the end of the experiment (28 days post-injury). Exercise combined with administration of ADSCs partially improved early motor function (7, 14, and 21 days post-injury), mechanical allodynia, mechanical hypoalgesia, thermal hyperalgesia, and thermal hypoalgesia. Administration of ADSCs reduced white and gray matter loss at the lesion site. In addition, fewer microglia and astrocytes (as identified by expression of ionized calcium-binding adapter molecule 1 and glial fibrillary acidic protein, respectively) were present in the lumbar dorsal horn in the SCI + ADSCs and SCI + exercise + ADSCs groups compared with the sham group. Our findings suggest that exercise combined with administration of ADSCs is beneficial for the early recovery of motor function and could partially ameliorate SCI-induced neuropathic pain.

Comparison of the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells and adipose-derived stem cells on erectile dysfunction in a rat model of bilateral cavernous nerve injury.

Background: Cavernous nerve injury (CNI) is the leading cause of erectile dysfunction (ED) after radical prostatectomy and pelvic fracture. Transplantation of human adipose-derived stem cells (ASCs) has been widely used to restore erectile function in CNI-ED rats and patients. Umbilical cord blood-derived MSCs (CBMSCs) are similarly low immunogenic but much primitive compared to ASCs and more promising in large-scale commercial applications due to the extensive establishment of cord blood banks. However, whether CBMSCs and ASCs have differential therapeutic efficacy on CNI-ED and the underlying mechanisms are still not clear. Materials and methods: A bilateral cavernous nerve injury (BCNI) rat model was established by crushing the bilateral cavernous nerves. After crushing, ASCs and CBMSCs were intracavernously injected immediately. Erectile function, Masson staining, and immunofluorescence analyses of penile tissues were assessed at 4 and 12 weeks. PKH-26-labeled ASCs or CBMSCs were intracavernously injected to determine the presence and differentiation of ASCs or CBMSCs in the penis 3 days after injection. In vitro experiments including intracellular ROS detection, mitochondrial membrane potential assay, EdU cell proliferation staining, cell apoptosis assay, and protein chip assay were conducted to explore the underlying mechanism of CBMSC treatment compared with ASC treatment. Results: CBMSC injection significantly restored erectile function, rescued the loss of cavernous corporal smooth muscles, and increased the ratio of smooth muscle to collagen. PKH-26-labeled CBMSCs or ASCs did not colocalize with endothelial cells or smooth muscle cells in the corpus cavernosum. Moreover, the conditioned medium (CM) of CBMSCs could significantly inhibit the oxidative stress and elevate the mitochondria membrane potential and proliferation of Schwann cells. Better therapeutic effects were observed in the CBMSC group than the ASC group both in vivo and in vitro. In addition, the content of neurotrophic factors and matrix metalloproteinases in CBMSC-CM, especially NT4, VEGF, MMP1, and MMP3 was significantly higher than that of ASC-CM. Conclusion: Intracavernous injection of CBMSCs exhibited a better erectile function restoration than that of ASCs in CNI-ED rats owing to richer secretory factors, which can promote nerve regeneration and reduce extracellular matrix deposition. CBMSC transplantation would be a promising therapeutic strategy for CNI-ED regeneration in the future.

Therapeutic Efficacy of Adipose-Derived Stem Cells Versus Bone Marrow Stromal Cells for Irradiated Mandibular Fracture Repair.

Mesenchymal stem cells have immense potential in applications of bone healing and regeneration. However, few studies have evaluated the therapeutic efficacy of adipose-derived stem cells (ASCs) and bone marrow stromal cells (BMSCs) in irradiated bone. The purpose of this study is to compare the ability of ASCs versus BMSCs to enhance healing outcomes in a murine model of irradiated mandibular fracture repair. Forty-eight isogenic male Lewis rats underwent radiation therapy followed by mandibular osteotomy with intraoperative placement of either ASCs or BMSCs. Animals were killed on postoperative day 40. Mandibles were analyzed for union rate, biomechanical strength, vascularity, and mineralization. Groups were compared at P < 0.05 significance. The ASC and BMSC groups demonstrated 92% and 75% union rates. Compared with the BMSC group, the ASC group demonstrated a trending increase in maximum load ( P = 0.095) on biomechanical strength analysis and a significant increase in vessel number ( P = 0.001), vessel thickness ( P = 0.035), and vessel volume fraction ( P = 0.007) on micro-computed tomography angiography analysis. No significant differences in bone mineralization were identified on micro-computed tomography analysis. This study demonstrates the superior therapeutic efficacy of ASCs over BMSCs in irradiated fracture healing as evidenced by union rate, vascular morphometry, and a trend in biomechanical strength. We posit that the robust vascular response induced by ASCs better recapitulates the sequence and synchronicity of physiologic bone healing compared with BMSCs, thereby improving the reliability of irradiated fracture repair.

Regeneration Using Adipose-Derived Stem Cell Sheets in a Rabbit Meniscal Defect Model Improves Tensile Strength and Load Distribution Function of the Meniscus at 12 Weeks

The purpose of this study was to evaluate the mechanical properties, such as the tensile strength and load distribution function, of the meniscus tissue regenerated using adipose-derived stem cell (ADSC) sheets in a rabbit meniscal defect model. ADSC sheets were prepared from adipose tissue of rabbits. The anterior half of the medial meniscus was removed from both knees. One knee was transplanted with an ADSC sheet; the contralateral knee was closed without transplantation. Mechanical tests were performed at 4 and 12 weeks posttransplantation. In the tensile test, tensile force was applied to the entire medial meniscus, including the normal area (n= 10/group). Compression tests were performed on the entire knee, with soft tissues other than the ligament removed. A pressure-sensitive film was inserted under the medial meniscus and a 40-N load was applied (n= 5/group). In the tensile test, the elastic modulus in ADSC-treated knees was higher at 12 weeks (ADSC: 70.30 ± 18.50 MPa, control: 43.71 ± 7.11 MPa, P= .009). The ultimate tensile strength (UTS) in ADSC-treated knees at 12 weeks was also higher (ADSC: 22.69 ± 5.87 N, control: 15.45 ± 4.08 N, P= .038). In the compression test, the contact area was larger in the ADSC group at 4 weeks (ADSC: 31.60 ± 8.17 mm2, control: 20.33 ± 2.86 mm2, P= .024) and 12 weeks (ADSC: 41.07 ± 6.09 mm2, control: 30.53 ± 5.47 mm2, P= .04). Peak pressure was significantly lower in ADSC-treated knees at 12 weeks (ADSC: 11.91 ± 1.03 MPa, control: 15.53 ± 2.3 MPa, P= .002). The regenerated meniscus tissue, 12 weeks after transplantation of the ADSC sheets into the meniscal defect area, had high elastic modulus and UTS. In the meniscus-tibia compartment, the contact area was large and the peak pressure was low. ADSC sheets promoted regeneration of meniscus. ADSC sheet transplantation for meniscal defects could be an effective regenerative therapy.

Low-Intensity Pulsed Ultrasound Enhanced Adipose-Derived Stem Cell-Mediated Angiogenesis in the Treatment of Diabetic Erectile Dysfunction through the Piezo-ERK-VEGF Axis.

Objectives Erectile dysfunction is a major comorbidity of diabetes. Stem cell transplantation is a promising method to treat diabetic erectile dysfunction. In this study, we evaluated whether low-intensity pulsed ultrasound (LIPUS) could enhance the efficacy of adipose-derived stem cells (ADSCs) and investigated the underlying molecular mechanism. Materials and methods. Sixty 8-week-old male Sprague–Dawley rats were randomly divided into the normal control (NC) cohort or the streptozocin-induced diabetic ED cohort, which was further subdivided into DM, ADSC, LIPUS, and ADSC+LIPUS groups. Rats in the ADSC or ADSC+LIPUS group received ADSC intracavernosal injection. Rats in the LIPUS or ADSC+LIPUS group were treated with LIPUS. The intracavernous pressure (ICP) and mean arterial pressure (MAP) were recorded at Day 28 after injection. The corpus cavernosum tissues were harvested and subjected to histologic analysis and ELISA. The effects of LIPUS on proliferation and cytokine secretion capacity of ADSCs were assessed in vitro. RNA sequencing and bioinformatic analysis were applied to predict the mechanism involved, and western blotting and ELISA were used for verification. Results Rats in the ADSC+LIPUS group achieved significantly higher ICP and ICP/MAP ratios than those in the DM, ADSC, and LIPUS groups. In addition, the amount of cavernous endothelium and cGMP level also increased significantly in the ADSC+LIPUS group. In vitro experiments demonstrated that LIPUS promoted proliferation and cell cycle progression in ADSCs. The excretion of cytokines such as CXCL12, FGF2, and VEGF was also enhanced by LIPUS. Bioinformatic analysis based on RNA sequencing indicated that LIPUS stimulation might activate the MAPK pathway. We confirmed that LIPUS enhanced ADSC VEGF secretion through the Piezo-ERK pathway. Conclusion LIPUS enhanced the curative effects of ADSCs on diabetic erectile dysfunction through the activation of the Piezo-ERK-VEGF pathway. ADSC transplantation combined with LIPUS could be applied as a synergistic treatment for diabetic ED.

Drug-free in vitro activation combined with 3D-bioprinted adipose-derived stem cells restores ovarian function of rats with premature ovarian insufficiency

BackgroundEmerging drug-free in vitro activation (IVA) technique enables patients with premature ovarian insufficiency (POI) to restore ovarian function and conceive their own genetic offspring. However, various issues have greatly restricted its clinical application. Transplantation of adipose-derived stem cells (ADSCs) has promising roles in restoring ovarian function of rats with POI, but insufficient retention has greatly hampered their efficiency. Here, we designed a 3D-bioprinted engineering ovary composed of drug-free IVA and ADSCs, which may prolong the retention of ADSCs and construct an early vascular microenvironment, thus compensating for the disadvantages of drug-free IVA to some extent and ameliorating impaired ovarian function in the POI rats.MethodsAfter intraperitoneal injection of cyclophosphamide, the POI model rats were randomized into 5 groups: (1) POI group; (2) ovarian fragments group; (3) 3D scaffold combined with ovarian fragments group; (4) ovarian fragments combined with ADSCs group; (5) 3D scaffold with ADSCs combined with ovarian fragments as 3D-bioprinted engineering ovary group. Normal rats were identified as the control group. The localization of CM-Dil-labeled ADSCs and co-localization with CD31 were observed to examine the distribution and underlying mechanism of differentiation. Histomorphological and immunohistochemical analyses were performed to calculate follicle number and assess proliferation and apoptosis of granulosa cells (GCs). Immunofluorescence staining was used to evaluate angiogenesis. Hormone levels were measured to evaluate the restoration of endocrine axis. Western blot analysis and RT-PCR were conducted to explore the potential mechanism.ResultsCM-Dil-labeled ADSCs were distributed in the interstitium of ovaries and had significantly higher retention in the 3D-bioprinted engineering ovary group. Several regions of the co-staining for CM-Dil and CD31 were in the area of vascular endothelial cells. Meanwhile, the follicle counts, GCs proliferation, neoangiogenesis, and hormone levels were significantly improved in the 3D-bioprinted engineering ovary group, as compared with other groups. Furthermore, the ovarian function was ameliorated and angiogenesis was promoted through regulating the PI3K/AKT pathway.ConclusionOur results suggested that 3D-bioprinted engineering ovary had great potential for restoring impaired ovarian function of rats with POI, which could compensate for the disadvantages of drug-free IVA to some extent.

Effect of Adipose-Derived Stem Cell on Collagen Deposition in Nude Mouse Model of Scleroderma

Systemic sclerosis is an autoimmune disease featured by uncontrolled fibrosis and collagen deposition. This study aimed to investigate the impact of adipose-derived stem cells (ADSCs) transplantation on collagen in scleroderma. After establishment of scleroderma model using Bleomycin, the animals received subcutaneous injection of 0.3 ml fat (group B) or 0.3 ml fat+1×106 ADSCs (group C), or 1×106 ADSCs alone (group D). Mice treated with PBS were taken as control group (group A). One month after operation, the skin tissue at injection site was collected for H&amp;E staining, and immunohistochemistry to determine the content of TGF-β1 and type III collagen. Compared to control group, group B, C, and D exhibited improvement in collagen deposition and reduction in TGF-β1 content and type III collagen. Combined graft of fat and ADSCs exerted more significant effect compared to single ADSCs treatment. In conclusion, fat and ADSCs transplantation improves collagen deposition in nude mice with scleroderma and the combined treatment exerts a higher efficacy, suggesting that ADSCs need adipose carrier and microenvironment. These findings provide a novel insight into the treatment of scleroderma.

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

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

Recovery of erectile dysfunction after administration of hydrogen sulfide donor sodium hydrogen sulfide and transplantation of adipose-derived stem cells in rats with bilateral cavernous nerve injury

ABSTRACT Introduction Radical prostatectomy causes nerve injury-related erectile dysfunction (ED), which results in a poor response to phosphodiesterase 5 inhibitors. Hydrogen sulfide (H2S), an endogenous gasotransmitter, is produced by cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (3-MST) and participates in erectile physiology. Adipose-derived stem cells (ADSCs) represent a promising cell source for cell replacement therapy in several degenerative diseases because of its self-renewal properties and multi-differentiation potential with low immunogenicity. Objective The current study was to evaluate the effects of H2S donor, sodium hydrogen sulfide (NaHS), and transplantation of ADSCs per se and their combination treatment on bilateral cavernous nerve injury (BCNI)-induced erectile dysfunction (ED). Methods A total of 35 Sprague-Dawley rats were divided into 5 groups, including sham-operated (Sham), BCNI (4-wk), BCNI treated with NaHS (5.6 mg/kg/day, i.p.), BCNI treated with ADSCs (1x106 cells by intracavernosal injection) and BCNI treated with NaHS and ADSCs groups. Rats in the ADSCs treatment groups were subjected to intracavernosal injection after BCNI. Erectile function was evaluated by the ratio of intracavernous pressure (ICP)/mean arterial pressure (MAP) and total ICP. The relaxant and contractile responses of corpus cavernosum (CC) were measured from in vitro studies. Protein expression and localization of nitric oxide synthases (NOS) and H2S synthesis enzymes were analyzed by Western blotting and immunohistochemistry. The smooth muscle/collagen ratio was determined using Masson trichrome staining. Results Rats with BCNI displayed significantly reduced ICP/MAP and total ICP (p&amp;lt;0.001 vs controls) when were partially restored by NaHS and ADSCs treatments alone. Interestingly, combined treatment completely enhanced decreased in vivo erectile responses (p&amp;lt;0.001 vs the NC group). Relaxation responses to acetylcholine and electrical field stimulation (EFS) as well as contractile responses to phenylephrine and EFS in the CN group were less than in controls (p&amp;lt; 0.05), which was normalized after all treatments groups. The combined treatment modulated changes of protein expressions and reduced ratio of smooth muscle to collagen in rats with BCNI. Conclusions The current data indicated that the combination treatment with NaHS and ADSCs is more successful than monotherapies in reversing BCNI-induced ED. H2S and cell-based therapy most probably may have a synergistic effect on erectile function via controlling molecular regulation and morphological penile alterations, which supports combined treatment for expanding clinical outcomes radical prostatectomy-induced ED. Disclosure Work supported by industry: no.

Adipose-derived stem cells alleviate liver injury induced by type 1 diabetes mellitus by inhibiting mitochondrial stress and attenuating inflammation

BackgroundType 1 diabetes mellitus (T1D) is a worldwide health priority due to autoimmune destruction and is associated with an increased risk of multiorgan complications. Among these complications, effective interventions for liver injury, which can progress to liver fibrosis and hepatocellular carcinoma, are lacking. Although stem cell injection has a therapeutic effect on T1D, whether it can cure liver injury and the underlying mechanisms need further investigation.MethodsSprague–Dawley rats with streptozotocin (STZ)-induced T1D were treated with adipose-derived stem cell (ADSC) or PBS via the tail vein formed the ADSC group or STZ group. Body weights and blood glucose levels were examined weekly for 6 weeks. RNA-seq and PCR array were used to detect the difference in gene expression of the livers between groups.ResultsIn this study, we found that ADSCs injection alleviated hepatic oxidative stress and injury and improved liver function in rats with T1D; potential mechanisms included cytokine activity, energy metabolism and immune regulation were potentially involved, as determined by RNA-seq. Moreover, ADSC treatment altered the fibroblast growth factor 21 (FGF21) and transforming growth factor β (TGF-β) levels in T1D rat livers, implying its repair capacity. Disordered intracellular energy metabolism, which is closely related to mitochondrial stress and dysfunction, was inhibited by ADSC treatment. PCR array and ingenuity pathway analyses suggested that the ADSC-induced suppression of mitochondrial stress is related to decreased necroptosis and apoptosis. Moreover, mitochondria-related alterations caused liver inflammation, resulting in liver injury involving the T lymphocyte-mediated immune response.ConclusionsOverall, these results improve our understanding of the curative effect of ADSCs on T1D complications: ADSCs attenuate liver injury by inhibiting mitochondrial stress (apoptosis and dysfunctional energy metabolism) and alleviating inflammation (inflammasome expression and immune disorder). These results are important for early intervention in liver injury and for delaying the development of liver lesions in patients with T1D.

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

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

MiR-20a Overexpression in Adipose-Derived Mesenchymal Stem Cells Promotes Therapeutic Efficacy in Murine Lupus Nephritis by Regulating Autophagy

Objective Lupus nephritis is the most common and severe complication of systemic lupus erythematosus. The aim of our study was to investigate the efficacy of miR-20a overexpressing adipose-derived stem cell (ADSC) transplantation in murine lupus nephritis (LN) and explore potential molecular mechanisms. Methods Mouse ADSCs were transfected with a miR-20a lentiviral vector to obtain miR-20a overexpression ADSCs (miR-20a-ADSCs). We first observed the influence of miR-20a on ADSC viability and apoptosis in vitro. B6.MRL/lpr mice were administered ADSC/miR-20a-ADSC intravenously every week from age 30 to 33 weeks, and the lupus and normal control groups received PBS on the same schedule. Results miR-20a expression increased in miR-20a-ADSC-derived exosomes, and miR-20a overexpression promoted ADSC proliferation and inhibited apoptosis. Compared with ADSCs, miR-20a-ADSC treatment significantly improved serologic and histologic abnormalities, as evidenced by reduced serum creatinine, anti-dsDNA antibody, 24 h urine protein levels, nephritis scores, and C3/IgG deposits. Furthermore, miR-20a-ADSC treatment resulted in downregulated Akt, mTOR, and p62 expression and upregulated miR-20a, Beclin 1, and LC3 II/I expression compared with ADSC treatment. After treatment with miR-20a-ADSC, a significant increase in the number of autophagosomes within podocytes was observed, along with upregulated expression of podocin and nephrin, compared with the ADSC group. Conclusions miR-20a-ADSC transplantation prevents the development of lupus nephritis and significantly ameliorates already-established disease, and its mechanism is related to autophagy by targeting the miR-20a-regulated mTOR pathway.

LINC00370 modulates miR-222-3p-RGS4 axis to protect against osteoporosis progression

BackgroundWe aimed to determine the role of the LINC00370/miR-222-3p/RGS4 axis in modulating the process of adipose-derived stem cell (ADSC) osteogenic differentiation. MethodsWe first evaluated the differential expression of LINC00370, miR-222-3p and RGS4 between normal and osteogenically induced ADSCs. Moreover, we transfected ADSCs with LINC00370 siRNA and an miR-222-3p inhibitor to determine the role of LINC00370 in modulating the process of ADSC osteogenic differentiation. Finally, we analyzed the dual-luciferase reporter gene to identify the relationship between LINC00370 and miR-222-3p. We first created osteoporotic rat models by ovariectomy (OVX) and treated with pcDNA-LINC00370. HE and immunohistochemical staining of OCN were performed to assess the changes in bone microarchitecture. ResultsLINC00370 and RGS4 expression was remarkably upregulated in the osteogenic ADSC group compared with the normal medium group. On the other hand, miR-222-3p expression was remarkably decreased in the osteogenic group compared with the normal medium group. Knockdown of LINC00370 reduced the osteogenic differentiation of ADSCs. Moreover, the inhibitor of miR-222-3p partially reversed the reduction of osteogenic differentiation by LINC00370 knockdown. Knockdown of LINC00370 reduced the expression of p-Akt and p-PI3K. The inhibitor of miR-222-3p partially reversed the reduction of the expression of p-Akt and p-PI3K by LINC00370 knockdown. A dual luciferase reporter assay indicated that LINC00370 can directly bind miR-222-3p. LINC00370 suppressed OP progression in OVX and partially upregulated OCN protein expression. ConclusionCollectively, the above results confirm that LINC00370 promotes the process of ADSC osteogenic differentiation via the miR-222-3p/RGS4 axis. Moreover, LINC00370 could protect against OVX-induced OP.

Adipose-derived stem cells alleviate radiation-induced dermatitis by suppressing apoptosis and downregulating cathepsin F expression

BackgroundRadiation-induced dermatitis is a serious side effect of radiotherapy, and very few effective treatments are currently available for this condition. We previously demonstrated that apoptosis is an important feature of radiation-induced dermatitis and adipose-derived stem cells (ADSCs) are one of the most promising types of stem cells that have a protective effect on acute radiation-induced dermatitis. Cathepsin F (CTSF) is a recently discovered protein that plays an important role in apoptosis. In this study, we investigated whether ADSCs affect chronic radiation-induced dermatitis, and the underlying mechanisms involved.MethodsADSCs were isolated from male Sprague-Dawley (SD) rats and characterized. For in vivo studies, rats were randomly divided into control and ADSC-treated groups, and cultured ADSCs were transplanted into radiation-induced dermatitis model rats. The effects of ADSC transplantation were determined by skin damage scoring, histopathological analysis, electron microscopy, immunohistochemical staining, and western blotting analysis of apoptosis-related proteins. To evaluate the effects of ADSCs in vitro, radiation-induced apoptotic cells were treated with ADSC culture supernatant, and apoptosis-related protein expression was investigated by TUNEL staining, flow cytometry, and western blotting.ResultsIn the in vivo studies, skin damage, inflammation, fibrosis, and apoptosis were reduced and hair follicle and sebaceous gland regeneration were enhanced in the ADSC group compared with the control group. Further, CTSF and downstream pro-apoptotic proteins (Bid, BAX, and caspase 9) were downregulated, while anti-apoptotic proteins (Bcl-2 and Bcl-XL) were upregulated. In vitro, ADSCs markedly attenuated radiation-induced apoptosis, downregulated CTSF and downstream pro-apoptotic proteins, and upregulated anti-apoptotic proteins.ConclusionADSCs protect against radiation-induced dermatitis by exerting an anti-apoptotic effect through inhibition of CTSF expression. ADSCs may be a good therapeutic candidate to prevent the development of radiation-induced dermatitis.

Simultaneous Treatment of Photobiomodulation and Demineralized Bone Matrix With Adipose-Derived Stem Cells Improve Bone Healing in an osteoporotic bone defect.

Introduction: The ability of simultaneous treatment of critical-sized femoral defects (CSFDs) with photobiomodulation (PBM) and demineralized bone matrix (DBM) with or without seeded adipose-derived stem cells (ASCs) to induce bone reconstruction in ovariectomized induced osteoporotic (OVX) rats was investigated. Methods: The OVX rats with CSFD were arbitrarily separated into 6 groups: control, scaffold (S, DBM), S + PBM, S + alendronate (ALN), S + ASCs, and S + PBM + ASCs. Each group was assessed by cone beam computed tomography (CBCT) and histological examinations. Results: In the fourth week, CBCT and histological analyses revealed that the largest volume of new bone formed in the S + PBM and S + PBM + ASC groups. The S + PBM treatment relative to the S and S + ALN treatments remarkably reduced the CSFD (Mann-Whitney test, P = 0.009 and P = 0.01). Furthermore, S + PBM + ASCs treatment compared to the S and S + ALN treatments significantly decreased CSFD (Mann Whitney test, P = 0.01). In the eighth week, CBCT analysis showed that extremely enhanced bone regeneration occurred in the CSFD of the S + PBM group. Moreover, the CSFD in the S + PBM group was substantially smaller than S, S + ALN and S + ASCs groups (Mann Whitney test, P = 0.01, P = 0.02 and P = 0.009). Histological observations showed more new bone formation in the treated CSFD of S + PBM + ASCs and S + PBM groups. Conclusion: The PBM plus DBM with or without ASCs significantly enhanced bone healing in the CSFD in OVX rats compared to control, DBM alone, and ALN plus DBM groups. The PBM plus DBM with or without ASCs significantly decreased the CSFD area compared to either the solo DBM or ALN plus DBM treatments.

Epigenetic modifications underlie the differential adipogenic potential of preadipocytes derived from human subcutaneous fat tissue.

Ceiling culture-derived preadipocytes (ccdPAs) and adipose-derived stem cells (ASCs) can be harvested from human subcutaneous fat tissue using the specific gravity method. Both cell types possess a similar spindle shape without lipid droplets. We previously reported that ccdPAs have a higher adipogenic potential than ASCs, even after a 7-wk culture. We performed a genome-wide epigenetic analysis to examine the mechanisms contributing to the adipogenic potential differences between ccdPAs and ASCs. Methylation analysis of cytosines followed by guanine (CpG) using a 450-K BeadChip was performed on human ccdPAs and ASCs isolated from three metabolically healthy females. Chromatin immunoprecipitation sequencing was performed to evaluate trimethylation at lysine 4 of histone 3 (H3K4me3). Unsupervised machine learning using t-distributed stochastic neighbor embedding to interpret 450,000-dimensional methylation assay data showed that the cells were divided into ASC and ccdPA groups. In Kyoto Encyclopedia of Genes and Genomes pathway analysis of 1,543 genes with differential promoter CpG methylation, the peroxisome proliferator-activated receptor (PPAR) and adipocytokine signaling pathways ranked in the top 10 pathways. In the PPARγ gene, H3K4me3 peak levels were higher in ccdPAs than in ASCs, whereas promoter CpG methylation levels were significantly lower in ccdPAs than in ASCs. Similar differences in promoter CpG methylation were also seen in the fatty acid-binding protein 4 and leptin genes. In conclusion, we analyzed the epigenetic status of adipogenesis-related genes as a potential mechanism underlying the differences in adipogenic differentiation capability between ASCs and ccdPAs.

Hypoxia Promotes Adipose-Derived Stem Cells to Protect Human Dermal Microvascular Endothelial Cells Against Hypoxia/Reoxygenation Injury

Microcirculation is important for regulating ischemia-reperfusion (I/R) injury associated with skin flap transplantation surgery. We investigated whether co-culture with adipose-derived stem cells (ADSCs) could protect human dermal microvascular endothelial cells (HDMECs) from I/R injury by inhibiting cell apoptosis and enhancing cell proliferation. We also investigated the effects of hypoxic preconditioning on ADSCs. HDMECs were divided into four groups, control, HDMECs in normoxic culture conditions; hypoxia/reoxygenation (H/R), HDMECs in a hypoxic incubator for 8 h then in saturated aerobic culture medium for 24 h; H/R+ADSC(N), HDMECs treated similar to the H/R group then co-cultured with normoxic ADSCs; and H/R+ADSC(H), HDMECs treated similar to the H/R group then co-cultured with hypoxia preconditioned ADSCs. The rate of HDMECs apoptosis significantly increased in the H/R group, but decreased upon co-culture with ADSCs for 24 h, especially in the H/R + ADSC(H) group. Co-culture with ADSCs, especially hypoxia preconditioned ADSCs, significantly enhanced cell proliferation ability compared with that of the H/R group after 48 h and 72 h, but not after 24 h. Vascular endothelial growth factor levels were significantly higher in the H/R+ADSC(N) and H/R+ADSC(H) groups than in the H/R group. ADSCs attenuated H/R injury in endothelial cells by promoting proliferation ability and reducing apoptosis, with an increase in Vascular endothelial growth factor level, especially in the context of hypoxic preconditioning. This approach suggests the potential for an easy and safe method to reduce I/R injury associated with skin flap transplantation surgery.

Using 3D-bioprinting scaffold loaded with adipose-derived stem cells to burns wound healing.

Three dimensional (3D) printing has recently expanded in popularity and has become an effective approach for tissue engineering. Advances in tissue engineering have increased the effectiveness of cell-based therapies. Indeed, the ultimate goal of such treatment is the development of conditions similar to fetal wound regeneration. In this context, technology of 3D printing also allows researchers to more effectively compose multi-material and cell-laden scaffolds with less effort. In this study, we explored a synthetic gel scaffold derived from 3D bioprinter with or without stem cells to accelerate wound healing and skin defects. Adipose-derived stem cells (ADSCs) were isolated and seeded into 3D bioprinter derived-gel scaffold. Morphological and cell adherence properties of 3D scaffold were assessed by hemotoxylin& eosin (H&E) staining and scanning electron microscopy and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide assay. In vivo assessment of the scaffold was done using H&E staining in the full-thickness burn rat model. The experimental groups included; (a) untreated (control), (b) 3D bioprinter derived-gel scaffold (Trial 1), and (c) 3D bioprinter derived-gel scaffold loaded with ADSC (Trial 2). Our results represented 3D bioprinter derived-gel scaffold with or without ADSCs accelerated wound contraction and healing compared to control groups. Epithelization was completed until 21days after operation in scaffold alone. In scaffold with ADSCs group, epithelization was faster and formed a multi-layered epidermis with the onset of cornification. In conclusion, 3D bioprinter derived-gel scaffold with or without ADSCs has the potential to be used as a wound graft material in skin regenerative medicine.

Facilitatory effects of artificial nerve filled with adipose-derived stem cell sheets on peripheral nerve regeneration: An experimental study

BackgroundWe evaluated how artificial nerves filled with adipose-derived stem cell (ADSC) sheets could facilitate peripheral nerve regeneration. MethodsWe prepared ADSC sheets following previously described protocols. We transected the sciatic nerve in 12-week-old Wistar rats, fixed the nerve ends to the artificial conduit, and prepared three groups: (1) conduits alone (control group); (2) conduits filled with ADSCs (ADSCs group), and (3) conduits filled with ADSC sheets (ADSC sheet group). We assessed the subjects 4 and 12 weeks post-transplantation (n = 24). We investigated βIII-tubulin and anti-S100 expression at 4 and 12 weeks post-transplantation, in longitudinal- and cross-sections of the central portion in the regenerated tissues. The vascular endothelial growth factor A (VEGFA) and neuregulin-1 expressions were analyzed using real-time reverse-transcription polymerase chain reaction (real-time RT-PCR). We evaluated the tibialis anterior muscle wet weight (affected/healthy sides, %) and sciatic function index (SFI) 12 weeks post-transplantation. ResultsThe ADSC sheet group comprised more S100-positive cells than the other groups. The regenerated axon length in the ADSC sheet group was markedly the longest among the studied groups. The immunostaining revealed a positive area in the regenerated tissue center in all groups, tending to be the largest in the ADSC sheet group. The muscle wet weight indicated that the ADSC sheet group exhibited significantly higher weight than the control. The mean SFI showed that the ADSC sheet group exhibited significantly better results than the control. The VEGFA expression was higher both in the ADSC and the ADSC sheet group than in the control. The neuregulin-1 expression was higher both in the ADSC and the ADSC sheet group than in the control. ConclusionsThe ADSC sheets could potentially support transplanting an adequate number of ADSCs at the target site. Compared with the conventional method of attaching ADSCs, the use of ADSC sheets promotes accelerated nerve regeneration.