Injection Of Bone Marrow Mesenchymal Stem Cells Research Articles

Mechanism underlying the anti-diabetic potential of bee venom as compared to bone marrow mesenchymal stem cells in the diabetic rat tongue.

Diabetes mellitus (DM) is a critical chronic metabolic disease. Several treatment modalities are currently under investigation. Both bee venom (BV) and bone marrow mesenchymal stem cells (BMSCs) can possibly offer an approach for treating type I diabetes. The aim of the present study was to investigate the mechanism underlying the anti-diabetic effect of BV as compared to BMSCs on the tongue mucosa of diabetic rats. A total of 52 male albino rats were used in the current study. The rats were randomly assigned into 4 groups: group 1 (control); group 2 (streptozocin (STZ)); group 3 (BV-treated); and group 4 (BMSC-treated). Diabetes mellitus was induced via an intraperitoneal (IP) injection of STZ in the rats from groups 2, 3 and 4. Following the diagnosis of DM, the rats in group 3 were injected with a daily dose of 0.5 mg/kg of BV, while the rats in group 4 were treated with a single injection of BMSCs. All rats were euthanized after 4 weeks, and their tongues were dissected and divided into halves. The right halves of the tongues were utilized for the histological examination, followed by morphometric analysis. In contrast, the left halves were used to detect the local gene expression of transforming growth factor beta 1 (TGF-β1) and vascular endothelial growth factor (VEGF). Group 2 revealed marked disruption in the morphology of the fungiform and filiform papillae, and atrophic epithelial changes in both dorsal and ventral surface epithelium as compared to other groups. Group 4 showed a significantly larger number of taste buds, and a higher gene expression of TGF-β1 and VEGF as compared to groups 2 and 3. Additionally, BV and BMSCs effectively increased the thickness of dorsal and ventral surface epithelium as compared to group 2. Treatment with BMSCs was associated with significant improvement in the morphology and number of lingual epithelial cells and taste buds in the tongues of diabetic rats as compared to BV-treated rats, which was due to the local upregulation of TGF-β1 and VEGF gene expression.

Therapeutic use of hematopoietic mesenchymal stem cells in osteoarthritis: a meta-analysis study

Millions of people worldwide suffer from osteoarthritis, which is a major health problem. The accessible medicines are used in changing the side effects of osteoarthritis however missing illness alterations, not with-standing drug after-effects, and care expenses without adequacy of side effects help. This study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. We look for relevant articles in all articles published between December 30, 2007, and December 30, 2022, in the Cochrane Library, PubMed, Embase, Wanfang Database, CNKI, and Web of Science databases. With an average follow-up of 18.33 months, this study included 364 patients with knee joint osteoarthritis from 12 completely published articles. Data from seven studies using the Visual Analog Scale (VAS) score. In Bone Marrow Mesenchymal Stem Cells (BM-MSCs), the baseline score on the Visual Analogue Scale decreased. These results suggested that there was no significant relationship between the BM-MSCs at three months (Weighted Mean Difference (WMD) = 1.07, 95% CI: − 2.44, − 0.39, p= 0.074); and the greatest decrease in VAS score compared to the control at 18.33 months, WMD = 6.04, 95 percent CI: − 10.34, − 0.38, p = 0.017. In clinical applications, intra-articular injection of BM-MSCs has advantages for treating OA because it reduces pain index, improves knee movement, and increases cartilage volume.

Effects of mesenchymal stem cells on Treg cells in rats with colitis.

The aim was to investigate the therapeutic effect of bone marrow mesenchymal stem cells (BM-MSC) on dextran sulfate sodium (DSS) induced colitis in rats and its effect on regulatory T cells (Treg). A model of DSS-induced colitis was established. BM-MSC was isolated and cultured to observe the efficacy of BM-MSC on colitis, including general vital signs, weight changes, colonic length changes, colonic histopathological changes, and colonic tissue MPO activity. The expression of inflammatory factors (IFN-γ, IL-4, IL-17, TGF-β) in colonic tissues was measured by real-time PCR. The amount of CD4 + CD25 + Treg was detected by flow cytometry. Real-time PCR was used to detect Foxp3+mRNA in CD4 + CD25 + Treg, western to detect Foxp3+protein expression in CD4 + CD25 + Treg, and ELISA was used to detect IL-35 and IL-10 cytokines in CD4 + CD25 + Treg culture supernatant. Results show that intravenous injection of BM-MSC significantly improved the clinical manifestations and histopathological changes in rats with experimental DSS colitis; significantly down-regulated the expression of inflammatory factors IFN-γ, IL-4, and IL-17 and up-regulated the expression of TGF-β in colon tissues; BM-MSC also increased the number of CD4+CD25+Foxp3+Treg and enhanced the function of CD4+CD25+Foxp3+Treg in colon tissues, and up-regulated the expression of IL-35. In conclusion, BM-MSC has a certain therapeutic effect on DSS-induced colitis. It can improve the general signs of colitis rats and reduce intestinal injury and inflammatory response. The immunoregulatory effect of BM-MSC is achieved by enhancing the function of CD4+CD25+Foxp3+Treg and up-regulating the secretion of immunosuppressive inflammatory factors.

Effect of PKH-26-labeled exosomes derived from bone marrow mesenchymal stem cells on corneal epithelium regeneration in diabetic mice.

It is known that bone marrow mesenchymal stem cells (BM-MSCs) could speed up the regeneration of diabetic corneal epithelium. To investigate the effect of exosomes derived from mouse BM-MSCs on corneal epithelium regeneration in diabetic mice. Diabetic mouse models were established using streptozotocin (STZ), and their central corneal epithelium was scratched under a microscope. The diabetic mice were randomly divided into three groups: the control group was injected with subconjunctival phosphate buffer saline; the exosomes group was treated with a subconjunctival injection of exosomes derived from BM-MSCs; and the BM-MSCs group was treated with a subconjunctival injection of BM-MSCs. The corneal epithelium repair rates in the three groups were compared, and the distribution of the exosomes derived from BM-MSCs labeled with PKH-26 was observed by immunofluorescence. Hematoxylin-eosin staining of the corneal tissue was observed 72 h after the treatments in the three groups. The diabetic mice were successfully established by a blood glucose level >16.7 mmol/L after 8 weeks. The corneal epithelium healing rates in experimental groups 1 and 2 were significantly higher than those of the control group at 24, 48, and 72 h (P<0.05). However, there was no significant difference in the corneal epithelial healing rate between experimental groups 1 and 2 (P>0.05). The exosomes derived from BM-MSCs were found in the superficial corneal stroma in experimental groups 1 and 2, with the majority of the exosomes distributed in the limbal epithelium at the edge of the injury area. The proliferation of corneal epithelial cells in experimental groups 1 and 2 was more obvious than that in the control group. The exosomes derived from BM-MSCs labeled with PKH-26 significantly promoted the repair of corneal epithelial injury in diabetic mice. These exosomes might be a substitute for BM-MSCs in the repair of diabetic keratopathy, suggesting a new idea for the repair of diabetic keratopathy with "cell-free" stem cell therapy, which will require a clinical study.

The potential therapeutic efficacy of intravenous versus subconjunctival mesenchymal stem cells on experimentally ultraviolet-induced corneal injury in adult male albino rats.

The X-rays and the visible light are the main source of ultraviolet radiation (UVR). About 90% of ultraviolet B (UVB) is absorbed by the cornea which may promote corneal inflammation, oedema and damage of its epithelial layer. Bone marrow mesenchymal stem cells (BM-MSCs) have been demonstrated to ameliorate the injured corneal tissue and accelerate its wound healing. This study aimed to compare the healing effect of intravenous (IV) versus subconjunctival (SC) BM-MSCs on the rats' corneas subjected to UVB-irradiation. Ten rats were used as donors for BM-MSCs and the other 40 were allocated into four equal groups: group I (control group), group II (ultraviolet-irradiated group), group III (ultraviolet-irradiated + IV BM-MSCs-treated group) and group IV (ultraviolet-irradiated +SC BM-MSCs-treated group). Rats of all groups were euthanized after 3 weeks and the corneal specimens were processed for histopathological, immunohistochemical and electron microscopy assessment. Ultraviolet-irradiated group showed remarkable thinning of epithelial thickness, wide partial epithelial separation, and desquamation. Neovascularisation of the disorganised stroma and disrupted Descemet's membrane were observed. The superficial and basal epithelial cells appeared irregular and separated by wide intercellular spaces and inflammatory cells. Immunohistochemical examination showed a significant decrease in proliferating cell nuclear antigen immunoreaction. In contrast, minimal changes were observed in rats treated with BM-MSCs with more improvement associated with the subconjunctival administration compared to IV route. Local SC injection of BM-MSCs has an amazing regenerative efficacy on the corneal injury compared to the systemic IV route.

Role of TGF-β3 and bone marrow mesenchymal stem cells on regeneration of myometrial injury in rats

ObjectiveThis study aimed to explore therapeutic effect of bone marrow mesenchymal stem cells (BMSCs) and TGF-β3 in regenerative injuries in the myometrium of the uterus in rats. MethodsTwenty-four pregnant rats were randomly assigned into four groups and their pregnancy was terminated on day 19 of pregnancy: 1. Blank scar uterus control group injected with the physiological saline, 2. TGF-β3 group injected with TGF-β3, 3. BMSCs group injected with BMSCs, 4. BMSCS + TGF-β3 injected with TGF-β3 and BMSCs. ResultsMSCs expressing specific surface antigens had multidirectional differentiation potential and showed a tendency to home to the site of injury, and the macroscopic appearance of the cell-treated uterus was smoother and tougher in the three groups than in the control group. The uterus improved by the TGF-β3+MSCs group had the best recovery showing significant glandular and vascular nature, with better repair effect and promoted endometrial cell proliferation. Proteins related to endometrial cell proliferation showed that the expression of Ki-67 was significantly increased in three intervention groups, and a positive signal of waveform protein was observed in the uterine stromal cells. The levels of waveform protein and CK-19 protein were significantly lower in the control group compared to their levels in the three interventional groups. Injection of BMSCs and TGF-β3 resulted in the repair of whole damaged myometrium and significant improvement in function compared to other groups, whereas TGF-β3 significantly reduced collagen fiber production. ConclusionsBMSCs can differentiate into uterine smooth muscle cells, repair uterine scar tissue and promote uterine scar healing.

Comparison of Bone Marrow Mesenchymal Stem Cells-Derived Exosomes, Microvesicles, and Soluble Proteins on Their Chondroprotective Effects and Therapeutic Efficacy of Osteoarthritis Treatment

As a disabling joint disorder, osteoarthritis (OA) is characterized by the degeneration of articular cartilage. Bone marrow mesenchymal stem cells (BMSCs) is considered and used as a tool that is regulated by paracrine for the treatment of OA. Herein, we explored the potential of exosomes (Exos), microvesicles (MVs), and soluble proteins (SPs) produced and isolated from rat BMSCs. First, we observed the biological effects of Exos, MVs, and SPs on IL-1β treated rat chondrocytes. Then, we intra-articularly injected Exos, MVs, SPs, and BMSCs in anterior cruciate ligament transection (ACLT)-induced rat OA model, and observed their therapeutic efficacy. SPs rather than Exos and MVs isolated from rat BMSCs exerted a chondroprotective effect in vitro; MVs and SPs showed comparable effects to the intra-articular injection of BMSCs, whereas the chondroprotective effects of Exos were less potent in vivo. In conclusion, the BMSCs secretome might have inconsistent effects in vivo with in vitro, and Exos might not be superior than other secretome in posttraumatic OA prevention.

Bone marrow mesenchymal stem cells therapy on bilateral pelvic nerve crush-induced voiding dysfunction in rats.

Neurogenic voiding dysfunction can be induced after radical pelvic surgery and severely affects patients' quality of life. This study aims to investigate the effects of bone marrow mesenchymal stem cells (BMSCs) on neurogenic voiding dysfunction in male rats and explore the underlying mechanisms. Thirty 4-week-old male Sprague-Dawley rats were randomly divided into three groups: (1) sham-operated (sham, n = 10), (2) intrabladder wall injection of phosphate buffer solution (PBS) after bilateral pelvic nerve crush (BPNC+PBS, n = 10), and (3) intrabladder wall injection of BMSCs after bilateral pelvic nerve crush (BPNC+BMSCs, n = 10). Four weeks postoperatively, functional and morphological examinations were performed. Compared to the sham group, BPNC rats manifested significant augmentation in the frequency of non-voiding contractions and postvoid residual and bladder capacity, and they had decreases in intravesical pressure and voiding efficiency. However, they were markedly improved after BMSC injection. Masson's trichrome staining showed that the ratio of collagen area in bladder wall tissue significantly increased in the BPNC+PBS group but was reduced following BMSC injection. BPNC increased the protein expression of TGF-β1, Smad2/3, and collagen I/III but decreased the expression of α-SMA. BMSC injection stimulated higher expression levels of α-SMA and lower expression levels of the other target proteins. The expression levels of vesicular acetylcholine transporters were reduced at 4weeks post-BPNC, whereas injection of BMSCs boosted the expression quantity. BMSC therapy suppressed detrusor fibrosis, improved intravesical pressure and voiding efficiency, and partially restored voiding function in male rats after BPNC.

Transplantation of Bone Marrow Mesenchymal Stem Cells (BMSCs) Overexpressing Circular Zinc Finger Protein 609 Alleviates Diabetic Retinopathy

Diabetic retinopathy (DR) is a chronic complications and its pathogenesis remains unclear. This study aims to elucidate the underlying mechanism by how bone marrow mesenchymal stem cells (BMSCs) affects DR development in a rat model. A rat model of DR was established and injected with BMSCs overexpressing Cir-ZNF609 and shRNA Cir-ZNF609 to vitreous body followed by analysis of the retinal vascular permeability and macular retinal layers thickness, and the levels of HIF-1α, ICAM-1 and VEGF in rat retina by ELISA and immunohistochemistry. Injection of BMSCs overexpressing Cir-ZNF609 resulted in decreased HIF-1α ICAM-1 and VEGF expression, amelioration of retinal ganglion choriocapillaris injury and reducing ganglion cells. Twelve weeks after treatment, neovascularization took place and fibroblasts appeared with some nucleus disappearing and pigment taking off. Besides, permeability also elevated in the presence of overexpressing Cir-ZNF609 and penetration rate for Evans blue (16.36+3.25, 15.45±3.46 μg/g) was lower than healthy rats (28.66±2.08, 32.24±4.36 μg/g) and controls (26.93±3.03, 33.49±5.02 μg/g) (p &lt; 0.01). Moreover, upregulation of Cir-ZNF609 decreased retinal thickness and macular volume in DR rats (p &lt; 0.05). In conclusion, intravitreal injection of mouse BMSCs overexpressing Cir-ZNF609 alleviates retinal injury and decreases retinal thickness and macular volume, and enhances neovascularization. These evidence provides a novel insight into gene therapy for DR.

Kartogenin Promotes the BMSCs Chondrogenic Differentiation in Osteoarthritis by Down-Regulation of miR-145-5p Targeting Smad4 Pathway.

Transplantation of mesenchymal stem cells (MSCs) is a potential therapeutic strategy for cartilage degeneration of osteoarthritis (OA). But controlling chondrogenic differentiation of the implanted MSCs in the joints remains a challenge. The role of kartogenin (KGN) for chondrogenesis of MSCs has been widely reported, however, the mechanism of chondrogenesis has not been elucidated in OA. In this study, we investigated the miR-145-5p, TGF-β, Samd4, and p-stat3/stat3 expression in cartilage of OA patients and bone marrow mesenchymal stem cells (BMSCs) treated with KGN or miR-145-5p inhibitor. In addition, the cell proliferation and chondrogenic differentiation in vitro and in vivo of BMSCs treated with KGN was also detected. In OA patients, the expression of miR-145-5p was up-regulated, and the expression of TGF-β, Samd4, and p-stat3/stat3 was inhibited. When the BMSCs treated with miR-145-5p inhibitor, the expression of TGF-β, Samd4, and p-stat3/stat3 was also significantly up-regulated. KGN-treated BMSCs had better proliferation and chondrogenic differentiation by up-regulating the expression of Sox 9, Col-2a1, aggrecan in vitro and in OA by down-regulation of miR-145-5p targeting Smad4 pathway. Moreover, intra-articular injection of KGN-treated BMSCs had a better pain relief effect in OA. The double effect on cartilage protection and pain relief indicates a great potential of intra-articular injection of KGN-treated BMSCs for the treatment of OA.

Intra-Articular Injection of miR-29a-3p of BMSCs Promotes Cartilage Self-Repairing and Alleviates Pain in the Rat Osteoarthritis.

Stem cells intra-articular injection stagey indicated a potential therapeutic effect on improving the pathological progress of osteoarthritis (OA). However, the long-term effect of stem cells intra-articular injection on the cartilage regeneration remains unclear. miR-29a-3p is predicted to be a critical target for inhibiting insulin-like growth factor-1 expression and may aggravate the progression of OA. In this study, we investigated the therapeutic efficacy of intra-articular injection of bone marrow mesenchymal stem cells (BMSCs) transfected with miR-29a-3p inhibitor in OA. miR-29a-3p inhibitor transfection did not influence cell viability of BMSCs, while the chondrogenic differentiation potential of BMSCs was significantly improved. Interestingly, intra-articular injection of BMSCs with miR-29a-3p inhibition significantly prevented articular cartilage degeneration by up-regulating the expression of Sox 9, Col-2a1, aggrecan and down-regulating the expression of matrix metalloproteinase, as well as relieved pain in OA. The double effects on cartilage protection and pain relief indicated a great potential of intra-articular injection of miR-29a-3p inhibitor-transfected BMSCs for the treatment of OA.

Therapeutic effects of bone marrow mesenchymal stem cells-derived exosomes on osteoarthritis.

Mesenchymal stem cells (MSCs) have shown chondroprotective effects in clinical models of osteoarthritis (OA). However, effects of MSC‐derived exosomes on OA remain unclear. The study aimed to investigate the therapeutic potential of exosomes from human bone marrow MSCs (BM‐MSCs) in alleviating OA. The anterior cruciate ligament transection (ACLT) and destabilization of the medial meniscus (DMM) surgery were performed on the knee joints of a rat OA model, followed by intra‐articular injection of BM‐MSCs or their exosomes. In addition, BM‐MSC‐derived exosomes were administrated to primary human chondrocytes to observe the functional and molecular alterations. Both of BM‐MSCs and BM‐MSC‐derived exosomes alleviated cartilage destruction and subchondral bone remodelling in OA rat model. Administration of BM‐MSCs and exosomes could reduce joint damage and restore the trabecular bone volume fraction, trabecular number and connectivity density of OA rats. In addition, in vitro assays showed that BM‐MSCs‐exosomes could maintain the chondrocyte phenotype by increasing collagen type II synthesis and inhibiting IL‐1β–induced senescence and apoptosis. Furthermore, exosomal lncRNA MEG‐3 also reduced the senescence and apoptosis of chondrocytes induced by IL‐1β, indicating that lncRNA MEG‐3 might partially account the anti‐OA effects of BM‐MSC exosomes. The exosomes from BM‐MSCs exerted beneficial therapeutic effects on OA by reducing the senescence and apoptosis of chondrocytes, suggesting that MSC‐derived exosomes might provide a candidate therapy for OA treatment.

Stromal Cell-Derived Factor (SDF)-1-Pretreated Bone Marrow Mesenchymal Stem Cells (BMSCs) Ameliorates Acute Kidney Injury in Mice

To explore the effects of stromal cell-derived factor (SDF-1) pretreatment of bone marrow mesenchymal stem cells (BMSCs) on acute kidney injury (AKI) in mice. BMSCs were cultured and treated with SDF-1 to detect osteogenic and adipogenic ability. Cisplatin (20 mg/kg) was used to establish AKI model and then divided into blank group, control group 2 (BMSCs injection), and experimental group (intraperitoneal injection of BMSCs treated with SDF-1 (80 ng/ml)) followed by analysis of serum cytokines (Toll like receptor 4 (TLR4), tumor necrosis factor-α (TNF-α), and interleukine-6 (IL-6)) by enzyme linked immunosorbent assay (ELISA). In cultured BMSCs, positive rates of CD29, CD44, CD45, and CD11b were 98.2%, 97.6%, 2.5% and 2.1%, respectively. When the concentration of SDF-1 was within 80 ng/mL, the chemotaxis and proliferation ability was dose-dependent (p &lt; 0.05). SDF-1 pretreatment did not affect BMSCs adipogenic and osteogenic abilities. The creatinine and serum cytokines (TLR4, TNF-α, and IL-6) level in experimental group showed statistical significance (p &lt; 0.05). At 24 h, thrombosis and tubular dilatation in the mesangial region of control group 2 and experimental group under light microscope were similar without difference of inflammatory cell infiltration and fibrosis. At 72 h, the glomerular mesangium widened in control group 2 with focal segmental sclerosis, renal tubules dilated, and protein casts and inflammatory cell infiltration and fibrosis. Experimental group showed a small amount of cell proliferation in the glomerular mesangium with few inflammatory cell infiltration and fibrosis. SDF-1 can enhance the migration and proliferation activity of BMSCs, reduce extracellular matrix precipitation, improve renal fibrosis, and alleviate AKI.

Bone marrow mesenchymal stem cells and their derived exosomes resolve doxorubicin-induced chemobrain: critical role of their miRNA cargo

BackgroundDoxorubicin (DOX), a widely used chemotherapeutic agent, can cause neurodegeneration in the brain, which leads to a condition known as chemobrain. In fact, chemobrain is a deteriorating condition which adversely affects the lives of cancer survivors. This study aimed to examine the potential therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) and their derived exosomes (BMSCs-Exo) in DOX-induced chemobrain in rat models.MethodsChemobrain was induced by exposing rats to DOX (2 mg/kg, i.p) once weekly for 4 consecutive weeks. After 48 h of the last DOX dose, a subset of rats was supplied with either an intravenous injection of BMSCs (1 × 106) or a single dose of 150 μg of BMSCs-Exo. Behavioral tests were conducted 7 days post injection. Rats were sacrificed after 14 days from BMSCs or BMSCs-Exo injection.ResultsBMSCs and BMSCs-Exo successfully restored DOX-induced cognitive and behavioral distortion. These actions were mediated via decreasing hippocampal neurodegeneration and neural demyelination through upregulating neural myelination factors (myelin%, Olig2, Opalin expression), neurotropic growth factors (BDNF, FGF-2), synaptic factors (synaptophysin), and fractalkine receptor expression (Cx3cr1). Halting neurodegeneration in DOX-induced chemobrain was achieved through epigenetic induction of key factors in Wnt/β-catenin and hedgehog signaling pathways mediated primarily by the most abundant secreted exosomal miRNAs (miR-21-5p, miR-125b-5p, miR-199a-3p, miR-24-3p, let-7a-5p). Moreover, BMSCs and BMSCs-Exo significantly abrogate the inflammatory state (IL-6, TNF-α), apoptotic state (BAX/Bcl2), astrocyte, and microglia activation (GFAP, IBA-1) in DOX-induced chemobrain with a significant increase in the antioxidant mediators (GSH, GPx, SOD activity).ConclusionsBMSCs and their derived exosomes offer neuroprotection against DOX-induced chemobrain via genetic and epigenetic abrogation of hippocampal neurodegeneration through modulating Wnt/β-catenin and hedgehog signaling pathways and through reducing inflammatory, apoptotic, and oxidative stress state.Graphical abstractProposed mechanisms of the protective effects of bone marrow stem cells (BMSCs) and their exosomes (BMSCs-Exo) in doxorubicin (DOX)-induced chemobrain. Blue arrows: induce. Red arrows: inhibit.

Therapeutic effect of mesenchymal stem cells on histopathological, immunohistochemical, and molecular analysis in second-grade burn model

Background and aimDeleterious cutaneous tissue damages could result from exposure to thermal trauma, which could be ameliorated structurally and functionally through therapy via the most multipotent progenitor bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to induce burns and examine the effect of BM-MSCs during a short and long period of therapy.Material and methodsNinety albino rats were divided into three groups: group I (control); group II (burn model), the animals were exposed to the preheated aluminum bar at 100°C for 15 s; and group III (the burned animals subcutaneously injected with BM-MSCs (2×106 cells/ ml)); they were clinically observed and sacrificed at different short and long time intervals, and skin samples were collected for histopathological and immunohistochemical examination and analysis of different wound healing mediators via quantitative polymerase chain reaction (qPCR).ResultsSubcutaneous injection of BM-MSCs resulted in the decrease of the wound contraction rate; the wound having a pinpoint appearance and regular arrangement of the epidermal layer with thin stratum corneum; decrease in the area percentages of ADAMs10 expression; significant downregulation of transforming growth factor-β (TGF-β), interleukin-6 (IL-6), tumor necrotic factor-α (TNF-α), metalloproteinase-9 (MMP-9), and microRNA-21; and marked upregulation of heat shock protein-90α (HSP-90α) especially in late stages.ConclusionBM-MSCs exhibited a powerful healing property through regulating the mediators of wound healing and restoring the normal skin structures, reducing the scar formation and the wound size.

Antioxidant and antiapoptotic paracrine effects of mesenchymal stem cells on spermatogenic arrest in oligospermia rat model

BackgroundOligospermia is one of the common causative factors of male infertility. Some medical and hormonal therapy for male infertility is typically with unsatisfactory outcome. Stem cell therapy has become a new therapeutic strategy for restoring function in addition to inducing proliferation and differentiation of malfunctioning germ cells. This work aims at investigating the potential ability of BM-MSCs to repair the spermatogenic arrest in oligospermic rat model. MethodsIn this work, a rat model of oligospermia was induced using two intraperitoneal injections of busulfan (15 mg/kg) with two weeks interval. Rats were divided into (i) donor group [source of the bone marrow mesenchymal stem cells (BM-MSCs) that were labelled and transfected with green fluorescent protein (GFP)] and (ii) experimental groups that were subdivided into: GpI (control), GpII (spermatogenic arrest model), GpIII (untreated rats), and GpIV (BM-MSCs treated rats). Estimation of the testicular weight, sperm count and motility % were performed. Histological and immunohistochemical staining for inducible nitric oxide synthase (iNOS) and caspase-3 (Cas-3) were conducted. Besides, the level of the testicular malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α) and testicular testosterone were estimated by ELISA. ResultsOligospermic rats illustrated hypospermatogenesis of the seminiferous tubule with spermatocyte and spermatid arrest, focal thickening of the basement membrane and significant increase in germ cells apoptosis and testicular oxidative stress. Compared with the control, MDA and TNF-α were markedly elevated with marked suppression of the testicular testosterone. Intra-testicular injection of BM-MSCs substantially ameliorated these changes and effectively improved the sperm count and motility %. ConclusionsBM-MSCs improved the induced-spermatogenic arrest in the rat model mainly through anti-apoptotic and antioxidant paracrine effects.

Bone Marrow-Derived Mesenchymal Stem Cells Reverse Radiotherapy-Induced Premature Ovarian Failure: Emphasis on Signal Integration of TGF-β, Wnt/β-Catenin and Hippo Pathways

Radiotherapy is an indispensable cancer treatment approach. However, it is associated with hazardous consequences on multiple organs characterized by insidious worsening severity over time. This study aimed to examine the potential therapeutic effects of bone marrow mesenchymal stem cells (BM-MSCs) in radiation-induced premature ovarian failure (POF). Exposing female rats to 3.2Gy whole-body ϒ-rays successfully induced POF. One week later, a single intravenous injection of BM-MSCs (2*106) cells was administered. BM-MSCs perfectly home to the damaged ovaries, enhanced ovarian follicle pool, and preserved the ovarian function manifested by restoring serum estradiol and follicle stimulating hormone levels, besides, rescuing the fertility outcomes of irradiated rats. These events have been associated with inhibiting ovarian apoptosis (Bax/Bcl2, caspase 3) and enhancing proliferation (PCNA). Interestingly, BM-MSCs reversed the inhibition of ovarian FOXO3 expression induced by radiation which resulted in increased primordial follicles stock. Moreover, BM-MSCs recovered the suppressed folliculogenesis process induced by radiation through upregulating FOXO1, GDF-9, and Fst genes expression accompanied by downregulating TGF-β which enhanced granulosa cells proliferation and secondary follicle development. Mechanistically, BM-MSCs miRNAs epigenetically upregulate Wnt/β-catenin and Hippo signaling pathways which are implicated in ovarian follicles growth and maturation. Therefore, BM-MSCs presented a ray of hope in the treatment of radiation-associated POF through genetic and epigenetic modulation of the integrated TGF-β, Wnt/β-catenin, and Hippo pathways which control apoptosis, proliferation, and differentiation of ovarian follicles.

Histological, Histochemical and Immunohistochemical Evaluation of the Role of Bone Marrow-Derived Mesenchymal Stem Cells on the Structure of Periodontal Tissues in Carbimazole-Treated Albino Rats

ObjectiveTo elucidate the role of bone marrow-mesenchymal stem cells (BM-MSCs) on the structure of periodontal tissues in carbimazole (antithyroid drug) treated rats at different durations. Design28 albino rats were divided into: Group I: received distilled water. Group II: received therapeutic dose of carbimazole. Group III: received carbimazole then single injection of BM-MSCs by the end of 3rd week. Group IV: received carbimazole and single injection of BM-MSCs at the beginning of the experiment. Specimens were examined by light microscope. New collagen and β-catenin-immunoreactivity area% were assessed histomorphometrically, and statistically using ANOVA test. ResultsHistological examination revealed normal periodontal tissues structure in Groups I & IV. Group II showed disorganized periodontal ligament fibers and different stainability of cementum and alveolar bone. Group III illustrated dense periodontal ligament fibers, normal stainability of cementum and most of alveolar bone. Masson's trichrome results of Groups I & IV illustrated large areas of new collagen in periodontal ligament, old collagen in cementum and intermingled old and new collagen in alveolar bone. Group II showed old collagen. Group III revealed only new collagen. β-catenin-immunoreactivity was strong in Groups I & IV, negative in Group II and moderate in Group III. Statistically, Group III showed highest mean of new collagen area% followed by Groups I, IV and II respectively. Highest mean of β-catenin-immunoreactivity area% was for Group I followed by Groups IV, III and II respectively. ConclusionsCarbimazole has damaging effects and BM-MSCs are capable to mend these destructive outcomes in time dependent manner.

Effects of local injection and intravenous injection of allogeneic bone marrow mesenchymal stem cells on the structure and function of damaged anal sphincter in rats.

Anal sphincter injury leads to damage to the anal structure and functions and has been identified as a major risk factor for fecal incontinence. Bone marrow mesenchymal stem cells (BMSCs) with capacities of multidifferentiation, paracrine, and low immunogenicity have been widely used in tissue repair and regeneration. The primary objective of this research was to compare the effects of different injection therapies of BMSCs on the injured anal sphincters. Ninety-six Sprague-Dawley female rats were randomly divided into four groups (n = 24 each): intravenous injection, local injection, sham operation, and normal control. For the first three groups, 25% removal of the anal sphincter complex was performed and 0.3-ml phosphate-buffered saline (PBS) (containing 107 green fluorescent protein-labeled allogeneic BMSCs) was given accordingly to the treatment group 24 h after operation for 7 consecutive days. The sham operation group was injected with 0.3-ml PBS only. All cases had undergone evaluation in the 1st, 7th, 14th, and 28th postoperative days. The rats were sacrificed on the 28th postoperative day, and the anal sphincters were dissected to be analyzed by morphological examination. At 14 days postoperatively, local injection of BMSC significantly improved the peak contraction pressure, electromyography amplitude, and frequency of the injured anal sphincter compared with tail vein, but there was no significant difference in resting pressure until 28 days after sphincterectomy. Masson staining results confirmed that the local injection group had significantly more new muscles on the wound. BMSC could remarkably improve peak contraction pressure, electromyography amplitude, and muscle fibers on the wound, and local injection is superior to intravenous injection.

Anti-inflammatory protein TSG-6 secreted by bone marrow mesenchymal stem cells attenuates neuropathic pain by inhibiting the TLR2/MyD88/NF-\u03baB signaling pathway in spinal microglia

BackgroundNeuroinflammation plays a vital role in the development and maintenance of neuropathic pain. Recent evidence has proved that bone marrow mesenchymal stem cells (BMSCs) can inhibit neuropathic pain and possess potent immunomodulatory and immunosuppressive properties via secreting a variety of bioactive molecules, such as TNF-α-stimulated gene 6 protein (TSG-6). However, it is unknown whether BMSCs exert their analgesic effect against neuropathic pain by secreting TSG-6. Therefore, the present study aimed to evaluate the analgesic effects of TSG-6 released from BMSCs on neuropathic pain induced by chronic constriction injury (CCI) in rats and explored the possible underlying mechanisms in vitro and in vivo.MethodsBMSCs were isolated from rat bone marrow and characterized by flow cytometry and functional differentiation. One day after CCI surgery, about 5 × 106 BMSCs were intrathecally injected into spinal cerebrospinal fluid. Behavioral tests, including mechanical allodynia, thermal hyperalgesia, and motor function, were carried out at 1, 3, 5, 7, 14 days after CCI surgery. Spinal cords were processed for immunohistochemical analysis of the microglial marker Iba-1. The mRNA and protein levels of pro-inflammatory cytokines (IL-1β, TNFα, IL-6) were detected by real-time RT-PCR and ELISA. The activation of the TLR2/MyD88/NF-κB signaling pathway was evaluated by Western blot and immunofluorescence staining. The analgesic effect of exogenous recombinant TSG-6 on CCI-induced mechanical allodynia and heat hyperalgesia was observed by behavioral tests. In the in vitro experiments, primary cultured microglia were stimulated with the TLR2 agonist Pam3CSK4, and then co-cultured with BMSCs or recombinant TSG-6. The protein expression of TLR2, MyD88, p-p65 was evaluated by Western blot. The mRNA and protein levels of IL-1β, TNFα, IL-6 were detected by real-time RT-PCR and ELISA. BMSCs were transfected with the TSG-6-specific shRNA and then intrathecally injected into spinal cerebrospinal fluid in vivo or co-cultured with Pam3CSK4-treated primary microglia in vitro to investigate whether TSG-6 participated in the therapeutic effect of BMSCs on CCI-induced neuropathic pain and neuroinflammation.ResultsWe found that CCI-induced mechanical allodynia and heat hyperalgesia were ameliorated by intrathecal injection of BMSCs. Moreover, intrathecal administration of BMSCs inhibited CCI-induced neuroinflammation in spinal cord tissues. The analgesic effect and anti-inflammatory property of BMSCs were attenuated when TSG-6 expression was silenced. We also found that BMSCs inhibited the activation of the TLR2/MyD88/NF-κB pathway in the ipsilateral spinal cord dorsal horn by secreting TSG-6. Meanwhile, we proved that intrathecal injection of exogenous recombinant TSG-6 effectively attenuated CCI-induced neuropathic pain. Furthermore, in vitro experiments showed that BMSCs and TSG-6 downregulated the TLR2/MyD88/NF-κB signaling and reduced the production of pro-inflammatory cytokines, such as IL-1β, IL-6, and TNF-α, in primary microglia treated with the specific TLR2 agonist Pam3CSK4.ConclusionsThe present study demonstrated a paracrine mechanism by which intrathecal injection of BMSCs targets the TLR2/MyD88/NF-κB pathway in spinal cord dorsal horn microglia to elicit neuroprotection and sustained neuropathic pain relief via TSG-6 secretion.