Multipotent adult progenitor cells improve healing of mouse burn wounds

Chronic intestinal pseudo-obstruction (CIPO) is caused by a plethora of primary and secondary causes, dominantly involving the neuromuscular tissue, interstitial cells of Cajal, or the connective tissue framework. The lack of the connective tissue framework, known as desmosis, is evaluated by Masson's trichrome (MT) or picrosirius red stains, both of which are recommended in the London classification. We evaluated the orcein stain in detecting desmosis in comparison to the MT stain. We performed both orcein and MT stains in six previously published cases of complete or partial desmosis along with six age-matched controls. Our results showed comparable results of the orcein stain as compared to the MT stain. Additional advantages of lower cost and a clearer background in orcein stain were noteworthy, whereas MT stain can be used for the detection of additional pathology. We believe that orcein stain can be used as a cheap alternative in resource-limited settings.

Comparison of the transcriptomes of pluripotent embryonic stem cells, multipotent adult progenitor cells and lineage restricted mesenchymal stem cells identified a unique gene expression profile of multipotent adult progenitor cells.

What's known on the subject? and what does the study add?: A decrease in bladder compliance is known to be correlated with deterioration of renal function after bladder outlet obstruction (BOO). Increased deposition of extracellular matrix (ECM) in the detrusor layer is the primary reason for decreased compliance. In the bladder, as in other organs, ECM deposition is dependent on the balanced activity of proteolytic enzymes, including matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs). The imbalance between MMPs and TIMPs is a key regulator in ECM turnover. It has been shown that an altered proteolytic balance between MMP-1 and TIMP-1 favours accumulation of ECM and decreases tissue compliance in an ovine fetal BOO model. Also, MMP-1 was significantly down-regulated, while TIMP-1 levels were increased, in a time- and pressure-dependent manner in a smooth muscle cell (SMC) mechanical strain model. In the present study we measured the bladder compliance of control, sham-operated and partial BOO (PBOO) rabbits using a UDS-600 urodynamic testing machine. Collagen deposition between and within the detrusor SM bundles was evaluated using Masson's Trichrome stain and transmission electron microscopy. The expression levels of MMP-1 and TIMP-1 were evaluated by Western blot. We found that the imbalance between MMP-1 and TIMP-1 favours accumulation of extracellular collagen, the structural components associated with decreased bladder compliance after PBOO. To investigate the underlying mechanisms of bladder compliance after partial bladder outlet obstruction (PBOO) and the role of the collagen degradation enzyme matrix metalloproteinase-1 (MMP-1) and its endogenous inhibitor tissue inhibitor of metalloproteinase-1 (TIMP-1) during this process. Bladder compliance of control, sham-operated and PBOO rabbits was measured using a UDS-600 urodynamic testing machine. Collagen deposition between and within the detrusor smooth muscle bundles was evaluated using Masson's Trichrome stain and transmission electron microscopy. The expression levels of MMP-1 and TIMP-1 were evaluated by Western blot. Bladder compliance decreased after PBOO. Collagen deposition increased both between and within the detrusor smooth muscle bundles, and had an inverse relationship with bladder compliance. MMP-1 and TIMP-1 expression negatively correlated with bladder compliance. These findings indicate that the imbalance between MMP-1 and TIMP-1 favours accumulation of extracellular collagen, the structural components associated with decreased bladder compliance after PBOO.

The host immune response is essential for the beneficial effect of adult stem cells after myocardial ischemia

IntroductionWe have demonstrated previously that the intravenous delivery of multipotent adult progenitor cells (MAPC) after traumatic brain injury affords neuroprotection via interaction with splenocytes, leading to an increase in systemic anti-inflammatory cytokines. We hypothesize that the observed modulation of the systemic inflammatory milieu is related to T regulatory cells and a subsequent increase in the locoregional neuroprotective M2 macrophage population.MethodsC57B6 mice were injected with intravenous MAPC 2 and 24 hours after controlled cortical impact injury. Animals were euthanized 24, 48, 72, and 120 hours after injury. In vivo, the proportion of CD4+/CD25+/FOXP3+ T-regulatory cells were measured in the splenocyte population and plasma. In addition, the brain CD86+ M1 and CD206+ M2 macrophage populations were quantified. A series of in vitro co-cultures were completed to investigate the need for direct MAPC:splenocyte contact as well as the effect of MAPC therapy on M1 and M2 macrophage subtype apoptosis and proliferation.ResultsSignificant increases in the splenocyte and plasma T regulatory cell populations were observed with MAPC therapy at 24 and 48 hours, respectively. In addition, MAPC therapy was associated with an increase in the brain M2/M1 macrophage ratio at 24, 48 and 120 hours after cortical injury. In vitro cultures of activated microglia with supernatant derived from MAPC:splenocyte co-cultures also demonstrated an increase in the M2/M1 ratio. The observed changes were secondary to an increase in M1 macrophage apoptosis.ConclusionsThe data show that the intravenous delivery of MAPC after cortical injury results in increases in T regulatory cells in splenocytes and plasma with a concordant increase in the locoregional M2/M1 macrophage ratio. Direct contact between the MAPC and splenocytes is required to modulate activated microglia, adding further evidence to the central role of the spleen in MAPC-mediated neuroprotection.

27 - Multipotent Adult Progenitor Cells

Effects of Octreotide on Activated Pancreatic Stellate Cell-Induced Pancreas Graft Fibrosis in Rats

Perivascular CD45−:Sca-1+:CD34− Cells Are Derived from Bone Marrow Cells and Participate in Dystrophic Skeletal Muscle Regeneration.

The healing of diabetic wounds is poor due to a collagen deposition disorder. Matrix metalloproteinase-9 (MMP-9) is closely related to collagen deposition in the process of tissue repair. Many studies have demonstrated that extracellular vesicles derived from adipose-derived stem cells (ADSC-EVs) promote diabetic wound healing by enhancing collagen deposition. In this study, we explored whether ADSC-EVs could downregulate the expression of MMP-9 in diabetic wounds and promote wound healing by improving collagen deposition. The potential effects of ADSC-EVs on MMP-9 and diabetic wound healing were tested both in vitro and in vivo. We first evaluated the effect of ADSC-EVs on the proliferation and MMP-9 secretion of HaCaT cells treated with advanced glycation end product-bovine serum albumin (AGE-BSA) using CCK-8, western blot and MMP-9 enzyme-linked immunosorbent assay(ELISA). Next, the effects of ADSC-EVs on healing, re-epithelialisation, collagen deposition, and MMP-9 concentration in diabetic wound fluids were evaluated in an immunodeficient mouse model via MMP-9 ELISA and haematoxylin and eosin, Masson's trichrome, and immunofluorescence staining for MMP-9. In vitro, ADSC-EVs promoted the proliferation and MMP-9 secretion of HaCaT cells. In vivo, ADSC-EVs accelerated diabetic wound healing by improving re-epithelialisation and collagen deposition and by inhibiting the expression of MMP-9. ADSC-EVs possess the potential of healing of diabetic wounds in a mouse model by inhibiting downregulating MMP-9 and improving collagen deposition. Thus, ADSC-EVs are a promising candidate for the treatment of diabetic wounds.

BackgroundSystemic scleroderma (SSc) is an acquired disorder characterized by excessive deposition of extracellular matrix in the skin and internal organs. So far, the molecular mechanisms underpinning the pathogenesis of SSc have remained unknown. Collagen triple helix repeat containing-1 (CTHRC1) has been indicated to be a cell type-specific inhibitor of transforming growth factor-β (TGF-β), which could have the potential for extensive clinical application owing to its ability to reduce collagen deposition. Our previous studies showed that CTHRC1 inhibited TGF-β1-induced collagen type I synthesis in keloid fibroblasts. In our present research, we attempted to probe the role of CTHRC1 in dermal fibrosis in bleomycin (BLM)-treated mice.MethodsCTHRC1 and TGF-β1 expression was detected in dermal tissues from patients with SSc and BLM-treated mice by immunohistochemistry. A 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay was used to assess TGF-β1-induced proliferation of human dermal fibroblasts. Collagen expression and fibroblast synthesis were evaluated by quantitative real-time polymerase chain reaction and the 3H-proline incorporation. Masson’s trichrome staining and western blotting were carried out to analyze the deposits and protein levels of type I collagen, respectively.ResultsCompared with those in normal tissues, the levels of CTHRC1 and TGF-β1 were elevated in dermal tissues from patients with SSc and in skin tissues from BLM-treated mice, respectively. Furthermore, recombinant CTHRC1 was found to inhibit TGF-β1-stimulated collagen deposition by fibroblasts. Finally, the in vivo experiments showed that CTHRC1 alleviated BLM-induced dermal fibrotic changes.ConclusionsCTHRC1 can inhibit human dermal fibroblast collagen deposition and can also exert protective effects against BLM-induced dermal fibrosis in mice. This research provides an indication that CTHRC1 may be a promising treatment choice for dermal fibrosis in SSc patients.

Objective: The primary objective of this study was to observe the impact of chronic sleep deprivation on the histomorphological features of the rat prostate gland and to evaluate the potential protective effects of omega-3 fatty acid supplementation. Materials & Methods: Thirty male Sprague-Dawley rats (200-300g) were randomly divided into three groups (n=10 per group): Group A (control, maintained on a normal sleep-wake cycle), Group B (sleep deprivation, subjected to 16 hours of sleep deprivation followed by an 8-hour sleep opportunity daily for 60 days), and Group C (sleep deprivation + omega-3, subjected to the same sleep deprivation regimen as Group B, supplemented with 260 mg/kg/day omega-3 fatty acid via oral gavage). Post-euthanasia and after dissection, prostate tissues were processed for histological observation using H&E and Masson's trichrome staining to assess epithelial apoptosis, collagen deposition, and smooth muscle architecture. Statistical analyses were performed using SPSS software version 21, with p<0.05 considered statistically significant. Results: Group A (control) showed normal histological structure. Group B (sleep deprivation) exhibited a significantly higher frequency of epithelial apoptosis (p<0.05), increased interacting collagen deposition (p<0.001), and marked irregularities in the smooth muscle layer surrounding the acini (p=0.002). Group C (sleep deprivation + omega-3) demonstrated reduced epithelial apoptosis, collagen deposition, and smooth muscle distortion relative to Group B. Conclusion: The present study observed that sleep deprivation caused significant changes in the histomorphology of the rat prostate gland, including epithelial apoptosis, collagen fibre deposition in the interacting space, and irregular arrangement of the smooth muscle fibres around the acini. Additionally, omega-3 fatty acid supplementation had an ameliorative effect on these histomorphological alterations induced by sleep deprivation.

Acute respiratory distress syndrome (ARDS) patients currently have relatively high mortality, which is associated with early lung fibrosis. This study aimed to investigate whether miR-17 suppression could alleviate ARDS-associated lung fibrosis by regulating Mfn2. A mouse model of ARDS-related lung fibrosis was constructed via intratracheal instillation of bleomycin. The expression level of miR-17 in lung tissues was detected via quantitative real time polymerase chain reaction (qRT-PCR). In the ARDS mouse model of lung fibrosis, the mitigating effects of miR-17 interference were evaluated via tail vein injection of the miR negative control or the miR-17 antagomir. The pathological changes in the lung tissue were examined via HE staining and Masson's trichrome staining, and the underlying molecular mechanism was investigated via ELISA, qRT-PCR and Western blotting. Bleomycin-induced pulmonary fibrosis significantly increased collagen deposition and the levels of hydroxyproline (HYP) and miR-17. Interfering with miR-17 significantly reduced the levels of HYP and miR-17 and upregulated the expression of Mfn2. The intravenous injection of the miR-17 antagomir alleviated lung inflammation and reduced collagen deposition. In addition, interference with miR-17 could upregulate LC3B expression, downregulate p62 expression, and improve mitochondrial structure. Interfering with miR-17 can improve pulmonary fibrosis in mice by promoting mitochondrial autophagy via Mfn2.

The endovascular technique of mechanochemical ablation (MOCA) has become popular in treating patients with saphenous reflux. We reported the histopathological findings in human ex-vivo incompetent great saphenous veins following treatment with saline, polidocanol, mechanical ablation and MOCA using ClariVein device. Twenty-four vein GSV specimens were obtained via traditional surgery and treated with four methods: Group A: 0.9% normal saline (NS); Group B: 3% polidocanol; Group C: mechanical ablation + 0.9% NS; Group D: mechanical ablation + 3% polidocanol (MOCA). Hematoxylin and eosin (HE), Masson's trichrome and immunohistochemical staining were performed on each specimen and integrated optical densities were measured with vWF and a-SMA stains and statistically evaluated. vWF staining was used to assess endothelial damage and a a-SMA staining was used to assess media injury. HE and Masson's trichrome staining of Groups C and D revealed severe damage to the endothelium and media compared to Groups A and B. The statistical result of vWF staining showed the damage of endothelium was significantly increased by Group D compared to Groups A, B and C. The statistical result of a-SMA staining showed the damage of media was significantly increased by Groups C and D compared to Groups A and B. The mechanism of MOCA was caused by both endothelium damage and media tearing. The damage of endothelium was significantly increased by MOCA when compared with mechanical ablation alone.

Altered smooth muscle development and innervation in the lower genitourinary and gastrointestinal tract of the male human fetus with myelomeningocele.

We have derived from normal human, mouse, and rat postnatal bone marrow primitive, multipotent adult progenitor cells (MAPCs) that can differentiate into most mesodermal cells and neuroectodermal cells in vitro and into all embryonic lineages in vivo.Here, we show that MAPCs can also differentiate into hepatocyte-like cells in vitro.Human, mouse, and rat MAPCs, cultured on Matrigel with FGF-4 and HGF, differentiated into epithelioid cells that expressed hepatocyte nuclear factor-3 (HNF-3), GATA4, cytokeratin 19 (CK19), transthyretin, and -fetoprotein by day 7, and expressed CK18, HNF-4, and HNF-1 on days 14-28.Virtually all human, as well as a majority of rodent cells stained positive for albumin and CK18 on day 21; 5% (rodent) to 25% (human) cells were binucleated by day 21.These cells also acquired functional characteristics of hepatocytes: they secreted urea and albumin, had phenobarbital-inducible cytochrome p450, could take up LDL, and stored glycogen.MAPCs, which can be expanded in vitro and maintained in an undifferentiated state for more than 100 population doublings, can thus differentiate into cells with morphological, phenotypic, and functional characteristics of hepatocytes.MAPCs may therefore be an ideal cell for in vivo therapies for liver disorders or for use in bioartificial liver devices.