Histology and histopathology Vol.33,nº12 (2018)
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- PublicationOpen AccessTreatment of osteoarthritis with collagen-based scaffold: A porcine animal model with xenograft mesenchymal stem cells(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Tseng, Wo Jan; Huang, Shu Wei; Fang, Chih Hsiang; Hsu, Lih Tao; Chen, Chih Yu; Shen, Hsin Hsin; Zwei Chieng Chang, Jenny; Sun, Jui Sheng; Lin, Feng HueiObjective. With the goal to explore a new approach to treat the early degenerative lesions of hyaline cartilage, we implanted in a porcine OA model a collagen-based scaffold containing chondroprogenitor cells derived from human bone marrow mesenchymal stem cells (hBM-MSCs). Experimental design. Porcine knee joints were subjected to anterior cruciate ligament (ACL) transection to surgically induce OA. After 4 months, the time necessary for the development of cartilage surface damage, animals were treated either with trephination bone plug wrapped with the chondroprogenic hBM-MSCs-embedded collagen scaffold or microfractures alone. Histological and histomorphometric evaluations were performed at 5 months after surgery. Results. All animals subjected to ACL transection showed osteoarthritic changes including mild lateral femoral condyle or moderate medial femoral condyle ulcerations. After 14 days’ chondrogenic induction, hBM-MSCs seeded onto the scaffold showed expression of chondroprogenitor markers such as SOX9 and COMP. At 5 months after the implantation, significant differences in the quality of the regenerated tissue were found between the hBM-MSCsembedded scaffold group and the control group. Newly generated tissue was only observed at the site of implantation with the hBM-MSCs-embedded scaffolds. Furthermore, histological examination of the generated tissue revealed evidence of cartilage-like tissue with lacuna formation. In contrast, fibrous layers or fissures were formed on the surface of the control knee joint. Conclusions. This study shows that xenogenic hBMMSC derived chondroprogenitor scaffolds can generate new cartilage tissue in porcine articular cartilage and have the potential as a useful treatment option for osteoarthritis.
- PublicationOpen AccessPhenytoin activates SMAD3 phosphorylation and periostin expression in drug-induced gingival enlargement(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Kim, Shawna S.; Nikoloudaki, Georgia; Darling, Mark; Rieder, Michael J.; Hamilton, Douglas W.Drug-induced gingival enlargement (DIGE) is a fibrotic condition associated with systemic administration of the anti-epileptic drug, phenytoin. We have previously demonstrated that periostin, which is transforming growth factor-beta (TGF-β) inducible gene, is upregulated in various fibrotic conditions including gingival enlargement associated with nifedipine. The objective of this study was to assess periostin expression in phenytoin-induced gingival enlargement (PIGE) tissues and to investigate the mechanisms underlying periostin expression. Human PIGE tissues were assessed using Masson’s trichrome, with cell infiltration and changes in extracellular matrix composition characterized through labeling with antibodies to periostin, phospho-SMAD 3, TGF-β, as well as the macrophage markers CD68 and RM3/1. Using human gingival fibroblasts (HGFs) in vitro, we examined the pathways through which phenytoin acts on fibroblasts. In PIGE tissues, which demonstrate altered collagen organization and increased inflammatory cell infiltration, periostin protein was increased compared with healthy tissues. p-SMAD2/3, the transcription factor associated with canonical TGF-β signaling, is localized to the nuclei in both gingival fibroblasts and oral epithelial cells in PIGE tissues, but not in healthy tissue. In vitro culture of HGFs with 15 and 30 μg/ml of phenytoin increased periostin protein levels, which correlated with p-SMAD3 phosphorylation. Inhibition of canonical TGF-β signaling with SB431542 significantly reduced phenytoin induction of SMAD3 phosphorylation and periostin expression in HGFs. Analysis of PIGE tissues showed a subset of CD68 stained macrophages were TGF-β positive and that RM1/3 regenerative macrophages were present in the tissues. Our results demonstrate that phenytoin up-regulates periostin in HGFs in a TGF-β-dependent manner.
- PublicationOpen AccessMesenchymal stem cells: from the perivascular environment to clinical applications(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Vezzani, Bianca; Pierantozzi, Enrico; Sorrentino, VincenzoAdult stem cells represent a fundamental biological system that has fascinated scientists over the last decades, and are currently the subject of a large number of studies aimed at better defining the properties of these cells, with a prominent focus on improving their application in regenerative medicine. One of the most used adult stem cells in clinical trials are mesenchymal stem cells (MSCs), which are multipotent cells able to differentiate into mature cells of mesodermal lineages. Following the initial studies on MSCs isolated from bone marrow, similar cells were also isolated from a variety of fetal and adult human tissues. Initially considered as identical and equipotent, MSCs from tissues other than bone marrow actually display differences in terms of their plastic abilities, which can be ascribed to the tissue of origin and/or to the procedures used for their isolation. Moreover, results from additional studies suggest that cultured MSCs represent the in vitro version of a subset of in vivo resident cells localized in the perivascular environment. In this review, we will focus our attention on MSCs from tissues other than bone marrow, their in vivo localization and their current applications in clinics.
- PublicationOpen AccessCancer stem cells: A new target for cancer therapy(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Li, Qianhui; Wu, Jiayu; Chen, Shuo; Zhao, YangThe introduction of the theory of cancer stem cells (CSCs) has provided a new direction and perspective for our understanding of the nature and origin of tumors. Cancer stem cells are believed to be responsible for the treatment failure, drug tolerance, metastasis, and recurrence of tumors. However, it remains a challenge to identify or isolate tumor stem cells and determine their regulatory mechanisms. Therefore, further understanding of the biological characteristics and functions of CSCs is of great practical significance and value to develop new methods of tumor diagnosis and treatment, and may bring new hope patients with cancer
- PublicationOpen AccessInterplay between metalloproteinases and cell signalling in blood brain barrier integrity(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Pla Navarro, Irene; Bevan, Damon; Hajihosseini, Mohammad K.; Lee, Martin; Gavrilovic, JelenaThe Blood-Brain Barrier (BBB) is a highly specialised interface separating the Central Nervous System (CNS) from circulating blood. Dysregulation of the BBB is a key early event in pathological conditions such as inflammation, in which the entry of activated leukocytes into the CNS is facilitated by BBB breakdown. The metzincin family of metalloproteinases (MPs) is one of the major contributors to BBB permeability as they cleave endothelial cell-cell contacts and underlying basal lamina components. However, the mechanisms by which MPs regulate BBB integrity has not yet been fully elucidated. The aim of this review is to provide an overview of pathways by which MPs could regulate the BBB in the context of neuroinflammation.
- PublicationOpen AccessThe multilayered structure of the human corpus spongiosum(2018) De Graaf, P.; Ramadan, R.; Linssen, E.C.; Staller, N.A.; Hendrickx, A.P.A.; Pigot, G.L.S.; Meuleman, E.J.H.; Bouman, M.; Özer, M.; Bosch, J.L.H.R.; de Kort, L.M.O.Purpose. Urethral reconstruction is performed in patients with urethral strictures or for correction of congenital disorders. In the case of shortage of tissue, engineered tissue may enhance urethral reconstruction. As the corpus spongiosum (CS) is important in supporting the function of the urethra, tissue engineering of the urethra should be combined with reconstruction of a CS. For that purpose, detailed knowledge of the composition of the CS, more specifically its extracellular matrix (ECM) and vascularization is needed for scaffold design. The objective of this study is to analyze the microarchitecture of the CS through (immuno) histology and scanning electron microscopy (SEM). Methods. The CS including the urethra of patients undergoing male-to-female genital confirming surgery was harvested. This CS was fixed and processed for either (immuno) histology or for SEM. Results. Four layers could be distinguished in the CS; first a transition zone from urethra epithelium to a collagen rich layer, which was highly vascularized, followed by a second, elastin rich layer. The third layer was formed by veins, arteries and vascular spaces and the last layer showed the transition from this vascular rich region to the collagen rich tunica albuginea. In this layer collagen bundles intertwined with elastic fibres. In the CS different components of the ECM were visible and distinguishable. Conclusion. This study provides novel and detailed information on the microarchitecture of the CS and the distribution of vascularization, which is important for scaffold design in tissue engineering.
- PublicationOpen AccessHistopathological alterations in mice under sub-acute treatment with Hintonia latiflora methanolic stem bark extract(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Flores Jiménez, Nancy G.; Rojas Lemus, Marcela; Fortoul, Teresa I.; Zepeda Rodríguez, Armando; López Camacho, Perla Y.; Anacleto Santos, Jhony; Malagón Gutiérrez, Filiberto; Basurto Islas, Gustavo; Rivera Fernández, NormaThe indiscriminate use of herbal products is increasingly growing worldwide; nonetheless consumers are not warned about the potential health risks that these products may cause. Hintonia latiflora (Hl) is a tree native to the American continent belonging to the Rubiaceae family and its stem bark is empirically used mainly to treat diabetes and malaria; supplements containing Hl are sold in America and Europe without medical prescription, thus scientific information regarding its toxicity as a consequence of a regular consumption is needed. In the present study, the histopathological effect of 200 and 1000 mg/kg of Hl methanolic stem bark extract (HlMeOHe) was evaluated in the small bowel, liver, pancreas, kidneys and brain of CD-1 male mice after oral sub-acute treatment for 28 days. No histopathological alterations were observed in the brain of the treated animals; however, mice presented diarrhea from day 2 of treatment with both doses. No histological changes were observed in the tissues collected from the animals treated with 200 mg/kg, except for the liver that depicted periportal hepatitis. Animals treated with the higher dose showed in the liver sections hydropic degeneration, hepatitis and necrosis small bowel sections showed dilated mucosal vessels, kidney sections depicted tubular necrosis and in pancreas sections, hydropic degeneration of the pancreatic islets was observed. In conclusion, HlMeOHe damaged the liver with an oral dose of 200 mg/kg, and at 1000 mg/kg injured the kidneys and pancreas of the CD1 male mice.
- PublicationOpen AccessAltered myelination in the Niemann-Pick type C1 mutant mouse(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Qiao, Liang; Yang, Enhui; Luo, Jiankai; Lin, Juntang; Yan, XinNiemann–Pick type C1 (NPC1) disease is a lysosomal storage disorder caused by mutation of Npc1 or Npc2 gene, resulting in various progressive pathological features. Myelin defection is a major pathological problem in Npc1 mutant mice; however, impairment of myelin proteins in the developing brain is still incompletely understood. In this study, we showed that the expression of myelin genes and proteins is strongly inhibited from postnatal day 35 onwards including reduced myelin basic protein (MBP) expression in the brain. Furthermore, myelination characterized by MBP immunohistochemistry was strongly perturbed in the forebrain, moderately in the midbrain and cerebellum, and slightly in the hindbrain. Our results demonstrate that mutation of the Npc1 gene is sufficient to cause severe and progressive defects in myelination in the mouse brain.
- PublicationOpen AccessPRP and MSCs on tenocytes artificial wound healing: an in vitro study comparing fresh and frozen PRP(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Veronesi, Francesca; Pagani, Stefania; Torricelli, Paola; Filardo, Giuseppe; Cavallo, Carola; Grigolo, Brunella; Fini, MilenaTendon tissue has poor regenerative capacity due to its low vascularization, cell density and extracellular matrix (ECM) production. Therefore, tendon injuries are an increasing clinical problem because of the formation of scar tissue with traditional therapies. Regenerative medicine aims at triggering a healing response through the use of biological treatments such as mesenchymal stromal cells (MSCs) and growth factors (GFs). MSCs show several advantages in tendon clinical setting, while platelet rich plasma (PRP) has gained popularity because of its high GF concentration, although its applications in the tendon clinical setting are still controversial. The aim of the present study was to evaluate a combined treatment of MSCs and PRP in an in vitro microwound model of tendon injuries. In addition, fresh and frozen PRP were compared. Single human tenocytes cultures or co-cultures with bone marrow derived MSCs (BMSCs) were set up with or without human PRP, fresh or frozen. After 24 hours of culture, it was observed that MSCs alone significantly increased tenocyte migration speed, microwound healing rate, fibronectin, collagen I and aggrecan production. These effects were enhanced by the combination with PRP, fresh being more effective than frozen PRP. In addition, the number of MSCs and tenocytes inside the microwound was significantly increased, especially with fresh PRP. In conclusion, the combination of MSCs and PRP, especially the fresh one, increases tenocytes and MSC migration speed, as well as ECM protein production compared to the use of MSCs alone