Multinucleated giant cells of bladder mucosa are modified telocytes:Diagnostic and immunohistochemistry algorithm and relation to PDL1 expression score

BACKGROUND Multinucleated giant cells(MGCs)in bladder carcinomas are poorly studied.AIM To describe the function,morphogenesis,and origin of mononuclear and MGCs in urothelial carcinoma(UC)of the bladder in Bulgarian and French patients.METHODS Urothelial bladder carcinomas(n=104)from 2016-2020 were analyzed retrospectively using immunohistochemical(IHC)and histochemical stain examination.Giant cells in the bladder stroma were found in 35.6%of cases,more often in highgrades.RESULTS We confirm that MGCs in the mucosa in UC of the bladder were positive for both mesenchymal and myofibroblast markers(vimentin,smooth muscle actin,Desmin,and CD34)and the macrophage marker CD68.Furthermore,IHC studies revealed the following profile of these cells:Positive for p16;negative for epithelial(CK AE1/AE3 and GATA-3),vascular(CD31),neural(PS100 and CKIT),cambial,blastic(CD34-blasts and C-KIT),and immune markers(IG G,immunoglobulin G4,and PD-L1);no proliferative activity,possess no specific immune function,and cannot be used to calculate the Combined Positive Score scale.CONCLUSION In conclusion,the giant stromal cells in non-tumor and tumor bladder can be used as a characteristic and relatively constant,although nonspecific,histological marker for chronic bladder damage,reflecting the chronic irritation or inflammation.Likewise,according to the morphological and IHC of the mono-and multinucleated giant cells in the bladder,they are most likely represent telocytes capable of adapting their morphology to the pathology of the organ.

Human endogenous retroviral syncytin exerts inhibitory effect on invasive phenotype of B16F10 melanoma cells

Objective; Fusogenic endogenous retroviral syncytin plays an important role in the formation of syncytiotrophoblasts in human placenta.Apart from its expression in placenta,brain and testis,syncytin has also been found in many cancers.Although syncytin has been proposed to serve as a positive prognostic marker in some cancers,the underlying mechanism is unclear.The aim of this study is to evaluate the effects of syncytin expression on the invasive phenotype of melanoma cells.Methods：The eukaryotic expression plasmid for syncytin-EGFP was constructed and transfected into B16F10 melanoma cells.The effect of syncytin on the invasion potential of rumor cells was evaluated in B16F10 subline cells that stably expressed syncytin-EGFP fusion protein or EGFP alone.Results：The B16F10 sublines that stably expressed syncytin-EGFP or EGFP alone were established respectively and confirmed by immunofluorescent and immtmoblotting assay.Syncytin expression in B16F10 cells was associated with decreased cell proliferation,migration and invasion.Multinucleated giant cells that contained as many as five nuclei were induced in syncytin-expressing cells.In addition,syncytin expression did not alter the sensitivity of B16F10 cells to trichosanthin,a toxin that damages syncytiotrophoblasts more efficiently than other tissues.Conclusions：These results suggest that syncytin expression in some cancers may confine their invasion potential and thus serve as a positive prognostic factor.

Analysis of the in vitro degradation and the in vivo tissue response to bi-layered 3D-printed scaffolds combining PLA and biphasic PLA/bioglass components--Guidance of the inflammatory response as basis for osteochondral regeneration

The aim of the present study was the in vitro and in vivo analysis of a bi-layered 3D-printed scaffold combining a PLA layer and a biphasic PLA/bioglass G5 layer for regeneration of osteochondral defects in vivo Focus of the in vitro analysis was on the(molecular)weight loss and the morphological and mechanical variations after immersion in SBF.The in vivo study focused on analysis of the tissue reactions and differences in the implant bed vascularization using an established subcutaneous implantation model in CD-1 mice and established histological and histomorphometrical methods.Both scaffold parts kept their structural integrity,while changes in morphology were observed,especially for the PLA/G5 scaffold.Mechanical properties decreased with progressive degradation,while the PLA/G5 scaffolds presented higher compressive modulus than PLA scaffolds.The tissue reaction to PLA included low numbers of BMGCs and minimal vascularization of its implant beds,while the addition of G5 lead to higher numbers of BMGCs and a higher implant bed vascularization.Analysis revealed that the use of a bi-layered scaffold shows the ability to observe distinct in vivo response despite the physical proximity of PLA and PLA/G5 layers.Altogether,the results showed that the addition of G5 enables to reduce scaffold weight loss and to increase mechanical strength.Furthermore,the addition of G5 lead to a higher vascularization of the implant bed required as basis for bone tissue regeneration mediated by higher numbers of BMGCs,while within the PLA parts a significantly lower vascularization was found optimally for chondral regeneration.Thus,this data show that the analyzed bi-layered scaffold may serve as an ideal basis for the regeneration of osteochondral tissue defects.Additionally,the results show that it might be able to reduce the number of experimental animals required as it may be possible to analyze the tissue response to more than one implant in one experimental animal.

Covalent linkage of sulfated hyaluronan to the collagen scaffold Mucograft® enhances scaffold stability and reduces proinflammatory macrophage activation in vivo

Sulfated glycosaminoglycans(sGAG)show interaction with biological mediator proteins.Although collagen-based biomaterials are widely used in clinics,their combination with high-sulfated hyaluronan(sHA3)is unexplored.This study aims to functionalize a collagen-based scaffold(Mucograft®)with sHA3 via electrostatic(sHA3/PBS)or covalent binding to collagen fibrils(sHA3+EDC/NHS).Crosslinking without sHA3 was used as a control(EDC/NHS Ctrl).The properties of the sHA3-functionalized materials were characterized.In vitro growth factor and cytokine release after culturing with liquid platelet-rich fibrin was performed by means of ELISA.The cellular reaction to the biomaterials was analyzed in a subcutaneous rat model.The study revealed that covalent linking of sHA3 to collagen allowed only a marginal release of sHA3 over 28 days in contrast to electrostatically bound sHA3.sHA3+EDC/NHS scaffolds showed reduced vascular endothelial growth factor(VEGF),transforming growth factor beta 1(TGF-β1)and enhanced interleukin-8(IL-8)and epithelial growth factor(EGF)release in vitro compared to the other scaffolds.Both sHA3/PBS and EDC/NHS Ctrl scaffolds showed a high proinflammatory reaction(M1:CD-68+/CCR7+)and induced multinucleated giant cell(MNGC)formation in vivo.Only sHA3+EDC/NHS scaffolds reduced the proinflammatory macrophage M1 response and did not induce MNGC formation during the 30 days.SHA3+EDC/NHS scaffolds had a stable structure in vivo and showed sufficient integration into the implantation region after 30 days,whereas EDC/NHS Ctrl scaffolds underwent marked disintegration and lost their initial structure.In summary,functionalized collagen(sHA3+EDC/NHS)modulates the inflammatory response and is a promising biomaterial as a stable scaffold for full-thickness skin regeneration in the future.