BACKGROUND: Inflammation-induced testicular damage is a significant contributing factor to the increasing incidence of infertility. Traditional treatments during the inflammatory phase often fail to achieve the desired fertility outcomes, necessitating innovative interventions such as cell therapy. METHODS: We explored the in vivo properties of intravenously administered Sertoli cells (SCs) in an acute lipopolysaccharide (LPS)-induced inflammatory mouse model. Infiltrating and resident myeloid cell phenotypes were assessed using flow cytometry. The impact of SC administration on testis morphology and germ cell quality was evaluated using computer-assisted sperm analysis (CASA) and immunohistochemistry. RESULTS: SCs demonstrated a distinctive migration pattern, importantly they preferentially concentrated in the testes and liver. SC application significantly reduced neutrophil infiltration as well as preserved the resident macrophage subpopulations. SCs upregulated MerTK expression in both interstitial and peritubular macrophages. Applied SC treatment exhibited protective effects on sperm including their motility and kinematic parameters, and maintained the physiological testicular morphology. CONCLUSION: Our study provides compelling evidence of the therapeutic efficacy of SC transplantation in alleviating acute inflammation-induced testicular damage. These findings contribute to the expanding knowledge on the potential applications of cell-based therapies for addressing reproductive health challenges and offer a promising approach for targeted interventions in male infertility.

• Klíčová slova
• Inflammation, Macrophages, Sertoli cells, Sperm, Testes,
• MeSH
• lipopolysacharidy toxicita   MeSH
• makrofágy metabolismus   MeSH
• motilita spermií MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• Sertoliho buňky * metabolismus   MeSH
• spermie * metabolismus   MeSH
• testis MeSH
• tyrosinkinasa c-Mer metabolismus   genetika   MeSH
• zánět * patologie   terapie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• lipopolysacharidy MeSH
• Mertk protein, mouse MeSH Prohlížeč
• tyrosinkinasa c-Mer MeSH

Background. The treatment of middle ear cholesteatoma requires surgical treatment and the reconstruction of the temporal bone, which represents an ongoing problem. Otologists have focused on the research of materials allowing an airy middle ear and the preservation of hearing function to reconstruct the temporal bone. Methods. This study evaluated the effect of cyclosporin A (CsA) and a combined biomaterial in the healing process of postoperative temporal bone defects in an animal model. Cultured human Bone Marrow Mesenchymal Stromal Cells (hBM-MSCs) were mixed with hydroxyapatite (Cem-Ostetic®), and subsequently applied as a bone substitute after middle ear surgery, showing that the therapeutic potential of hBM-MSCs associated with bone regeneration and replacement is directly influenced by CsA, confirming that it promotes the survival of MSCs in vivo. Results. The therapeutic efficacy of the combination of MSCs with CsA is greater than the sole application of MSCs in a hydroxyapatite carrier. Conclusion. The reconstruction of a temporal bone defect using hBM-MSCs requires an immunosuppressant to improve the results of treatment.

• Klíčová slova
• cholesteatoma, cyclosporin A, mesenchymal stem cells, osteogenesis, temporal bone,
• Publikační typ
• časopisecké články MeSH

It is becoming increasingly evident that selecting an optimal source of mesenchymal stromal cells (MSCs) is crucial for the successful outcome of MSC-based therapies. During the search for cells with potent regenerative properties, Sertoli cells (SCs) have been proven to modulate immune response in both in vitro and in vivo models. Based on morphological properties and expression of surface markers, it has been suggested that SCs could be a kind of MSCs, however, this hypothesis has not been fully confirmed. Therefore, we compared several parameters of MSCs and SCs, with the aim to evaluate the therapeutic potential of SCs in regenerative medicine. We showed that SCs successfully underwent osteogenic, chondrogenic and adipogenic differentiation and determined the expression profile of canonical MSC markers on the SC surface. Besides, SCs rescued T helper (Th) cells from undergoing apoptosis, promoted the anti-inflammatory phenotype of these cells, but did not regulate Th cell proliferation. MSCs impaired the Th17-mediated response; on the other hand, SCs suppressed the inflammatory polarisation in general. SCs induced M2 macrophage polarisation more effectively than MSCs. For the first time, we demonstrated here the ability of SCs to transfer mitochondria to immune cells. Our results indicate that SCs are a type of MSCs and modulate the reactivity of the immune system. Therefore, we suggest that SCs are promising candidates for application in regenerative medicine due to their anti-inflammatory and protective effects, especially in the therapies for diseases associated with testicular tissue inflammation.

• Klíčová slova
• Mesenchymal stem cells. Sertoli cells. Immunomodulation. Mitochondrial transfer,
• MeSH
• antiflogistika MeSH
• imunita MeSH
• lidé MeSH
• mezenchymální kmenové buňky * MeSH
• mitochondrie MeSH
• Sertoliho buňky * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antiflogistika MeSH

PURPOSE: The present study aims to prepare poly(D,L-lactic acid) (PLA) nanofibers loaded by the immunosuppressant cyclosporine A (CsA, 10 wt%). Amphiphilic poly(ethylene glycol)s (PEG) additives were used to modify the hydrophobic drug release kinetics. METHODS: Four types of CsA-loaded PLA nanofibrous carriers varying in the presence and molecular weight (MW) of PEG (6, 20 and 35 kDa) were prepared by needleless electrospinning. The samples were extracted for 144 h in phosphate buffer saline or tissue culture medium. A newly developed and validated LC-MS/MS method was utilized to quantify the amount of released CsA from the carriers. In vitro cell experiments were used to evaluate biological activity. RESULTS: Nanofibers containing 15 wt% of PEG showed improved drug release characteristics; significantly higher release rates were achieved in initial part of experiment (24 h). The highest released doses of CsA were obtained from the nanofibers with PEG of the lowest MW (6 kDa). In vitro experiments on ConA-stimulated spleen cells revealed the biological activity of the released CsA for the whole study period of 144 h and nanofibers containing PEG with the lowest MW exhibited the highest impact (inhibition). CONCLUSIONS: The addition of PEG of a particular MW enables to control CsA release from PLA nanofibrous carriers. The biological activity of CsA-loaded PLA nanofibers with PEG persists even after 144 h of previous extraction. Prepared materials are promising for local immunosuppression in various medical applications.

• Klíčová slova
• LC-MS/MS, cyclosporine A, drug release kinetics, poly(D,L-lactic acid) nanofibers, poly(ethylene glycol),
• MeSH
• buněčné linie MeSH
• cyklosporin aplikace a dávkování   chemie   MeSH
• hydrofobní a hydrofilní interakce MeSH
• imunosupresiva aplikace a dávkování   chemie   MeSH
• kinetika MeSH
• kultivační média MeSH
• lidé MeSH
• nanovlákna chemie   MeSH
• nosiče léků MeSH
• polyestery chemie   MeSH
• polyethylenglykoly chemie   MeSH
• povrchové vlastnosti MeSH
• slezina cytologie   MeSH
• techniky tkáňových kultur MeSH
• uvolňování léčiv MeSH
• velikost částic MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyklosporin MeSH
• imunosupresiva MeSH
• kultivační média MeSH
• nosiče léků MeSH
• poly(lactide) MeSH Prohlížeč
• polyestery MeSH
• polyethylenglykoly MeSH

Immunosuppressive drugs are widely used to treat undesirable immune reaction, however their clinical use is often limited by harmful side effects. The combined application of immunosuppressive agents with mesenchymal stem cells (MSCs) offers a promising alternative approach that enables the reduction of immunosuppressive agent doses and simultaneously maintains or improves the outcome of therapy. The present study aimed to determinate the effects of immunosuppressants on individual T cell subpopulations and to investigate the efficacy of MSC-based treatment combined with immunosuppressive drugs. We tested the effect of five widely used immunosuppressants with different action mechanisms: cyclosporine A, mycophenolate mofetil, rapamycin, and two glucocorticoids - prednisone and dexamethasone in combination with MSCs on mouse CD4+ and CD8+ lymphocyte viability and activation, Th17 (RORγt+), Th1 (T-bet+), Th2 (GATA-3+) and Treg (Foxp3+) cell proportion and on the production of corresponding key cytokines (IL-17, IFNγ, IL-4 and IL-10). We showed that MSCs modulate the actions of immunosuppressants and in combination with immunosuppressive drugs display distinct effect on cell activation and balance among different T lymphocytes subpopulations and exert a suppressive effect on proinflammatory T cell subsets while promoting the functions of anti-inflammatory Treg lymphocytes. The results indicated that MSC-based therapy could be a powerful strategy to attenuate the negative effects of immunosuppressive drugs on the immune system.

• Klíčová slova
• Immunomodulation, Immunosuppressive drugs, Mesenchymal stem cells, Stem cell therapy, T cells,
• MeSH
• aktivace lymfocytů účinky léků   imunologie   MeSH
• cyklosporin farmakologie   MeSH
• cytokiny metabolismus   MeSH
• dexamethason farmakologie   MeSH
• glukokortikoidy farmakologie   MeSH
• imunosupresiva farmakologie   MeSH
• kokultivační techniky MeSH
• kultivované buňky MeSH
• kyselina mykofenolová farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   imunologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• prednison farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• průtoková cytometrie MeSH
• sirolimus farmakologie   MeSH
• T-lymfocyty - podskupiny cytologie   účinky léků   imunologie   MeSH
• transplantace mezenchymálních kmenových buněk metody   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyklosporin MeSH
• cytokiny MeSH
• dexamethason MeSH
• glukokortikoidy MeSH
• imunosupresiva MeSH
• kyselina mykofenolová MeSH
• prednison MeSH
• sirolimus MeSH