Efekti interleukina-17 na funkcionalna svojstva humanih mezenhimskih matičnih ćelija periodoncijuma

In the last few decades regenerative and modulatory properties of mesenchymal stem cells (MSCs) have gained the attention of scientists since their characteristics indicated the possibility of their use in regenerative medicine and cell therapy. Although MSCs isolated from various adult and neonatal tissues exhibit the potential of self-renewal and multipotent differentiation (towards cells of mesodermal, ectodermal and endodermal origin), many aspects concerning their functional characteristics are yet to be explored. One of the recently discovered sources of MSCs is easily accessible tissue of periodontal ligament, obtained after tooth extraction for orthodontic reasons. Periodontal ligament tissue strengthens the root of the tooth in the alveolar bone. The regenerative potential of periodontal ligament MSCs (PDLSCs) is reflected in self-renewal and multipotent differentiation ability, as well as potency for the formation of tissues which support the teeth, including the periodontal ligament and cement. These features make the PDLSCs good candidates for use in the regeneration/reconstruction of supporting tissue of teeth in periodontal disease, a chronic inflammatory disease caused by bacterial plaque. The development of chronic periodontal disease includes the process of bone tissue remodeling governed by a number of factors, including osteoblasts and osteoclasts, which reciprocally interact with cellular and humoral compartments of the immune system, thus regulating inflammation, degradation and renewal of bone tissue. The role of the local microenvironment in the periodontal disease is one of the key factors in the context of periodontal ligament tissue response to persistent infection. The presence of T helper cells17 (Th17) cells, important regulators of tissue destruction during inflammatory processes has been confirmed in periodontal lesions. Th17 cells produce proinflammatory cytokine interleukin (IL)-17. In the context of periodontal disease, as well as in other inflammatory diseases, IL-17 stimulates the activation and migration of neutrophils to the site of inflammation. Additionally, IL-17 directly stimulates differentiation of osteoclasts and contributes to the alveolar bone resorption...Istraživanja regenerativnih i modulatornih uloga mezenhimskih matičnih ćelija (MMĆ) u poslednjih nekoliko decenija su zaokupirala pažnju naučnika s obzirom da ove ćelije ispoljavaju karakteristike koje ukazuju na mogućnosti njihove primene u regenerativnoj medicini, kao i na polju ćelijske terapije. Iako MMĆ izolovane iz različitih adultnih i neonatalnih izvora ispoljavaju potencijal samoobnove i multipotentne diferencijacije (u ćelije mezodermalnog, ali i ektodermalnog i endodermalnog porekla), mnoga istraživanja koja se tiču funkcionalnih karakteristika ovih ćelija tek predstoje kako bi njihova primena bila efikasna i bezbedna. Jedan od novije otkrivenih izvora MMĆ je i tkivo periodoncijuma koje predstavlja bogat i lako dostupan izvor MMĆ, s obzirom da se ovo tkivo dobija nakon ekstrakcije zuba iz ortodontskih razloga. Periodoncijum je vezivno tkivo čija je glavna uloga uspostavljanje čvrste veze između korena zuba i alveolarne kosti. Regenerativni potencijal MMĆ periodoncijuma (PD-MMĆ) ogleda se u njihovom potencijalu samoobnove i multipotentne diferencijacije, kao i sposobnosti formiranja tkiva sličnih potpornom tkivu zuba, uključujući periodoncijum i cement. Ove osobine čine PD-MMĆ dobrim kandidatima za primenu u regeneraciji potpornog tkiva zuba u parodontopatiji, hroničnom inflamatornom oboljenju uzrokovanom bakterijama zubnog plaka. Razvoj hronične parodontopatije podrazumeva procese remodelovanja koštanog tkiva pod uticajem brojnih faktora u koje su uključeni osteoblasti i osteoklasti koji ostvaruju recipročne interakcije sa ćelijama imunskog odgovora regulišući inflamaciju i degradaciju koštanog tkiva. Uloga lokalne mikrosredine u parodontopatiji predstavlja jedan od ključnih faktora u kontekstu ćelijskog odgovora tkiva periodoncijuma na perzistentnu infekciju. U okviru parodontalnih lezija potvrđeno je prisustvo Th17 (od engl. T helper cells17) ćelija, značajnih regulatora tkivnog razaranja u toku zapaljenskih procesa, koje produkuju proinflamatorni citokin interleukin (IL)-17. U kontekstu parodontopatije, do sada je pokazano da IL-17 ostvaruje svoju ulogu stimulišući privlačenje neutrofila na mesto inflamacije kao i direktnim stimulatornim delovanjem Th17 ćelija na diferencijaciju osteoklasta..

Effects of interleukin-17 on functional properties of human periodontal ligament mesenchymal stem cells

Istraživanja regenerativnih i modulatornih uloga mezenhimskih matičnih ćelija (MMĆ) u poslednjih nekoliko decenija su zaokupirala pažnju naučnika s obzirom da ove ćelije ispoljavaju karakteristike koje ukazuju na mogućnosti njihove primene u regenerativnoj medicini, kao i na polju ćelijske terapije. Iako MMĆ izolovane iz različitih adultnih i neonatalnih izvora ispoljavaju potencijal samoobnove i multipotentne diferencijacije (u ćelije mezodermalnog, ali i ektodermalnog i endodermalnog porekla), mnoga istraživanja koja se tiču funkcionalnih karakteristika ovih ćelija tek predstoje kako bi njihova primena bila efikasna i bezbedna. Jedan od novije otkrivenih izvora MMĆ je i tkivo periodoncijuma koje predstavlja bogat i lako dostupan izvor MMĆ, s obzirom da se ovo tkivo dobija nakon ekstrakcije zuba iz ortodontskih razloga. Periodoncijum je vezivno tkivo čija je glavna uloga uspostavljanje čvrste veze između korena zuba i alveolarne kosti. Regenerativni potencijal MMĆ periodoncijuma (PD-MMĆ) ogleda se u njihovom potencijalu samoobnove i multipotentne diferencijacije, kao i sposobnosti formiranja tkiva sličnih potpornom tkivu zuba, uključujući periodoncijum i cement. Ove osobine čine PD-MMĆ dobrim kandidatima za primenu u regeneraciji potpornog tkiva zuba u parodontopatiji, hroničnom inflamatornom oboljenju uzrokovanom bakterijama zubnog plaka. Razvoj hronične parodontopatije podrazumeva procese remodelovanja koštanog tkiva pod uticajem brojnih faktora u koje su uključeni osteoblasti i osteoklasti koji ostvaruju recipročne interakcije sa ćelijama imunskog odgovora regulišući inflamaciju i degradaciju koštanog tkiva. Uloga lokalne mikrosredine u parodontopatiji predstavlja jedan od ključnih faktora u kontekstu ćelijskog odgovora tkiva periodoncijuma na perzistentnu infekciju. U okviru parodontalnih lezija potvrđeno je prisustvo Th17 (od engl. T helper cells17) ćelija, značajnih regulatora tkivnog razaranja u toku zapaljenskih procesa, koje produkuju proinflamatorni citokin interleukin (IL)-17. U kontekstu parodontopatije, do sada je pokazano da IL-17 ostvaruje svoju ulogu stimulišući privlačenje neutrofila na mesto inflamacije kao i direktnim stimulatornim delovanjem Th17 ćelija na diferencijaciju osteoklasta...In the last few decades regenerative and modulatory properties of mesenchymal stem cells (MSCs) have gained the attention of scientists since their characteristics indicated the possibility of their use in regenerative medicine and cell therapy. Although MSCs isolated from various adult and neonatal tissues exhibit the potential of self-renewal and multipotent differentiation (towards cells of mesodermal, ectodermal and endodermal origin), many aspects concerning their functional characteristics are yet to be explored. One of the recently discovered sources of MSCs is easily accessible tissue of periodontal ligament, obtained after tooth extraction for orthodontic reasons. Periodontal ligament tissue strengthens the root of the tooth in the alveolar bone. The regenerative potential of periodontal ligament MSCs (PDLSCs) is reflected in self-renewal and multipotent differentiation ability, as well as potency for the formation of tissues which support the teeth, including the periodontal ligament and cement. These features make the PDLSCs good candidates for use in the regeneration/reconstruction of supporting tissue of teeth in periodontal disease, a chronic inflammatory disease caused by bacterial plaque. The development of chronic periodontal disease includes the process of bone tissue remodeling governed by a number of factors, including osteoblasts and osteoclasts, which reciprocally interact with cellular and humoral compartments of the immune system, thus regulating inflammation, degradation and renewal of bone tissue. The role of the local microenvironment in the periodontal disease is one of the key factors in the context of periodontal ligament tissue response to persistent infection. The presence of T helper cells17 (Th17) cells, important regulators of tissue destruction during inflammatory processes has been confirmed in periodontal lesions. Th17 cells produce proinflammatory cytokine interleukin (IL)-17. In the context of periodontal disease, as well as in other inflammatory diseases, IL-17 stimulates the activation and migration of neutrophils to the site of inflammation. Additionally, IL-17 directly stimulates differentiation of osteoclasts and contributes to the alveolar bone resorption..

Inflammatory niche: Mesenchymal stromal cell priming by soluble mediators

Mesenchymal stromal/stem cells (MSCs) are adult stem cells of stromal origin that possess self-renewal capacity and the ability to differentiate into multiple mesodermal cell lineages. They play a critical role in tissue homeostasis and wound healing, as well as in regulating the inflammatory microenvironment through interactions with immune cells. Hence, MSCs have garnered great attention as promising candidates for tissue regeneration and cell therapy. Because the inflammatory niche plays a key role in triggering the reparative and immunomodulatory functions of MSCs, priming of MSCs with bioactive molecules has been proposed as a way to foster the therapeutic potential of these cells. In this paper, we review how soluble mediators of the inflammatory niche (cytokines and alarmins) influence the regenerative and immunomodulatory capacity of MSCs, highlighting the major advantages and concerns regarding the therapeutic potential of these inflammatory primed MSCs. The data summarized in this review may provide a significant starting point for future research on priming MSCs and establishing standardized methods for the application of preconditioned MSCs in cell therapy

The Roles of Mesenchymal Stromal/Stem Cells in Tumor Microenvironment Associated with Inflammation

State of tumor microenvironment (TME) is closely linked to regulation of tumor growth and progression affecting the final outcome, refractoriness, and relapse of disease. Interactions of tumor, immune, and mesenchymal stromal/stem cells (MSCs) have been recognized as crucial for understanding tumorigenesis. Due to their outstanding features, stem cell-like properties, capacity to regulate immune response, and dynamic functional phenotype dependent on microenvironmental stimuli, MSCs have been perceived as important players in TME. Signals provided by tumor-associated chronic inflammation educate MSCs to alter their phenotype and immunomodulatory potential in favor of tumor-biased state of MSCs. Adjustment of phenotype to TME and acquisition of tumor-promoting ability byMSCs help tumor cells inmaintenance of permissive TME and suppression of antitumor immune response. Potential utilization of MSCs in treatment of tumor is based on their inherent ability to home tumor tissue that makes them suitable delivery vehicles for immune-stimulating factors and vectors for targeted antitumor therapy. Here, we review data regarding intrusive effects of inflammatory TME on MSCs capacity to affect tumor development through modification of their phenotype and interactions with immune system

Adipogenesis in Different Body Depots and Tumor Development

Adipose tissue (AT) forms depots at different anatomical locations throughout the body, being in subcutaneous and visceral regions, as well as the bone marrow. These ATs differ in the adipocyte functional profile, their insulin sensitivity, adipokines' production, lipolysis, and response to pathologic conditions. Despite the recent advances in lineage tracing, which have demonstrated that individual adipose depots are composed of adipocytes derived from distinct progenitor populations, the cellular and molecular dissection of the adipose clonogenic stem cell niche is still a great challenge. Additional complexity in AT regulation is associated with tumor-induced changes that affect adipocyte phenotype. As an integrative unit of cell differentiation, AT microenvironment regulates various phenotype outcomes of differentiating adipogenic lineages, which consequently may contribute to the neoplastic phenotype manifestations. Particularly interesting is the capacity of AT to impose and support the aberrant potency of stem cells that accompanies tumor development. In this review, we summarize the current findings on the communication between adipocytes and their progenitors with tumor cells, pointing out to the co-existence of healthy and neoplastic stem cell niches developed during tumor evolution. We also discuss tumor-induced adaptations in mature adipocytes and the involvement of alternative differentiation programs

Dental mesenchymal stromal/stem cells in different microenvironments — implications in regenerative therapy

Current research data reveal microenvironment as a significant modifier of physical functions, pathologic changes, as well as the therapeutic effects of stem cells. When comparing regeneration potential of various stem cell types used for cytotherapy and tissue engineering, mesenchymal stem cells (MSCs) are currently the most attractive cell source for bone and tooth regeneration due to their differentiation and immunomodulatory potential and lack of ethical issues associated with their use. The microenvironment of donors and recipients selected in cytotherapy plays a crucial role in regenerative potential of transplanted MSCs, indicating interactions of cells with their microenvironment indispensable in MSC-mediated bone and dental regeneration. Since a variety of MSC populations have been procured from different parts of the tooth and tooth-supporting tissues, MSCs of dental origin and their achievements in capacity to reconstitute various dental tissues have gained attention of many research groups over the years. This review discusses recent advances in comparative analyses of dental MSC regeneration potential with regards to their tissue origin and specific microenvironmental conditions, giving additional insight into the current clinical application of these cells

Tumorigenic Aspects of MSC Senescence—Implication in Cancer Development and Therapy

As an organism ages, many physiological processes change, including the immune system. This process, called immunosenescence, characterized by abnormal activation and imbalance of innate and adaptive immunity, leads to a state of chronic low-grade systemic inflammation, termed inflammaging. Aging and inflammaging are considered to be the root of many diseases of the elderly, as infections, autoimmune and chronic inflammatory diseases, degenerative diseases, and cancer. The role of mesenchymal stromal/stem cells (MSCs) in the inflammaging process and the age-related diseases is not completely established, although numerous features of aging MSCs, including altered immunomodulatory properties, impeded MSC niche supporting functions, and senescent MSC secretory repertoire are consistent with inflammaging development. Although senescence has its physiological function and can represent a mechanism of tumor prevention, in most cases it eventually transforms into a deleterious (para-)inflammatory process that promotes tumor growth. In this review we are going through current literature, trying to explore the role of senescent MSCs in making and/or sustaining a microenvironment permissive to tumor development and to analyze the therapeutic options that could target this process

Platelet-poor plazma sportista kao potencijalni induktor miogene diferencijacije C2C12 mioblasta

Introduction. Blood products, i.e. platelet rich plasma (PRP), leukocyte-poor plasma (PRP) and platelet poor plasma (PPP), have previously been used to improve muscle regeneration. In this study, six months' frozen-stored PPP of individuals who practiced different types of physical exercise was analysed; it could steer mouse C2C12 myoblast cells towards proliferation, migration and myogenic differentiation, and it could affect the morphology/shape of myotubes. Materials and Methods. PPP of male Olympic weightlifters, football players and professional folk dancers, aged 15-19, was collected 12 h post-training and stored for 6 months at -20°C. C2C12 cell proliferation was assessed by MTT test, motility by scratch assay, myogenic differentiation by myotube formation and gelatinase activity by gel-zymography. Results and Conclusions. PPP induced proliferation and migration of C2C12 cells. Proliferative capacity was as follows: weightlifters gt dancers gt football players; mean migratory capacity was: weightlifters = dancers gt football players. PPP induced formation of myotubes; significant inter-individual variations were detected: PPP from weightlifters induced formation of round myotubes, and PPP from football players and dancers induced formation of elongated myotubes. The mean myotube area was as follows: football players gt dancers gt weightlifters. PPP gelatinolytic activity was observed; it was negatively correlated with C2C12 myoblast proliferation. These results provide general but distinct evidence that PPP of individuals practicing certain types of exercise can specifically modify myoblast morphology/function. This is significant for explaining physiological responses and adaptations to exercise. In conclusion, longterm, frozen-stored PPP preserves its potential to modify myoblast morphology and function.Uvod. Krvna plazma obogaćena leukocitima, plazma sa niskim sadržajem leukocita i plazma sa niskim sadržajem trombocita (platelet poor plasma; PPP) su produkti krvi koji se koriste za stimulaciju regeneracije mišića. U ovom radu smo ispitivali da li zamrzavana PPP osoba koje se bave različitim tipovima fizičke aktivnosti, usmerava C2C12 myoblaste u pravcu povećane proliferacije, migracije i miogene diferencijacije, i da li utiče na morfologiju/izgled miotuba. Materijal i metode. PPP osoba muškog pola starih 15-19 godina je izolovana iz krvi dizača tegova, fudbalera i profesionalnih igrača folklora, 12 sati nakon treninga. Uzorci PPP su čuvani šest meseci na -20ºC. Uticaj PPP na proliferaciju C2C12 ćelija je analiziran MTT testom, na migraciju "scratch" testom, a uticaj na miogenu diferencijaciju je analiziran na osnovu sposobnosti PPP da indukuju formiranje miotuba. Želatinolitička aktivnost PPP je analizirana gel-zimografijom. Rezultati i zaključak. Uzorci PPP su indukovali proliferaciju i migraciju C2C12 ćelija, a kapacitet da stimulišu proliferaciju je bio: dizači tegova gt igrači gt fudbaleri. Kapacitet PPP da utiču na migraciju C2C12 ćelija je bio: dizači tegova = igrači gt fudbaleri. Svi uzorci PPP su indukovali formiranje miotuba, ali su zapažene značajne interindividualne varijacije. PPP dizača tegova su indukovali formiranje okruglih miotuba, dok su miotube formirane u prisustvu PPP igrača i fudbalera bile izdužene. Površina miotuba se, zavisno od tipa fizičke aktivnosti, menjala po sledećem rasporedu: fudbaleri gt igrači gt dizači tegova. Želatinolitička aktivnost PPP je nagativno korelirala sa proliferacijom C2C12 ćelija. Rezultati ove studije pokazuju da PPP osoba koje se bave određenim tipom fizičke aktivnosti mogu da na specifičan način modulišu morfologiju/funciju mioblasta. Ovaj rezultat je od značaja za objašnjnje fiziološkog odgovora i adaptacije na vežbanje. On pokazuje i da PPP nakon dugotrajnog zamrzavanja imaju očuvanu spospbnost modifikovanja morfologije i funkcije mioblasta

Mesenchymal stem cells isolated from human periodontal ligament

Mesenchymal stem cells (MSCs) were isolated from human periodontal ligament (hPDL-MSCs) and characterized by their morphology, clonogenic efficiency, proliferation and differentiation capabilities. hPDL-MSCs, derived from normal impacted third molars, possessed all of the properties of MSC, including clonogenic ability, high proliferation rate and multi-lineage (osteogenic, chondrogenic, adipogenic, myogenic) differentiation potential. Moreover, hPDL-MSCs expressed a typical MSC epitope profile, being positive for mesenchymal cell markers (CD44H, CD90, CD105, CD73, CD29, Stro-1, fibronectin, vimentin, alpha-SMA), and negative for hematopoietic stem cell markers (CD34, CD11b, CD45, Glycophorin-CD235a). Additionally, hPDL-MSCs, as primitive and highly multipotent cells, showed high expression of embryonic markers (Nanog, Sox2, SSEA4). The data obtained provided yet further proof that cells with mesenchymal properties can be obtained from periodontal ligament tissue. Although these cells should be further investigated to determine their clinical significance, hPDL-MSCs are believed to provide a renewable and promising cell source for new therapeutic strategies in the treatment of periodontal defects

Immunomodulatory capacity of human mesenchymal stem cells isolated from adipose tissue, dental pulp, peripheral blood and umbilical cord Wharton's jelly

Mesenchymal stem cells (MSCs), beside regenerative potential, possess immunomodulatory properties and their use in managing immune-mediated diseases is intensively studied. We analyzed the effects of MSCs isolated from human adipose tissue (AT-MSCs), dental pulp (DP-MSCs), peripheral blood (PB-MSCs) and umbilical cord Wharton's jelly (UC-MSCs), on the proliferation of allogeneic peripheral blood mononuclear cells (PBMCs). While only AT-MSCs functioned as alloantigen presenting cells, proliferation of PBMCs in response to a phytohemagglutinin (PHA) and alloantigens in mixed lymphocytes reaction (MLR) was inhibited by all MSCs in a cell concentration-dependent manner. Conditioned medium (CM) derived from DP-MSCs, PB-MSCs and UC-MSCs, suppressed the baseline, PHA- and alloantigens-mediated proliferation of PBMC, whereas AT-MSCs-derived CM inhibited MLR, but failed to suppress the spontaneous and PHA-induced PBMCs proliferation. Differences between MSC types were observed in expression of genes related to immunomodulation, including human leukocyte antigens (HLA)-A, HLA-DR, HLA-G5, interleukin 6 (IL)-6, transforming growth factor (TGF)-beta, cyclooxygenase-2 (COX-2) and indoleamine 2,3-dioxygenase (IDO-1), under basal conditions, as well as in response to proinflammatory cytokines, interferon (IFN)-gamma and tumor necrosis factor alpha (TNF)-alpha. While AT-MSCs showed a positive constitutive expression of almost all tested genes that was augmented in response to IFN-gamma and TNF-alpha, only combined cytokine treatment increased HLA-A, COX2 and IL-6 mRNA expression in DP-MSCs and slightly stimulated the expression of HLA-G and TGF-beta in UC-MSCs. Although MSCs from different tissues showed similar potential to suppress proliferation of PBMCs, heterogeneity in the expression of genes related to immunomodulation emphasizes the importance of investigating the role of specific molecular mechanisms in the regulation of immunomodulatory activity of MSCs