Mesenchymal Stromal/Stem Cells-Derived Exosomes as an Antimicrobial Weapon for Orodental Infections

The oral cavity as the second most various microbial community in the body contains a broad spectrum of microorganisms which are known as the oral microbiome. The oral microbiome includes different types of microbes such as bacteria, fungi, viruses, and protozoa. Numerous factors can affect the equilibrium of the oral microbiome community which can eventually lead to orodental infectious diseases. Periodontitis, dental caries, oral leukoplakia, oral squamous cell carcinoma are some multifactorial infectious diseases in the oral cavity. In defending against infection, the immune system has an essential role. Depending on the speed and specificity of the reaction, immunity is divided into two different types which are named the innate and the adaptive responses but also there is much interaction between them. In these responses, different types of immune cells are present and recent evidence demonstrates that these cell types both within the innate and adaptive immune systems are capable of secreting some extracellular vesicles named exosomes which are involved in the response to infection. Exosomes are 30–150 nm lipid bilayer vesicles that consist of variant molecules, including proteins, lipids, and genetic materials and they have been associated with cell-to-cell communications. However, some kinds of exosomes can be effective on the pathogenicity of various microorganisms and promoting infections, and some other ones have antimicrobial and anti-infective functions in microbial diseases. These discrepancies in performance are due to the origin of the exosome. Exosomes can modulate the innate and specific immune responses of host cells by participating in antigen presentation for activation of immune cells and stimulating the release of inflammatory factors and the expression of immune molecules. Also, mesenchymal stromal/stem cells (MSCs)-derived exosomes participate in immunomodulation by different mechanisms. Ease of expansion and immunotherapeutic capabilities of MSCs, develop their applications in hundreds of clinical trials. Recently, it has been shown that cell-free therapies, like exosome therapies, by having more advantages than previous treatment methods are emerging as a promising strategy for the treatment of several diseases, in particular inflammatory conditions. In orodental infectious disease, exosomes can also play an important role by modulating immunoinflammatory responses. Therefore, MSCs-derived exosomes may have potential therapeutic effects to be a choice for controlling and treatment of orodental infectious diseases

Ekspresija mRNA agutiju srodnog peptida i mRNA melanokortin-4 receptora u arkuatnoj jezgri za vrijeme gravidnosti i laktacije štakorica.

Pregnancy is associated with a range of physiological adjustments to adapt the body to the demands of the growth of the fetus and subsequent lactation. It has been observed that agouti-related peptide (AGRP) and melanocortin-4 receptor (MC4R) are involved in energy homeostasis. A randomized controlled experimental study was planned to investigate the expression of AGRP and MC4R mRNAs in the stages of pregnancy and lactation in rat arcuate nucleus (ARC) of the hypothalamus. Thirty-two adult female rats were randomly divided into six groups. Pregnant rats were assigned into three groups (n = 6) of 7, 14, and 21 days of pregnancy. Two more groups were also assigned of non-suckling rats (n = 5), immediately separated from their pups after parturition, and suckling rats (n = 5), allowed to suckle five pups until day 8 (increasing milk). The sixth group consisted of four ovariectomized rats, which were assigned two weeks after surgery and served as control. Using real-time PCR, the relative expressions (compared to controls) of MC4R and AGRP mRNAs in ARC were calculated in the pregnant, suckling and non-suckling rats. Expression of AGRP mRNAs in pregnant rats on days 14 and 21 was higher than that observed in suckling and non-suckling rats (P<0.05). Expression of MC4R mRNAs in pregnant rats on days 14 and 21 was lower than that observed in suckling rats, and was higher on day 7 than that observed in both suckling and non-suckling rats (P<0.05). In conclusion, expression of AGRP in pregnancy and MC4R in lactation in ARC of rats controls energy homeostasis.Gravidnost je povezana s nizom fizioloških prilagodbi kojima tijelo odgovara zahtjevima povezanima s rastom fetusa i kasnije laktacije. Uočeno je da su agutiju srodan peptid (AGRP) i melanokortin-4 receptor (MC4R) uključeni u energetsku homeostazu. Randomiziranim kontroliranim pokusom planirano je u arkuatnoj jezgri (ARC) hipotalamusa štakorica istražiti ekspresiju mRNA AGRP i mRNA MC4R tijekom gravidnosti i laktacije. Trideset dvije odrasle štakorice nasumično su bile podijeljene u šest skupina. Gravidne su štakorice podijeljene u tri skupine (n = 6), s obzirom na 7., 14. i 21. dan gravidnosti. Još dvije skupine (n = 5) činile su nedojne štakorice koje su odvojene od svoje mladunčadi odmah nakon porođaja te dojne štakorice kojima je dozvoljeno dojenje 5 mladunaca do osmog dana rastuće laktacije. Četiri ovarijektomizirane štakorice, dva tjedna nakon operacije, dodijeljene su u 6. kontrolnu skupinu. Koristeći PCR u stvarnom vremenu, relativna ekspresija (usporedba s kontrolama) mRNA MC4R i mRNA AGRP u ARC izračunata je za gravidne, nedojne i dojne štakorice. Ekspresija mRNA AGRP kod gravidnih štakorica 14. i 21. dan bila je veća od one opažene kod dojnih i nedojnih štakorica (P<0,05). Ekspresija MC4R mRNA u gravidnih štakorica 14. i 21. dan bila je manja u odnosu na dojne štakorica i veća 7. dan u odnosu na skupine dojnih i nedojnih štakorica (P<0,05). Zaključno, ekspresija AGRP u ARC tijekom gravidnosti i MC4R u ARC tijekom laktacije kontrolira energetsku homeostazu štakorica

Histomorphometric Evaluation of Superovulation Effect on Follicular Development after Autologous Ovarian Transplantation in Mice

The effect of superovulation by pregnant mare serum gonadotropin (PMSG) on autologous transplanted ovaries in the lumbar muscles of mice was histomorphometrically evaluated using the indices of number and volume of different kind of follicles and volume of corpora lutea, ovary, and stroma. Angiogenesis was observed after mouse ovarian transplantation on days 14 and 21 after ovarian grafting. After transplantation, the total number and volume of primary and secondary follicles reduced, while PMSG superovulation increased the total number and total volume of tertiary follicles and also the ovarian volume after transplantation. Transplantation increased the average size of primary, secondary, and tertiary follicles. Therefore, primary and secondary follicles can survive after autologous transplantation but their reservations diminished by increasing the time of transplantation. However, number of tertiary follicles and their response to superovulation increased over time after transplantation

Expression of melanocortin-4 receptor and agouti-related peptide mRNAs in arcuate nucleus during long term malnutrition of female ovariectomized rats

Objective: Melanocortin-4 receptor (MC4R) and agouti-related peptide (AgRP) are involved in energy homeostasis in the rat. The aim of the present study was to evaluate the expression of MC4R and AgRP mRNAs in arcuate nucleus (ARC) during long term malnutrition of female ovariectomized rats. Materials and Methods: Ten female ovariectomized rats were divided into two equal groups (n=6) of normal and restricted diet groups. Using real-time PCR, the relative expressions (compared to controls) of MC4R and AgRP mRNAs were compared between both diet groups. Results: The relative expression of MC4R and AgRP mRNA in the ARC of female ovariectomized rats during long term malnutrition was higher than those with normal diet (

Ekspresija mRNA agutiju srodnog peptida i mRNA melanokortin-4 receptora u arkuatnoj jezgri za vrijeme gravidnosti i laktacije štakorica.

Pregnancy is associated with a range of physiological adjustments to adapt the body to the demands of the growth of the fetus and subsequent lactation. It has been observed that agouti-related peptide (AGRP) and melanocortin-4 receptor (MC4R) are involved in energy homeostasis. A randomized controlled experimental study was planned to investigate the expression of AGRP and MC4R mRNAs in the stages of pregnancy and lactation in rat arcuate nucleus (ARC) of the hypothalamus. Thirty-two adult female rats were randomly divided into six groups. Pregnant rats were assigned into three groups (n = 6) of 7, 14, and 21 days of pregnancy. Two more groups were also assigned of non-suckling rats (n = 5), immediately separated from their pups after parturition, and suckling rats (n = 5), allowed to suckle five pups until day 8 (increasing milk). The sixth group consisted of four ovariectomized rats, which were assigned two weeks after surgery and served as control. Using real-time PCR, the relative expressions (compared to controls) of MC4R and AGRP mRNAs in ARC were calculated in the pregnant, suckling and non-suckling rats. Expression of AGRP mRNAs in pregnant rats on days 14 and 21 was higher than that observed in suckling and non-suckling rats (P<0.05). Expression of MC4R mRNAs in pregnant rats on days 14 and 21 was lower than that observed in suckling rats, and was higher on day 7 than that observed in both suckling and non-suckling rats (P<0.05). In conclusion, expression of AGRP in pregnancy and MC4R in lactation in ARC of rats controls energy homeostasis.Gravidnost je povezana s nizom fizioloških prilagodbi kojima tijelo odgovara zahtjevima povezanima s rastom fetusa i kasnije laktacije. Uočeno je da su agutiju srodan peptid (AGRP) i melanokortin-4 receptor (MC4R) uključeni u energetsku homeostazu. Randomiziranim kontroliranim pokusom planirano je u arkuatnoj jezgri (ARC) hipotalamusa štakorica istražiti ekspresiju mRNA AGRP i mRNA MC4R tijekom gravidnosti i laktacije. Trideset dvije odrasle štakorice nasumično su bile podijeljene u šest skupina. Gravidne su štakorice podijeljene u tri skupine (n = 6), s obzirom na 7., 14. i 21. dan gravidnosti. Još dvije skupine (n = 5) činile su nedojne štakorice koje su odvojene od svoje mladunčadi odmah nakon porođaja te dojne štakorice kojima je dozvoljeno dojenje 5 mladunaca do osmog dana rastuće laktacije. Četiri ovarijektomizirane štakorice, dva tjedna nakon operacije, dodijeljene su u 6. kontrolnu skupinu. Koristeći PCR u stvarnom vremenu, relativna ekspresija (usporedba s kontrolama) mRNA MC4R i mRNA AGRP u ARC izračunata je za gravidne, nedojne i dojne štakorice. Ekspresija mRNA AGRP kod gravidnih štakorica 14. i 21. dan bila je veća od one opažene kod dojnih i nedojnih štakorica (P<0,05). Ekspresija MC4R mRNA u gravidnih štakorica 14. i 21. dan bila je manja u odnosu na dojne štakorica i veća 7. dan u odnosu na skupine dojnih i nedojnih štakorica (P<0,05). Zaključno, ekspresija AGRP u ARC tijekom gravidnosti i MC4R u ARC tijekom laktacije kontrolira energetsku homeostazu štakorica

Transplantation of Adult Monkey Neural Stem Cells into A Contusion Spinal Cord Injury Model in Rhesus Macaque Monkeys

Objective: Currently, cellular transplantation for spinal cord injuries (SCI) is the subject of numerous preclinical studies. Among the many cell types in the adult brain, there is a unique subpopulation of neural stem cells (NSC) that can self-renew and differentiate into neurons. The study aims, therefore, to explore the efficacy of adult monkey NSC (mNSC) in a primate SCI model. Materials and Methods: In this experimental study, isolated mNSCs were analyzed by flow cytometry, immunocytochemistry, and RT-PCR. Next, BrdU-labeled cells were transplanted into a SCI model. The SCI animal model was confirmed by magnetic resonance imaging (MRI) and histological analysis. Animals were clinically observed for 6 months. Results: Analysis confirmed homing of mNSCs into the injury site. Transplanted cells expressed neuronal markers (TubIII). Hind limb performance improved in transplanted animals based on Tarlov’s scale and our established behavioral tests for monkeys. Conclusion: Our findings have indicated that mNSCs can facilitate recovery in contusion SCI models in rhesus macaque monkeys. Additional studies are necessary to determine the improvement mechanisms after cell transplantation