This study evaluates the balance between cellular removal and extracellular matrix (ECM) preservation in cardiac tissue engineering by comparing chemical and physical decellularization methods. Cardiac tissues were treated with chemical agents (sodium dodecyl sulfate and Triton X-100) and physical methods (freeze-thawing and ultrasound). These methods were assessed based on residual cellular content, DNA quantification, ECM structural integrity, and preservation of key ECM components like collagen and glycosaminoglycan (GAG). The results revealed that while chemical methods, particularly SDS, achieved more complete cell removal, they significantly compromised ECM integrity. In contrast, physical methods, such as freeze-thawing, preserved ECM structure more effectively, despite moderate cellular removal. The findings underscore the importance of tailoring decellularization techniques to specific cardiac tissue engineering needs, with chemical methods excelling in cell removal and physical methods offering superior ECM preservation. Future research should aim to optimize these methods to achieve a better balance between decellularization efficiency and ECM integrity.
Background: Despite its anticancer activity, cisplatin exhibits severe testicular toxicity when used in chemotherapy. Owing to its wide application in cancer therapy, the reduction of damage to normal tissue is of imminent clinical need. In this study, we evaluated the effects of catechin hydrate, a natural flavon-3-ol phytochemical, on cisplatin-induced testicular injury. Methods: Type 2 mouse spermatogonia (GC-1 spg cells) were treated with 0-100 μM catechin and cisplatin. Cell survival was estimated using a cell proliferation assay and Ki-67 immunostaining. Apoptosis was assessed via flow cytometry with the Dead Cell Apoptosis assay. To determine the antioxidant effects of catechin hydrate, Nrf2 expression was measured using qPCR and CellROX staining. The anti-inflammatory effects were evaluated by analyzing the gene and protein expression levels of iNOS and COX2 using qPCR and immunoblotting. Results: The 100 μM catechin hydrate treatment did not affect healthy GC-1 spg cells but, prevented cisplatin-induced GC-1 spg cell death via the regulation of anti-oxidants and inflammation-related molecules. In addition, the number of apoptotic cells, cleaved-caspase 3 level, and BAX gene expression levels were significantly reduced by catechin hydrate treatment in a cisplatin-induced GC-1 spg cell death model. In addition, antioxidant and anti-inflammatory marker genes, including Nrf2 , iNOS, and COX2 were significantly downregulated by catechin hydrate treatment in cisplatintreated GC-1 cells. Conclusions: Our study contributes to the opportunity to reintroduce cisplatin into systemic anticancer treatment, with reduced testicular toxicity and restored fertility.
Cellular myxoma is an uncommon type of myxoid benign tumor, predominantly occurring in adult female patients aged >40 years. This report aims to document a case of cellular myxoma that occurred in the buccal mucosa. Compared to intramuscular myxomas, cellular myxoma demonstrates hypercellularity and vascularity. Its manifestation in the soft tissue of the head and neck area is exceptionally rare. Generally, cellular myxoma manifests as a firm and immovable mass covered with normal oral mucosa, with no associated clinical symptoms. Homogenous low signal intensity on T1-weighted scans and high-signal intensity on T2-weighted magnetic resonance scans reveal cellular myxoma, as most lesions show well-defined margins and heterogeneous contrast enhancement. The significant histologic features include a focal or diffuse increase in cellularity with fibroblast-like cells and vascularity with an abundant collagenous matrix. Our presented case reflects these facts examinations, based on which a final diagnosis of cellular myxoma was made. Immunohistochemistry revealed locally and diffusely positive SMA and CD34. The clinical tendency of cellular myxoma with hypercellularity may affect the production of myxoid and collagenous substrates, and if complete resection is not performed, the possibility of local recurrence in the primarily affected region remains. Hence, complete surgical excision was performed under general anesthesia, and follow-up until a year after treatment revealed no observed recurrence. To achieve precise diagnosis and complete treatment without local recurrence, several diagnostic examinations should be considered.
The cellular communication network factor (CCN) family proteins regulate many biological events such as angiogenesis, tumor growth, placentation, implantation, and embryogenesis. The expression and function of CCN1, CCN2, and CCN3 at the maternal-conceptus interface are established in humans and rodents, but little is known about the role of CCN4 to CCN6 in the reproductive organs in any other species. Several studies in transcriptome analysis in pigs have shown that the expression of CCN4 and CCN6 increases in the endometrium during early pregnancy. However, their expression, regulation, and function in the endometrium throughout the estrous cycle and pregnancy have not been fully understood in pigs. Thus, we determined the expression, localization, and regulation of CCN4 and CCN6 during the estrous cycle and at the maternal-conceptus interface in pigs. We found that the levels of CCN4, but not CCN6, changed during the estrous cycle. The levels of CCN4 were greater during mid- to late pregnancy than in the early stage, and the levels of CCN6 were greatest on Day 15 of pregnancy. CCN4 and CCN6 were detected in conceptus tissues during early pregnancy and in chorioallantoic tissues during the later stage of pregnancy. CCN4 mRNA was mainly localized to epithelial cells, CCN6 mRNAs to epithelial and stromal cells in the endometrium. In endometrial explant cultures, CCN4 expression was increased by progesterone, and CCN6 expression by interferon-γ. These results suggest that CCN4 and CCN6 may play roles in the establishment and maintenance of pregnancy by regulating the endometrial epithelial cell functions in pigs.
Protein and peptide candidates are screened to apply therapeutic application as a drug. Ensuring that these candidates are delivered and maximized effectiveness is still challenging and a variety of studies are ongoing. As drug delivery system vehicles, cell-penetrating peptide (CPP) can deliver various kinds of cargo into the cell cytosol. In a previous study, we developed Ara27 CPP, which are a zinc knuckle family protein of Arabidopsis, and confirmed internalization in human dermal fibroblasts and human dental pulp stem cells at low concentration with short time treatment condition without any toxicity. Ara27, an amphipathic CPP, could be modified and utilized in the biomedical field excluding the risk of toxicity. Therefore, we would like to confirm the non-toxic induced penetrating ability of Ara27 in various cell lines. The purpose of this study was to screen the cell internalization ability of Ara27 in various cell lines and to confirm Ara27 as a promising core CPP structure. First, Ara27 was screened to confirm non-toxicity concentration. Then, fluorescence-labeled Ara27 was treated on human normal cell lines, cancer cell lines and animal cell lines to identify the cellular internalization of Ara27. Ara27 was well intracellular localized in all cell lines and the intensity of fluorescence was remarkably increased in time pass manner. These results indicate that Ara27 has the potential as a core structure for applications in various drug delivery systems.
Spirulina platensis is a unique photosynthetic cyanobacterium that is used as a commercial feed for animals and a dietary supplement for humans. S. platensis has significant auto-flotation activity due to its gas vesicles. The natural flotation activity of these cells provides an effective method for harvesting of Spirulina biomass. The present research found that hydrodynamic pressure, light level and bicarbonate concentration affected the natural flotation of S. platensis. In particular, auto-flotation increased as light level decreased (5.5-times greater at no supplemental light than at 300 μ㏖ photons/㎡/s), as bicarbonate level increased (5-times higher at 0.6 than 0.2 M) and as hydrodynamic pressure decreased (indicated by depth of the culture). These results should be considered when designing photobioreactors for direct harvesting by auto-flotation.
Autophagy is an evolutionarily well-conserved cellular homeostasis program that responds to various cellular stresses and degrades unnecessary or harmful intracellular materials in lysosomes. Accumulating evidence has shown that autophagy dysfunction often results in various human pathophysiological conditions, including metabolic disorders, cancers, and neurodegenerative diseases. The discovery of an autophagy machinery protein network has revealed underlying molecular mechanisms of autophagy, and advances in the understanding of its regulatory mechanism have provided novel therapeutic targets for treating human diseases. Recently, reports have emerged on the involvement of autophagy in oral squamous cell carcinoma (OSCC). Although the role of autophagy in cancer therapy is controversial, the beneficial use of the induction of autophagic cell death in OSCC has drawn significant attention. In this review, the types of autophagy, mechanism of autophagosome biogenesis, and modulating molecules and therapeutic candidates affecting the induction of autophagic cell death in OSCC are briefly described.
PURPOSES : In general, a high-performance concrete construction method is a method of loading mixed cement and other materials on a mobile mixer equipped with a concrete plant. However, the construction cost is high because the unit cost of the mixed cement is very high and the equipment usage fee of the mobile mixer is also high. To compensate for these drawbacks, a cellular spray concrete method was developed. This is a highly functional customized concrete construction method in which admixtures are added to high-flow concrete with cellular (air bubbles) added at the site and constructed through spraying.
METHODS : Field workability evaluation using cellular concrete was carried out using a spray method in 2017 and an anti-foaming agent in 2018. The test construction section was set as a new road-pavement construction site. After construction, strength, durability characteristics, and void analysis were performed to compare the construction methods. The results of laboratory tests under optimal conditions were also compared to the on-field results. By comparing the indoor mixing and the on-field results, we analyzed whether there were any problems concerning the performance expression. The economic feasibility at the initial construction stage was analyzed by comparing the existing and the cellular concrete construction methods in terms of material cost and equipment usage fee.
RESULTS : In the case of cellular spray concrete, construction through spraying is cumbersome, and the need to additionally use large equipment such as a pump car and compressor constitutes a disadvantage. However, the most relevant feature is that it can be constructed not only on the floor, but also on a slope. In the case of cellular concrete with an antifoaming agent, it was possible to quickly produce high-performance concrete using on-site remixing equipment. By supplying materials to the pouring surface using a conveyor, smooth construction and construction in narrow spaces were also possible.
CONCLUSIONS : The cellular concrete method allows the immediate on-site production of high-performance concrete, and it is possible to selectively apply spray construction, antifoaming agent construction, etc.
Several factors, including genetic and environmental insults, impede protein folding and secretion in the endoplasmic reticulum (ER). Accumulation of unfolded or mis-folded protein in the ER manifests as ER stress. To cope with this morbid condition of the ER, recent data has suggested that the intracellular event of an unfolded protein response plays a critical role in managing the secretory load and maintaining proteostasis in the ER. Tauroursodeoxycholic acid (TUDCA) is a chemical chaperone and hydrophilic bile acid that is known to inhibit apoptosis by attenuating ER stress. Numerous studies have revealed that TUDCA affects hepatic diseases, obesity, and inflammatory illnesses. Recently, molecular regulation of ER stress in tooth development, especially during the secretory stage, has been studied. Therefore, in this study, we examined the developmental role of ER stress regulation in tooth morphogenesis using in vitro organ cultivation methods with a chemical chaperone treatment, TUDCA. Altered cellular events including proliferation, apoptosis, and dentinogenesis were examined using immunostaining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. In addition, altered localization patterns of the formation of hard tissue matrices related to molecules, including amelogenin and nestin, were examined to assess their morphological changes. Based on our findings, modulating the role of the chemical chaperone TUDCA in tooth morphogenesis, especially through the modulation of cellular proliferation and apoptosis, could be applied as a supporting data for tooth regeneration for future studies.
Hydrogen peroxide (H2O2) is originally an endogenous small molecule which is reduced into water in cells. In order to know the H2O2-induced oxidative stress in RAW 264.7 cells, first of all, the optimum concentration of exogenous H2O2 which show reactive cellular responses was determined as 40 μM by MTT assay, and followed by 40 μM H2O2 application in RAW 264.7 cells for 30 min, 1, or 2 hours. The expressional changes of essential proteins for cellular proliferation, epigenetic modification, inflammation, apoptosis, survival, and protection were assessed by immunoprecipitation high performance liquid chromatography (IP-HPLC) using 51 antisera. 40 μM H2O2 treatment down-regulated proliferation-related proteins, Ki-67, PCNA, CDK4, cyclin D2, cMyc, and PLK4, induced histone methylation/ deacetylation and DNA methylation by increasing levels of HDAC10 and DMAP1 and by decreasing levels of DNMT1 and KDM4D, activated inflammatory reaction by increasing levels of MCP-1, COX-2, CD68, LTA4H, CXCR4, and lysozyme, and dramatically up-regulated cellular apoptosis-, survival-, and protection-related proteins, AIF, PARP-1, caspase 9, c-caspase 9, pAKT1/2/3, SOD-1, HO-1, NF-kB, NRF2, and GSTO1 in RAW 264.7 cells. These observations suggest exogenous 40 μM H2O2-induced oxidative stresses which resulted global cellular responses including not only antioxidant, inflammation, and apoptosis but also proliferation and epigenetic modification. Particularly, 40 μM H2O2-induced apoptosis was mainly derived from PARP-1/AIF signaling leading parthanatos, and 40 μM H2O2-induced suppression of cMyc/MAX/MAD network was relevant to reduction of RAW 264.7 cell proliferation. Accordingly, H2O2 appears to affect RAW 264.7 macrophages in several ways eliciting not only oxidative stresses but also genome-wide DNA damage.
Gonadotropin releasing hormone (GnRH) centrally plays a role in control of the hypothalamicpituitary- gonadal axis-related hormone secretions in the reproductive neuroendocrine system. In addition, hormone receptors like luteinizing hormone receptor (LHR) are important element for hormones to take effect in target organ. However, ageing-dependent changes in terms of the distribution of GnRH neurons in the brain and LHR expression in the acyclic ovary have not been fully understood yet. Therefore, we comparatively investigated those ageing-dependent changes using young (1-5 months), middle (11-14 months) and old (21-27 months) aged female mice. Whereas a number of GnRH positive fibers and neurons with monopolar or bipolar morphology were abundantly observed in the brain of the young and middle aged mice, a few GnRH positive neurons with multiple dendrites were observed in the old aged mice. In addition, acyclic ovary without repeated development and degeneration of the follicles was shown in the old aged mice than others. LHR expression was localized in theca cells, granulosa cell, corpora lutea and atretic follicle in the ovaries from young and middle aged mice, in contrast, old aged mice had few positive LHR expression on the follicles due to acyclic ovary. However, the whole protein level of LHR was higher in the ovary of old aged mice than others. These results are expected to be used as an important basis on the relationship between GnRH and LHR in old aged animals as well as in further research for reproduction failure.
The expression of MMPs in the development of the fertilized egg has a very important role in cell configuration. Objective To evaluate the clinical, the effect of differentially expressed MMPs on serum and serum - free medium on the maturation of blastocysts. The expression patterns of MMPs in serum and serum-free medium were compared at 6 h, 18 h and at the blastocyst stage using real-time PCR, ELISA and immunofluorescence. The results showed that the expression of MMPs was increased in the embryos of the serum medium, as a result of analysis of MMPs and TIMPs, MMP-2 was expressed in the cytoplasm of embryos in the serum-free medium, And it was found to be higher in expression than MMP-9. The serum medium was different from the bloodless badge: overall, TIMPs showed a higher expression in the ovarian cells than cyanosis, and TIMP-3 was more pronounced. Development rate of blastocyst according to in vitro culture method was higher than that of serum - free medium (61.22% 60/98) and serum - free medium (48.28% 28/58). Analysis of the protein release locations of MMPs and TIMPs showed that MMPs and TIMPs are highly expressed in serum mediums, focusing on the inner cell mass. However, very low expression appeared in the tropoblast. On the other hand, serum - free medium showed different expression from serum medium and TIMPs expression was generally low.
Therefore, in the case of serum media, the expression of MMPs is highly expressed in the cytoplasm of the fertilized egg, increasing the reconstruction of cells.
Transplantation of stem cells, such as mesenchymal stem cells (MSCs), is a promising strategy for treating several types of intractable disorders. Mechanistically, it could not only replace damaged cells by direct contribution, but also establish an anti-inflammatory or immunomodulatory microenvironment. However, the cellular mechanisms underlying molecular and biological properties of stem cells during ex vivo expansion and also after transplantation in pathological environments remain largely elusive. We recently developed the cyanoacrylamide-based coumarin derivatives (Fluorescent real-time thiol tracer; FreSHtracer*) reversibly react with glutathione for monitoring of glutathione levels in living stem cells. These probes revealed that glutathione levels are heterogeneous among subcellular organelles and among individual cells and show dynamic changes and heterogeneity in repopulating stem cells depending on oxidative-stress or culture conditions. Importantly, a subpopulation of stem cells with high-glutathione levels exhibited increased self-renewal and migration activities in vitro and showed improved therapeutic efficiency in treating asthma. Furthermore, employing a novel combination of longitudinal intravital confocal fluorescence imaging and microcystoscopy in living animals, we investigated the distributions and properties of transplanted multipotent MSCs derived from human embryonic stem cells at single-cell resolution in real-time by performing confocal imaging of bladder tissues in a rat model of IC/BPS for up to 6 months post-transplantation. These novel real-time monitoring strategies demonstrate the novel molecular insight for maintaining stem cell functions and also enhance understanding of the in vivo behaviors of the engrafted stem cells, which is crucial to determine the efficacy and safety of stem cell-based therapies. This strategy may facilitate the translation of various stem cell-based approaches into clinical practice.
We investigated the effects of two Brucella proteins expressed in a pMAL expression system, RocF and EF-Ts, as subunit vaccines on immune modulation and protective efficacy using a mouse model. Mice vaccinated with MBP-RocF and MBP-EF-Ts displayed increased production of TNF, IFN-, MCP-1, IL-10 and IL-6, and TNF and MCP-1, respectively. Furthermore, mice vaccinated with MBP-EF-Ts showed decreased induction of IFN- and Th2-related cytokines, IL-10 and IL-6. Higher proportions of CD4+ and CD8+ T cells were observed in the blood of mice vaccinated with MBP-RocF than in the PBS-vaccinated group, although the increases were not significant. Furthermore, significantly reduced Brucella proliferation in the spleens of the MBP-RocF and MBP-EF-Ts groups were observed, but inflammation of these organs was not attenuated. Overall, these results indicate that RocF and EF-Ts could be potential subunit vaccine candidates against animal brucellosis.
Phospholipase A2 (PLA2) hydrolyzes fatty acids from phospholipids at sn-2 position. Two immune-associated PLA2s are known in Spodoptera exigua. Both are calcium-independent cellular PLA2 (iPLA2A and iPLA2B). These iPLA2s are inducible to immune challenge using different epitopes such as virus, Gram+, Gram- bacteria, and fungi. However, their expression profiles are different between these two iPLA2s. Especially, immune challenge with Gram+ bacteria and fungi significantly up-regulated these iPLA2 expression, suggesting their expression under Toll signal pathway. RNA interferences of Toll or IMD signal components are being investigated.
To elucidate the effect of cellular phone electromagnetic wave (EMW) exposure on the developing cerebellar cortex of neonatal Sprague-Dawley rats, animals were exposed to cellular phone electromagnetic waves for 1 hr per day for 3 weeks. At the end of the experimental period, animals were sacrificed by cardiac perfusion, after which histological samples were prepared and observed microscopically. In the EMW exposure group, external granule cells were remained partially in the external granular layer without migrating into the internal granular layer. In addition, dark stained shrunken Purkinje cells with pyknotic nuclei increased and the outline of cells became irregular and showed degenerative signs, such as mitochondrial swelling and disrupted cristae. Moreover, the cisternae of rough endoplasmic reticula and Golgi complex were severely swollen. Bergmann glial cells adjacent to the dark stained Purkinje cells were swollen and cytoplasmic organelles were scant. Dark stained shrunken granule cells were also observed and the outline of cells was irregular. The results of the present study suggest that cellular phone EMW exposure to neonatal Sprague-Dawley rats leads to a partial delay of early migration of cerebellar cortical cells and degenerative changes in Purkinje cells, Bergmann glial cells and granule cells.
곤충의 면역반응에 대한 연구는 곤충 체내 침입한 미생물들과 직접 반응하는 기작들을 중심으로 연구되었다. 그러나 미생물들이 곤충 체내에 침입 한 후 발생되는 다양한 미생물 분비물질에 의한 곤충 면역반응의 시작여부 등에 대한 연구는 거의 없는 실정이다. 이를 위하여 흰점박이 꽃무지(Protaetia brevitarsis seulensis) 유충의 장내에 존재하는 공생균과 체외 병원균을 동일한 조건에서 배양 하고 다양한 분비물질들이 존재 할 거라 예상되는 배양액만을 분리, 유충에 주사하여 면역반응 여부를 조사하였다. 공생균 배양액을 주입한 유충들은 비교적 건강하고 면역반응도 발생하지 않았으나 병원균 배양액을 주입한 유충의 경우 150시간 후 60% 이상 사망하였고 주사된 자리도 짙은 갈색의 멜라닌화가 관찰 되었다. 이러한 면역반응은 과립혈구세포의 리소좀(Lysosomes) 활성화 여부로 재확인 하였다. 병원균 배양액이 주입된 유충들의 경우 12시간 후 리소좀 이 ~50% 이상 활성화 되었으나 공생균 배양액이 주입된 유충들의 경우 ~5% 미만으로 활성화 되는 것으로 나타났다. 따라서 공생균 배양액내에 는 기주면역반응을 유도하는 물질들이 없거나 량이 매우 적게 존재하는 것을 추측 할 수 있었다.