Alcian Blue Research Articles

Effects of hydroxychloroquine on the mucosal barrier and gut microbiota during healing of mice colitis.

The present study aimed to evaluate the impact of hydroxychloroquine (HCQ) on the mucosal barrier and gut microbiota during the healing of mice colitis. The body weight, colon length, colon Hematoxylin-Eosin (H&E) staining, occult blood in feces and serum inflammatory factor levels were measured to evaluate the function of HCQ on inflammatory process in colitis mice. The Alcian blue staining, immunohistochemistry, immunofluorescence and serum FITC-Dextran assay were performed to assess the intestinal mucosal permeability. And the composition and expression differences of intestinal microorganisms in feces were analyzed with 16S rDNA sequencing for exploration of HCQ impact on gut microbiota in colitis. The results showed that the administration of HCQ did not significantly alter the body weight, colon length, or fecal occult blood of the mice. However, HCQ treatment did lead to recovery of the structure and morphology of the intestinal mucosa, increased expression of tight junction proteins (E-cadherin and Occludin), decreased permeability of the intestinal mucosal barrier, increased serum IL-10, and decreased level of tumor necrosis factor-alpha (TNF-α). Additionally, HCQ was found to increase the abundance of Euryarchaeota, Lactobacillus_murinus and Clostridium_fusiformis, while decreasing the abundance of Oscillibacter, uncultured_Odoribacter, Bacterioidetes and Muribaculum. These findings support that HCQ plays a role in the treatment of mice colitis possibly by altering the gut microbiota.

Morphogenesis of the ostrich (Struthio camelus) trachea and lung in different embryonic and fetal stages.

The ostrich (Struthio camelus) is an important wild species highlighted in national and international livestock industry. This research was conducted to analyze the development of the ostrich respiratory system during fetal and embryonic stages. A total of 50 fertile ostrich eggs were collected from commercial farms and then incubated at 36.00 - 37.00 ˚C and 25.00 ± 2.00% humidity for 40 days. Sections were taken on days 13, 22, 26, 30, 36, and 42 of incubation from the lung and the cranial, middle, and caudal parts of the neck after decapitation of ostrich embryos and blood drainage. After fixation, processing, blocking, and sectioning, all samples were stained by Hematoxylin and Eosin, Alcian Blue (AB), Van Gieson, and Periodic acid-Schiff (PAS) techniques. It was concluded that the trachea in the 13-day-old embryo and goblet cells (PAS-positive and AB-positive) had incomplete rings of hyaline cartilage and differentiation of mesenchymal to the loose connective tissue. The bronchial stage of the lung was observed in the 22-day-old embryo, pseudoglandular stage in the 26-day-old embryo, and parabrachial and air capillary stage in the 30-day-old embryo. The information obtained from this study will be useful for diagnosing pathologies affecting this vital system and results in improving industrial breeding management.

Quantitation of Mucin by Densitometry of an Alcian Blue-Stained Membrane.

It is a challenging task to quantify mucin using conventional protein quantification methods due to the large number of glycans attached to the peptide, which make up approximately 50-90% of its molecular weight. To address this issue, we propose a simple quantification method that involves spotting mucins onto a membrane and staining them with Alcian blue.

Effective separation of dyes/salts by sulfonated covalent organic framework membranes based on phenolamine network conditioning.

This study developed a modified polyacrylonitrile (PAN) membrane controlled by a phenol-amine network and enhanced with a sulfonated covalent organic framework (SCOF), aimed at improving the efficiency of textile wastewater treatment. Utilizing a phenol-amine network control strategy allows for precise manipulation of interfacial reactions in the synthesis of SCOF, achieving highly uniform modification on the surface of the PAN membrane. This modified membrane demonstrated high rejection of over 98% for various water-soluble dyes, including Alcian blue 8GX, Coomassie Brilliant Blue G250, methyl blue, congo red, and rose bengal, and also exhibited specific selectivity in processing salt-containing wastewater. By adjusting the deposition time of the phenol-amine and the concentration of SCOF monomers, optimal retention performance and permeate flux were achieved, effectively separating dyes and salts. This research provides a new and effective solution for treating textile wastewater, especially in separating and recovering dyes and salts, offering broad application prospects in environmental management and water resource management, and highlighting its significant practical implications.

A pilot study for decellularizing human and porcine cornea for future use in corneal regeneration

Purpose: To effectively decellularize human and porcine corneas for future use in corneal regeneration by using an already established decellularization technique being efficient in decellularizing blood vessels.Methods: One human and 4 porcine corneas were decellularized by using various washing steps with 3‐[(3‐cholamidopropyl) dimethylammonio]‐1‐propanesulfonate (CHAPS) detergent, Benzonase, Ethylenediaminetetraacetic acid (EDTA), 5% Dextran or 5% D‐Mannitol. The quality of the decellularization process was assessed by quantitative and qualitative measurement of DNA content, glycosaminoglycans (GAGs), immunofluorescent staining for Collagen I, V and keratocan, and Transmission Electron Microscopy (TEM) to observe the structure of the collagen fibrils was used.Results: The decellularization process showed 99,94% and 99,52% DNA content reduction in the human and porcine corneas when using Dextran, respectively, while more DNA remained in the porcine corneas when D‐Mannitol was used. Similar pattern was observed in the preservation of GAGs, with the highest amount being found in the human cornea, and less in the porcine corneas. H&E and Immunofluorescent staining showed no presence of cell nuclei when compared to the control native corneas, while Alcian blue staining qualitatively confirmed the presence of GAGs. Collagen I, V and keratocan were still present in the decellularized corneas when compared to the native ones, while TEM microscopy further confirmed the similar patterns of the collagen fibrils in the decellularized‐, compared to the native‐ corneas.Conclusions: This pilot study showed that our decellularization method is effective in decellularizing human and porcine corneas, with very high amount of DNA being removed from the corneas, while the GAGs were preserved to an acceptable extent, and the structure and pattern of the collagen fibrils maintained.

An unusually large superficial angiomyxoma with delayed presentation due to COVID-19: a case report

Background: Superficial angiomyxoma is a rare, benign, soft-tissue tumor. It has a non-specific presentation and there is no clear diagnostic investigation. Therefore, it is usually not suspected pre-operatively and managed incorrectly with a biopsy or narrow margin excision, which increases the recurrence rate. Case Presentation: A case of an unusually large superficial angiomyxoma is described with a delayed presentation to a plastic surgery clinic due to the COVID-19 pandemic. The patient was a 53-year-old male with an atraumatic 8 × 4 cm subcutaneous, mobile swelling to the left thigh. Excision was performed with no evidence of recurrence at 6 months. The mass was an uniloculated cyst with gelatinous myxoid stroma. Immunohistochemistry was positive for Alcian blue and CD34. Conclusion: This case report raises awareness of this rare lesion and provides a review of diagnostic aids as well as clear intra-operative photography of what a surgeon might expect when this tumor is encountered.

An unusually large superficial angiomyxoma with delayed presentation due to COVID-19: a case report

Background: Superficial angiomyxoma is a rare, benign, soft-tissue tumour. It has a non-specific presentation and there is no clear diagnostic investigation. Therefore, it is usually not suspected pre-operatively and managed incorrectly with a biopsy or narrow margin excision, which increases the recurrence rate. Case presentation: A case of an unusually large superficial angiomyxoma is described with a delayed presentation to a plastic surgery clinic due to the COVID-19 pandemic. The patient was a 53-year-old male with an atraumatic 8x4 cm subcutaneous, mobile swelling to the left thigh. Excision was performed with no evidence of recurrence at six months. The mass was an uniloculated cyst with gelatinous myxoid stroma. Immunohistochemistry was positive for Alcian blue and CD34. Conclusion: This case report raises awareness of this rare lesion and provides a review of diagnostic aids as well as clear intra-operative photography of what a surgeon might expect when this tumour is encountered.

Succinylation-Alcian Blue Staining of Mucins on Polyvinylidene Difluoride Membrane.

Mucins are often stained with the basic dye Alcian blue, but mucins with a low acidic glycan content cannot be stained with it. Succinylation-Alcian blue staining is a method that temporarily modifies glycans with succinic acid to visualize mucins with low acidic glycan content. This method can be used to stain mucins on polyvinylidene difluoride (PVDF) membranes separated via supported molecular matrix electrophoresis (SMME) and mucins blotted onto PVDF membranes from gel electrophoreses. The succinyl groups of the modified glycans can be easily and completely removed by releasing O-glycan from the stained mucin bands. Therefore, the glycans can be analyzed using the same methods as those used for mucins with a high acidic glycan content.

Regeneration of amputated mice digit tips by including Wnt signaling pathway with CHIR99021 and IWP-2 chemicals in limb organ culture system.

Mammals have limited limb regeneration compared to amphibians. The role of Wnt signaling pathways in limb regeneration has rarely been studied. So, this study aimed to investigate the effect of Wnt-signaling using chemicals CHIR99021and IWP-2on amputated mice digit tips regeneration in aninvitroorgan culture system. The distal phalanx of paws from C57BL/6J mouse fetuses at E14.5, E16.5, and E18.5 was amputated.Then, the hands were cultured for 7 days.Subsequently, paws were treated with 1-50 µg/ml concentration of CHIR99021 and 5-10 µg/ml concentration of IWP-2. Finally, the new tissue regrowth was assessed by histological analysis, immunohistochemistry for BC, TCF1, CAN, K14, and P63 genes, and beta-catenin and Tcf1 genes were evaluated with RT-qPCR. The paws of E14.5 and E16.5 days were shrinkaged and compressed after 7 days, so the paws of 18.5E that were alive were selected.As a result, newly-grown masses at digit tips were observed in 25 and 30 µl/ml concentrations of the CHR99021 group but not in the IWP2 treatment (*P<0.05; **P<0.01). qRT-PCR analysis confirmed the significant up-regulation of beta-catenin and Tcf1 genes in CHIR99021 group in comparison to the IWP-2 group (P<0.05). Moreover, Alcian-blue staining demonstrated the presence of cartilage-like tissue at regenerated mass in the CHIR group. In immunohistochemistry analysis beta-catenin, ACN, Keratin-14, and P63 protein expression were observed in digit tips in the CHIR-treated group. By activating the Wnt signaling pathway, cartilage-like tissue formed in the blastema-like mass in the mouse's amputated digit tips.

Role of Alcian Blue Stain in Differentiating Prostatic Adenocarcinoma from Nodular Hyperplasia of Prostate

Objective: Enlargement of prostate is one of the leading cause of morbidity in men. Alcian Blue stain demonstrates that the acidic mucin is present in prostatic adenocarcinoma but absent in nodular hyperplasia of prostste. So this histochemical marker can be used in distinguishing them. This study aims to determine the role of Alcian Blue stain in differentiating prostatic adenocarcinoma from the nodular hyperplasia of prostate. Materials and Methods: This cross-sectional study was conducted in the Department of Pathology, Rajshahi Medical College, Rajshahi and in the Department of Pathology, Bangabandhu Sheikh Mujib Medical University (BSMMU) during the period of March 2018 to February 2020. Specimens were processed routinely for Haematoxilin and Eosin stain. The Alcian Blue stain was performed to demonstrate the presence of acidic mucin and the findings were categorized as positive or negative staining. Statistical analyses was carried out by using the Statistical package for Social Sciences (SPSS) version 20 for Windows. The components of accuracy test were computed by respective formulae for the tests. Result: The study involved a total 60 specimens of prostatic tissue among which 30 were histopathologically diagnosed cases of prostatic adenocarcinoma and another 30 were histopathologically diagnosed cases of nodular hyperplasia. Among the 30 cases of prostatic adenocarcinoma the acidic mucin was positive in 25 (83.3 %) cases and negative in 5 (16.7%) cases but in nodular hyperplasia it was positive in 8 (26.6%) cases and negative in 22 (73.4%) cases. The sensitivity of Alcian Blue stain in favour of malignancy was 83.3% and specificity was 73.3%. The overall diagnostic accuracy of Alcian Blue stain was 78.3% and it showed a statistically significant difference in staining between the cases of adenocarcinoma and nodular hyperplasia, indicated by p&lt;0.05. Conclusion: Alcian Blue stain has a valuable role in differentiating prostatic adenocarcinoma from the nodular hyperplasia. So it can be used in the diagnosis of prostatic adenocarcinoma. TAJ 2023; 36: No-2: 57-63

Comparison of Different Methods of Semiquantitative Assessment and Subjective Scores for Retropatellar Articular Cartilage Evaluation in Advancing Osteoarthritis.

cartilage layer thickness, composition of the extracellular matrix (ECM), geometry and configuration of retropatellar cartilage partially differ significantly from those found at other locations and are essential for patellofemoral biomechanics. 119 serial medial and lateral patella facet samples of patients undergoing implantation of a total knee endoprosthesis of areas showing mild (Group A, macroscopically ICRS Grade 1b) respectively advanced (Group B, macroscopically ICRS Grade 3a/3b) (38 each) osteoarthritis according to the histological-histochemical grading system (HHGS) were compared with 22 healthy biopsies by means of immunohistochemistry and histology investigations. We quantified our results on the gene expression of collagen type I and II and aggrecan with real-time (RT)-PCR rsp using colourimetry for proteoglycan content. The digitized images of histology and immunohistochemistry staining with Safranin O, Alcian blue, PAS, Masson Goldner, immunostaining, e.g. for collagen I and II were also analyzed with Photoshop software. The most significant decrease in staining intensity was revealed for Safranin-O staining in Group B at the lateral patellar facet, and the most relevant increase was for Col I staining at the same site. The lateral patella site in Group B also showed the highest increase in the ratio of expression indices for the genes Col1A1 and the reference gene following the equation 2-ΔCt with a quotient of 29.6. 1. Comparisons of our retropatellar cartilage analysis with femoral and tibial studies utilizing similar techniques show significant differences. 2. Cartilage layer thickness, ECM composition, geometry and configuration are essential for patellofemoral biomechanics. 3. Consequently, there is a need for diversified approaches towards retropatellar surface during TKA as well as for advanced cartilage restoration techniques.

Regenerative potential of platelet derived growth factor in nicotine induced intervertebral disc degenerative model – In vivo study

Intervertebral disc (IVD) degeneration is an age-related disease affecting the elderly population worldwide. The disease is contributed by degeneration of the cartilage tissue in the vertebral column. The current study explores the possibility of exploiting platelet derived growth factor (PDGF), a growth factor secreted by platelets, as a therapeutic agent against the IVD degeneration. BALB/c mice were exposed to nicotine at higher concentrations to induce IVD degeneration. Three different concentrations of PDGF (1 ng/mL, 2 ng/mL, 3 ng/mL) were used in the study. Oxidative stress parameters were assessed through Lipid peroxidation (LPO), Myeloperoxidase (MPO) and Nitric oxide (NO). Histopathology was done for assessing the extent of degeneration, collagen II, mucin, and calcium deposition by Haematoxylin and Eosin staining, immunohistochemistry, Alcian blue and Alizarin red staining respectively. Gene expression studies were carried out by quantitative real-time Polymerase chain reaction (RT-PCR) for Aggrecan, alkaline phosphatase, RUNX2, Collagen I alpha and osteocalcin. IVD degeneration was prominent and PDGF treatment restored the tissue as evidenced by H&E staining. Oxidative stress was induced by the nicotine treatment and all the biomarkers of oxidative stress were restored back to normal by PDGF in a dose dependent manner. Biomarkers of endochondral bone formation (aggrecan, alkaline phosphatase, RUNX2, Collagen I alpha and osteocalcin) was observed to be normalized by exposing to PDGF in mRNA level though RT-PCR. Mucin, calcium contents were also brought back to normal by PDGF treatment. In conclusion, current work strongly supports that PDGF could be used as a therapeutic agent for the treatment of IVD degeneration.

Do fossorial water voles have a functional vomeronasal organ? A histological and immunohistochemical study.

The fossorial water vole, Arvicola scherman, is an herbivorous rodent that causes significant agricultural damages. The application of cairomones and alarm pheromones emerges as a promising sustainable method to improve its integrated management. These chemical signals would induce stress responses that could interfere with the species regular reproductive cycles and induce aversive reactions, steering them away from farmlands and meadows. However, there is a paucity of information regarding the water vole vomeronasal system, both in its morphological foundations and its functionality, making it imperative to understand the same for the application of chemical communication in pest control. This study fills the existing gaps in knowledge through a morphological and immunohistochemical analysis of the fossorial water vole vomeronasal organ. The study is primarily microscopic, employing two approaches: histological, using serial sections stained with various dyes (hematoxylin-eosin, Periodic acid-Schiff, Alcian blue, Nissl), and immunohistochemical, applying various markers that provide morphofunctional and structural information. These procedures have confirmed the presence of a functional vomeronasal system in fossorial water voles, characterized by a high degree of differentiation and a significant expression of cellular markers indicative of active chemical communication in this species.

Effects of diquafosol sodium in povidone iodine-induced dry eye model.

To investigate the effects of diquafosol sodium (DQS) for dry eye model induced with povidone-iodine (PI) solution. Ten Sprague Dawley rats as the control group. Thirty Sprague Dawley rats were used to establish the dry eye model with stimulation of 10 g/L PI for 14d, then divided rats into three groups: dry eye group with no treatment (DED group, n=10); phosphate buffer saline treated group (PBS group, n=10); diquafosol treated group (DQS group, n=10). Clinical changes were observed by tear production test, fluorescein staining, tear break-up time (TBUT) test, corneal confocal microscope and ocular surface comprehensive analyzer. Eyeballs were collected on day 10 of treatment for hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and alcian blue staining. TUNEL assay, polymorphonuclear (PMN) and mucin 1 (MUC1) immunofluorescence were performed and corneal ultrastructural changes were detected by electron microscopy. Compared with DED and PBS groups, tear production (7.26±0.440 vs 4.07±0.474 mm; 7.26±0.440 vs 3.74±0.280 mm; all P<0.01) and TBUT (7.37±0.383s vs 1.49±0.260s; 7.37±0.383s vs 1.42±0.437s; all P<0.01) were significantly increased in DQS group. HE, PAS, and alcian blue staining and MUC1 immunofluorescence showed mucins and conjunctival goblet cells density (8.45±0.718 vs 5.21±0.813 cells/0.1 mm2; 8.45±0.718 vs 5.36±0.615 cells/0.1 mm2; all P<0.01) increased in DQS group. Confocal microscopy, PMN immunofluorescence and TUNEL staining showed inflammatory infiltration and corneal epithelial cells apoptosis decreased in DQS group. The increased number of microvilli in corneal epithelial and the recovered cell junction were observed in DQS group. PI instillation can induce goblet cells and mucin loss, epithelial cell apoptosis and inflammation, which are consistent with the pathological manifestations of dry eye. Diquafosol can repair the ocular surface damage caused by PI, reduce corneal inflammation, inhibit corneal epithelial cell apoptosis, promote mucin secretion and maintain tear film stability.

The enteroprotective effect of nifuroxazide against methotrexate-induced intestinal injury involves co-activation of PPAR-γ, SIRT1, Nrf2, and suppression of NF-κB and JAK1/STAT3 signals

Methotrexate (MTX) has long manifested therapeutic efficacy in several neoplastic and autoimmune disorders. However, MTX-associated intestinal toxicity restricts the continuation of treatment. Nifuroxazide (NIF) is an oral antibiotic approved for gastrointestinal infections as an effective antidiarrheal agent with a high safety profile. The current study was designed to explore the potential efficacy of NIF in alleviating intestinal toxicity associated with MTX chemotherapy with the elucidation of the proposed molecular mechanisms. Rats were administered NIF (50 mg/kg; p.o.) for ten days. On day five, a single i.p. injection of MTX (20 mg/kg) was given to induce intestinal intoxication. At the end of the experiment, duodenal tissue samples were isolated for biochemical, Western blotting, immunohistochemical (IHC), and histopathological analysis via H&E, PSA, and Alcian blue stains. NIF showed antioxidant enteroprotective effects against MTX intestinal intoxication through enhanced expression of the redox-sensitive signals of PPAR-γ, SIRT1, and Nrf2 estimated by IHC. Moreover, NIF down-regulated the pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), NF-κB protein expression, and the phosphorylation of JAK1/STAT3 proteins, leading to mitigation of intestinal inflammation. In accordance, the histological investigation revealed that NIF ameliorated the intestinal pathological changes, preserved the goblet cells, and reduced the inflammatory cells infiltration. Therefore, NIF could be a promising candidate for adjunctive therapy with MTX to mitigate the associated intestinal injury and increase its tolerability.

Positive Regulation of S-Adenosylmethionine on Chondrocytic Differentiation via Stimulation of Polyamine Production and the Gene Expression of Chondrogenic Differentiation Factors.

S-adenosylmethionine (SAM) is considered to be a useful therapeutic agent for degenerative cartilage diseases, although its mechanism is not clear. We previously found that polyamines stimulate the expression of differentiated phenotype of chondrocytes. We also found that the cellular communication network factor 2 (CCN2) played a huge role in the proliferation and differentiation of chondrocytes. Therefore, we hypothesized that polyamines and CCN2 could be involved in the chondroprotective action of SAM. In this study, we initially found that exogenous SAM enhanced proteoglycan production but not cell proliferation in human chondrocyte-like cell line-2/8 (HCS-2/8) cells. Moreover, SAM enhanced gene expression of cartilage-specific matrix (aggrecan and type II collagen), Sry-Box transcription factor 9 (SOX9), CCN2, and chondroitin sulfate biosynthetic enzymes. The blockade of the methionine adenosyltransferase 2A (MAT2A) enzyme catalyzing intracellular SAM biosynthesis restrained the effect of SAM on chondrocytes. The polyamine level in chondrocytes was higher in SAM-treated culture than control culture. Additionally, Alcian blue staining and RT-qPCR indicated that the effects of SAM on the production and gene expression of aggrecan were reduced by the inhibition of polyamine synthesis. These results suggest that the stimulation of polyamine synthesis and gene expression of chondrogenic differentiation factors, such as CCN2, account for the mechanism underlying the action of SAM on chondrocytes.

Carbon quantum dot-nanocomposite hydrogel as Denovo Nexus in rapid chondrogenesis

The incapability of cartilage to naturally regenerate and repair chronic muscular injuries urges the development of competent bionic rostrums. There is a need to explore faster strategies for chondrogenic engineering using mesenchymal stem cells (MSCs). Along these lines, rapid chondrocyte differentiation would benefit the transplantation demand affecting osteoarthritis (OA) and rheumatoid arthritis (RA) patients. In this report, a de novo nanocomposite was constructed by integrating biogenic carbon quantum dot (CQD) filler into synthetic hydrogel prepared from dimethylaminoethyl methacrylate (DMAEMA) and acrylic acid (AAc). The dominant structural integrity of synthetic hydrogel along with the chondrogenic differentiation potential of garlic peel derived CQDs led to faster chondrogenesis within 14 days. By means of extensive chemical and morphological characterization techniques, we illustrate that the hydrogel nanocomposite possesses lucrative features to influence rapid chondrogenesis. These results were further corroborated by bright field imaging, Alcian blue staining and Masson trichome staining. Thus, this stratagem of chondrogenic engineering conceptualizes to be a paragon in clinical wound care for the rapid manufacturing of chondrocytes.

Development and Characterization of Decellularized Lung Extracellular Matrix Hydrogels.

The use of extracellular matrix (ECM)-derived hydrogels in tissue engineering has become increasingly popular, as they can mimic cells' natural environment in vitro. However, maintaining the native biochemical content of the ECM, achieving mechanical stability, and comprehending the impact of the decellularization process on the mechanical properties of the ECM hydrogels are challenging. Here, a pipeline for decellularization of bovine lung tissue using two different protocols, downstream characterization of the effectiveness of decellularization, fabrication of reconstituted decellularized lung ECM hydrogels and assessment of their mechanical and cytocompatibility properties were described. Decellularization of the bovine lung was pursued using a physical (freeze-thaw cycles) or chemical (detergent-based) method. Hematoxylin and Eosin staining was performed to validate the decellularization and retention of major ECM components. For the evaluation of residual collagen and sulfated glycosaminoglycan (sGAG) content within the decellularized samples, Sirius red and Alcian blue staining techniques were employed, respectively. Mechanical properties of the decellularized lung ECM hydrogels were characterized by oscillatory rheology. The results suggest that decellularized bovine lung hydrogels can provide a reliable organotypicalternative to commercial ECM products by retaining most native ECM components. Furthermore, these findings reveal that the decellularization method of choice significantly affects gelation kinetics as well as the stiffness and viscoelastic properties of resulting hydrogels.

Liquid platelet-rich fibrin produced via horizontal centrifugation decreases the inflammatory response and promotes chondrocyte regeneration in vitro.

Objective: Recently, liquid platelet-rich fibrin (PRF), a rich source of concentrated platelets and growth factors, has emerged as a promising agent for stimulating tissue regeneration. However, its specific efficacy in chondrocyte proliferation and cartilage regeneration remains underexplored. To address this question, we investigated liquid PRF obtained through horizontal centrifugation and compared its effects with hyaluronic acid (HA), a high molecular weight glucosamine supplement widely used in clinical practice to safeguard against chondral damage. Materials and Methods: Liquid PRF, produced using horizontal centrifugation (liquid H-PRF) at 500g for 8min, served as our experimental agent. We conducted cell viability and proliferation assays using PRF-conditioned medium. We assessed the chondrocyte phenotype of ATDC5 cells through toluidine blue and alcian blue staining, real-time polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence staining. Furthermore, we examined the expression of genes involved in inflammation through RT-PCR and Western blot analysis. Results: Liquid H-PRF exerted notable effects on chondrocytes, influencing proliferation, inflammatory responses, and chondrogenic differentiation. The H-PRF group displayed significantly higher expression of chondrogenic markers, including Col2a1, compared to HA-treated cells, whereas aggrecan expression was significantly higher in the HA group. PRF also demonstrated the ability to reduce inflammatory levels in chondrogenic ATDC5 cells, and this effect was further enhanced when PRF from the buffy coat zone was added. In comparison, chondrocytes cultured in the HA group produced significantly fewer inflammatory factors than those in the PRF group, as confirmed qualitatively by Western blot analysis. Conclusion: Liquid H-PRF emerged as a potent stimulator for chondrogenesis and a regulator of the inflammatory response, achieving levels similar to HA. Moreover, liquid H-PRF exhibited strong potential for enhancing the production of cartilage extracellular matrix and promoting chondrogenic regeneration with notably increased Col2a1 levels. Future research should encompass animal studies and human trials to further evaluate the comparative effectiveness of liquid PRF versus HA, potentially as an alternative or complementary strategy for future clinical applications.

Alzheimer’s disease induces pathological changes in the lungs of humans and the AppNL‐G‐F mouse model

AbstractBackgroundAlzheimer’s disease (AD) is an age‐related neurodegenerative disorder characterized by brain accumulation of amyloid beta (Aβ) plaques, neurofibrillary tangles, neuron death, robust gliosis, and neuroinflammation, affecting around 35 million individuals worldwide. It is well known that brain immune changes are a component of the disease, but immune dysfunction in secondary organs has also been reported. To assess peripheral immune changes in AD, we asked whether lung dysfunction and inflammatory change are present during the disease.MethodMale and female C57BL/6 wild type and AppNL‐G‐F mice (8‐9 months old) were sacrificed, and broncho‐alveolar lavage fluid (BALF) and lungs were collected. BALF was used to quantify immune cell phenotype by flow cytometry, and the BALF supernatant was used to determine Aβ and cytokine concentrations and total protein content. Lungs were sectioned and stained with hematoxylin and eosin, Alcian blue and Masson’s trichrome. In addition, lung sections from wild‐type and AppNL‐G‐F mice and human AD patient and healthy controls were immunostained for APP, Aβ and claudin 4.ResultAD mice showed significantly increased pulmonary airway thickening and mucus and collagen production as indicated by hematoxylin and eosin, Alcian blue and Masson’s trichrome staining, respectively, compared to the wild‐type controls. Female but not male AppNL‐G‐F mice also demonstrated reduced B cell and neutrophil numbers and elevated IL‐1β in their BALF compared to controls. Although there were no differences in BALF supernatant total protein content between wild‐type and AppNL‐G‐F mice, lungs from both sexes of AppNL‐G‐F mice had decreased claudin‐4 immunoreactivity suggesting altered epithelial permeability. BALF supernatant from male and female AppNL‐G‐F mice also contained soluble Aβ1‐40 and Aβ1‐42 correlating with lung APP immunoreactivity. Human AD male and female lungs had reduced claudin‐4 immunoreactivity and increased APP and Aβ staining compared to controls verifying lung changes are also present in human disease.ConclusionThese data demonstrate disease‐associated changes in the lungs of both human AD patients and the AppNL‐G‐F mouse model. Further characterization of lung dysfunction and its contribution to brain changes may define a novel lung‐to‐brain contribution to disease.