Articular Cartilage Formation Research Articles

An Advanced Bioconjugation Strategy for Covalent Tethering of TGFβ3 with Silk Fibroin Matrices and its Implications in the Chondrogenesis Profile of Human BMSCs and Human Chondrocytes: A Paradigm Shift in Cartilage Tissue Engineering.

The transforming growth factor-β class of cytokines plays a significant role in articular cartilage formation from mesenchymal condensation to chondrogenic differentiation. However, their exogenous addition to the chondrogenic media makes the protocol expensive. It reduces the bioavailability of the cytokine to the cells owing to their burst release. The present study demonstrates an advanced bioconjugation strategy to conjugate transforming growth factor-β3 (TGFβ3) with silk fibroin matrix covalently via a cyanuric chloride coupling reaction. The tethering and change in secondary conformation are confirmed using various spectroscopic analyses. To assess the functionality of the chemically modified silk matrix, human bone marrow-derived mesenchymal stem cells (hBMSCs) and chondrocytes are cultured for 28 days in a chondrogenic differentiation medium. Gene expression and histological analysis reveal enhanced expression of chondrogenic markers with intense Safranin-O and Alcian Blue staining in TGFβ3 conjugated silk matrices than where TGFβ3 is exogenously added to the media for both hBMSCs and chondrocytes. Therefore, this study successfully recapitulates the native niche of TGFβ3 and the role of the silk as a growth factor stabilizer. When cultured over TGFβ3 conjugated silk matrices, hBMSCs display increased proteoglycan secretion and maximum chondrogenic trait with attenuation of chondrocyte hypertrophy over human chondrocytes.

Selenium Deficiency Can Promote the Expression of VEGF and Inflammatory Factors in Cartilage Differentiation and Mediates Cartilage Injury.

Selenium plays a crucial role as a micronutrient, primarily exerting its biological functions through selenoproteins. It has been established that selenium deficiency adversely impacts cartilage development, leading to alterations in chondrocyte function. In regions with low selenium intake, endemic osteochondrosis has been documented, characterized by compromised growth plate and articular cartilage formation. Vascular endothelial growth factor (VEGF) stands out as a pivotal angiogenic factor, with elevated levels contributing significantly to vascular invasion into chondrocytes. This VEGF-mediated invasion serves as a key signal, prompting morphological changes in the growth plate and initiating cartilage remodeling. In animal models, the selenium deficiency group exhibited heightened levels of the cartilage damage marker matrix metalloproteinases 13 (MMP13). This resulted in articular cartilage degeneration, accompanied by a substantial increase in VEGF expression within the growth plate and articular cartilage, as compared to the normal group. In a chondrogenic progenitor cell (CPC) differentiation model, insufficient selenium induced chondrocyte damage and upregulated inflammatory factors such as inducible NO synthase (iNOS) and cyclooxygenase-2 (COX2). The selenium-deficient groups showed elevated expressions of VEGF, VEGFR2, MMP13, Collagen X, and Angiopoietin 1, accelerating the degradation of the extracellular matrix (ECM), which further promoted the development of cartilage-related diseases. Taken together, these findings provide novel insights for a better understanding of the role of low selenium in cartilage degeneration and angiogenesis. They shed light on the intricate influence of low selenium levels on the development of articular cartilage, emphasizing the interconnected pathways and processes involved.

Morphohistological study of regeneration of knee joint cartilage defects in an experimental model under the influence of nuclear magnetic resonance therapy

Despite the variety of modern methods of treatment, the problem of hyaline cartilage regeneration is still relevant. Purpose of the study: to determine the effect of nuclear magnetic resonance therapy on the regenerative ability of simulated defects in the cartilage tissue of the knee joint in rats, to assess the dynamics of microscopic changes in articular cartilage in the main and control groups. The study was performed on 60 sex-mature rats. The defects were created in the area of the articular surfaces of the left knee joint – 30 defects in the main group and 30 defects in the control group. The right knee joint of both study groups was used as the norm. Medical (antibiotics, anti-inflammatory, analgesic) therapy was carried out for 3 days after the surgery. On the 4th day after surgery, rats of the main group were given nuclear magnetic resonance therapy for 60 minutes, for 7 days. After 7, 14, 21, 28 days after therapy, histological analysis of cartilage regenerate was performed. 28 days after the use of nuclear magnetic resonance therapy, the height of cartilage regenerate in rats was 82.12±8.89 μm in the intervention group and 56.34±7.82 μm in the control group. Cartilage regenerate in rats after nuclear magnetic resonance therapy was close to the structure of intact hyaline cartilage. However, complete regeneration did not occur, as evidenced by the smaller thickness of the articular cartilage compared to that in the right knee joint. In the control group, the formation of the regeneration had pronounced signs of dysregeneration. The cartilage tissue in the area of the defect, was predominantly fibrous in the nature with areas of necrosis. Nuclear magnetic resonance therapy contributes to the formation of articular cartilage in the defect – cartilage regenerate, which in its histological structure approaches hyaline cartilage.

A New Bioactive Fibrin Formulation Provided Superior Cartilage Regeneration in a Caprine Model

The effective and long-term treatment of cartilage defects is an unmet need among patients worldwide. In the past, several synthetic and natural biomaterials have been designed to support functional articular cartilage formation. However, they have mostly failed to enhance the terminal stage of chondrogenic differentiation, leading to scar tissue formation after the operation. Growth factors substantially regulate cartilage regeneration by acting on receptors to trigger intracellular signaling and cell recruitment for tissue regeneration. In this study, we investigated the effect of recombinant insulin-like growth factor 1 (rIGF-1), loaded in fibrin microbeads (FibIGF1), on cartilage regeneration. rIGF-1-loaded fibrin microbeads were injected into full-thickness cartilage defects in the knees of goats. The stability, integration, and quality of tissue repair were evaluated at 1 and 6 months by gross morphology, histology, and collagen type II staining. The in vivo results showed that compared to plain fibrin samples, particularly at 6 months, FibIGF1 improved the functional cartilage formation, confirmed through gross morphology, histology, and collagen type II immunostaining. FibIGF1 could be a promising candidate for cartilage repair in the clinic.

A Bibliometric Analysis of Exosomes Therapy in the Treatment of Osteoarthritis from 2012 to 2022.

Osteoarthritis (OA) is a common clinical disease characterized by the destruction of articular cartilage, subchondral ossification, cystic degeneration and osteophyte formation. Recently, more and more scholars draw attention to exosomes in the field of OA, and exciting breakthroughs have been achieved in recent years. However, bibliometric analysis of the literature in this research field is lacking. Considering its potential in treatment of OA, this article aimed to analyze the research status and identify future hotspots of exosomes in osteoarthritis in recent 10 years by bibliometrics tools. Relevant publications in this field from 2012 to 2022 was retrieved from the Web of Science core collection database (WOSSCC). And we used Vosviewers, CiteSpace, an online analysis platform and the R package "Bibliometrix" for bibliometric analysis. A total of 484 publications (including 319 articles and 165 reviews) from 51 countries, 720 institutions, were included in this study. IRCCS Ist Ortoped Galeazzi, Shanghai Jiao Tong University, and Sun Yat-sen University are the leading research institutions in this field. International Journal of Molecular Sciences contributed the largest number of articles, and Osteoarthritis and Cartilage is the most co-cited journal. Of the 2664 scholars who participated in the study, Ragni E, De Girolamo L, Orfei CP, and Colombini A had the largest number of articles. Zhang, SP is the most co-cited author. "Mesenchymal stem cell", "biomaterials", "Inflammation" and "regenerative medicine" are the keywords in the research. This is the first bibliometric analysis of exosomes in osteoarthritis. We explored current research status in recent years and identified frontiers and hot spots in this research field. We highlight the significant roles of mesenchymal stem cell-derived exosomes (MSC-Exos) in the treatment of osteoarthritis, and identified exosomal biomaterials as frontier in this research domain, which can provide reference for the researchers who focus on this research field.

Preparation of mesoporous silica nanocarriers targeting glucose-6-phosphate isomerase inhibition and application in the treatment of rheumatoid arthritis.

Glucose 6-phosphate isomerase (G6PI) is an indicator to assist in diagnosis of rheumatoid arthritis (RA) and monitor the disease. It also plays a key role in proliferating RA synovial tissues, pannus formation, and invasion and destruction of articular cartilage. In this study, we synthesized nanoparticles targeting G6PI (siG6PI-MSN) using mesoporous silica nanocarriers (MSN) and small interfering RNA (siRNA), followed by identifying the characteristics and functions, and preliminarily exploring their application in the treatment of RA in vivo with a type II collagen-induced arthritis (CIA) rat model. It showed that the synthetic functionalized carrier had a regular pore structure and a specific volume and surface area. No obvious hemolysis or toxicity of the carrier was found when its concentration was below 100 µg/ml. Cytological results in vitro suggested that siG6PI-MSN significantly inhibited G6PI expression and reduced the ability of proliferation, migration, and invasion of FLSs, compared with the siNC-MSN group. In vivo results in the CIA rat model showed that the arthritis index and degree of joint swelling among rats in the siG6PI-MSN-treatment group were significantly lower than those in the control group. Moreover, the number of FLSs in Synovium and the levels of TNF α and IL-1 β were also significantly decreased in the siG6PI-MSN group. Histopathology of the synovial tissue and cartilage revealed siG6PI-MSN treatment significantly reduced the pathological manifestations of arthritis. In conclusion, siG6PI-MSN effectively suppresses the proliferation and invasive growth of synovial tissue and improve joint swelling and inflammatory infiltration, thereby preventing joint damage in RA. This carrier may be a new therapeutic measure for RA, with potential social and economic benefits.

Screening and identification of potential hub genes and immune cell infiltration in the synovial tissue of rheumatoid arthritis by bioinformatic approach

BackgroundRheumatoid arthritis (RA) is an autoimmune disease that affects individuals of all ages. The basic pathological manifestations are synovial inflammation, pannus formation, and erosion of articular cartilage, bone destruction will eventually lead to joint deformities and loss of function. However, the specific molecular mechanisms of synovitis tissue in RA are still unclear. Therefore, this study aimed to screen and explore the potential hub genes and immune cell infiltration in RA. MethodsThree microarray datasets (GSE12021, GSE55457, and GSE55235), from the Gene Expression Omnibus (GEO) database, have been analyzed to explore the potential hub genes and immune cell infiltration in RA. First, the LIMMA package was used to screen the differentially expression genes (DEGs) after removing the batch effect. Then the clusterProfiler package was used to perform functional enrichment analyses. Second, through weighted coexpression network analysis (WGCNA), the key module was identified in the coexpression network of the gene set. Third, the protein-protein interaction (PPI) network was constructed through STRING website and the module analysis was performed using Cytoscape software. Fourth, the CIBERSORT and ssGSEA algorithm were used to analyze the immune status of RA and healthy synovial tissue, and the associations between immune cell infiltration and RA-related diagnostic biomarkers were evaluated. Fifth, we used the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) to validate the expression levels of the hub genes, and ROC curve analysis of hub genes for discriminating between RA and healthy tissue. Finally, the gene-drug interaction network was constructed using DrugCentral database, and identification of drug molecules based on hub genes using the Drug Signature Database (DSigDB) by Enrichr. ResultsA total of 679 DEGs were identified, containing 270 downregulated genes and 409 upregulated genes. DEGs were primarily enriched in immune response and chemokine signaling pathways, according to functional enrichment analysis of DEGs. WGCNA explored the co-expression network of the gene set and identified key modules, the blue module was selected as the key module associated with RA. Seven hub genes are identified when PPI network and WGCNA core modules are intersected. Immune infiltration analysis using CIBERSORT and ssGSEA algorithms revealed that multiple types of immune infiltration were found to be upregulated in RA tissue compared to normal tissue. Furthermore, the levels of 7 hub genes were closely related to the relative proportions of multiple immune cells in RA. The results of the qRT-PCR demonstrated that the relative expression levels of 6 hub genes (CD27, LCK, CD2, GZMB, IL7R, and IL2RG) were up-regulated in RA synovial tissue, compared with normal tissue. Simultaneously, ROC curves indicated that the above 6 hub genes had strong biomarker potential for RA (AUC >0.8). ConclusionsThrough bioinformatics analysis and qRT-PCR experiment, our study ultimately discovered 6 hub genes (CD27, LCK, CD2, GZMB, IL7R, and IL2RG) that closely related to RA. These findings may provide valuable direction for future RA clinical diagnosis, treatment, and associated research.

Bijadushti Janya Asthi Dhatu Kshaya with Special Reference to Progressive Pseudo-Rheumatoid Arthropathy (Dysplasia) of Childhood – A Case Report

Progressive pseudo-rheumatoid arthropathy dysplasia of childhood PPAC PPD is a genetic disorder characterized by poly articular non-inflammatory arthropathy due to impaired articular cartilage formation and maintenance caused by mutation in WISP3 WNT1-inducible signalling pathway protein 3 gene. Osteoarthritis joint space reduction and contracture formation results in severe handicap and incapacitation. Although few preventive measures are described to avert genetic disorders Ayurveda generally considers genetic disorders incurable. Hence palliative care becomes very important in managing such conditionsA North Indian 11.5 years old male child with Bijadushti Janya Vyadhi genetic disorder diagnosed as Progressive pseudo-rheumatoid arthropathy dysplasia of childhood PPAC PPD approached for pain and swelling of multiple joints gait and postural problems. Bijadushti was responsible for Asthi Dhatu Kshyaya diminution of bone tissue Sandhi Shotha joint swelling Sandhi Sankoch contractures Karma Hani loss of action and complications like lameness kyphosis and short stature in this case.He was managed with Ayurvedic treatment and Yoga and showed encouraging improvements in a short period. The treatment plan included Deepana-Pachana improving the digestion and metabolism Abhyanga therapeutic massage Swedana sudation and Basti therapeutic enema along with Asthidhatu Vardhana Chikitsa measures to supplement bone tissue Rasayana rejuvenation and range of motion ROM exercises in the form of Sukshma Vyayama gentle joint movements synchronized with breathing. If the etiopathogenesis of incurable genetic disorders is understood in the light of Ayurvedic tenets and palliative treatment is planned accordingly the patientrsquos life can be improved by providing symptomatic relief.

Runx1 is a key regulator of articular cartilage homeostasis by orchestrating YAP, TGFβ, and Wnt signaling in articular cartilage formation and osteoarthritis

Runt-related transcription factor 1 (Runx1) plays a key role in cartilage formation, but its function in articular cartilage formation is unclear. We generated non-inducible and inducible Runx1-deficient mice (Runx1f/fCol2α1-Cre and Runx1f/fCol2α1-CreER mice) and found that chondrocyte-specific Runx1-deficient mice developed a spontaneous osteoarthritis (OA)-like phenotype and showed exacerbated articular cartilage destruction under OA, characterized by articular cartilage degradation and cartilage ossification, with decreased Col2α1 expression and increased Mmp13 and Adamts5 expression. RNA-sequencing analysis of hip articular cartilage from the Runx1f/fCol2α1-Cre mice compared to that from wild-type mice and subsequent validation analyses demonstrated that Runx1 is a central regulator in multiple signaling pathways, converging signals of the Hippo/Yap, TGFβ/Smad, and Wnt/β-catenin pathways into a complex network to regulate the expression of downstream genes, thereby controlling a series of osteoarthritic pathological processes. RNA-sequencing analysis of mutant knee joints showed that Runx1’s role in signaling pathways in articular cartilage is different from that in whole knee joints, indicating that Runx1 regulation is tissue-specific. Histopathologic analysis confirmed that Runx1 deficiency decreased the levels of YAP and p-Smad2/3 and increased the levels of active β-catenin. Overexpression of Runx1 dramatically increased YAP expression in chondrocytes. Adeno-associated virus-mediated Runx1 overexpression in the knee joints of osteoarthritic mice showed the protective effect of Runx1 on articular cartilage damaged in OA. Our results notably showed that Runx1 is a central regulator of articular cartilage homeostasis by orchestrating the YAP, TGFβ, and Wnt signaling pathways in the formation of articular cartilage and OA, and targeting Runx1 and its downstream genes may facilitate the design of novel therapeutic approaches for OA.

MiR-200b-3p/ERG/PTHrP axis mediates the inhibitory effect of ethanol on the differentiation of fetal cartilage into articular cartilage

PurposeThis study aims to further explore cartilage development in prenatal ethanol exposure (PEE) offspring at different times to explore the specific time points and mechanism of ethanol-induced fetal cartilage dysplasia. MethodsOn gestational day (GD)14, GD17, and GD20, PEE fetal cartilage was evaluated by morphological analysis. RT-qPCR, immunohistochemistry, and immunofluorescence were used to detect the expression of cartilage marker genes and their regulatory factors. Bone marrow mesenchymal stem cells (BMSCs) were used to explore the effect of ethanol on the differentiation of chondrocytes. Additionally, we used inhibitors, overexpression plasmids and a luciferase reporter assay on GD17 chondrocytes to verify the mechanism. ResultsPEE significantly reduced cartilage matrix content and the expression of marker genes on GD17 and GD20 but had no effect on GD14. The inhibition of chondrogenic differentiation by PEE mainly occurred on GD14-17. Furthermore, the expression of miR-200b-3p was increased, while that of ERG and PTHrP was markedly reduced in PEE fetal cartilage. In vitro, ethanol (30–120 mM) inhibited the differentiation of BMSCs into chondrocytes in a concentration-dependent manner, accompanied by strong expression of miR-200b-3p and low expression of ERG and PTHrP. Moreover, PTHLH and ERG overexpressed, as well as a miR-200b-3p inhibitor reversed the inhibitory effect of ethanol on the differentiation of fetal chondrocytes. Furthermore, miR-200b-3p could target and negatively regulate ERG. ConclusionsPEE can significantly inhibit the development of articular cartilage, especially during articular cartilage formation. The mechanism is related to the decreased differentiation of fetal cartilage into articular cartilage mediated by the miR-200b-3p/ERG/PTHrP axis.

The synovial microenvironment suppresses chondrocyte hypertrophy and promotes articular chondrocyte differentiation

During the development of the appendicular skeleton, the cartilaginous templates undergo hypertrophic differentiation and remodels into bone, except for the cartilage most adjacent to joint cavities where hypertrophic differentiation and endochondral bone formation are prevented, and chondrocytes instead form articular cartilage. The mechanisms that prevent hypertrophic differentiation and endochondral bone formation of the articular cartilage have not been elucidated. To explore the role of the synovial microenvironment in chondrocyte differentiation, osteochondral allografts consisting of articular cartilage, epiphyseal bone, and growth plate cartilage from distal femoral epiphyses of inbred Lewis rats expressing enhanced green fluorescent protein from a ubiquitous promoter were transplanted either in inverted or original (control) orientation to matching sites in wildtype littermates, thereby allowing for tracing of transplanted cells and their progenies. We found that no hypertrophic differentiation occurred in the growth plate cartilage ectopically placed at the joint surface. Instead, the transplanted growth plate cartilage, with time, remodeled into articular cartilage. This finding suggests that the microenvironment at the articular surface inhibits hypertrophic differentiation and supports articular cartilage formation. To explore this hypothesis, rat chondrocyte pellets were cultured with and without synoviocyte-conditioned media. Consistent with the hypothesis, hypertrophic differentiation was inhibited and expression of the articular surface marker lubricin (Prg4) was dramatically induced when chondrocyte pellets were exposed to synovium- or synoviocyte-conditioned media, but not to chondrocyte- or osteoblast-conditioned media. Taken together, we present evidence for a novel mechanism by which synoviocytes, through the secretion of a factor or factors, act directly on chondrocytes to inhibit hypertrophic differentiation and endochondral bone formation and promote articular cartilage formation. This mechanism may have important implications for articular cartilage development, maintenance, and regeneration.

A novel prostaglandin E receptor 4 (EP4) small molecule antagonist induces articular cartilage regeneration

Articular cartilage repair and regeneration is an unmet clinical need because of the poor self-regeneration capacity of the tissue. In this study, we found that the expression of prostaglandin E receptor 4 (PTGER4 or EP4) was largely increased in the injured articular cartilage in both humans and mice. In microfracture (MF) surgery-induced cartilage defect (CD) and destabilization of the medial meniscus (DMM) surgery-induced CD mouse models, cartilage-specific deletion of EP4 remarkably promoted tissue regeneration by enhancing chondrogenesis and cartilage anabolism, and suppressing cartilage catabolism and hypertrophy. Importantly, knocking out EP4 in cartilage enhanced stable mature articular cartilage formation instead of fibrocartilage, and reduced joint pain. In addition, we identified a novel selective EP4 antagonist HL-43 for promoting chondrocyte differentiation and anabolism with low toxicity and desirable bioavailability. HL-43 enhanced cartilage anabolism, suppressed catabolism, prevented fibrocartilage formation, and reduced joint pain in multiple pre-clinical animal models including the MF surgery-induced CD rat model, the DMM surgery-induced CD mouse model, and an aging-induced CD mouse model. Furthermore, HL-43 promoted chondrocyte differentiation and extracellular matrix (ECM) generation, and inhibited matrix degradation in human articular cartilage explants. At the molecular level, we found that HL-43/EP4 regulated cartilage anabolism through the cAMP/PKA/CREB/Sox9 signaling. Together, our findings demonstrate that EP4 can act as a promising therapeutic target for cartilage regeneration and the novel EP4 antagonist HL-43 has the clinical potential to be used for cartilage repair and regeneration.

Reduction of knee joint load suppresses cartilage degeneration, osteophyte formation, and synovitis in early-stage osteoarthritis using a post-traumatic rat model.

The purpose of this study was to clarify the histological effect of reducing the loading to knee on cartilage degeneration, osteophyte formation, and synovitis in early-stage osteoarthritis (OA) using a post-traumatic rat model. Ten male rats were randomly allocated into two experimental groups: OA induction by surgical destabilization of medial meniscus (DMM, OA group) and hindlimb suspension after OA induction by DMM (OAHS group). The articular cartilage, osteophyte formation, and synovial membrane in the medial tibiofemoral joint were analyzed histologically and histomorphometrically at 2 and 4 weeks after surgery. The histological scores and changes in articular cartilage and osteophyte formation were significantly milder and slower in the OAHS group than in the OA group. At 2 and 4 weeks, there were no significant differences in cartilage thickness and matrix staining intensity between both the groups, but chondrocytes density was significantly lower in the OA group. Synovitis was milder in OAHS group than in OA group at 2 weeks. Reducing knee joint loading inhibited histological OA changes in articular cartilage, osteophyte formation, and synovial inflammation. This result supports the latest clinical guidelines for OA treatment. Further studies using biochemical and mechanical analyses are necessary to elucidate the mechanism underlying delayed OA progression caused by joint-load reduction.

Remifentanil repairs cartilage damage and reduces the degradation of cartilage matrix in post-traumatic osteoarthritis, and inhibits IL-1β-induced apoptosis of articular chondrocytes via inhibition of PI3K/AKT/NF-κB phosphorylation.

BackgroundRemifentanil (RFT) is an opioid analgesic with a unique pharmacokinetic profile, and plays an important role in the intra- and post-operative periods. Post-traumatic osteoarthritis (PTO) is a particular type of osteoarthritis (OA) that occurs secondary to a traumatic injury. In the present study, we investigated the effects of RFT both in vivo and in vitro.MethodsIn vivo, 50 Sprague Dawley (SD) rats (7 weeks old) were randomly divided into five groups. Four groups of rats received RFT (0.2, 0.5, and 1 µg) or vehicle (PTO group), while the remaining group served as the control. A PTO model in rats was established using the Hulth method. The cartilage damage, articular cartilage formation, and the degradation of cartilage matrix were evaluated. The effects of RFT on cell proliferation, apoptosis, and nuclear factor (NF)-κB phosphorylation were also examined.ResultsThe results indicated that RFT improved cartilage damage, enhanced articular cartilage formation, and inhibited the degradation of cartilage matrix in PTO model rats. Compared with the control group, the protein levels of Osterix (OSX), Collagen type I alpha 1 (COL1A1), and osteocalcin (OC) were down-regulated in PTO model rats. RFT also inhibited the interleukin-1β (IL-1β)-induced apoptosis of chondrocytes in vitro. Furthermore, the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/NF-κB pathway was inhibited both in vitro and in vitro.ConclusionsRFT has significant potential as a therapeutic intervention to ameliorate PTO and provides a foundation for further clinical studies.

Synergistic interaction of hTGF-\u03b23 with hBMP-6 promotes articular cartilage formation in chitosan scaffolds with hADSCs: implications for regenerative medicine

BackgroundHuman TGF-β3 has been used in many studies to induce genes coding for typical cartilage matrix components and accelerate chondrogenic differentiation, making it the standard constituent in most cultivation media used for the assessment of chondrogenesis associated with various stem cell types on carrier matrices. However, in vivo data suggests that TGF-β3 and its other isoforms also induce endochondral and intramembranous osteogenesis in non-primate species to other mammals. Based on previously demonstrated improved articular cartilage induction by a using hTGF-β3 and hBMP-6 together on hADSC cultures and the interaction of TGF- β with matrix in vivo, the present study investigates the interaction of a chitosan scaffold as polyanionic polysaccharide with both growth factors. The study analyzes the difference between chondrogenic differentiation that leads to stable hyaline cartilage and the endochondral ossification route that ends in hypertrophy by extending the usual panel of investigated gene expression and stringent employment of quantitative PCR.ResultsBy assessing the viability, proliferation, matrix formation and gene expression patterns it is shown that hTGF-β3 + hBMP-6 promotes improved hyaline articular cartilage formation in a chitosan scaffold in which ACAN with Col2A1 and not Col1A1 nor Col10A1 where highly expressed both at a transcriptional and translational level. Inversely, hTGF-β3 alone tended towards endochondral bone formation showing according protein and gene expression patterns.ConclusionThese findings demonstrate that clinical therapies should consider using hTGF-β3 + hBMP-6 in articular cartilage regeneration therapies as the synergistic interaction of these morphogens seems to ensure and maintain proper hyaline articular cartilage matrix formation counteracting degeneration to fibrous tissue or ossification. These effects are produced by interaction of the growth factors with the polysaccharide matrix.

The use of antifreeze proteins to modify pore structure in directionally frozen alginate sponges for cartilage tissue engineering

It is thought that osteoarthritis is one of the world’s leading causes of disability, with over 8.75 million people in the UK alone seeking medical treatment in 2013. Although a number of treatments are currently in use, a new wave of tissue engineered structures are being investigated as potential solutions for early intervention. One of the key challenges seen in cartilage tissue engineering is producing constructs that can support the formation of articular cartilage, rather than mechanically inferior fibrocartilage. Some research has suggested that mimicking structural properties of the natural cartilage can be used to enhance this response. Herein directional freezing was used to fabricate scaffolds with directionally aligned pores mimicking the mid-region of cartilage, anti-freeze proteins were used to modify the porous structure, which in turn effected the mechanical properties. Pore areas at the tops of the scaffolds were 180.46 ± 44.17 μm2 and 65.66 ± 36.20 μm2 for the AFP free and the AFP scaffolds respectively, and for the bases of the scaffolds were 91.22 ± 19.05 μm2 and 69.41 ± 21.94 μm2 respectively. Scaffolds were seeded with primary bovine chondrocytes, with viability maintained over the course of the study, and regulation of key genes was observed.

Fibrodysplasia ossificans progressiva (FOP): A disorder of osteochondrogenesis.

Fibrodysplasia ossificans progressiva (FOP) is an ultra-rare genetic disorder of extraskeletal bone formation, but could appropriately be viewed as a seminal disorder of osteochondrogenesis. Many, if not most, of the musculoskeletal features of FOP are related to dysregulated chondrogenesis including abnormal articular cartilage formation, abnormal diarthrodial joint specification, growth plate dysplasia, osteochondroma formation, heterotopic endochondral ossification (HEO), and precocious arthropathy. In FOP, causative activating mutations of Activin receptor A type I (ACVR1), a bone morphogenetic protein (BMP) type I receptor, are responsible for the osteochondrodysplasia that impacts developmental phenotypes as well as postnatal features of this illustrative disorder. Here, we highlight the myriad developmental and postnatal effects on osteochondrogenesis that emanate directly from mutant ACVR1 and dysregulated bone morphogenetic protein (BMP) signaling in FOP.

Stiffness Matters: Fine-Tuned Hydrogel Elasticity Alters Chondrogenic Redifferentiation.

Biomechanical cues such as shear stress, stretching, compression, and matrix elasticity are vital in the establishment of next generation physiological in vitro tissue models. Matrix elasticity, for instance, is known to guide stem cell differentiation, influence healing processes and modulate extracellular matrix (ECM) deposition needed for tissue development and maintenance. To better understand the biomechanical effect of matrix elasticity on the formation of articular cartilage analogs in vitro, this study aims at assessing the redifferentiation capacity of primary human chondrocytes in three different hydrogel matrices of predefined matrix elasticities. The hydrogel elasticities were chosen to represent a broad spectrum of tissue stiffness ranging from very soft tissues with a Young’s modulus of 1 kPa up to elasticities of 30 kPa, representative of the perichondral-space. In addition, the interplay of matrix elasticity and transforming growth factor beta-3 (TGF-β3) on the redifferentiation of primary human articular chondrocytes was studied by analyzing both qualitative (viability, morphology, histology) and quantitative (RT-qPCR, sGAG, DNA) parameters, crucial to the chondrotypic phenotype. Results show that fibrin hydrogels of 30 kPa Young’s modulus best guide chondrocyte redifferentiation resulting in a native-like morphology as well as induces the synthesis of physiologic ECM constituents such as glycosaminoglycans (sGAG) and collagen type II. This comprehensive study sheds light onto the mechanobiological impact of matrix elasticity on formation and maintenance of articular cartilage and thus represents a major step toward meeting the need for advanced in vitro tissue models to study both re- and degeneration of articular cartilage.

Theobromine mitigates IL-1β-induced oxidative stress, inflammatory response, and degradation of type II collagen in human chondrocytes

Osteoarthritis is one of the major causes of disability in elderly adults. Chondrocytes are responsible for the formation and remodeling of articular cartilage in joint tissue. The dysfunction of chondrocytes is a significant factor in the development of osteoarthritis. In the current study, we found that theobromine, a constituent of the cacao plant, possesses a preventive effect against interleukin (IL)-1β-induced chondrocyte dysfunction. Theobromine ameliorates IL-1β-induced production of cellular reactive oxygen species (ROS) and inflammatory mediators including cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2). The presence of theobromine suppresses IL-1β-induced inducible nitro oxide synthase (iNOS) expression and cellular nitro oxide (NO) production. Theobromine also suppresses IL-1β-induced production of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1), as well as matrix metalloproteinases (MMP)-3 and MMP-13. Additionally, theobromine mitigates IL-1β-induced type II collagen degradation. Mechanistically, we show that theobromine inhibits IL-1β-induced IκBα activation, nuclear factor-κB (NF-κB) protein p65 accumulation, and transfected NF-κB promoter activity, indicating that theobromine suppresses the NF-κB pathway in chondrocytes. Collectively, our study demonstrates that the natural molecule theobromine has a protective effect to counter cytokine-induced chondrocyte dysfunction, implying its beneficial effect in the prevention of osteoarthritis.

The efficacy of intraarticular PRP, ozone and PRP + ozone injections in patients with osteoarthritis [abstract

BACKGROUND: Osteoarthritis (OA) is the most common form of arthritis and some joints are predisposed more than others. The prevalence of OA increases with age and it represents the major problem for functional impairment in older patients.
 
 PURPOSE: This study was performed to determine efficiency of ozone + PRP (platelet rich plasma) injections on pain in patients with knee osteoarthritis.
 
 MATERIAL and METHODS: 102 patients with the diagnosis of OA according to the criteria of the American College of Rheumatology were enrolled in this study. Patients were randomized into three equal groups (34 patients each group).
 Patients in the first group were treated with intra-articular injections of ozone 2 times/week, with 10 ?g/ml ozone, 5ml in volume for a total of 12 times .
 Patients in the second group were treated with intra-articular injections of PRP + ozone (10 ?g/ml ozone, 5 ml volume) once a week for a total of 3 times.
 Patients in the third group were treated only with PRP injections once a week for a total of 3 times.
 The pain levels of patients were measured with visual analog scale (VAS).We chose the patients with VAS score 5 and above.
 The improvement strength has been evaluated according to Clinical Global image scale.
 
 RESULTS: In a retrospective study, Evaluation of the Clinical Global impression scale in the intra-articular injection of all patients (102 patients in total); 36 males and 66 females of 102 patients were found to have an average age of 56.11.
 The stress subscale of the 34 patients who had only ozone injections decreased from 5.17 to the healing scale of 2.91
 In the 34 patients who received Ozone + PRP combination, the stress subscale decreased from 5,.14 to the healing scale 1.52.
 In the 34 patients who had only PRP, the subscale of stress subscale decreased from 4.9 to the healing scale 1.85.
 When the mean recovery scores were evaluated, it was 2.26 in the ozone alone group, 3.62 in the ozone + PRP combination and 3.06 in the PRP patients only.
 The recovery scores of the patients who received the ozone + PRP combination were significantly better than ozone or PRP injections alone.
 No side effects were observed during and after the treatments.
 
 DISCUSSION: OA is a degenerative joint disease that increases with age and causes pain and disability to significantly impair an individual's quality of life.
 It is considered to be an organ disease that affects cartilage destruction and changes in the subchondral bone, and all the joints and tissues around the joint. Pathologically, destruction and loss of articular cartilage, subchondral sclerosis and osteophyte formation are often accompanied by synovial inflammation and destruction of other structures that support joint. The loss of pain and functioning impairs the patient to a wide variety of treatment seeking, but unfortunately there are no medical or physical methods that have proven scientifically and proven to be effective in humans to prevent joint cartilage degradation and therefore all treatment approaches are aimed at relieving symptomatic pain and minimizing functional deficits . The goal of treatment in knee OA should be to control pain, to preserve and correct joint functions, to provide functional independence and to improve quality of life. In order to achieve these goals, knee OA therapy should include non pharmacologic, pharmacologic and if necessary, surgical methods. Treatment should be tailored to each patient. Non-steroidal anti-inflammatory (NSAID) drugs are thought to be the standard treatment for OA in many cases but many patients are not able to tolerate these drugs or are exposed to side effects, while only analgesics are adequate in some patients.
 As we have seen in the study above; ozone or PRP injections alone can be used for the patients with knee OA but the results of Ozone + PRP injections in knee OA reduced the pain more than the ozone and PRP injections alone.
 This might be because of the anti inflammatory effect of ozone has been supported by the release of the growth factors from the platelets in PRP.
 Therefore we can add ozone + PRP injections to patients who has OA without hesitation in additional to the classical treatment .
 For further studies we can use Major Autohemotherapy for regeneration in addition to the injections of ozone, PRP, Ozone + PRP.
 
 CONCLUSION: The combination of ozone + PRP offers the best results compared with PRP or ozone alone.
 We could not get any biopsy from the cartilage so we could not show the healing or changes of the cartilage according to the decrease of pain and the increase of quality of life.
 For further studies we could include biopsy and the measurement of inflammatory and anti inflammatory markers from the cartilage tissue before and after treatment.