In our previous study, we reported the fabrication and characterization of a novel tricalcium phosphatefucoidan-chitosan (TCP-Fu-Ch) biocomposite scaffold. However, the previous report did not show whether the biocomposite scaffold can exhibit osteogenic differentiation of human bone marrow stromal cells in osteogenic media and normal media supplemented with platelet-derived growth factor (PDGF-BB). On day 15, the release of osteocalcin, was significant in the TCP-Fu-Ch scaffold, when compared with that in the TCP-Ch scaffold, and the level of release was approximately 8 and 6 ng/ml in osteogenic and normal media supplemented with PDGF-BB, respectively. Scanning electron microscopy of the TCP-Fu-Ch scaffold demonstrated mineralization and apatite layer formation on day 14, while the addition of PDGF-BB also improved the osteogenic differentiation of the scaffold. An array of gene expression analysis demonstrated that TCP-Fu-Ch scaffold cultured in osteogenic and normal media supplemented with PDGF-BB showed significant improvement in the expression of collagen 1, Runt-related transcription factor 2, osteonectin, bone gamma-carboxyglutamate protein, alkaline phosphatase, and PPA2, but a decline in the expression of integrin. Altogether, the present study demonstrated that fucoidan-incorporated TCP-Ch scaffold could be used in the differentiation of bone marrow stromal cells and can be a potential candidate for the treatment of bone-related ailments through tissue engineering technology.
In this paper we report the differentiating properties of platelet-rich plasma releasates (PRPr) on human chondrocytes within elastomeric polycaprolactone triol-citrate (PCLT-CA) porous scaffold. Human-derived chondrocyte cellular content of glycosaminoglycans (GAGs) and total collagen were determined after seeding into PCLT-CA scaffold enriched with PRPr cells. Immunostaining and real time PCR was applied to evaluate the expression levels of chondrogenic and extracellular gene markers. Seeding of chondrocytes into PCLT-CA scaffold enriched with PRPr showed significant increase in total collagen and GAGs production compared with chondrocytes grown within control scaffold without PRPr cells. The mRNA levels of collagen II and SOX9 increased significantly while the upregulation in Cartilage Oligomeric Matrix Protein (COMP) expression was statistically insignificant. We also report the reduction of the expression levels of collagen I and III in chondrocytes as a consequence of proximity to PRPr cells within the scaffold. Interestingly, the pre-loading of PRPr caused an increase of expression levels of following extracellular matrix (ECM) proteins: fibronectin, laminin and integrin b over the period of 3 days. Overall, our results introduce the PCLT-CA elastomeric scaffold as a new system for cartilage tissue engineering. The method of PRPr cells loading prior to chondrocyte culture could be considered as a potential environment for cartilage tissue engineering as the differentiation and ECM formation is enhanced significantly. Keywords Cartilage tissue engineering Á PCLT-CA scaffolds Á PRPr cells Á Human chondrocytes Á ECM formation 1 Introduction Cartilage healing is a slow and sensitive process that often leads to a permanent cartilage defect. Cartilage repair is commonly suppressed by factors such as avascular condition of articular cartilage, limited chondrocytes in mature and aged tissue and inability of chondrocytes to migrate to the damaged site. In most cases, the fibro-cartilage would replace the damaged cartilage, which does not provide sustained repair and desirable mechanical support & Hussin A. Rothan
Clinical investigations have shown a significant relationship between osteoarthritis (OA) and estrogens levels in menopausal women. Therefore, treatment with exogenous estrogens has been shown to decrease the risk of OA. However, the effect estrogen has not been clearly demonstrated in the chondrocytes using phytoestrogens, which lack the specific side-effects of estrogens, may provide an alternative therapy. This study was designed to examine the possible effects of phytoestrogen (daidzein) on human chondrocyte phenotype and extracellular matrix formation. Phytoestrogens which lack the specific side-effects of estrogens may provide beneficial effect without causing hormone based side effect. Human chondrocytes cells were cultured in 2D (flask) and 3D (PCL-CA scaffold) systems. Daidzein cytotoxic effect was determined by MTT assay. Chondrocyte cellular content of glycosaminoglycans (GAGs), total collagen and chondrogenic gene expression were determined in both culture systems after treatment with daidzein. Daidzein showed time-dependent and dose-independent effects on chondrocyte bioactivity. The compound at low doses showed significant (p \ 0.05) increase in total collagen and GAGs production at similar levels in 2D and 3D culture environment. The mRNA levels of Collagen II and Sox9 were increased significantly (p \ 0.01) after the treatment while the upregulation in COMP expression was statistically insignificant (p [ 0.05). The expression levels of Fibronectin, Laminin and Integrin b1 were significantly increased especially in 3D culture system. This study was illustrated the potential positive effects of daidzein on maintenance of human chondrocyte phenotype and extracellular matrix formation suggesting an attractive and viable alternative therapy for OA. Keywords Phytoestrogens Á Human chondrocyte Á Extracellular matrix Á PCL-CA scaffold Á Osteoarthritis Abbreviations CTX-II C-terminal crosslinked telopeptide type II collagen CA Citric acid COMP Cartilage oligomeric matrix protein COX Cyclooxygenase DMEM Dulbecco's Modification of Eagle's Medium DMMB Dimethylmethylene blue & Thamil Selvee Ramasamy
In this in vivo study, Sprague Dawley (SD) rats were used to investigate the bioactivity as well as the microstructural and mechanical properties of Ti-6Al-4V samples embedded with hydroxyapatite (HA) using two different coating methods-superplastic embedment (SPE) and superplastic deformation (SPD). The HA layer thickness for the SPE and SPD samples increased from 249.1 ± 0.6 nm to 874.8 ± 13.7 nm, and from 206.1 ± 5.8 nm to 1162.7 ± 7.9 nm respectively, after 12 weeks of implantation. The SPD sample exhibited much faster growth of newly formed HA compared to SPE. The growth of the newly formed HA was strongly dependent on the degree of HA crystallinity in the initial HA layer. After 12 weeks of implantation, the surface hardness value of the SPE and SPD samples decreased from 661 ± 0.4 HV to 586 ± 1.3 HV and from 585 ± 6.6 HV to 425 ± 86.9 HV respectively. The decrease in surface hardness values was due to the newly formed HA layer that was more porous than the initial HA layer. However, the values were still higher than the substrate surface hardness of 321 ± 28.8 HV. Wear test results suggest that the original HA layers for both samples were still strongly intact, and to a certain extent the newly grown HA layers also were strongly bound with the original HA layers. This study confirms the bioactivity and mechanical stability of the HA layer on both samples in vivo.
Background There is scarce information about the relationship between periodontal disease and osteoarthritis. This study investigated the effect of surgically induced osteoarthritis on alveolar bone loss in experimental periodontitis in rats. Methods 12 rats were divided into test and control groups. On day 1, the animals were anaesthetized, and silk ligatures were ligated around 6 maxillary posterior teeth in each animal from both groups. Surgical induction of osteoarthritis was performed on the left knees in the test group. No knee surgeries were performed in the control group. The ligatures were kept in place for 30 days, at which time the animals were euthanatized, and the maxillae and knee joints were harvested and processed for histological analysis. The alveolar bone loss was assessed using a zoom stereomicroscope. Results The knee joint histologic sections of the control group showed normal joint features, whereas in the test group there were substantial changes typical of osteoarthritis, including wide joint spaces, prominent monocytic infiltration of the synovium, invasion of periarticular bone, and decreased chondrocyte density. Comparison of the bone height between the groups showed a significantly higher bone loss in the test than in the control group The marginal mean bone height, adjusted for covariates and the intraclass correlation between sites, was 1.19 and 0.78 mm in the test and control groups, respectively (p < .0001). Conclusions Surgically induced osteoarthritis leads to greater alveolar bone loss in the experimental periodontitis model in rats.
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