Introduction:The purpose of the present study seeks to determine the signal timing of BMP–7 and TGF-β1 from a novel chitosan based hydrogel system that may affect chondrocyte proliferation resulting in the presence of a synergism seen conspicuously in consecutive controlled delivery.Methods:Four groups of cultured chondrocytes were seeded on a novel designed chitosan based hydrogel. The hydrogel was left empty (control) in one group and loaded with BMP–7, TGF-β1 and their combination in the other groups, respectively. Hydrogel structure was analyzed with scanning electron microscope. The release kinetics of Growth Factors (GFs) was determined with ELISA. Chondrocyte viability and toxicity after being tested with MTS and collagen type II synthesis, were quantified with western blotting. Canonical regression analysis was used for measuring statistical evaluation.Results:Chitosan based hydrogel allowed controlled release of GFs in different time intervals for BMP–7 and TGF-β1. Double peak concentration gradient was found to be present in the group loaded with both GFs. In this group, substantially higher chondrocyte growth and collagen synthesis were also detected. Conclusions:We concluded that, chitosan based hydrogel systems may be adjusted to release GFs consecutively during biodegradation at the layers of surface, which may increase the cell number and enhance collagen type II synthesis.
A AB BS ST TR RA AC CT T O Ob bj je ec ct ti iv ve e: : Both Bone Morphogenetic Protein 7 (BMP-7) and Transforming Growth Factor Beta 1 (TGF-β1) have stimulating effects on chondrocyte proliferation. These growth factors act at different time points of the biological repair process. We aimed to design a hydrogel, which would enable consecutive controlled release of the growth factors. M Ma at te er ri ia al l a an nd d M Me et th ho od ds s: : We designed a chitosan particle impregnated Poly (vinylalcohol) (PVA)-borax hydrogel (CPBH). CPBH allowed controlled release of BMP-7 and TGF-β1 at different time points. Hydrogel structure was analyzed by Scanning Electron Microscope (SEM). The release kinetics of Growth Factors (GFs) were determined with enzyme-linked immunosorbent assay (ELISA) and UV-spectrophotometer. Chondrocyte viability and toxicity were tested through CellTiter 96 ® Non-Radioactive Cell Proliferation (MTS) ELISA. R Re es su ul lt ts s: : The designed hydrogel showed high swelling and mucoadhesion characteristics under acidic conditions. CPBH released BMP-7 first and rapidly and TGF-β1 consecutively and slowly. It also allowed controlled release of protein/peptide based drugs for 21 days without altering their bioactive properties. At the end of 21 days, 82.62% of BMP-7 and 98.34% of TGF-β1 were consecutively released. The difference between the groups was significant (for TGF-β1, p<0.05 and for BMP-7, p<0.001). C Co on nc cl lu us si io on n: : The controlled and slow release of growth factors has been shown to be beneficial on cartilage regeneration. As it is not cytotoxic, we suggest that this hydrogel might be used in medical and pharmaceutical applications areas. K Ke ey y W Wo or rd ds s: : Delayed-action preparations; hydrogels; TGF-β1 protein, human; bone morphogenetic protein 7 Ö ÖZ ZE ET T A Am ma aç ç: : Kemik Morfojenik Proteini-7 (BMP-7) ve Transforme edici Büyüme Faktörü Beta-1 (TGF-β1)'in kıkırdak hücreleri üzerinde prolifere edici etkileri bulunmaktadır. Bu büyüme faktörlerinin etkisi biyolojik tamir sürecinin farklı zamanlarında gerçekleşmektedir. Bu çalışmada, büyüme faktörlerinin ardışık ve kontrollü olarak salınımını gerçekleştirecek bir hidrojel tasarlamayı amaçladık. G Ge er re eç ç v ve e Y Yö ön nt te em ml le er r: : Bu amaçla kitosan emdirilmiş polivinil alkol (PVA)-boraks hidrojeli (CPBH) tasarlandı. Bu hidrojel BMP-7 ve TGF-β1'in farklı zamanlarda kontrollü salınımını sağladı. Tasarladığımız hidrojelin yüzey yapısı taramalı elektron mikroskobu (SEM) ile değerlendirildi. Büyüme faktörlerinin salım kinetiği enzim aracılı immün deney (ELISA) ve ultraviyole (UV)spektrofotometre ile saptandı. Kondrosit canlılığı ve toksisite testi "CellTiter 96 ® Non-Radioactive Cell Proliferation (MTS)" ELISA yöntemiyle değerlendirildi. B Bu ul lg gu ul la ar r: : Tasarlanan hidrojel, asidik ortam koşullarında büyük oranda şişme ve mukoadezyon özellikleri sergiledi. CPBH, önce hızlı bir biçimde BMP-7'nin, sonra da yavaş olarak TGF-β1'in salınımını ardışık olarak gerçekleştirdi. İçerisindeki p...
Sonographically guided corticosteroid injection enables real-time imaging of the plantar fascia during needle insertion. A caudocranial oblique injection beneath the fascia may have an incremental value in the treatment response in patients with plantar fasciosis.
The shortening of the tendon occurs as a result of adherence in the distal part of the tendon. It would appear that it is this shortening that causes the difficulties encountered during arthroplasty surgery of osteotomy patients, and not patella infera.
Purpose. This study has researched the affect of different methodologies of harvesting and analysing the samples in determining the mediators emerging after the rat articular cartilage injury. Materials and Methods. One hundred and forty-four male wistar rats were divided into 2 groups. Synovial fluid samples were taken from all of the rats. We entered into the right knees of the rats in group I (n = 36) under anaesthesia and took cartilage tissue samples from their distal femur. Samples were taken as reference values for enzyme linked immunosorbent assay (ELISA) and histopathological evaluations. We entered into the right knees of the rats in group II (n = 108) and formed complete layer of cartilage injury in their medial femoral condyles. At the end of the 15th day, the rats were sacrificed after taking synovial fluid samples from their right knees creating defect in the rats in group II. The molecular markers in the synovial fluid and cartilage tissue samples which were taken from the experimental and control groups (MMP-9, MMP-13, TIMP-1, TNF-α, and NO) were analysed by direct or indirect methodologies. SPSS 18.0 Package program was used in the statistical evaluation. Students t-test where the measurement variables between the experimental and control groups were compared was applied. Receiver Operating Characteristics (ROC) curves were used in the determination of the diagnostic sufficiency from the tissue. Results. No difference was found between TIMP-1 (P = 0.67) and MMP-9 (P = 0.28) levels in synovial fluid and cartilage tissue. From the molecular markers, when MMP-9, MMP-13, NO, TIMP-1, TNF-α′, the area under ROC curve, and P values were examined, MMP-13 (P < 0.0001, 95% CI: 0.70–0.85), NO (P < 0.0001, 95% CI: 0.72–0.86), and TNF-α (P < 0.0001, 95% CI: 0.91–0.98) results were found to be statistically significant. Inferences. The indirect ELISA protocol which we apply for the cartilage tissue as an alternative to synovial lavage fluid is a reliable method which can be used in the determination of articular cartilage injury markers.
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