BackgroundOsteochondral allograft transplantation has a good clinical outcome, however, there is still debate on optimization of allograft storage protocol. Storage temperature and nutrient medium composition are the most critical factors for sustained biological activity of grafts before implantation. In this study, we performed a time-dependent in vitro experiment to investigate the effect of various storage conditions on electromechanical, histological and histochemical properties of articular cartilage.MethodsOsteochondral grafts derived from goat femoral condyles were frozen at −70 °C or stored at 4 °C and 37 °C in the medium supplemented with or without insulin-like growth factor-1 (IGF-1). After 14 and 28 days the cartilage samples were quantitatively analysed for electromechanical properties, glycosaminoglycan distribution, histological structure, chondrocyte viability and apoptosis. The results were compared between the experimental groups and correlations among different evaluation methods were determined.ResultsStorage at −70 °C and 37 °C significantly deteriorated cartilage electromechanical, histological and histochemical properties. Storage at 4 °C maintained the electromechanical quantitative parameter (QP) and glycosaminoglycan expression near the normal levels for 14 days. Although hypothermic storage revealed reduced chondrocyte viability and increased apoptosis, these parameters were superior compared with the storage at −70 °C and 37 °C. IGF-1 supplementation improved the electromechanical QP, chondrocyte viability and histological properties at 37 °C, but the effect lasted only 14 days. Electromechanical properties correlated with the histological grading score (r = 0.673, p < 0.001), chondrocyte viability (r = −0.654, p < 0.001) and apoptosis (r = 0.416, p < 0.02). In addition, apoptosis correlated with glycosaminoglycan distribution (r = −0.644, p < 0.001) and the histological grading score (r = 0.493, p = 0.006).ConclusionsOur results indicate that quality of allografts is better preserved at currently established 4 °C storage temperature. Storage at −70 °C or at 37 °C is unable to maintain cartilage function and metabolic activity. IGF-1 supplementation at 37 °C can enhance chondrocyte viability and improve electromechanical and histological properties of the cartilage, but the impact persists only 14 days. The correlations between cartilage electromechanical quantitative parameter (QP) and metabolic activity were detected. Our findings indicate that non-destructive assessment of cartilage by Arthro-BST is a simple and reliable method to evaluate allograft quality, and could be routinely used before implantation.
Polycaprolactone (PCL) is a non-cytotoxic, completely biodegradable biomaterial, ideal for cartilage tissue engineering. Despite drawbacks such as low hydrophilicity and lack of functional groups necessary for incorporating growth factors, it provides a proper environment for different cells, including stem cells. In our study, we aimed to improve properties of scaffolds for better cell adherence and cartilage regeneration. Thus, electrospun PCL–scaffolds were functionalized with ozone and loaded with TGF-β3. Together, human-muscle-derived stem cells (hMDSCs) were isolated and assessed for their phenotype and potential to differentiate into specific lineages. Then, hMDSCs were seeded on ozonated (O) and non-ozonated (“naïve” (NO)) scaffolds with or without protein and submitted for in vitro and in vivo experiments. In vitro studies showed that hMDSC and control cells (human chondrocyte) could be tracked for at least 14 days. We observed better proliferation of hMDSCs in O scaffolds compared to NO scaffolds from day 7 to day 28. Protein analysis revealed slightly higher expression of type II collagen (Coll2) on O scaffolds compared to NO on days 21 and 28. We detected more pronounced formation of glycosaminoglycans in the O scaffolds containing TGF-β3 and hMDSC compared to NO and scaffolds without TGF-β3 in in vivo animal experiments. Coll2-positive extracellular matrix was observed within O and NO scaffolds containing TGF-β3 and hMDSC for up to 8 weeks after implantation. These findings suggest that ozone-treated, TGF-β3-loaded scaffold with hMDSC is a promising tool in neocartilage formation.
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We report the case of a 15-year-old patient who underwent concomitant autologous chondrocyte implantation and osteochondral grafting for the treatment of a massive osteochondritis dissecans defect in the left knee and autologous chondrocyte implantation in the right knee joint. Magnetic resonance imaging showed large osteochondral defects in both the knee joints measuring 8–9 cm2. Both defects were located in the weight-bearing areas of the medial femoral condyles. Therefore, simultaneous autologous chondrocyte implantation (ACI) and osteochondral autograft transplantation (OAT) for the left knee defect and ACI for the right knee joint were performed. Osteochondral plugs were harvested from the patellofemoral joint of the same left knee and grafted into the most dorsal regions of the large osteochondral defect of the left knee. The remaining osteochondral defect was covered with ACI using collagen type I and III membrane and chondrocyte cells. The membrane was implanted into more proximal part of the osteochondral defect of the left knee. Time interval between operations of the left and right knee joints was 6 months. Magnetic resonance imaging at 6 months after each knee surgery showed good preservation of the OAT and ACI grafts. The most recent follow-up examination, performed 12 months after surgeries, has shown excellent results with an International Knee Documentation Committee score of 95.59±4.64 and 96.88±4.69 for the right and left knee joints, respectively, and full range of knee motions with no symptoms. In this clinical case, the combination of ACI and OAT methods in a one-step procedure produced a good reconstruction of the joint surface with excellent clinical outcomes in the both knee joints of the same patient. Autologous osteochondral grafting and autologous chondrocyte implantation can be combined for the treatment of large osteochondral defects of the knee.
Background/Aim: To evaluate the association between TRIB1(rs6987702) and IL-9(rs1859430, rs2069870) genotypes with the development and manifestation of pituitary adenoma (PA). Materials and Methods: The study group included 141 patients with PA and the control group consisted of 287 healthy people. The genotyping of rs6987702, rs1859430 and rs2069870 was carried out using a real-time polymerase chain reaction. Results: Statistically significant results were obtained regarding the rs1859430, but there were no significant results regarding rs6987702. We found that the rs1859430 A/A genotype increased the odds of having recurrent PA six times (p=0.006) under the co-dominant model and four times (p=0.021) under the recessive model. Furthermore, the analysis showed that the G/A genotype increased the odds of having recurrent PA 2.3 times (p=0.003) under the co-dominant model, while G/A and A/A genotypes increased the odds 2.7 times (p=0.011) under the over-dominant model. Conclusion: Certain genotypes of rs1859430 can be associated with PA recurrence. Pituitary adenoma (PA) is a benign pituitary gland tumor, which starts from the cells of adenohypophysis (1). PAs are mainly inactive, but certain types of PAs secrete hormones and become clinically apparent (2). They can also be classified according to their size: microadenomas, which are more commonly found in female and <60 years old patients, and macroadenomas, which are more prevalent in male and >60 years old patients (3-5). According to the latest statistical report of Central Brain Tumor Registry of the United States (CBTRUS), the incidence rate of benign pituitary tumors has increased, and currently they present 16.4% of all primary brain and central nervous system tumors (6). The report of Brain Tumor Register of Japan published in 2017 showed that pituitary gland was a primary location for 22.6% of all brain tumors and 91.1% of them were benign (7). Meta-analysis of 143 studies showed that the recurrence rate depends on the type of PA (8) and varies from 5% to 16% (7). Although the exact mechanism of PA development is still unknown, it may include various immunogenetic factors. Many potential molecular markers, which might be involved in the tumorigenesis of PAs, are currently under investigation. In the present study, one TRIB1 gene polymorphism and two IL-9 gene polymorphisms were selected for further investigation. Tribbles-1 (TRIB1) is a protein, encoded by the TRIB1 gene, most commonly found in the nucleus of a cell. This protein belongs to a group of pseudokinases, which consist of 3 different domains: N-terminal PEST domain, pseudokinase domain and C-terminal COP1 binding peptide domain. TRIB kinases have unique sequences located at the C-terminal end (9). Tribbles-1 is mostly found in myeloid cells. It is thought to be important in intracellular signaling, and thereby, to participate in the management of cell cycle, differentiation, metabolism and proliferation, migration and invasion (9-11). TRIB1 is thought to have oncogenic properties. Miyajima et a...
Thermography is a relatively new contact-free method used in experimental and clinical studies and in cardiovascular surgery to investigate the myocardium and coronary artery function. Objects of complex study included mongrel dogs and patients with coronary artery disease who underwent cardiac surgery. For active dynamic thermography, we used a thermovision camera “A20V” (FLIR Systems, USA). Our data indicate that both experimental and clinical study performed on beating hearts could be an important approach to interoperation inspection of autovenous graft function. An infrared camera also can be successfully used to determine the extent of ischemic damage to the myocardium, heart, and blood vessels during surgery as a significant prognostic tool for evaluating outcome after cardiac operation.
Objective To determine the applicability of a minimally invasive diagnostic device to evaluate the quality of articular cartilage following autologous (OAT) and allogeneic (OCA) osteochondral graft transplantation in goat model. Design OAT grafts were harvested from lateral femoral condyles (LFCs) and transplanted into osteochondral defects created in medial femoral condyles (MFCs) of contralateral knees. OCA grafts were transplanted into MFC condyles after in vitro storage. Autologous platelet-rich plasma (PRP) was administered intraarticularly after the surgery and at 1 and 2 months postoperatively. OAT and OCA grafts were evaluated macroscopically (Oswestry arthroscopy score [OAS]), electromechanically (quantitative parameter, QP), and histologically (O'Driscoll score, safranin O staining intensity) at 3 and 6 months after transplantation. Results were compared with preoperative graft evaluation. Results Transplanted cartilage deteriorated within 6 months in all groups. Cartilage quality was better retained in OAT group compared with a decline in OCA group. QP and OAS scores were comparable in OAT and OCA groups at 3 months, but superior in OAT group at 6 months, according to all the methods applied. PRP injections significantly improved QP and OAS score at 6 months compared with 3 months in OAT group. QP moderately correlated with OAS, O'Driscoll score, and safranin O staining intensity. Conclusions Grafts did not retain preoperative quality parameters at 6 months follow-up; however, OAT were superior to OCA grafts. PRP may have a beneficial effect on macroscopic and electromechanical properties of cartilage; however, histological improvement is yet to be proved. Electromechanical diagnostic device enables reliable assessment of transplanted cartilage.
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