Is tranexamic acid safe for the hip joint?

To show the effects of tranexamic acid, which is a drug frequently used to control bleeding, on the hip joint and sciatic nerve in animal experiments. There were 15 rats in each of the 3 groups, with a total of 45 rats. Topical saline injections were applied to the first group, topical TXA injections to the second group, and intravenous (IV) TXA injections to the third group. In the samples taken from the hip joint 3 weeks later, femoral head cartilage, sciatic nerve, and joint capsule thicknesses were analyzed histologically. Statistically significantly more cartilage degradation was detected in the femoral head cartilage in both the IV and intraarticular TXA group when compared to the control group. The groups were also compared in terms of acetabular cartilage; however, no histological difference was found between the groups. It was seen that when the femoral head cartilage thickness (the average of the measurements made from 3 different points were used) was examined, the cartilage thickness in the topical TXA group was less when compared to the other 2 groups. However, this difference was determined to not be statistically significant. The data of the hip joint capsule thickness measurement, it was found that the capsule thickness in the topical TXA applied group was less when compared to the other 2 groups. However, this difference was not statistically significant. When the sciatic nerves in all 3 groups were compared, no different staining characteristics were found in the immunofluorescence examination. TXA, which is frequently used in orthopedic practice, shows negative effects on hip joint cartilage in both topical and intravenous application.

Tranexamic acid administered intraarticularly to the knee is safer for the articular cartilage and anterior cruciate ligament compared to intravenous administration: Histological analysis of an experimental rat model.

In this study, the effects of tranexamic acid (TXA) on the knee's articular cartilage, anterior cruciate ligament (ACL), and joint capsule were assessed histologically. There were 15 rats in each of the 3 groups, totaling 45 rats. Intraarticular (IA) saline injections were applied for the first group, IA TXA injections for the second group, and intravenous (IV) TXA injections for the third group. Using samples taken from the knee joint 3 weeks later, the medial/lateral femoral condyle and medial/lateral tibial plateau articular cartilages were evaluated with Osteoarthritis Research Society International (OARSI) scoring, while ACL diameter and joint capsule thickness were analyzed histologically. In comparisons of OARSI scores for the medial/lateral femoral condyle and medial/lateral tibial plateau cartilage regions, the scores obtained for the IV TXA group were significantly higher than those of the IA saline group (P < 0.001, P = 0.001, P = 0.003, P = 0.011). In comparisons of medial/lateral femoral condyle and medial/lateral tibial plateau OARSI scores, the scores obtained for the IV TXA group were again significantly higher than those of the IA TXA group (P < 0.001, P < 0.001, P < 0.001, P = 0.002). When ACL diameters were compared, a significant decrease was observed in the ACL diameters of the IV TXA group compared to the IA saline and IA TXA groups (P < 0.001, P = 0.039). Histologically, IV TXA damages the articular cartilage and ACL more than IA TXA. IA administration of TXA is more protective when the articular cartilage and ACL are preserved.

Investigation the Effect of Human Recombinant Epidermal Growth Factor on Rotator Cuff Healing: An Experimental Model.

Objective To investigate the effectiveness of human recombinant epidermal growth factor in the healing of rotator cuff tear in the rabbit shoulder. Methods Rotator cuff tears (RCTs) were experimentally created on both shoulders of 20 New Zealand rabbits. The rabbits were divided into the following groups: RCT (sham group; n = 5), RCT + EGF (EGF group; n = 5), RCT + transosseous repair (repair group; n = 5), and RCT + EGF + transosseous repair (combined repair + EGF group; n = 5). All rabbits were then observed for 3 weeks, and biopsies were taken from the right shoulders in the third week. After three more weeks of observation, all rabbits were sacrificed, and a biopsy removed from their left shoulders. All biopsy material was stained with haematoxylin & eosin (H&E) and vascularity, cellularity, the proportion of fibers and the number of fibrocartilage cells were evaluated under light microscope. Results The highest collagen amount and the most regular collagen sequence was detected in the combined repair + EGF group. The repair group and the EGF group showed higher fibroblastic activity and capillary formation when compared with the sham group, but the highest fibroblastic activity and capillary formation with highest vascularity was detected in the combined repair + EGF group ( p < 0.001). EGF seems to improve wound healing in the repair of RCT. The EGF application alone, even without repair surgery, seems to be beneficial to RCT healing. Conclusion In addition to rotator cuff tear repair, application of human recombinant epidermal growth factor has an effect on rotator cuff healing in rabbit shoulders.

Investigation the Effect of Human Recombinant Epidermal Growth Factor on Rotator Cuff Healing: An Experimental Model / Investigação do efeito do fator de crescimento epidérmico recombinante humano na cicatrização do manguito rotador: Um modelo experimental

Abstract Objective To investigate the effectiveness of human recombinant epidermal growth factor in the healing of rotator cuff tear in the rabbit shoulder. Methods Rotator cuff tears (RCTs) were experimentally created on both shoulders of 20 New Zealand rabbits. The rabbits were divided into the following groups: RCT (sham group; n = 5), RCT + EGF (EGF group; n = 5), RCT + transosseous repair (repair group; n = 5), and RCT + EGF + transosseous repair (combined repair + EGF group; n = 5). All rabbits were then observed for 3 weeks, and biopsies were taken from the right shoulders in the third week. After three more weeks of observation, all rabbits were sacrificed, and a biopsy removed from their left shoulders. All biopsy material was stained with haematoxylin & eosin (H&E) and vascularity, cellularity, the proportion of fibers and the number of fibrocartilage cells were evaluated under light microscope. Results The highest collagen amount and the most regular collagen sequence was detected in the combined repair + EGF group. The repair group and the EGF group showed higher fibroblastic activity and capillary formation when compared with the sham group, but the highest fibroblastic activity and capillary formation with highest vascularity was detected in the combined repair + EGF group (p < 0.001). EGF seems to improve wound healing in the repair of RCT. The EGF application alone, even without repair surgery, seems to be beneficial to RCT healing. Conclusion In addition to rotator cuff tear repair, application of human recombinant epidermal growth factor has an effect on rotator cuff healing in rabbit shoulders.

Resumo Objetivo Investigar a eficácia do fator de crescimento epidérmico (EGF) recombinante humano na cicatrização da lesão do manguito rotador no ombro de coelhos. Métodos As rupturas do manguito rotador (RMRs) foram criadas experimentalmente em ambos os ombros de 20 coelhos Nova Zelândia. Os coelhos foram divididos nos seguintes grupos: RMR (grupo controle; n = 5), RMR + EGF (grupo EGF; n = 5), RMR + reparo transósseo (grupo reparo; n = 5) e RMR + EGF + reparo transósseo (grupo reparo combinado+ EGF; n = 5). Todos os coelhos foram observados por 3 semanas, e amostras de biópsias foram coletadas do ombro direito na 3ª semana. Após mais 3 semanas de observação, todos os coelhos foram submetidos à eutanásia, e uma amostra de biópsia foi coletada dos ombros esquerdos. Todo o material de biópsia foi corado com hematoxilina e eosina (H&E) para avaliação de vascularidade, celularidade, proporção de fibras e número de células fibrocartilaginosas à microscopia óptica. Resultados O grupo reparo combinado + EGF apresentou a maior quantidade e a sequência mais regular de colágeno. O grupo reparo e o grupo EGF apresentaram maior atividade fibroblástica e formação capilar em comparação ao grupo controle, mas a maior atividade fibroblástica e a formação capilar com maior vascularidade foram detectadas no grupo reparo combinado + EGF (p < 0,001). O EGF parece melhorar a cicatrização da ferida no reparo da RMR. A aplicação isolada de EGF, mesmo sem cirurgia reparadora, parece melhorar a cicatrização da RMR. Conclusão Além do reparo da RMR, a aplicação de EGF recombinante humano auxilia a cicatrização do manguito rotador dos ombros de coelhos.

The determination of the relationship between p97/VCP, small VCP-interacting protein, two ERAD proteins and steroidogenesis in Leydig cell lines.

BACKGROUND: In recent studies, it was shown that Endoplasmic reticulum-associated degradation (ERAD) is regulated by androgens and small VCP-interacting protein (SVIP) is an ERAD inhibitor. There is no data available about the interactions of ERAD proteins with proteins involved in steroidogenesis. The aim of the study was to investigate the expressions of SVIP, p97/VCP, StAR, CYP17A1 and 3ß-HSD in human and mouse. METHODS AND RESULTS: HLC, TM3 and MA-10 Leydig cell lines were used to determine roles of ERAD proteins in steroidogenesis based on immunofluorescence, Western blot, qRT-PCR, ELISA. Findings showed that StAR, CYP17A1 and 3ß-HSD were colocalized with SVIP and p97/VCP in Leydig cells. A decrease in CYP17A1, 3ß-HSD and StAR expressions was observed as a result of suppression of SVIP siRNAs and p97/VCP siRNAs expressions in MA10, TM3 and HLC. When siSVIP transfected cells were compared with siSVIP transfected with hCG-exposed cells, SVIP protein expression was significantly increased as compared to the SVIP transfected group in human Leydig cells. CONCLUSION: We suggest that the suppression of protein expressions by p97/VCP and SVIP siRNAs in Leydig cells, the effects of proteins involved in steroidogenesis (StAR, CYP17A1 and 3ß-HSD) have proven to be originating from p97/VCP and SVIP which were playing a role in the steroidogenesis process. Additionally, it was demonstrated that testosterone levels decreased after transfection with p97/VCP siRNA and SVIP siRNA, p97/VCP and SVIP created an effect on testosterone synthesis while taking place in the steps of testosterone synthesis. Further, it was determined in the study that the SVIP was affected by hCG stimulations.

Three dimensional nanofibrous and compressible poly(L-lactic acid) bone grafts loaded with platelet-rich plasma.

In this study, nanofibrous matrices of poly(L-lactic acid)-hydroxyapatite (PLLA-HAp) were successfully fabricated by three-dimensional (3D) electrospinning for use in the treatment of irregular bone damages. Compressibility analysis showed that 3D nanofibrous grafts occupied at least 2-fold more volume than their 2D form and they can easily take shape of the defect zone with irregular geometry. Moreover, the compression moduli of the PLLA and PLLA-HAp grafts were calculated as 8.0 ± 3.0 kPa and 11.8 ± 3.9 kPa, respectively, while the strain values of the same samples at the maximum load of 600 kPa were 164 ± 28% and 130 ± 20%, respectively. Treatment of the grafts with aqueous sodium hydroxide solution increased the surface roughness and thus the alloplastic graft materials (PLLA-HAp/M) protecting the fiber morphology were produced successfully. Then, platelet-rich plasma (PRP) was loaded into the surface modified grafts and activated with 10% calcium chloride. The efficiency of the activation was evaluated with flow cytometry and it was found that after activation the percentages of CD62 (P-selectin) and CD41/61 (glycoprotein IIb/IIIa) proteins increased approximately 4-fold. Surface hydrophilicity and biological activity of the PLLA-HAp grafts were enhanced by fibrin coating after PRP activation. Thein vitrocell culture studies which were carried out by using mouse pre-osteoblasts (MC3T3-E1) showed that graft materials supported by PRP increased cellular proliferation and osteogenic differentiation significantly. Thein vivoresults demonstrated that compared with bare PLLA-HAp/M grafts, the PRP loaded grafts (PRP-PLLA-HAp/M) induced significantly greater bone formation based on computed tomography, histological and immunohistochemical analyses. Our findings suggest that 3D PLLA nanofibrous matrices can be used as a graft material for irregular bone defects especially when combined with PRP as an osteogenic induction agent.

Ubiquitin proteasome system and autophagy associated proteins in human testicular tumors.

Ubiquitin proteasome sytem (UPS) and autophagy govern protein quality control by degradation and clearance of damaged proteins. Many proteins working in these pathways such as p97/VCP, Ubiquitin (Ub), Jab1/CSN5, p62, LC3B and Beclin 1 are known to be essential for different pathological conditions, especially in cancer, but their expression in human testicular tumors has not been characterized yet. In the present study, we aimed to investigate the expression of UPS (p97/VCP, Ubiquitin, Jab1/CSN5) and autophagic (p62, LC3B, Beclin 1) proteins in human testicular tumors and cancer adjacent normal testicular tissues. We used an immunohistochemical staining technique. 120 cases of testicular germ and non-germ cell tumors, which are 42 seminomas, 31 embryonal carcinomas, 11 yolk sac tumors, 25 intratubular germ cell neoplasms, 6 Leydig cell tumors, 5 Sertoli cell tumors, were collected and evaluated on tissue microarray. For the first time, the expression of p97/VCP, Ub, Jab1/CSN5, p62, LC3B and Beclin 1 in different type of human testicular tumors has been confirmed. We found that p97/VCP, Ub and Jab1/CSN5 were frequently expressed at higher levels in testicular tumours. In contrast to UPS markers, p62, LC3B and Beclin 1 showed significantly diminished expressions in testicular tumors. Accordingly, a negative correlation between p97/VCP and autophagic markers (p62 and LC3B) was found, suggesting a relationship between UPS and autophagy in different type of testicular tumors. The current results displayed elevated level of p97/VCP, Ub and Jab1/CSN5 expressions in contrast to the diminished expression of p62, LC3B and Beclin 1 in human testicular tumors, thereby supporting a correlation between p97/VCP and autophagic markers in testicular tumors.

Surgical and Immunohistochemical Outcomes of Scleral Reconstruction with Autogenic, Allogenic and Xenogenic Grafts: An Experimental Rabbit Model.

Purpose: Choukroun's platelet-rich fibrin (PRF), a second-generation platelet concentrate, has unique morphological and chemical features and may be considered as a scaffold for scleral reinforcement and regeneration. The purpose of this study was to compare the use of xenogenic human-derived amniotic membrane (HAM), allogenic sclera, and autogenic PRF in rabbit lamellar scleral defect model with respect to both anatomical and immunohistochemical improvement. Methods: A total of 45 adult New Zealand rabbits were randomized into five groups: normal control; without surgical procedure, negative control; scleral defect model (SDM), xenogenic HAM; SDM+HAM graft, allogenic sclera; SDM+allogenic sclera graft, autogenic PRF; SDM+autogenic PRF graft. Clinical findings, Hematoxylin&Eozin (HE), Masson Trichrome, Verhoeff Acid Fuchsin, Transforming Growth Factor ß Receptor 1, Fibroblast Growth Factor, Bone Morphogenetic Protein 2, collagen type 1, aggrecan, and Matrix Metalloproteinase 2 were evaluated. Results: Ocular surface inflammation was significantly lower in normal control and autogenic PRF groups (p < .001). Graft was avascular and not integrated to scleral wound area in 25% rabbits of allogenic sclera group (p = .02), was out of the scleral wound in 33.3% rabbits of xenogenic HAM group (p > .05), all the grafts were at the normal location and viable in autogenic PRF group. The inflammation and vascularization in autogenic PRF group was significantly lower than negative control and xenogenic HAM groups in HE (p < .001). The collagen score of negative control and xenogenic HAM groups were significantly lower than normal control (p < .001) and autogenic PRF (p < .001) groups. There were insignificant differences between allogenic sclera and autogenic PRF groups (p > .05). For immunohistochemistry, the closest values to normal control group were detected in autogenic PRF group for all immunomarkers. Conclusion: Autogenic PRF showed superior features via its excellent anatomical and chemical composition for scleral regeneration when compared to single-layered xenogenic HAM and allogenic sclera grafts.

Analysis of the developmental expression of small VCP-interacting protein and its interaction with steroidogenic acute regulatory protein in Leydig cells.

Small VCP-interacting protein (SVIP) is a 9-kDa protein that is composed of 76 amino acids, and it plays a role in the endoplasmic reticulum-associated protein degradation (ERAD) pathway. Recent studies have shown that SVIP is an androgen-responsive protein and its expression is regulated by androgens. Because no data are available regarding the cellular localization and expression of SVIP in the mouse testis, where androgens are highly expressed, immunohistochemistry and western blotting were performed. In the fetal testis, we found that moderate but consistent staining of SVIP is present in the cytoplasm of Leydig cells. In prepubertal and adult life, SVIP remains present in Leydig cells as well as in the cytoplasm of some peritubular and Sertoli cells. From postnatal day 15 onward, SVIP is strongly expressed in the cytoplasm of Leydig cells. Furthermore, TM3, MA-10 Leydig and Sertoli cell lines were also used to evaluate the expression of SVIP. To identify the interacting partners, such as steroidogenic acute regulatory (STAR) protein, colocalization studies were performed by fluorescence microscopy, showing that STAR colocalized with SVIP in the adult mouse testis. The expression changes of STAR were studied by using SVIP siRNAs in Leydig cell line cultures. Depletion of SVIP resulted in decreased expression of STAR. Additionally, the number and size of lipid droplets were significantly increased in SVIP-depleted Leydig cells. Taken together, our data identify SVIP as a marker of Leydig cell lineage and as a regulator of STAR protein expression and lipid droplet status in Leydig cells.

Effects of 1,25-Dihydroxy vitamin D3 on TNF-α induced inflammation in human chondrocytes and SW1353 cells: a possible role for toll-like receptors.

The purpose of this study is to evaluate anti-inflammatory and chondro-protective effects of 1,25(OH)2D3 in human chondrocytes and SW1353 cells via investigating expressions of MMPs, TIMPs, VDR, and intracellular signalling pathway mediators such as TLR-2 and -4. The HC and SW1353 cells were treated with 1,25(OH)2D3 at 10, 100, and 1000 nM concentrations in the absence/presence of TNF-α (20 ng/mL) for 48 h. The mRNA expressions of MMP-1, -2, -3, -9, and -13, TIMP-1 and -2, VDR, TLR-2 and -4 in HC and SW1353 cells were detected by qPCR after treatments. The cytotoxicity and cell proliferation analyses were assessed by LDH and WST-1 assay, respectively. Protein levels of MMPs, TIMPs, and VDR were analysed by immunocytochemistry and ELISA methods. TNF-α markedly increased cytotoxicity for 24, 48, 72 h (p < 0.05) and vitamin D treatment was shown to diminish the cytotoxic effect of TNF-α. Cell proliferations increased by Vitamin D in a dose-dependent manner. mRNA expressions of MMP-1, -2, -3, -9, and -13, TLR-2 and -4 genes decreased with 1,25(OH)2D3 treatment (p < 0.05). VDR, TIMP-1 and -2 levels elevated after TNF-α exposure compared with the control group in HC cells (p < 0.05). Protein expression levels were determined using Western blotting, ELISA and immunocytochemistry. 1,25(OH)2D3 via binding to VDR, reversed the effects of TNF-α by inhibiting TLR-2 and 4. Decreased levels of VDR, TIMP-1 and -2 after TNF-α treatment were elevated by 1,25(OH)2D3 proportional with increasing 1,25(OH)2D3 doses. 1,25(OH)2D3 and TNF-α co-treatment decreased MMP-1, -2, -3, -9, and -13 levels were after TNF-α exposure.