• Clinics in Shoulder and Elbow
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• 제21권1호
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• pp.3-14
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• 2018

Background: Platelet-rich plasma (PRP) stimulates cell proliferation and enhances matrix gene expression and synthesis. However, there have been no comparative study of the PRP effect on the normal and degenerative tenocytes. The purpose of this study was to compare the effect of PRP on tenocytes from normal and degenerative tendon. Methods: Tendon tissues were obtained from patients undergoing arthroscopic repair (n=9) and from healthy donors (n=3). Tenocytes were cultured with 10% (vol/vol) platelet-poor plasma, PRP activated with calcium, and PRP activated with calcium and thrombin. The total cell number was assessed at days 7 and 14. The expressions of type I and III collagen, decorin, tenascin-C, and scleraxis were evaluated by quantitative real-time reverse transcriptase polymerase chain reaction. The total collagen and glycosaminoglycan (GAG) synthesis was evaluated at days 7 and 14. Results: No differences were observed between the groups at day 7, but cell proliferation was remarkably increased in tenocytes from the degenerative tendon at day 14. In both tenocyte groups, the gene expressions of type I and III collagen were up-regulated. GAG synthesis was greater in the normal tendon, whereas the expressions of decorin and tenascin-C were increased in tenocytes from the degenerative tendon. Tenocytes from the degenerative tendon had higher fold-change of GAG synthesis and a lower collagen III/I ratio than normal tenocytes. Conclusions: PRP promoted the cell proliferation and enhanced the synthesis of tendon matrix in both groups. PRP has a greater positive effect on cell proliferation, matrix gene expression and synthesis in tenocytes from degenerative tendon.

• Clinics in Shoulder and Elbow
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• 제24권4호
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• pp.224-230
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• 2021

Background: Local anesthetics often are used in rotator cuff tears as therapeutic tools, although some cases have reported that they have detrimental effects. Neurotropin (NTP) is used widely in Japan as a treatment for various chronic pain conditions and is shown to have protective effects on cartilage and nerve cells. In this study, we investigated the protective effect of NTP against lidocaine-induced cytotoxicity. Methods: Tenocytes from rotator cuff tendons were incubated with lidocaine, NTP, lidocaine with NTP, and a control medium. Cell viability was evaluated using the WST-8 assay. Cell apoptosis was detected via annexin V staining using flow cytometry. The expression of BCL-2 and cytochrome c, which are involved in the intrinsic mitochondrial pathway of apoptosis, was evaluated via Western blotting and immunohistochemical staining. Results: In the cell viability assay, lidocaine decreased cell viability in a dose-dependent manner, and NTP did not affect cell viability. Moreover, NTP significantly inhibited the cytotoxic effect of lidocaine. The flow cytometry analysis showed that lidocaine significantly induced apoptosis in tenocytes, and NTP considerably inhibited this lidocaine-induced apoptosis. Western blotting experiments showed that lidocaine decreased the protein expression of BCL-2, and that NTP conserved the expression of BCL-2, even when used with lidocaine. Immunohistochemical staining for cytochrome c showed that 0.1% lidocaine increased cytochrome c-positive cells, and NTP suppressed lidocaine-induced cytochrome c expression. Conclusions: NTP suppresses lidocaine-induced apoptosis of tenocytes by inhibiting the mitochondrial apoptotic pathway. Intra-articular/bursal injection of NTP with lidocaine could protect tenocytes in rotator cuff tendons against lidocaine-induced apoptosis.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.297-303
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• 2019

Corticosteroids are commonly used for pain control in rotator cuff tear. Deregulated $NF-{\kappa}B$ activation is a hallmark of chronic inflammatory diseases and has been responsible for the pathogenesis of rotator cuff tear. The Dexamethasone(DEXA) is a synthetic corticosteroid. The purpose of this study was to examine the exact effect of dexamethasone on $NF-{\kappa}B$ signaling in rotator cuff tear. We measured $NF-{\kappa}B$ expression in four groups: control, $TNF-{\alpha}$-treated, DEXA-treated, and combined treatment with $TNF-{\alpha}$ and DEXA. Tenocytes were isolated from patients with rotator cuff tears and pre-incubated with $TNF-{\alpha}$ (10 ng/ml), DEXA ($1{\mu}M$), or both of them for 10 min, 1 h, and 2 h. Expression of p65, p50, and p52 in the nuclei and cytosol was analyzed by western blotting and immunofluorescence imaging using confocal microscopy. We also evaluated nucleus/cytosol (N/C) ratios of p65, p50, and p52. In our study, the combined treatment with DEXA and $TNF-{\alpha}$ showed increased N/C ratios of p65, p50, and p52 compared with those in the $TNF-{\alpha}$ group at all time points. Additionally, in the DEXA group, N/C ratios of p65, p50, and p52 gradually increased from 10 min to 2 h. In conclusion, DEXA promoted the nuclear localization of p65, p50, and p52, but was not effective in inhibiting the inflammatory response of $TNF-{\alpha}$-stimulated rotator cuff tear.

• Journal of Ginseng Research
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• 제46권4호
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• pp.526-535
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• 2022

Background: During the pathogenesis of tendinopathy, the chronic inflammation caused by the injury and apoptosis leads to the generation of scars. Ginsenoside Rg1 (Rg1) is extracted from ginseng and has anti-inflammatory effects. Rg1 is a unique phytoestrogen that can activate the estrogen response element. This research aimed to explore whether Rg1 can function in the process of tendon repair through the estrogen receptor. Methods: In this research, the effects of Rg1 were evaluated in tenocytes and in a rat model of Achilles tendinitis (AT). Protein levels were shown by western blotting. qRT-PCR was employed for evaluating mRNA levels. Cell proliferation was evaluated through EdU assay and cell migration was evaluated by transwell assay and scratch test assay. Results: Rg1 up-regulated the expression of matrix-related factors and function of tendon in AT rat model. Rg1 reduced early inflammatory response and apoptosis in the tendon tissue of AT rat model. Rg1 promoted tenocyte migration and proliferation. The effects of Rg1 on tenocytes were inhibited by ICI182780. Rg1 activates the insulin-like growth factor-I receptor (IGF1R) and MAPK signaling pathway. Conclusion: Rg1 promotes injured tendon healing in AT rat model through IGF1R and MAPK signaling pathway activation.

• Anatomy and Cell Biology
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• 제55권3호
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• pp.304-310
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• 2022

Anterior cruciate ligament is a commonly damaged ligament of the knee. Reconstruction of this ligament usually entails the use of graft harvested from the same subject (i.e., autografts). Several tendons, for example quadriceps, patellar or semitendinosus tendon can be used as an autograft. The composition of the tendons is similar to the anterior cruciate ligament but there is no data that directly compares the compositions of the quadriceps, patellar and semitendinosus tendons. This study quantified and compared the tenocyte distribution and collagen content of these tendons from cadavers of South Africans of European Ancestry. The tenocyte distribution and collagen content were assessed using the ImageJ software. The results showed similarities in the collagen content across the tendons in both sexes (P>0.05). The tenocyte distribution was significantly higher in the quadriceps (P=0.019) or semitendinosus (P=0.016) tendon than in the patellar tendon in the female but no difference was seen in the male (P=0.872). This shows that a large harvestable area may not be directly associated with a more abundant collagen content or tenocyte distribution in the tendon. However, sex-specific tenocyte distribution is an important observation that underpins the possible influence of underlying biological factors on the composition of each tendon and this requires further investigations. In all, this study will contribute to knowledge and assist orthopaedic surgeons in making an informed decision on the choice of graft.

• Applied Microscopy
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• 제36권2호
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• pp.93-99
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• 2006

Gemifloxacin은 대부분의 G(-) 세균들과 G(+) 세균들에 대해 강한 활성을 나타내며 미성숙동물에서 비교적 낮은 연골독성 가능성을 가진 합성 fluoroquinolone계 항생제이다. 본 연구는 미성숙 SD계 랫드의 아킬레스건에 gemifloxacin이 미치는 영향을 평가하기 위해 gemifloxacin을 생후 4주 후부터 0(vehicle), 600mg/kg body weight 용량으로 5일 동안 매일 한번씩 경구투여를 하여 ofloxacin과 비교하였다. 아킬레스건의 형태학적 변화를 투과전자현미경을 이용하여 관찰한 결과 부형제만 처치한 대조군과 비교하였을 때, gemifloxacin과 ofloxacin을 처치한 랫드로부터 채취한 샘플에서 초미세구조의 변화가 관찰되었다. quinolone을 처치한 군의 tenocyte에서는 다양한 퇴행성 변화가 관찰되었고, 세포외기질에서 분리된 세포들이 존재하였는데 이는 ofloxacin을 처치한 그룹에서 특히 높았고 gemifloxacin을 처치한 그룹에서는 매우 낮았다. 따라서, gemifloxacin이 아킬레스건과 같은 결합조직 구조물들의 퇴행성 변화를 보다 적게 야기한다는 것을 보여준다.