• Title/Summary/Keyword: PRP

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PRP4 Kinase Domain Loss Nullifies Drug Resistance and Epithelial-Mesenchymal Transition in Human Colorectal Carcinoma Cells

  • Ahmed, Muhammad Bilal;Islam, Salman Ul;Sonn, Jong Kyung;Lee, Young Sup
    • Molecules and Cells
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    • v.43 no.7
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    • pp.662-670
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    • 2020
  • We have investigated the involvement of the pre-mRNA processing factor 4B (PRP4) kinase domain in mediating drug resistance. HCT116 cells were treated with curcumin, and apoptosis was assessed based on flow cytometry and the generation of reactive oxygen species (ROS). Cells were then transfected with PRP4 or pre-mRNA-processing-splicing factor 8 (PRP8), and drug resistance was analyzed both in vitro and in vivo. Furthermore, we deleted the kinase domain in PRP4 using Gateway™ technology. Curcumin induced cell death through the production of ROS and decreased the activation of survival signals, but PRP4 overexpression reversed the curcumin-induced oxidative stress and apoptosis. PRP8 failed to reverse the curcumin-induced apoptosis in the HCT116 colon cancer cell line. In xenograft mouse model experiments, curcumin effectively reduced tumour size whereas PRP4 conferred resistance to curcumin, which was evident from increasing tumour size, while PRP8 failed to regulate the curcumin action. PRP4 overexpression altered the morphology, rearranged the actin cytoskeleton, triggered epithelial-mesenchymal transition (EMT), and decreased the invasiveness of HCT116 cells. The loss of E-cadherin, a hallmark of EMT, was observed in HCT116 cells overexpressing PRP4. Moreover, we observed that the EMT-inducing potential of PRP4 was aborted after the deletion of its kinase domain. Collectively, our investigations suggest that the PRP4 kinase domain is responsible for promoting drug resistance to curcumin by inducing EMT. Further evaluation of PRP4-induced inhibition of cell death and PRP4 kinase domain interactions with various other proteins might lead to the development of novel approaches for overcoming drug resistance in patients with colon cancer.

Measurement of Free Polysaccharide in Tetanus Toxoid-Conjugate Vaccine Using Antibody/Ammonium Sulfate Precipitation

  • Yoo, Tae-Hyeon;Kim, Hyun-Sung;Park, Sung-Sik;Bang, Eun-Young;Oh, Yong-K.;Kim, Li-Seop;Kim, Hun;Hur, Byung-Ki;Ryu, Yeon-Woo;Kim, Jong-Su
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.469-472
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    • 2003
  • A method that effectively precipitates capsular polysaccharide of Haemophilus influenzae type b (polyribosylribitol phosphate, PRP) conjugated to tetanus toxoid (TT), PRP TT in a liquid vaccine has been developed to measure free PRP present in TT-conjugate vaccine. The method involves adding anti-TT antibody and ammonium sulfate to precipitate PRP-TT conjugate and measuring free PRP in tile supernatant. This new method provides a complete precipitation of the total PRP-TT, and provides an accurate and reproducible measurements of free PRP. The accuracy of the assay was confirmed by spiking known amounts of unconjugated PRP to PRP-TT conjugate, and the new method was found to have no effect on free PRP while precipitating PRP-TT. The published acid precipitation method did not produce reproducible results due to incomplete precipitation of PRP-TT, especially when the vaccine is formulated in a salt-buffered solution.

Prospective Clinical Research of the Efficacy of Platelet-rich Plasma in the Outpatient-based Treatment of Rotator Cuff Tendinopathy

  • Lee, Ho-Won;Choi, Kyung-Ho;Kim, Jung-Youn;Yang, Ik;Noh, Kyu-Cheol
    • Clinics in Shoulder and Elbow
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    • v.22 no.2
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    • pp.61-69
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    • 2019
  • Background: The purpose of this study is to compare the clinical outcomes of the control group and platelet-rich plasma (PRP) group among the patients who failed to respond to conservative treatment as outpatient-based therapy for rotator cuff tendinopathy, and to compare the clinical results of leukocyte-poor (LP) PRP and leukocyte-rich (LR) PRP. Methods: Inclusion criteria are (1) over 18-year-old, (2) patients with rotator cuff tendinopathy, no rotator cuff tear by radiologic diagnosis (ultrasonography or magnetic resonance imaging) within the last 3 months, and (3) not effective to conservative treatment for more than 1 month. Of the final 60 subjects, 33 patients in the exercise treatment group and 27 patients in the PRP injection group (LP-PRP, 13; LR-PRP, 14) were included. Clinical evaluation was carried out by assessing the outcomes of treatment using the Numeric Rating Scale pain score, the American Shoulder and Elbow Society (ASES) score, and the Constant score at baseline and at 3 and 6 months after the procedure. Results: There was a statistically significant difference in ${\Delta}ASES_{3months}$ ($ASES_{3months}-ASES_{first}$) score between the control and PRP groups (p=0.006). However, there was no statistical significance between LP-PRP and LR-PRP groups (p>0.05). Conclusions: This study showed that PRP injection was more effective than exercise therapy for the first 3 months. However, there was no difference between the LP-PRP group and the LR-PRP group. Regardless of the type of PRP, clinical application of PRP injection in patients with rotator cuff tendinopathy seems to be effective in early treatment.

Multiscale Analysis on Vibration of the Photo Responsive Polymer (광변형 고분자의 동적 진동에 관한 멀티스케일 해석)

  • Yun, Jung-Hoon;Li, Chenzhe;Chung, Hayoung;Choi, Joonmyung;Cho, Maenghyo
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.29 no.6
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    • pp.571-575
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    • 2016
  • Photo responsive polymer(PRP) is well known for its photo deformation under UV light, and goes back to its original shape in visible light due to the photoisomerization of the azobenzene inside the PRP. In this paper, dynamic study of the vibration in PRP is discussed. In order to predict photo-deformation of the PRP a multiscale modeling is introduced which covers quantum level photo excitation, microscopic morphology, and macroscopic deformation of the PRP. A simple 1D beam model is introduced to model dynamic bending behavior of the PRP. Through fast Fourious transformation analysis, we identify that vibration frequency of the PRP can be controlled by light polarization angle.

Clinical Application of Autologous Platelet-Rich Plasma (PRP) on Delayed Wound Healing of a Dog with Burns

  • Lee, Shinho;Cheong, Jongtae;Lee, Joo-Myoung
    • Journal of Veterinary Clinics
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    • v.35 no.5
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    • pp.229-232
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    • 2018
  • A 9-year-old intact female Poodle (weighing 3.6 kg) was presented for the treatment of a partial and full thickness burn that covering 45% of body including bilateral thigh and sacrolumbar region. Autologous platelet-rich plasma (PRP) stimulates angiogenesis, promoting vascular in-growth and fibroblast proliferation. On the unrecovered right thigh of the dog, autologous PRP was injected into the lesion after application of micro-needles. For macroscopic evaluation digital, photographs were taken from wounds at days 0, 3, 5, 13, 21, and 28. On the $3^{rd}$ day after PRP application, epithelialization was accelerated. Application of autologous PRP accelerated wound-healing rate and healing time in full thickness burns as well as secondary complications originating from unrecovered wounds. The delayed lesion was completely healed on the $28^{th}$ day by autologous PRP treatment. In human, PRP was increasingly used in the treatment of a variety of soft tissue in the management of chronic non-healing wounds. This study has shown that PRP treatment can be a valuable and effective aid on intractable wound healing in the dog with burns.

FABRICATION OF PLATELET-RICH PLASMA IN A RAT MODEL AND THE EFFICACY TEST IN VITRO (백서에서 혈소판 풍부 혈장의 제작과 유효성에 대한 실험적 연구)

  • Lee, Sang-Hoon;Cho, Young-Uk;Chi, Hyun-Sook;Ahn, Kang-Min;Lee, Bu-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.2
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    • pp.113-122
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    • 2007
  • Purpose: Platelet-rich plasma (PRP) is known to accelerate and/or enhance hard and soft tissue healing and regeneration. As such, PRP has been used in various clinical fields of surgery. Recently there have been several attempts to use PRP in the field of tissue engineering. However, some controversies still exist on exact mechanism and benefits of PRP. Therefore various animal experiments are necessary to reveal the effect of the PRP. However, even if animal experiment is performed, the efficacy of the experiment could not be validated due to absence of an animal PRP model. The purpose of this study is to establish rat PRP model by comparing several PRP fabricating methods, and to assay growth factor concentration in the PRP. Materials and methods: Rat blood samples were collected from nine SD rat (body weight: 600-800g). PRP was prepared using three different PRP fabricating methods according to previously reported literatures. (Method 1: 800 rpm, 15 minute, single centrifuge; Method 2: 1000 rpm, 10 minute, double centrifuge; Method 3: 3000 rpm, 4min and 2500 rpm, 8 min, double centrifuge). Platelet counts were evaluated in an automated machine before and after PRP fabrications. In terms of growth factor assay, prepared PRP were activated with 100 unit thrombin and 10% calcium chloride. Growth factor (PDGF-BB, VEGF) concentrations on incubation time were determined by sandwich-ELISA technique. Results: An average of 3ml (via infraorbital venous plexus) to 15ml (via celiac axis) the rat blood could be collected. By using Method 3 (3000 rpm, 4 min and 2500 rpm, 8 min, double centrifugation), around 1.5ml of PRP could be prepared. This method allowed us to concentrate platelet 3.77-fold on average. PDGF-BB concentration (mean, 1942.10 pg/ml after 1 hour incubation) and VEGF concentration (mean, 952.71 pg/ml after 1 hour incubation) in activated PRP were higher than those in untreated blood. Also PDGF-BB showed constant concentration during 4-hour incubation, while VEGF concentration was decreased after 1 hour. Conclusion: Total 11,000 g minute separation and condensation double centrifuge method can produce efficient platelet-rich plasma. Platelet-rich plasma activated with thrombin has showed higher concentrations of growth factors such as PDGF-BB and VEGF, compared to the control group. Platelet-rich plasma model in a rat model was confirmed in this study.

Platelet-Rich Plasma Enhances Proliferation and Migration and Inhibits Inflammatory Processes in Canine Chondrocytes

  • Kim, Dongyub;Jeong, Seong Mok;Kwon, Youngsam;Yun, Sungho
    • Journal of Veterinary Clinics
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    • v.36 no.4
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    • pp.200-206
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    • 2019
  • This study was performed to assess the anti-inflammatory and cartilage regenerative effects of platelet-rich plasma (PRP) on canine chondrocytes. Proliferation and migration assays under both normal and lipopolysaccharide (LPS)-induced inflammatory conditions were performed with various concentrations of PRP (1% to 10%). The expression levels of genes related to osteoarthritis were evaluated in the following groups: PRP group, LPS group and LPS + PRP group. mRNA expression levels were detected using real-time polymerase chain reaction (RT-PCR). Proliferation assays showed significantly enhanced proliferation in all PRP-treated groups compared with the no serum group. Compared with 10% fetal bovine serum (FBS), PRP concentrations above 3% in the normal condition and 1% to 7% PRP in the LPS-induced inflammatory condition were found to significantly promote chondrocyte proliferation. In the normal condition, all PRP-treated groups showed significantly increased cell migration compared with the no serum group. Chondrocyte migration was decreased with LPS-induced inflammation, but PRP treatment resulted in significantly enhanced migration compared with the other groups in this condition. According to RT-PCR, the LPS + PRP group showed significantly higher levels of COL1A1, IL-6, aggrecan and lower levels of $TNF-{\alpha}$, MMP-1, MMP-3 mRNA expression compared to the LPS group. The results of this study suggest that PRP application can enhance the proliferation and migration of canine chondrocytes and improve canine articular cartilage regeneration.

Can platelet-rich plasma injections provide better pain relief and functional outcomes in persons with common shoulder diseases: a meta-analysis of randomized controlled trials

  • Barman, Apurba;Mishra, Archana;Maiti, Rituparna;Sahoo, Jagannatha;Thakur, Kaustav Basu;Sasidharan, Sreeja Kamala
    • Clinics in Shoulder and Elbow
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    • v.25 no.1
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    • pp.73-89
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    • 2022
  • Background: To evaluate the efficacy of autologous platelet-rich plasma (PRP) injections in the treatment of common shoulder diseases. Methods: The PubMed, Medline, and Central databases and trial registries were searched from their inception to October 2020 for randomized controlled trials of autologous PRP injections for shoulder diseases versus placebo or any control intervention. Preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were followed in the selection, analysis, and reporting of findings. The primary outcome was pain intensity (visual analog scale), and secondary outcomes were changes in function and quality of life (QoL). Results: A total of 17 randomized controlled trials of PRP versus control were analyzed. From 8-12 weeks to ≥1 year, PRP injections were associated with better pain relief and functional outcomes than control interventions. PRP injections were also associated with greater QoL, with an effect size of 2.61 (95% confidence interval, 2.01-14.17) at medium-term follow-up. Compared with placebo and corticosteroid injections, PRP injections provided better pain relief and functional improvement. In subgroup analyses, trials in which PRP was prepared by the double centrifugation technique, the platelet concentration in the PRP was enriched ≥5 times, leucocyte-rich PRP was used, or an activating agent was used before application reported the most effective pain relief at 6-7 months. Conclusions: PRP injections could provide better pain relief and functional outcomes than other treatments for persons presenting with common shoulder diseases. PRP injections have a greater capacity to improve shoulder-related QoL than other interventions.

Immunogenicity and Safety of a Haemophilus influenzae Type b Polysaccharide-Tetanus Toxoid Conjugate Vaccine (PRP-T: HiberixTM) in Korean Infants (우리나라 영아에서 PRP-T(HiberixTM)백신의 면역원성 및 안전성에 대한 연구)

  • Chung, Eun Hee;Kim, Yae Jean;Kim, Yun Kyung;Kim, Dong Ho;Seo, Jeong Wan;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.10 no.1
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    • pp.71-80
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    • 2003
  • Purpose : Four kinds of Haemophilus influenzae type b protein conjugate vaccines, PRPD, PRP-T, PRP-OMP and PRP-CRM197, have been developed, and PRP-T vaccines are currently produced by two manufacturer, $ActHib^{(R)}$ by Aventis and $Hiberix^{TM}$ by GlaxoSmith-Kline Biologicals. The purpose of this study is to evaluate the immunogenicity and safety of $Hiberix^{TM}$ in Korean infants. Methods : Seventy-three healthy infants(43 male infants) were recruited for this study after parental informed consent was obtained. Each infant was vaccinated at 2, 4 and 6 months of age with the study vaccine. At each visit, infants were also immunized with DTaP, trivalent oral polio vaccine and hepatitis B vaccine when indicated. The serum anti-PRP antibody was measured at prevaccination, 2 month later after the 2nd dose, and 1 month later after the 3rd dose by the ELISA method. The local and systemic adverse reactions of vaccination were monitored for 3 consecutive days after each immunization. Immunogenicity of vaccine was evaluated in infants who received all the scheduled immunization and the adverse reactions were evaluated for infants who received at least one dose of the study vaccine. Results : Among seventy three infants, enrolled in this study; sixty three(37 male infants) completed all the scheduled immunizations. The geometric mean titer(GMT) of anti-PRP antibodies at prevaccination was 0.17 ${\mu}g/mL$(95% confidence interval[CI]; 0.13~0.22). The GMT of anti-PRP antibodies increased to 4.14 ${\mu}g/mL$(95% CI; 2.65~6.48) at 2 month later after the 2nd dose of PRP-T and 14.65 ${\mu}g/mL$(95% CI; 10.83~19.81) at 1 month later after the 3rd dose. Anti-PRP antibody ${\geq}0.15$ ${\mu}g/mL$, was observed in 98.4%(95% CI; 91.8~100) after 2 doses and 100%(95% CI; 100~100) after 3 doses. Anti-PRP antibody ${\geq}1.0$ ${\mu}g/mL$, was obtained in 77.8%(95% CI; 67.5~88.0) after 2 doses, and 98.4%(95% CI; 95.3~100) after 3 doses. Most of the adverse reaction after vaccination were mild. Irritability, the most common systemic reaction, was observed in 45.5%, followed by drowsiness(30.5%), poor feeding(26.7%) and fever(5.6%). Among the local reactions tenderness was observed in 7.9%, redness(${\geq}5$ mm) in 2.8% and swelling(${\geq}5$ mm) in 1.8%. Conclusion : The PRP-T vaccine used in this study was highly immunogenic and safe in Korean young infants. The finding that high GMT and high frequency of infants with a protective titer achieved after 2 doses is consistent with the previous studies which were done with a PRP-T vaccine of other manufacturer. This study suggests that the immunization schedule of PRP-T vaccine for Korean infants may need re-evaluation.

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THE EFFECT OF GROWTH FACTORS IN PLATELET-RICH PLASMA ON THE ACTIVITY OF OSTEOBLAST CELL LINE (혈소판농축혈장 내의 성장요소가 조골세포주의 활성도에 미치는 영향)

  • Jung Tae-Wook;Kim Yung-Soo;Kim Chang-Whe;Jang Kyung-Soo;Lim Young-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.2
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    • pp.175-191
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    • 2004
  • Statement of problem: Platelet-rich plasma(PRP) is well known to be very effective method to stimulate and accelerate the healing of bone and soft tissue. However, there are few reports which deal with the mechanisms of the PRP on the activation of the osteoblasts. Purpose: This study was aimed to investigate the effect of growth factors in PRP on the activity of osteoblasts. Material and method: To evaluate the effect on human, human osteoblast cell line was cultured. PRP was extracted from the blood of a healthy volunteer. Using the recombinant growth factors of PDGF, $TGFT-\beta$, IGF-1, bFGF which are mainly found at bone matrix and their neutralizing antibody, the effect of PRP on the attachment and proliferation of osteoblasts was evaluated. To evaluate the autocrine and paracrine effects, conditioned media(CM) of PRP was made and compared with PRP. By the western blot analysis, the expression of growth factors in PRP, CM was examined. Cell morphology was compared by the light microscope. Results : 1) The effects of CM on osteoblast were similar to the effects of PRP. 2) PRP, CM, recombinant $TGF-\beta$, bFGF, IGF-1 showed significantly higher cellular attachment than control(p<0.05) in the cell attachment assay. In the cell proliferation assay, PRP, CM, recombinant $TGF-\beta$, IGF-1, bFGF, PDGF increased significantly cell proliferation(p<0.01). Among the recombinant growth factors, IGF-1 showed the highest cellular attachment and proliferation. 3) In the western blot assay, bFGF, IGF-1, PDGF weve equally expressed in PRP and CM. 4) The attachment of osteoblast cell decreased significantly after the addition of neutralizing antibody against $TGF-\beta$, IGF-1(p<0.05). In the cell proliferation assay, the addition of neutralizing antibody against $TGF-\beta$, bFGF, PDGF, IGF-1 decreased significantly the cellular proliferation(p<0.05). The amount of decreasing in the cell attachment and proliferation is the highest in at-lGF-1. 5) The cells in control group were flattened and elongated with a few cellular processes in the a light microscope. But, the cells appeared as spherical, plump cells with well developed cellular processes in experimental groups. The cells in PRP and CM had more prominent developed features than recombinant growth factor groups. Conclusions : These findings imply that PRP maximize the cellular activity in early healing period using the synergistic effect, autocrine, paracrine effects of growth factors and increase the rate and degree of bone formation.