scholarly journals A possible Injectable Tissue Engineered Nucleus Pulposus constructed with platelet-rich plasma and ADSCs in Vitro

2020 ◽  
Author(s):  
Zhicheng Zhang ◽  
Jian Ma ◽  
Dajiang Ren ◽  
Fang Li
Keyword(s):  
Nucleus Pulposus ◽  
Platelet Rich Plasma ◽  
Positive Staining ◽  
Adipose Derived Stromal Cells ◽  
Genes Expression ◽  
Significant Difference ◽  
Safranin O ◽  
Collagen Genes ◽  
Prp Gel

Abstract Background: Injectable tissue engineered nucleus pulposus is a new idea for minimally invasive repair for degenerative intervertebral disc. The platelet-rich plasma (PRP) and adipose derived stromal cells(ADSCs) could be harvested from autologous tissue easily. PRP has mixed autologous growth factors and fibrous reticulate structure which may have the potential to make ADSCs differentiated into nucleus pulposus-like cells. The goal of this study was to explore the feasibility of constructing a possible injectable tissue engineered nucleus pulposus with PRP gel scaffold and ADSCs. Methods: After the rabbit ADSCs were identified with flow cytometry, the ADSCs were seeded into PRP gel and cultured in vitro. At the 2nd, 4th and 8th week, the PRP gel/ADSCs complex was observed by macroscopy, histological staining, BrdU immunofluorescence, and scanning electron microscopy. The glycosaminoglycans (GAG) in the PRP gel/ADSCs complex were measured by safranin O staining with spectrophotometry. PRP gel/ADSCs complex genes expression of HIF-1α, Aggrecan, Type Ⅱ collagen were detected by RT-PCR. The injectability of this complex was tested. Results: Macroscopically, the complex was solidified into gel with smooth surface and good elasticity. The safranin O staining confirmed almost no positive staining at 2nd week, however, the positive staining of extracellular matrix was enhanced obviously at 4th, 8th week. The HE staining and SEM demonstrated that the cells were well-distributed in the reticulate scaffold. BrdU immunofluorescence showed that ADSCs can survive and proliferate in PRP gel at each time points. The level of GAG at 4th week was higher than those at 2nd week (P<0.05), and significant difference was also noted between 4th and 8th week (P<0.05). HIF-1α, Aggrecan, Type Ⅱ collagen genes expression at 4th week were much more than those at 2nd week (P<0.05), and significant differences were also noted between 4th and 8th week (P<0.05). The flow rate of complex was 0.287 mL/min when passed through the 19-gauge needle with the 100mmHg pressure. Conclusions: The PRP gel made it possible for rabbit ADSCs differentiated into nucleus pulposus-like cells after mixed culture in vitro. Maybe, it is a better feasible method for construction of autologous injectable tissue engineered nucleus pulposus.

scholarly journals Evaluation of platelet-rich plasma gel as an angiogenesis-inducing agent in canineadvancement skin flaps

2020 ◽  
Vol 40 (6) ◽  
pp. 474-478
Author(s):  
Grazielle A.S. Aleixo ◽  
Maria C.O.C. Coelho ◽  
Telga L.A. Almeida ◽  
Márcia F. Pereira ◽  
Miriam N. Teixeira ◽  
...  
Keyword(s):  
Blood Vessel ◽  
Skin Color ◽  
Platelet Rich Plasma ◽  
Skin Flap ◽  
Skin Flaps ◽  
Experimental Conditions ◽  
Significant Difference ◽  
Experimental Group ◽  
Prp Gel ◽  
Made In

ABSTRACT: This work aimed to evaluate the effect of platelet-rich plasma (PRP) on advancement skin flaps in dogs regarding improvement of vascularization, with focus on increasing its viable area, since there are reports that it is a potential angiogenesis stimulator. The experimental group was composed of eight adult bitches, in which two advancement skin flaps were made in the ventral abdominal region. No product was applied in the control flap (CF), while PRP was used in the contralateral flap, called treated flap (TF). The areas were clinically evaluated every two days until the 7th postoperative day regarding skin color and presence of necrosis. At 10 days, both flaps were removed and submitted to histological examination and blood vessel morphometry. The vessels counted in each group were statistically analyzed by the F-test at 1% probability. Results showed no significant difference in macroscopic changes in the wound, or CF and TF vascularization, thus suggesting that PRP gel did not improve advancement skin flap angiogenesis in bitches under the experimental conditions in which this research was developed.

Effect of transforming growth factor-beta 1 (TGF-ß1) released from a scaffold on chondrogenesis in an osteochondral defect model in the rabbit

2006 ◽  
Vol 1 (1) ◽  
pp. 43-60 ◽  
Author(s):  
Magali Cucchiarini ◽  
Jerome Sohier ◽  
Karin Mitosch ◽  
Gunter Kaul ◽  
David Zurakowski ◽  
...  
Keyword(s):  
Cartilage Repair ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Knee Joints ◽  
Polymeric Scaffold ◽  
Significant Difference ◽  
Membrane Defects ◽  
Safranin O

AbstractArticular cartilage repair might be stimulated by the controlled delivery of therapeutic factors. We tested the hypotheses whether TGF-ß1 can be released from a polymeric scaffold over a prolonged period of time in vitro and whether its transplantation modulates cartilage repair in vivo. Unloaded control or TGF-ß1 poly(ether-ester) copolymeric scaffolds were applied to osteochondral defects in the knee joints of rabbits. In vitro, a cumulative dose of 9 ng TGF-ß1 was released over 4 weeks. In vivo, there were no adverse effects on the synovial membrane. Defects treated with TGF-ß1 scaffolds showed no significant difference in individual parameters of chondrogenesis and in the average cartilage repair score after 3 weeks. There was a trend towards a smaller area (42.5 %) of the repair tissue that stained positive for safranin O in defects receiving TGF-ß1 scaffolds. The data indicate that TGF-ß1 is released from emulsion-coated scaffolds over a prolonged period of time in vitro and that application of these scaffolds does not significantly modulate cartilage repair after 3 weeks in vivo. Future studies need to address the importance of TGF-ß1 dose and release rate to modulate chondrogenesis.

BRCA1 protein expression as a predictor of outcome following chemotherapy in sporadic epithelial ovarian cancer (EOC)

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 5527-5527 ◽  
Author(s):  
J. E. Carser ◽  
J. E. Quinn ◽  
C. O. Michie ◽  
W. G. McCluggage ◽  
A. R. Williams ◽  
...  
Keyword(s):  
Protein Expression ◽  
Predictive Marker ◽  
Chemotherapy Response ◽  
Positive Staining ◽  
Brca1 Protein ◽  
Enhanced Sensitivity ◽  
Platinum Based Chemotherapy ◽  
Significant Difference ◽  
Taxane Chemotherapy

5527 Background: Reduced expression of BRCA1 is observed in a substantial proportion of sporadic EOC. First line management of EOC involves platinum and taxane based chemotherapy. In vitro studies demonstrate that reduction in BRCA1 protein expression leads to enhanced sensitivity to platinum but relative resistance to taxane chemotherapy. We investigated the relationship between BRCA1 protein expression by immunohistochemistry (IHC) and survival in EOC patients, correlating outcome with type of chemotherapy received. Methods: 97 archival formalin-fixed, paraffin embedded tumour samples were identified from the Edinburgh Cancer Centre ovarian database. All patients received platinum based chemotherapy (platinum only n = 50, platinum/taxane n = 47). Tumour sections were stained for BRCA1 protein and scored by two independent reviewers. Tumours were graded from 0 (no BRCA1 protein) to 4 (>90% cells positive). BRCA1 protein expression was correlated with progression-free (PFS) and overall survival (OS) by Kaplan-Meier analysis. Results: Overall, patients with no detectable BRCA1 protein had a significantly improved PFS compared to those with positive staining (17.4 months vs 12.2 months, p = 0.04). Specifically, patients receiving platinum only regimens had a significantly improved PFS if they had no detectable BRCA1 protein compared to those with positive staining (14.5 months vs 10.8 months, p = 0.035).In contrast, patients positive for BRCA1 protein (grades 1–4) had a significant increase in both OS (43.7 months vs 20.4 months, p = 0.03) and PFS (12.7 months vs 10.9 months p = 0.02) if they received platinum/taxane combination over platinum alone. Patients with no detectable BRCA1 protein (grade 0) had no significant difference in OS or PFS whether they were treated with combination or platinum only chemotherapy. Conclusions: This study provides initial evidence that BRCA1 protein expression may be a predictive marker of chemotherapy response in sporadic EOC. Specifically, patients with positive BRCA1 staining demonstrate a better clinical outcome with platinum/taxane based chemotherapy over platinum alone whereas no such difference was seen in those with no BRCA1. Further prospective clinical studies are required to validate these findings. No significant financial relationships to disclose.

scholarly journals Nondestructive Assessment of Articular Cartilage Electromechanical Properties after Osteochondral Autologous and Allogeneic Transplantation in a Goat Model

Cartilage ◽  
2018 ◽  
Vol 11 (3) ◽  
pp. 348-357
Author(s):  
Tomas Mickevicius ◽  
Alius Pockevicius ◽  
Audrius Kucinskas ◽  
Rimtautas Gudas ◽  
Justinas Maciulaitis ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Platelet Rich Plasma ◽  
Allogeneic Transplantation ◽  
Staining Intensity ◽  
Quality Parameters ◽  
Electromechanical Properties ◽  
Diagnostic Device ◽  
Safranin O ◽  
Femoral Condyles

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.

In-vitro production and pre-validation of lyophilized canine platelet-rich plasma for therapeutic use

2021 ◽  
Vol 41 ◽  
Author(s):  
Natália P.P. Freitas ◽  
Maria Márcia M.S. Maior ◽  
Beatriz A.P. Silva ◽  
Marcus R.L. Bezerra ◽  
José F. Nunes ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Proliferation ◽  
Growth Factors ◽  
Platelet Rich Plasma ◽  
Three Dimensional ◽  
In Vitro Production ◽  
Mesh Structure ◽  
Prp Gel

ABSTRACT: Platelet-rich plasma (PRP) has been considered a promising therapeutic alternative, since platelets are rich in growth factors that are used in the Regenerative Medicine field. However, fresh PRP cannot be stored for long periods. This study aimed to develop a protocol for obtaining lyophilized canine PRP capable of maintaining viability after its reconstitution. For that purpose, canine PRP extraction and lyophilization protocols were initially tested. Subsequently, assays were carried out to quantify the growth factors VEGF and TGF-β, before and after the lyophilization process, gelation test and the three-dimensional gel structure analysis of the reconstituted lyophilized PRP by electron microscopy, as well as in vitro cell proliferation test in lyophilized PRP gel. Additionally, the immunogenicity test was performed, using allogeneic samples of lyophilized PRP. The results showed that the lyophilized PRP had adequate therapeutic concentrations of growth factors VEGF and TGF-β (9.1pg/mL and 6161.6pg/mL, respectively). The reconstituted PRP gel after lyophilization showed an in vitro durability of 10 days. Its electron microscopy structure was similar to that of fresh PRP. In the cell proliferation test, an intense division process was verified in mesenchymal stem cells (MSCs) through the three-dimensional mesh structure of the lyophilized PRP gel. The immunogenicity test showed no evidence of an immune reaction. The findings were promising, suggesting the possibility of having a lyophilized canine PRP that can be marketed. New in vivo and in vitro studies must be carried out for therapeutic confirmation.

scholarly journals The Differential Effects of Leukocyte-Containing and Pure Platelet-Rich Plasma on Nucleus Pulposus-Derived Mesenchymal Stem Cells: Implications for the Clinical Treatment of Intervertebral Disc Degeneration

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Jun Jia ◽  
Shan-zheng Wang ◽  
Liang-yu Ma ◽  
Jia-bin Yu ◽  
Yu-dong Guo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Platelet Rich Plasma ◽  
Enzyme Linked Immunosorbent Assay ◽  
Stem Cell Markers

Background. Platelet-rich plasma (PRP) is a promising strategy for intervertebral disc degeneration. However, the potential harmful effects of leukocytes in PRP on nucleus pulposus-derived mesenchymal stem cells (NPMSCs) have seldom been studied. This study aimed at comparatively evaluating effects of pure platelet-rich plasma (P-PRP) and leukocyte-containing platelet-rich plasma (L-PRP) on rabbit NPMSCs in vitro. Methods. NPMSCs isolated from rabbit NP tissues were treated with L-PRP or P-PRP in vitro, and then cell proliferation and expression of stem cell markers, proinflammatory cytokines (TNF-α, IL-1β), production of ECM (extracellular matrix-related protein), and NF-κB p65 protein were validated by CCK-8 assay, real-time polymerase chain reaction, enzyme-linked immunosorbent assay, immunofluorescence, and western blot respectively. Results. NPMSCs differentiate into nucleus pulposus-like cells after treatment of PRPs (P-PRP and L-PRP), and NPMSCs exhibited maximum proliferation at a 10% PRP dose. L-PRP had observably higher concentration of leukocytes, TNF-α, and IL-1β than P-PRP. Furthermore, compared to P-PRP, L-PRP induced the differentiated NPMSCs to upregulate the expression of TNF-α and IL-1β, enhanced activation of the NF-κB pathway, increased the expression of MMP-1 and MMP-13, and produced less ECM in differentiated NPMSCs. Conclusions. Both P-PRP and L-PRP can induce the proliferation and NP-differentiation of NPMSCs. Compared to L-PRP, P-PRP can avoid the activation of the NF-κB pathway, thus reducing the inflammatory and catabolic responses.

scholarly journals Salinity Depression in Vitro Impress the Growth Potential and, Mediates the Antioxidants Pool and the Tolerance Genes Expression of Grapevine

2020 ◽  
Author(s):  
Mohammad Aazami ◽  
Mohammad Hassanpouraghdam
Keyword(s):  
Growth Parameters ◽  
Growth Potential ◽  
Salinity Treatment ◽  
Fresh Weight ◽  
Genes Expression ◽  
Significant Difference ◽  
Mda Content ◽  
Expression Rate ◽  
Growth Development

Abstract Undoubtedly, salinity is the major environmental stress affecting the crops growth, development and yield. Grapevine is a dominant horticulture crop and mildly sensitive to the salinity. Salinity triggers variations at the cellular and molecular levels and hence induces some specific genes expression. The responses of two grapevine cultivars (Fakhri and Sultanin) were evaluated for its tolerance to the salinity stress in vitro. The results for the explants exposed to the salinity revealed that the viability, fresh weight of the regenerated explant and the proliferation rate were declined compared to the control ones. The activity of SOD enzyme and MDA content were increased with salinity. However, protein content declined. There was no significant difference in CAT and APX activities with the salinity treatment. With salinity adding up, the DREB/CBFs genes expression pattern was significantly increased in both cultivars. ‘Fakhri’ was more responsive in growth parameters and the activity of antioxidant enzymes and higher expression rate of DREB/CBFs under salinity compared to the variety ‘Sultanin’.

scholarly journals CYTOTOXIC EFFECT OF FREEZE DRIED BOVINE CARTILAGE POWDER AND PLATELET RICH PLASMA (PRP) TO MESENCHYMAL STEM CELL (MSCs)

2019 ◽  
Vol 6 (2) ◽  
pp. 63
Author(s):  
Dwikora Novembri Utomo ◽  
Anthoni Yusbida
Keyword(s):  
Stem Cells ◽  
Platelet Rich Plasma ◽  
Clinical Problem ◽  
Physical Structure ◽  
Control Group ◽  
Control Group Design ◽  
Freeze Dried ◽  
Significant Difference ◽  
Bovine Cartilage

Cartilage repair is a challenging clinical problem because the damage is an irreversible condition. Many studies had been performed using several kinds of natural or synthetic scaffold. Attempts to repair articular cartilage using scaffold usually found many problems, lacks the physical structure and mechanical properties necessary to ensure long-term efficacy to cartilage defect. Furthermore, scaffold frequently cause toxicity to the host. Therefore, this study was performed in vitro to test the toxicity effect of scaffold freeze dried bovine cartilage powder and platelets Rich Plasma (PRP). This research was conducted using pure experimental research design in 4 groups of animal stem cells which being added with scaffold freeze dried bovine cartilage scaffold provided with platelet rich plasma. This study using posttest only control group design. The result being processed with MTT assay and spectrophotometer for counting the viable stem cells. There was no significant difference in the amount of macrophage between control and the freeze dried bovine cartilage scaffold provided with PRP (p=0,128). With this result in the number of macrophages between the control with freeze dried bovine cartilage scaffold provided PRP, it can be concluded that these biomaterials have biocompatibility.

scholarly journals Platelet-rich plasma inhibits Adriamycin-induced inflammation via blocking the NF-κB pathway in articular chondrocytes

2021 ◽  
Vol 27 (1) ◽  
Author(s):  
Haijun Zhao ◽  
Weijie Zhu ◽  
Wude Mao ◽  
Chengkai Shen
Keyword(s):  
Articular Chondrocytes ◽  
Platelet Rich Plasma ◽  
Cartilage Destruction ◽  
Chondrocyte Apoptosis ◽  
Related Factors ◽  
Protein Levels ◽  
Cell Matrix ◽  
Safranin O

Abstract Background Previous studies showed that doxorubicin could lead to osteoarthritis (OA) by inducing chondrocyte inflammation and apoptosis. Besides, it is reported that platelet-rich plasma (PRP) could suppress the activation of inflammatory NF-κB signaling. Here, we aimed to determine whether PRP was able to exert a protective effect against doxorubicin-induced chondrocyte damages. Methods To determine whether PRP protects chondrocytes against destabilization of the medial meniscus (DMM)-induced osteoarthritis, mice were treated with PRP and doxorubicin, and the cartilage destruction was observed through Safranin O-fast green staining and osteoarthritis scoring. ELISA assay was used to check the release of TNF-α and ILs. In vitro, we treated chondrocytes with doxorubicin and PRP; CCK-8 was used to measure cell viability. Western blot, real-time PCR, and ELISA were applied to check apoptosis-related signaling and inflammation-associated factors. Results The results from the mouse model suggested that PRP attenuated doxorubicin-induced cartilage destruction in vivo. Doxorubicin promoted chondrocyte apoptosis while PRP ameliorated this damage. PRP inhibited doxorubicin-induced dysregulation of cell matrix-related factors, including SOX9, Col2A1, Col10A1, and Aggrecan, reduced protein levels of doxorubicin-induced inflammatory markers, COX-2, and iNOS, and blocked doxorubicin-induced phosphorylation of IκB and NF-κB in articular chondrocytes. Conclusions PRP improved doxorubicin-induced damage on chondrocytes. This research might provide a new theoretical basis for the clinical treatment of osteoarthritis caused by doxorubicin.

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