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1.
J Biomater Appl ; 28(7): 1039-50, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23828781

RESUMO

OBJECTIVE: The purpose of this study was to determine whether the platelet-rich plasma-agarose gel scaffold could be a bioactive scaffold capable of growth factors release for cartilage repair. METHODS: Porcine chondrocytes were seeded in agarose gel and platelet-rich plasma-agarose gel. During the 28-days culture, microstructure of hydrogels and morphologies of chondrocytes seeded in the hydrogels were observed using scanning electron microscope; viability of chondrocytes in gels was examined by live/dead assay; qualitative and quantitative analysis of glycosaminoglycan, collagen and DNA were assessed by histological, immunohistochemical staining and biochemical assay; gene expression was measured by real-time polymerase chain reaction. In vitro cartilage ring models were used to evaluate the integration of the scaffolds, and the integration strength was analyzed by mechanical push-out tests. RESULTS: Scanning electron microscope revealed both scaffolds had highly uniform porous structure. Live/dead scaffolds showed 100% cells alive in both groups. After 28-days culture, glycosaminoglycan, collagen, DNA content and chondrocyte-related genes expression in platelet-rich plasma-agarose gel were significantly higher than pure agarose gel. Integration strength in platelet-rich plasma-agarose gel was also higher compared to pure agarose gel. CONCLUSION: Platelet-rich plasma showed a positive effect on chondrocytes proliferation, differentiation and integration between native cartilage and engineered tissue when combined with agarose gel. Our findings suggest that platelet-rich plasma-agarose gel scaffold is a promising bioactive scaffold for future cartilage tissue engineering and future clinical works.


Assuntos
Cartilagem Articular/citologia , Plasma Rico em Plaquetas/metabolismo , Sefarose/metabolismo , Alicerces Teciduais , Animais , Sequência de Bases , Cartilagem Articular/metabolismo , Colágeno/metabolismo , DNA/metabolismo , Primers do DNA , Glicosaminoglicanos/metabolismo , Microscopia Eletrônica de Varredura , Porosidade , Reação em Cadeia da Polimerase em Tempo Real , Suínos , Porco Miniatura
2.
Knee Surg Sports Traumatol Arthrosc ; 21(8): 1770-6, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22572868

RESUMO

PURPOSE: The aim of this study was to evaluate whether the microfracture combined with osteochondral paste implantation could promote the quality of the regenerated tissue in the knee joints of rabbits. METHODS: Sixty-six New Zealand white rabbits were used. Bilateral knee joints from the same rabbit were randomly divided into experimental group and microfracture group. An articular cartilage defect was established in the femoral trochlear groove. In the experimental group, the defect was microfractured and covered with osteochondral paste harvested from the intercondylar notch. The regenerated tissues were harvested for gross morphology, histology, biochemistry and gene expression analysis at 4, 8 and 12 weeks postoperatively. RESULTS: The regenerated tissue had a slowly mature process in both groups. At 12 weeks, the regenerated tissue in the experimental group appeared much more thicker and white with higher percentages of defect filling macroscopically. In histology, the experimental group found a majority of hyaline-like regenerate tissue with intense Safranin-O and collagen type II staining, while fibrocartilage-like tissue was mostly seen in the microfracture group with poor Safranin-O and collagen type II staining. The experimental group had higher Wakitani scores and narrower acellular zones than those in the microfracture group (P < 0.05). For biochemical analysis, both the GAG content and the DNA-normalized GAG content saw a time-dependent increase with a much higher value found in the experimental group at 8 and 12 weeks (P < 0.05). On the contrary, the total DNA content decreased with time in both groups, and the difference between the two groups was only found at 4 and 8 weeks (P < 0.05). For gene expression analysis, the experimental group had much higher expression levels than the microfracture group as for collagen type II and aggrecan, but not for collagen type I. CONCLUSION: Microfracture combined with paste implantation can result in improved quality of the reparative tissue and may have a positive effect on the integration to the surrounding cartilage in the rabbit model. The technique offers a promising treatment option for cartilage defects and improves the regeneration of articular cartilage for patients with painful chondral lesions.


Assuntos
Artroplastia Subcondral , Transplante Ósseo , Cartilagem Articular/lesões , Cartilagem Articular/cirurgia , Cartilagem/transplante , Agrecanas/genética , Agrecanas/metabolismo , Animais , Cartilagem Articular/fisiologia , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , DNA/metabolismo , Glicosaminoglicanos/metabolismo , Fenazinas/metabolismo , Coelhos , Distribuição Aleatória , Regeneração
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