Platelet-Rich Plasma (PRP) Stimulates the Extracellular Matrix Metabolism of Porcine Nucleus Pulposus and Anulus Fibrosus Cells Cultured in Alginate Beads

Akeda, Koji; An, Howard S.; Pichika, Rajeswari; Attawia, Mohamed; Thonar, Eugene J.‐M.A.; Lenz, Mary Ellen; Uchida, Atsushi; Masuda, Koichi

doi:10.1097/01.brs.0000214942.78119.24

Cited by 123 publications

(91 citation statements)

References 48 publications

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“…Alginate forms a gel consisting of negatively-charged and cross-linked 3D lattices in the presence of a divalent cation solution, such as calcium chloride (20). Because of its high biocompatibility and the low impact of the gelatin procedure on cell viability, alginate has been used as a scaffold to maintain the phenotype of articular chondrocytes (9,(21)(22)(23), as well as intervertebral disc cells (7,8,24), during 3D culture and as a dressing to promote wound healing (25,26). Alginate-encapsulated vector-producing cell lines have also been used for cancer gene therapy in vivo (27).…”

Section: Discussionmentioning

confidence: 99%

Three-dimensional alginate spheroid culture system of murine osteosarcoma

Akeda

Nishimura²,

Satonaka³

et al. 2009

Oncol Rep

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Abstract. Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.

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Section: Discussionmentioning

confidence: 99%

Three-dimensional alginate spheroid culture system of murine osteosarcoma

Akeda

Nishimura²,

Satonaka³

et al. 2009

Oncol Rep

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“…For example, proliferation of IVD cells has been shown in response to BMP-2 [42], BMP-7 [48], GDF-5 [7] and TGF-b1 [13], all of them belonging to the TGF-bsuperfamily. Furthermore, regarding the effects of classical potent mitogens such as PDGF, bFGF and IGF-I [50] on the proliferation of IVD cells in vitro, in some cases the data are indirect [1,8] or contradictory (e.g., regarding the effects of IGF-I on annulus fibrosus cells [14,41]), possibly due to the variety of the assay systems used. Hence, we aimed at a systematic study of these three mitogens under the same experimental conditions.…”

Section: Introductionmentioning

confidence: 99%

PDGF, bFGF and IGF-I stimulate the proliferation of intervertebral disc cells in vitro via the activation of the ERK and Akt signaling pathways

2007

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Intervertebral disc (IVD) degeneration is frequently characterized by increased cell proliferation, probably as a tissue regenerative response. Although many growth factors and their receptors have been shown to be expressed normally in the disc, and generally to be overexpressed during degeneration, not all of them have been thoroughly studied concerning their effects on IVD cell proliferation. In the present report, three potent mitogens, platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I) are examined regarding their capacity to induce proliferation in vitro of bovine coccygeal nucleus pulposus (NP) and annulus fibrosus (AF) cells, as well as to activate major intracellular signal transduction pathways. PDGF, bFGF and IGF-I were found to induce DNA synthesis in quiescent IVD cells in a dose-dependent manner. Maximum stimulation was induced by PDGF, while stimulation by all three factors simultaneously exceeded only slightly that caused by PDGF alone. All three growth factors were shown to phosphorylate immediately extracellular-signal regulated kinases (ERKs), while the stimulation by bFGF especially resulted in sustained ERK phosphorylation. Furthermore, all three growth factors induced phosphorylation of Akt in both Thr308 and Ser473 residues immediately after stimulation, although bFGF-induced phosphorylation was much weaker than that provoked by PDGF and IGF-I. In addition, the MEK inhibitor PD98059 and the PI 3-K inhibitor wortmannin were shown to block growth factor-induced ERK-and Akt-phosphorylation, respectively, in IVD cells. Inhibition of the MEK/ERK or the PI 3-K/Akt pathways provoked a significant decline of the proliferative effects of PDGF, bFGF or IGF-I on IVD cell cultures, while the simultaneous inhibition of both signaling pathways abolished completely the mitogenicity of these growth factors. The above effects of the three growth factors were reproduced similarly in both NP and AF cell cultures. Overall, the above results indicate that PDGF, bFGF and IGF-I stimulate the proliferation of IVD cells via the ERK and Akt signaling pathways.

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“…[16][17][18] During the centrifugation process used to make PRP, mononuclear white blood cells are also concentrated compared to whole blood; although to a lesser degree than platelets. 22 This may pose a concern in tissue regenerative efforts due to the pro-inflammatory mediators such as neutral proteases and acid hydrolases contained in white blood cells as well as thromboxane released from platelets.…”

Section: Introductionmentioning

confidence: 99%

“…[1][2][3][4][5][6][7] Platelet rich plasma (PRP) has been used for several years in oral and maxillofacial surgery to accelerate peri-implant soft tissue and bone healing, [8][9][10][11][12] and it has recently been investigated for regeneration of bone, [13][14][15][16] cartilage, 16,17 and ligament. 18 The main rationale for the use of PRP arises from the growth factors released from platelet a-granules, including platelet-derived growth factor (PDGF), transforming growth factor-b (TGF-b), fibroblastic growth factor (FGF), vascular endothelial growth factor (VEGF), insulin-like growth factor-I (IGF-I), and epidermal growth factor (EGF).…”

Section: Introductionmentioning

confidence: 99%

Platelet rich plasma (PRP) enhances anabolic gene expression patterns in flexor digitorum superficialis tendons

Schnabel

Mohammed

Miller

et al. 2006

Journal Orthopaedic Research

349

267

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Platelet rich plasma (PRP) has recently been investigated for use in tissue regeneration studies that seek to utilize the numerous growth factors released from platelet a-granules. This study examined gene expression patterns, DNA, and collagen content of equine flexor digitorum superficialis tendon (SDFT) explants cultured in media consisting of PRP and other blood products. Blood and bone marrow aspirate (BMA) were collected from horses and processed to obtain plasma, PRP, and platelet poor plasma (PPP). IGF-I, TGF-b1, and PDGF-BB were quantified in all blood products using ELISA. Tendons were cultured in explant fashion with blood, plasma, PRP, PPP, or BMA at concentrations of 100%, 50%, or 10% in serum-free DMEM with amino acids. Quantitative RT-PCR for expression of collagen type I (COL1A1), collagen type III (COL3A1), cartilage oligomeric matrix protein (COMP), decorin, matrix metalloproteinase-3 (MMP-3), and matrix metalloproteinase-13 (MMP-13) was performed as were DNA and total soluble collagen assays. TGF-b1 and PDGF-BB concentrations were higher in PRP compared to all other blood products tested. Tendons cultured in 100% PRP showed enhanced gene expression of the matrix molecules COL1A1, COL3A1, and COMP with no concomitant increase in the catabolic molecules MMP-3 and MMP-13. These findings support in vivo investigation of PRP as an autogenous, patient-side treatment for tendonitis. ß

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Platelet-Rich Plasma (PRP) Stimulates the Extracellular Matrix Metabolism of Porcine Nucleus Pulposus and Anulus Fibrosus Cells Cultured in Alginate Beads

Cited by 123 publications

References 48 publications

Three-dimensional alginate spheroid culture system of murine osteosarcoma

Three-dimensional alginate spheroid culture system of murine osteosarcoma

PDGF, bFGF and IGF-I stimulate the proliferation of intervertebral disc cells in vitro via the activation of the ERK and Akt signaling pathways

Platelet rich plasma (PRP) enhances anabolic gene expression patterns in flexor digitorum superficialis tendons

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