Effect Of Growth Differentiation Factor-5 Research Articles

Growth differentiation factor 5 inhibits lipopolysaccharide-mediated pyroptosis of nucleus pulposus mesenchymal stem cells via RhoA signaling pathway.

Degenerative disc disease(DDD)is one of the most important causes of low back pain (LBP). Programmed death of human nucleus pulposus mesenchymal stem cells (NPMSCs) plays an important role in the progression of DDD. Growth differentiation factor-5 (GDF-5) is a protein that promotes chondrogenic differentiation, and has been reported to slow the expression of inflammatory factors in nucleus pulposus cells. Compared with those in normal rats, MRI T2-weighted images show hypointense in the central nucleus pulposus region of the intervertebral disc in GDF-5 knockout rats. We aimed to evaluate the role of GDF-5 and Ras homolog family member A (RhoA) in NPMSCs. We used lipopolysaccharide (LPS) to simulate the inflammatory environment in degenerative disc disease, and performed related experiments on the effects of GDF-5 on NPMSCs, including the effects of pyroptosis, RhoA protein, and the expression of extracellular matrix components, and the effects of GDF-5, on NPMSCs. In addition, the effect of GDF-5 on chondroid differentiation of NPMSCs was included. The results showed that the addition of GDF-5 inhibited the LPS-induced pyroptosis of NPMSCs, and further analysis of its mechanism showed that this was achieved by activating the RhoA signaling pathway. These findings suggest that GDF-5 plays an important role in inhibiting the pyroptosis of NPMSCs and GDF-5 may have potential for degenerative disc disease gene-targeted therapy in the future.

Supplementation of growth differentiation factor-5 increases proliferation and size of chondrogenic pellets of human umbilical cord-derived perivascular stem cells

In recent years, perivascular stem cells (PVCs) have gained increasing attention as a promising source for regenerative medicine due to their greater differentiation potential compared to multipotent mesenchymal stem cells (MSCs). It has been reported that growth differentiation factor-5 (GDF-5) is involved in regulating proliferation and chondrogenic differentiation of MSCs. In this study, we investigated the effect of GDF-5 on the proliferation and chondrogenic differentiation of PVCs isolated from human umbilical cords. The supplementation of PVC cultures with GDF-5 (100 ng/mL) significantly enhanced their proliferative rate and augmented the size of pellets in micromass pellet cultures for chondrogenic induction. Although similar expression levels of chondrogenic-related genes were observed in chondrogenic pellets treated with GDF-5 compared to the pellets without GDF-5 treatment, these results indicate that supplementation of GDF-5 is able to acquire more chondrocytes when starting with equal amount of PVCs. Our study suggests that GDF-5 is an effective agent for the enhancement of PVC proliferation, thereby achieving a higher number of chondrocytes that are applicable in therapeutic doses for cartilage regeneration.

The Effect of Growth Differentiation Factor-5, 6, 7 in Chondrogenic Cell Differentiation of ATDC-5

The proteins known as growth differentiation factors (GDFs) are members of the BMP family; GDF-5, -6, and -7 play important roles in skeletogenesis, especially with regard to chondrogenesis. The functional differences among these GDFs in chondrogenesis and chondrocyte cell differentiation remains unclear. Here, we attempt to assay cell proliferation, the production of chondrogenic matrices indicated by alcian blue intensity, and the profile of a chondrogenic cell differentiation marker gene in a cell culture of the chondrogenic cell line ATDC5 with or without recombinant mouse GDF-5, -6, and -7. After 24h culture, the ATDC5 cell number was significantly decreased in the exogenous presence of each GDF compared with a control culture absent all GDFs. On the other hand, alcian blue staining of cell cultures after 2w culture showed significantly increased intensity compared with that of the control. The expression levels of the chondrogenic cell differentiation marker genes Sox9 and aggrecan were increased after 24h and 48h culture by all GDFs, but were significantly increased in the presence of GDF-5 compared with the presence of GDF-6 or -7. These findings suggest that GDF-5, -6, and -7 could all promote chondrogenic cell differentiation of ATDC5, but GDF-5 may induce chondrogenic genes more potently than GDF-6 and -7 do.

The Distinct Roles of Growth/Differentiation Factor-5 in Cell Proliferation and Odontoblast Differentiation from Dental Pulp Cells

Growth/differentiation factor-5 (GDF-5) belongs to the bone morphogenetic protein (BMP) family, which is expressed in dental pulp tissues. To date, various studies have reported that BMP-family growth factors can induce cells isolated from dental pulp tissues to differentiate into odontoblasts. Nevertheless, the effect of GDF-5 on odontoblast differentiation from dental pulp tissues remains unknown. In this study, we assayed the effect of exogenous mouse recombinant GDF-5 on cell proliferation of dental pulp cells and determined the expression levels of odontoblast differentiation marker genes in the cells of isolated mouse dental pulp tissues in the presence of GDF-5. There were no significant differences in dental pulp cell proliferation patterns related to the absence or presence of GDF-5 after seven days of culture. Odontoblast marker gene levels, on the other hand, were significantly elevated after seven days of culture in the presence of GDF-5. GDF-5 also increased the gene expression and enzyme activity of alkaline phosphatase. Furthermore, dental pulp cells cultured in the presence of GDF-5 formed alizarin redpositive nodules after 10 days of culture. These results indicate that, although GDF-5 may have no effect on cell proliferation in isolated dental pulp tissue, it could promote odontoblast differentiation.

The effects of growth and differentiation factor 5 on bone marrow stromal cell transplants in an in vitro tendon healing model

The effects of growth differentiation factor-5 (GDF-5) and bone marrow stromal cells (BMSCs) on tendon healing were investigated under invitro tissue culture conditions. BMSCs and GDF-5 placed in a collagen gel were interpositioned between the cut ends of dog flexor digitorum profundus tendons. The tendons were randomly assigned into four groups: 1) repaired tendon without gel; 2) repaired tendon with BMSC-seeded gel; 3) repaired tendon with GDF-5 gel without cells; and 4) repaired tendon with GDF-5 treated BMSC-seeded gel. At 2 and 4 weeks, the maximal strength of repaired tendons with GDF-5 treated BMSCs-seeded gel was significantly higher than in tendons without gel interposition. However, neither BMSCs nor GDF-5 alone significantly increased the maximal strength of healing tendons at 2 or 4 weeks. These results suggest that the combination of BMSCs and GDF-5 accelerates tendon healing, but either BMSCs or GDF-5 alone are not effective in this model.

Differential effects of growth differentiation factor-5 on porcine dental papilla- and follicle-derived cells

In this study, the effect of growth differentiation factor-5 (GDF-5) on the growth and differentiation of porcine dental papilla- and follicle-derived cells was investigated. Furthermore, the effect was compared with that of BMP-2. Recombinant mouse GDF-5 (rmGDF-5) enhanced alkaline phosphatase (ALP) activity in dental papilla-derived cells in a dose-dependent manner, while ALP activity in dental follicle-derived cells was reduced. In rmGDF-5 stimulated dental papilla-derived cells, the expressions of odontoblast-marker genes were up-regulated. Conversely, recombinant human BMP-2 (rhBMP-2) enhanced ALP activity dose-dependently in both dental papilla- and follicle-derived cells. When combined, GDF-5 did not further enhance BMP-2-induced ALP activities. Rather, GDF-5 reduced BMP-2-induced ALP activities in both dental papilla- and follicle-derived cells. This suggests that affinity of GDF-5 to the shared receptors may be higher than that of BMP-2 in both cell types. These observations indicate that GDF-5 regulates differentiation of both dental papilla and follicle during odontogenesis, co-operatively with other growth factors such as BMP-2.

GDF-5 is suppressed by IL-1β and enhances TGF-β3-mediated chondrogenic differentiation in human rheumatoid fibroblast-like synoviocytes

Objective To investigate the expression of growth differentiation factor-5 (GDF-5) in chondrocytes (HC) and fibroblast-like synoviocytes (FLS) from humans with rheumatoid arthritis (RA) when stimulated with proinflammatory cytokines and to explore whether GDF-5 plays a role in regulating the differentiation of FLS-RA into chondrocytes. Methods Expression of GDF-5 in synovium and cartilage in RA and osteoarthritis (OA) was assessed by immunohistochemistry. GDF-5 production in FLS-RA and HC-RA was examined through real-time quantitative RT-PCR (Q-PCR) and western blotting. Expressions of GDF-associated receptors on FLS-RA were determined by semiquantitative-PCR, and MTT assay was used to study the effects on FLS-RA proliferation. Effect of GDF-5 and TGF-β3 on in vitro chondrogenic ability of FLS-RA was investigated using pellet-culture system, Q-PCR and histological analysis. Results Immunohistochemical analysis demonstrated that GDF-5 expression in the synovium and cartilage from joints of RA patients was much lower than that of OA patients. Addition of IL-1β or TNF-α appeared to downregulate the expression of GDF-5 in HC-RA and FLS-RA. Inhibition of GDF-5 expression by IL-1β in RA-FLS was attenuated by pretreatment with MEK1/2 inhibitor. GDF-5-associated receptors were expressed in FLS-RA, but GDF-5 had no effect on FLS-RA proliferation. GDF-5 had a strong chondrogenic-promoting effect on TGF-β3-induced chondrocyte differentiation in FLS-RA. Conclusions GDF-5 is expressed in FLS-RA and HC-RA, and its expression is strongly downregulated by proinflammatory cytokines. MEK-ERK pathway is a negative regulator of GDF-5 expression in FLS-RA. In FLS-RA, synergy between GDF-5 and TGF-β3 enhances chondrogenesis. Anti-inflammatory drugs combined with GDF-5 might be a new therapeutic treatment for RA.

Intracavernous Growth Differentiation Factor‐5 Therapy Enhances the Recovery of Erectile Function in a Rat Model of Cavernous Nerve Injury

Neurogenic erectile dysfunction remains a serious complication in the postprostatectomy population. Effective protective and regenerative neuromodulatory strategies are needed. To determine the effect of growth differentiation factor-5 (GDF-5) on erectile function and its mechanism in a rat model of cavernous nerve (CN) injury. Erectile function was assessed by CN electrostimulation at 4 weeks. Penile tissues were examined by real-time polymerase chain reaction (PCR) and immunohistochemical analyses. Forty-eight male Sprague-Dawley rats were randomly divided into six equal groups: one group underwent sham operation (uninjured controls), while five groups underwent bilateral CN crush. Crush-injury groups were treated at the time of injury with intracavernous injection of a slow-release suspension of liquid microparticles containing no GDF-5 (vehicle), 0.4 microg (low concentration), 2 microg (intermediate concentration), or 10 microg GDF-5 (high concentration). One untreated group served as injured controls. GDF-5 enhanced erectile recovery and significantly increased intracavernous pressure in the low and intermediate-concentration groups vs. injured controls. Low-concentration GDF-5 demonstrated the best functional preservation, as the intracavernous pressure increase in this group did not differ significantly from uninjured controls. A dose-response relationship was confirmed for the effects of GDF-5 in penile tissue. Low-concentration GDF-5 showed better preservation of the penile dorsal nerves and antiapoptotic effects in the corpus cavernosum (P < 0.05 vs. injured controls). Although high concentration GDF-5 did not confer meaningful erectile recovery, this dose was more effective at decreasing transforming growth factor-beta than low-concentration GDF-5. Intracavernous injection of low (0.4 microg) or intermediate-concentration GDF-5 (2 microg) was effective in preserving erectile function in a rat model of neurogenic erectile dysfunction. The underlying mechanism appears to involve neuron preservation and antiapoptosis.

Differential effects of GDF5 on the medial and lateral rat ventral mesencephalon

Growth/differentiation factor 5 (GDF5) is a member of the transforming growth factor-beta (TGF-beta) superfamily that has potent neurotrophic and protective effects on dopaminergic neurones and is expressed in the developing rat substantia nigra (the ventral mesencephalon; VM). GDF5 has the potential to be used in the treatment of Parkinson's disease (PD), a neurodegenerative disorder characterised by the selective degeneration of nigrostriatal dopaminergic neurones. One therapy being explored for PD involves transplantation of fetal VM tissue into the striatum in order to replace lost dopaminergic neurones. The majority of transplantation studies have used transplants incorporating the whole VM. The principal location of dopaminergic neurones in the E14 rat VM is in the medial VM. In the present study, the effects of GDF5 on cultures prepared from medial, lateral and whole E14 rat VM tissue were compared. GDF5 treatment increased the number of dopaminergic neurones in whole and lateral, but not in medial, VM cultures, whereas it increased total cell number in medial, but not in whole or lateral, VM cultures. RT-PCR studies showed that the receptors for GDF5 were differentially expressed in E14 VM; the expression of BMPR-IB and Ror2 was low in medial but high in lateral VM tissue. This study suggests that GDF5 increases the number of dopaminergic neurones in whole VM cultures by acting on BMPR-IB and Ror2-expressing cells in the lateral VM.

Modulation of VEGF Expression in Rat Bone Marrow Stromal Cells by GDF-5

Angiogenesis is essential for bone formation and several bone morphogenetic proteins (BMPs) have been shown to induce angiogenesis through osteoblast-derived vascular endothelial growth factor (VEGF)-A. Growth differentiation factor-5 (GDF-5) is a member of the BMP family expressed in bone and known to induce angiogenesis in vivo. In this study, the effects of GDF-5 on osteogenic differentiation and expression of VEGF-related genes were determined using rat bone marrow stromal cells. GDF-5 stimulated osteogenic differentiation. It also upregulated the expression of VEGF-A after 3 hr, accompanied by a trend of decrease in its receptor VEGFR-2 at 6 and 24 hr. VEGF-D and its receptor VEGFR-3 showed peak expression at later time points. This regulation may be further controlled by neuropilin 2 that exhibited a parallel profile to VEGF-D. These observations indicate that GDF-5 stimulates osteogenic differentiation and has a potential to induce angiogenesis through osteoblast-derived VEGF-A in bone.