Recombinant Platelet-derived Growth factor-BB Research Articles

Macrophage-derived PDGF-BB modulates glycolytic enzymes expression and pyroptosis in nucleus pulposus cells via PDGFR-β/TXNIP pathway

ObjectiveIntervertebral Disc Degeneration (IVDD) is one of the leading causes of low back pain, significantly impacting both individuals and society. This study aimed to investigate the significance of macrophage infiltration and the role of macrophage-secreted platelet-derived growth factor-BB (PDGF-BB) in IVDD progression. MethodsTo confirm the protective function of macrophage-derived PDGF-BB on nucleus pulposus cells (NPCs), we employed Lysm-Cre transgenic mice to genetically ablate PDGF-B within the myeloid cells. Immunohistochemistry was utilized to detect the expression of glycolytic enzymes and pyroptosis-related proteins during the process of IVDD. Western blot, RT-PCR, ELISA and immunofluorescence were used to detect the protective effect of recombinant PDGF-BB on NPCs. ResultsMacrophage-derived PDGF-BB deficiency resulted in the loss of NPCs and the increased ossification of cartilage endplates during lumbar disc degeneration. Also, PDGF-BB deficiency triggered the inhibition of glycolytic enzymes’ expression and the activation of pathways related to pyroptosis in nucleus pulposus. Mechanistically, our results suggest that PDGF-BB predominantly conveys its protective influence on NPCs through the PDGF receptor- beta (PDGFR-β)/ thioredoxin-interacting protein (TXNIP) pathway. ConclusionsThe absence of PDGF-BB originating from macrophages expedites the advancement of IVDD, whereas the application of PDGF-BB treatment holds potential for retarding intervertebral disc degeneration in the human body.

PDGF-BB prevents destructive repair and promotes reparative osteogenesis of steroid-associated osteonecrosis of the femoral head in rabbits.

Destructive repair characterized by inadequate angiogenesis and osteogenesis is the main pathological progression in steroid-associated osteonecrosis of the femoral head (SONFH). Platelet-derived growth factor-BB (PDGF-BB) is an "angiogenesis and osteogenesis coupling" factor that has been used for the treatment of bone defects in clinic. This study was designed to analyze the ability of PDGF-BB for preventing destructive repair and promoting reparative osteogenesis in SONFH. Steroid-associated osteonecrosis (SAON) was induced and triggered destructive repair of the femoral head by repeated lipopolysaccharide (LPS) and methylprednisolone (MPS) injections in rabbits. At 2, 4, and 6 weeks after induction, recombinant human PDGF-BB, neutralizing PDGF-BB antibody, or saline was intramedullary injected into the proximal femora. At week 6 after SAON induction, the proximal femora were dissected for bone architecture and histological analysis. C3H10T1/2 cells and HUVECs were used for further mechanistic investigation. After PDGF-BB treatment, type H vessels and leptin receptor-positive (LepR+) mesenchymal stem cells (MSCs) increased in the affected femoral head, and more osteoblastic osteogenesis along the bone surfaces but scattered adipocytes in bone marrow tissue than that in the SAON group. PDGF-BB treatment prevented destructive repair progression and led to 50-70 % of osteonecrotic femoral heads undergoing reparative osteogenesis. In particular, we found that PDGF-BB could mediate MSC self-renewal and maintain their osteogenic potency by activating PDGFR/Akt/GSK3β/CERB signaling in the presence of steroids. Moreover, PDGF-BB also stabled the newly formed vascular tubes by recruiting MSCs for improving intraosseous vascular integration. PDGF-BB may be a candidate for the promotion of reparative osteogenesis in SONFH.

In situ regeneration of nasal septal defects using acellular cartilage enhanced with platelet-derived growth factor

Nasal septum defects can currently only be reconstructed using autologous cartilage grafts. In this study, we examine the reconstruction of septal cartilage defects in a rabbit model using porcine decellularized nasal septal cartilage (DNSC) functionalized with recombinant platelet-derived growth factor-BB (PDFG-BB). The supportive function of the transplanted DNSC was estimated by the degree of septum deviation and shrinkage using magnetic resonance imaging (MRI). The biocompatibility of the transplanted scaffolds was evaluated by histology according to international standards. A study group with an autologous septal transplant was used as a reference. In situ regeneration of cartilage defects was assessed by histological evaluation 4 and 16 weeks following DNSC transplantation. A study group with non-functionalized DNSC was introduced for estimation of the effects of PDFG-BB functionalization. DNSC scaffolds provided sufficient structural support to the nasal septum, with no significant shrinkage or septal deviations as evaluated by the MRI. Biocompatibility analysis after 4 weeks revealed an increased inflammatory reaction of the surrounding tissue in response to DNSC as compared to the autologous transplants. The inflammatory reaction was, however, significantly attenuated after 16 weeks in the PDGF-BB group whereas only a slight improvement of the biocompatibility score was observed in the untreated group. In situ regeneration of septal cartilage, as evidenced by the degradation of the DNSC matrix and production of neocartilage, was observed in both experimental groups after 16 weeks but was more pronounced in the PDFG-BB group. Overall, DNSC provided structural support to the nasal septum and stimulated in situ regeneration of the cartilage tissue. Furthermore, PDFG-BB augmented the regenerative potential of DNSC and enhanced the healing process, as demonstrated by reduced inflammation after 16 weeks.

Glucocorticoids Disrupt Skeletal Angiogenesis Through Transrepression of NF-κB-Mediated Preosteoclast Pdgfb Transcription in Young Mice.

In the growing skeleton, angiogenesis is intimately coupled with osteogenesis. Chronic, high doses of glucocorticoids (GCs) are associated with decreased bone vasculature and induce osteoporosis and growth failure. The mechanism of GC-suppression of angiogenesis and relationship to osteoporosis and growth retardation remains largely unknown. Type H vessels, which are regulated by preosteoclast (POC) platelet-derived growth factor-BB (PDGF-BB), are specifically coupled with bone formation and development. We determined the effect of GCs on POC synthesis of PDGF-BB in relation to type H vessel formation, bone mass, and bone growth in the distal femur of 2-week-old young mice receiving prednisolone or vehicle for 2, 4, or 6 weeks. After 2 weeks of prednisolone, the number of POCs were unchanged while POC synthesis of PDGF-BB was reduced. Longer treatment with prednisolone reduced POCs numbers and PDGF-BB. These changes were associated with a reduction in type H vessels, bone formation rate, bone mass, and bone length at each time point. In vitro, excessive concentrations of prednisolone (10-6 M) resulted in decreased PDGF-BB concentration and POC numbers. Conditioned medium from POC cultures treated with control concentration of prednisolone (10-7 M) or recombinant PDGF-BB stimulated endothelial tube formation, whereas conditioned medium from control concentration of prednisolone-treated POC cultures neutralized by PDGF-BB antibody or excessive prednisolone inhibited endothelial tube formation. Administration of excessive prednisolone attenuated the P65 subunit of nuclear factor kappa B (NF-κB) binding to the Pdgfb promoter, resulting in lower Pdgfb transcription. Co-treatment with excessive prednisolone and the glucocorticoid receptor (GR) antagonist (RU486), GR siRNA, or TNFα rescued NF-κB binding to the Pdgfb promoter and endothelial tube formation. These results indicate that PDGF-BB synthesis in POCs is suppressed by GCs through transrepression of GR/NF-κB, thus inhibiting type H vessel formation and associated osteoporosis and growth failure. © 2020 American Society for Bone and Mineral Research.

Abstract WMP31: Dedifferentiation of Smooth Muscle Cells and its Potential Contribution to Pathogenesis of Intracranial Aneurysms

Introduction: Intracranial aneurysm (IA) affects 1 to 5 % in general public and becomes the primary cause of subarachnoid hemorrhage, the most severe form of stroke. However, currently, no drug therapy is available for IAs to prevent progression and rupture of lesions. Elucidation of mechanisms underlying the disease is thus mandatory. Considering the important role of vascular smooth muscle cells (SMCs) in the maintenance of stiffness of arterial walls and also in the pathogenesis of atherosclerosis via mediating inflammatory responses, we in the present study analyzed morphological or phenotypical changes of SMCs during the disease development in the lesions. Methods: We subjected rats to an IA model in which lesions are induced by increase of hemodynamic force loading on intracranial arterial bifurcations and performed histopathological analyses of induced lesions including the electron microscopic examination. We then immunostained specimens from induced lesions to explore factors responsible for dedifferentiation or migration of SMCs. In vitro study was also done to examine effect of some candidate factors on dedifferentiation or migration of cultured SMCs. Results: We first found the accumulation of SMCs underneath the endothelial cell layer mainly at the neck portion of the lesion. These cells was positive for the embryonic form of myosin heavy chain, a marker for the dedifferentiated SMCs, and the expression of pro-inflammatory factors like TNF-α. In immunostaining to explore the potential factor regulating the dedifferentiation of SMCs, we found that Platelet-derived growth factor-BB (PDGF-BB) was expressed in endothelial cells at the neck portion of IA walls. Consistently, recombinant PDGF-BB could promote the dedifferentiate of SMCs and chemo-attracted them in in vitro. Finally, in the stenosis model of the carotid artery, PDGF-BB expression was induced in endothelial cells in which high wall shear stress was loaded and the dedifferentiation of SMCs occurred there. Conclusions: The findings from the present study imply the role of dedifferentiated SMCs partially recruited by PDGF-BB from endothelial cells in the formation of inflammatory microenvironment at the neck portion of IA walls, leading to the progression of the disease.

PDGF-BB exhibited therapeutic effects on rat model of bisphosphonate-related osteonecrosis of the jaw by enhancing angiogenesis and osteogenesis.

The mechanism and effective treatment of bisphosphonate-related osteonecrosis of the jaw (BRONJ) are still uncertain. Our previous study revealed that zoledronate (ZOL) preferentially inhibited osteoclasts formation and platelet-derived growth factor-BB (PDGF-BB) secretion, causing suppression of angiogenesis and osteogenesis in vitro. The present study aimed to elucidate whether PDGF-BB had therapeutic effects on rat model of BRONJ by enhancing angiogenesis and angiogenesis. Firstly, rat model of BRONJ was established by ZOL and dexamethasone administration, followed by teeth extraction. The occurrence of BRONJ was confirmed and detected dead bone formation by maxillae examination, micro-CT scan and HE staining (10/10). Compared to control rats (0/10), both angiogenesis and mature bone formation were suppressed in BRONJ-like rats, evidenced by enzyme-linked immunosorbent assay (ELISA) for VEGF (P < 0.01), immunohistochemistry of CD31 (P < 0.05) and OCN (P < 0.01). Moreover, in the early stage of bone healing, the number of preosteoclasts (P < 0.001) and PDGF-BB secretion (P < 0.05) were significantly decreased in bisphosphonates-treated rats, along with the declined numbers of microvessels (P < 0.05) and osteoblasts (P < 0.05). In vitro study, CCK8 assay, alizarin red S staining and western blot assay showed that mandible-derived bone marrow mesenchymal stem cells (BMMSCs) in BRONJ-like rats presented suppressed functions of proliferation, osteogenesis and angiogenesis. Interestingly, recombinant PDGF-BB was able to rescue the impaired functions of BMMSCs derived from BRONJ-like rats at more than 10 ng/ml. Then fibrin sealant with or without recombinant PDGF-BB were tamped into the socket after debridement in BRONJ rats. After 8 weeks, fibrin sealant containing PDGF-BB showed significant therapeutic effects on BRONJ-like rats (bone healing: 8/10 vs 3/10, P < 0.05) with enhancing microvessels and mature bone formation. Our study suggested that the inhibition of angiogenesis and osteogenesis, the potential mechanisms of BRONJ, might partly result from suppression of PDGF-BB secretion in the early stage of bone healing. PDGF-BB local treatment after debridement might avail the healing of BRONJ by increasing angiogenesis and osteogenesis.

Recombinant platelet derived growth factor-BB and hyaluronic acid effect in rat osteoarthritis models

Osteoarthritis (OA) arises from imbalance of cartilage metabolism between the synthesis and degradation of type II collagen by the chondrocyte. Collagen type II degradation is characterized by increase in the biomarker of C-telopeptide fragment of type II collagen (CTX-II), while the anabolic process of cartilage is characterized by an increase in the biosynthesis of procollagen amino terminal N-propeptide type IIA (PIIANP). Platelet derived growth factor (PDGF) with Hyaluronic Acid (HA) as a potent growth factor can be used to stimulate the higher formation of chondrocyte and PIIANP levels and lower CTX-II levels in mouse knee osteoarthritis model.

Platelet derived growth factor-BB levels in gingival crevicular fluid of localized intrabony defect sites treated with platelet rich fibrin membrane or collagen membrane containing recombinant human platelet derived growth factor-BB: A randomized clinical and biochemical study.

Development of autologous and recombinant growth factor/matrix combination products represent a new emerging trend in regenerative therapeutics and have gained increasing attention as a strategy to optimize tissue regeneration. The aim of the present study was to evaluate the levels of platelet derived growth factor-BB (PDGF-BB) in gingival crevicular fluid (GCF) during early healing period after the regenerative treatment of intrabony defects using beta tricalcium phosphate (β-TCP) as a bone regeneration material with either platelet rich fibrin (PRF) membrane or collagen membrane (CM) treated with recombinant human PDGF-BB (rhPDGF-BB). Twenty patients (13 males and 7 females) with chronic periodontitis participated in this prospective, randomized clinical and biochemical study. Each patient was randomly assigned to PRF membrane (group A) or CM incorporated with rhPDGF-BB (group B). GCF samples were obtained on days 3, 7, 14, and 30 for evaluation of PDGF-BB levels and alkaline phosphatase (ALP) levels. On days 3 and 7 following surgery, mean levels of PDGF-BB at sites treated with PRF membrane or CM incorporated with rhPDGF-BB as a barrier membrane were not significantly different. PDGF-BB levels decreased significantly in samples collected on days 14 and 30 with significant differences between both the groups. ALP levels significantly increased from day 3 to day 30 but there was no difference between two groups. Within the limitations of the study, both PRF membrane and CM incorporated with rhPDGF-BB showed comparable GCF levels of PDGF-BB initially with PRF showing more sustained levels throughout the study period.

Recombinant platelet-derived growth factor-BB and hyaluronic acid stimulates knee cartilage regeneration by forming higher chondrocytes count and lower YKL-40 level in rats model

ObjectiveOsteoarthritis is caused by cartilage degeneration arising from cartilage degradation of type II collagen which synthesis also deteriorated. Nowadays, osteoarthritis is still difficult to handle because of the irreversibility and progressivity of the disease. Regenerative therapy offers a great challenge and better result for osteoarthritis treatment. This study aims to prove that the administration of recombinant platelet-derived growth factor-BB (rrPDGF-BB) with hyaluronic acid (HA) can stimulate the higher forms of chondrocyte and lower the YKL-40 levels as a specific marker of cartilage degradation in mouse knee osteoarthritis model. MethodThis was an experimental study, post-test only control group design using white Wistar rats as subjects that were induced by monosodium iodoacetate (MIA) to create osteoarthritis (OA). The treatment group was given treatment group was given rrPDGF-BB and HA, while no such treatment was given to the control group. The chondrocyte cell count was examined with routine histopathology, and YKL-40 levels were calculated by ELISA. ResultStatistical analysis using independent t-test showed that the mean difference in the number of cell counts of chondrocytes was significantly higher than the control group. While the mean difference of YKL-40 level in the treatment group was significantly lower compared to the control group with. ConclusionWe conclude that the administration of rrPDGF-BB and HA in mouse knee joint osteoarthritis model have a regenerative effect on knee joint cartilage characterized by a higher number of chondrocytes and lower YKL-40 levels.

The effects of recombinant human platelet derived growth factor-BB on biological behaviours of human retinal vascular endothelial cells

Objective This study was to investigate the role of recombinant human platelet derived growth factor-BB(rhPDGF-BB) in the proliferation and migration of human retinal vascular endothelial cells (hRVECs). Methods hRVECs were cultured in DMEM with 10% fetal bovine serum.The rhPDGF-BB at the concentrations of 10, 50 and 200 ng/ml were added into the medium of exponential phase-growth cells for 24 and 48 hours, respectively, and no rhPDGF-BB was added in the normal control group.The proliferation of the cells (absorbancy) was assayed by cell counting kit 8(CCK8) method.Cell scratch test was employed to evaluate the relative migration area of cells (migrated acellular area/initial acellular area). The relative expression of rhPDGF-BB recepter (rhPDGF-BBR) mRNA in the cells was detected by reverse transcription PCR.The relative expression of VEGF mRNA and integrin mRNA in the cells was detected using real-time fluorescence quantitative PCR. Results hRVECs grew well and a expressing band according with rhPDGF-BBR prime was displayed.The absorbancy values of the cells were 1.01±0.05, 1.09±0.04, 1.10±0.02 and 1.13±0.05 in the normal control group and 10, 50, 200 ng/ml rhPDGF-BB groups at 24 hours after culture, and those in the 10, 50 and 200 ng/ml rhPDGF-BB groups were significantly increased in comparison with the normal control group (t=2.504, 3.430, 3.483, all at P<0.05). The relative migrated areas of the cells in the normal control group and 10, 50, 200 ng/ml rhPDGF-BB groups were 0.42±0.10, 0.38±0.09, 0.55±0.06 and 0.61±0.05 at 24 hours after culture, and those at 48 hours were 0.75±0.06, 0.81±0.02, 0.87±0.02 and 0.98±0.02, showing significant differences among the groups (Fgroup=16.283, P=0.000; Ftime=209.129, P=0.000), and the relative migrated areas was depended upon the rhPDGF-BB dose and time.The relative expressions of integrin mRNA were 1.06±0.02, 1.30±0.10, 1.20±0.16 and 1.27±0.08, and those of VEGF mRNA were 0.97±0.05, 1.06±0.16, 1.58±0.18 and 1.66±0.21 in the normal control group and 10, 50 ng/ml, 200 ng/ml rhPDGF-BB groups, respectively, and increased expressions of integrin mRNA and VEGF mRNA were found in the 50 and 200 ng/ml rhPDGF-BB groups compared with the normal control group (integrin mRNA: t=3.900, 4.014, both at P<0.05; VEGF mRNA: t=6.940, 7.210, both at P< 0.05). Conclusions rhPDGF-BB/rhPDGF-BBR signal promotes the proliferation and migration of hRVECs probably by up-regulating the expressions of integrin and VEGF. Key words: Endothelium, vascular/cytology; Neovascularization, pathologic/metabolism; Platelet-derived growth factor; Receptors, platelet-derived growth factor; Recombinant proteins; Humans; Cells, cultured; Integrin; Vascular endothelial growth factor

Transglutaminase type 2 affects cell migration through post-translational modification of platelet-derived growth factor-BB.

Migration is a key cellular function with important implications in cell physiology. Impairment of such function is observed in angiogenesis, cancer, central nervous system development, and many other physiological and pathological events. Serum is considered among the most potent physiological chemotactic stimuli. Transglutaminase 2 (TG2) is involved in most of the mentioned processes, suggesting the hypothesis that TG2 may modulate cell movement and chemotaxis by acting on serum factors. Cell biology and biochemistry studies confirmed this hypothesis, showing that human serum contains potent chemotactic signals significantly impaired by activated TG2. Bioinformatics studies indicated that one potent serum factor potential substrate of TG2-dependent transamidation is platelet-derived growth factor-BB (PDGF-BB). Cell biology and immunometric experiments carried out with U87MG human glioma cell line showed that human recombinant PDGF-BB pre-incubated with calcium-activated TG2 lost about 70% of its chemotactic activity and antigenicity. These data indicate that PDGF-BB is a substrate of TG2-transamidating activity, and such modification may play a key role in the modulation of PDGF's chemotactic features. Further, these findings suggest a novel point of view to study the extracellular functions of TG2 and to understand how protein signals, such as growth factors and cytokines, act in the extracellular space to reach their specific targets.

Platelet-derived growth factorBB enhances osteoclast formation and osteoclast precursor cell chemotaxis.

Enhanced osteoclast formation increases bone resorption, which triggers bone remodeling. Platelet-derived growth factor BB (PDGF-BB) enhances precursor cell homing, angiogenesis, and bone healing, and thereby could also treat osteoporosis. However, the effect of PDGF-BB on osteoclast formation is not fully understood. We investigated whether exogenous recombinant PDGF-BB directly affects osteoclast formation and osteoclast precursor cell chemotaxis. The murine monocyte-macrophage cell line RAW264.7 and bone-marrow-derived macrophages were cultured with recombinant mouse PDGF-BBwith or without aplatelet-derived growth factor receptorβ inhibitor (AG-1295) or a Janus kinase2 inhibitor (AG-490) to analyze the effect on osteoclastogenesis in vitro. PDGF-BB with or without AG-490 or AG-1295 was locally administrated in the mandibular fracture of 16-week-old Sprague Dawley rats (n=18) for 1-2weeks to analyze the effect on osteoclastogenesis in vivo. The effect of the treatments on osteoclast formation, osteoclast precursor cell migration, and expression of osteoclastogenic signaling molecules was analyzed. PDGF-BB enhanced osteoclast formation both in vitro and in vivo, but AG-490 and AG-1295 inhibited this effect. PDGF-BB enhanced phosphorylation of extracellular-signal-regulated kinase 1/2 (ERK1/2), Akt, and signal transducer and activator of transcription3 (STAT3) in RAW264.7 cells. AG-490 inhibited PDGF-BB-induced STAT3 phosphorylation. PDGF-BB enhanced RAW264.7 cell migration and gene expression of osteoclastogenic signaling molecules (i.e., nuclear factor of activated Tcells1, dendrocyte-expressed seven transmembrane protein, and B-cell lymphoma2), and treatment with AG-1295, AG-490, or S3I-201 (a STAT3 inhibitor) reduced this effect. PDGF-BB enhanced osteoclast formation, osteoclast precursor cell chemotaxis, and phosphorylation of STAT3, Akt, and ERK1/2. but AG-1295 and AG-490 reduced this effect. These findings reflect the complexity of PDGF-BB in bone biology.

Effect of recombinant PDGF-BB on bone formation in the presence of β-tricalcium phosphate and bovine bone mineral matrix: a pilot study in rat calvarial defects.

BackgroundSupplementation of bone substitutes with recombinant platelet-derived growth factor-BB (PDGF-BB) can enhance bone regeneration. The aim of the study was to evaluate the effect of PDGF-BB on bone formation in the presence of β-tricalcium phosphate and bovine bone mineral matrix in a rat calvaria defect model.MethodsThe authors examined 5 mm rat calvarial defects treated with β-tricalcium phosphate (TCP) or demineralized bovine bone mineral (DBBM) with and without 0.3 mg/ml recombinant PDGF-BB. Calvaria defects were randomly divided into the following treatment groups (n = 5); TCP; TCP plus PDGF-BB; DBBM; DBBM plus PDGF-BB; and untreated empty control. After 45 days, bone formation was evaluated by histomorphometry and fluorescence microscopy.ResultsThe authors report that the area of newly formed bone was similar between the empty controls and the two bone substitutes, TCP and DBBM. Supplementation of TCP and DBBM with PDGF-BB had no significant impact on bone formation. Fluorochrome staining revealed no visible changes in the pattern of bone formation in defects filled with TCP and DBBM, irrespective of PDGF-BB. Furthermore, supplementation with PDGF-BB did not influence biomaterial degradation.ConclusionsThe authors concluded that PDGF-BB had no impact on bone formation and degradation of bone substitutes in the respective rodent models. Thus, possible beneficial effects of PDGF-BB may require other model situations.

CD44 promotes the migration of bone marrow-derived mesenchymal stem cells toward glioma.

Previous in vivo and in vitro studies have shown that human mesenchymal stem cells (MSCs) exhibit tropism for gliomas. However, the mechanism underlying this directed migration remains unclear. The aim of the present study was to investigate the possible mechanism underlying platelet-derived growth factor-BB (PDGF-BB)-induced chemotactic migration of bone marrow-derived MSCs (BMSCs) toward glioma. Rat glioma C6 cell-conditioned medium was utilized to evaluate the chemotactic response of BMSCs toward glioma using an in vitro migration assay. Recombinant rat PDGF-BB was added to C6 cell-conditioned medium to assess its effect on the tropism of BMSCs. The effect of PDGF-BB on the expression levels of cluster of differentiation (CD)44 in BMSCs was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence assays. The results revealed that chemotactic migration was induced in BMSCs by rat glioma C6 cell-conditioned medium, which was enhanced by PDGF-BB treatment in a dose-dependent manner. Furthermore, RT-PCR and immunofluorescence assays showed that CD44 expression was upregulated in BMSCs following treatment with 40 ng/ml PDGF-BB for 12 h. Additionally, 3-h pretreatment with the anti-CD44 neutralizing antibody OX-50 was observed to attenuate the tropism of BMSCs toward glioma in the presence or absence of PDGF-BB. The results of the present study indicate that CD44 mediates the tropism of BMSCs toward glioma, and PDGF-BB promotes the migration of BMSCs toward glioma via the upregulation of CD44 expression in BMSCs. These findings suggest CD44 inhibition may be a potential therapeutic target for the treatment of glioma.

Consecutive Case Series Using a Composite Allograft Containing Mesenchymal Cells with an Amnion-Chorion Barrier to Treat Mandibular Class III/IV Furcations.

This retrospective case series describes a layered regenerative approach for five Class III and one borderline Class IV furcation, involving treatment consisting of root management that included conditioning with tetracycline solution followed by the topical application of recombinant platelet-derived growth factor BB. A composite allograft with mesenchymal stem cells was subsequently placed into the furcation and covered by a barrier derived from human amnion-chorion with flaps advanced to completely cover the site. Three furcations, including the one diagnosed as Class IV, had complete closure, two were converted to Class I, and in one instance, there was no improvement. This regenerative algorithm for mandibular Class III furcations may present the potential to save these teeth by altering the prognosis, which has traditionally been poor to hopeless, resulting in many of these teeth routinely being extracted.

The osteoregenerative effects of platelet‐derived growth factor BB cotransplanted with mesenchymal stem cells, loaded on freeze‐dried mineral bone block: A pilot study in dog mandible

Due to shortcomings associated with autogenous bone graft, the gold standard of craniofacial grafting, investigators seek alternatives that are accessible, efficient, and affordable. Accordingly, in the present pilot study, bone regeneration was induced using bone marrow-derived mesenchymal stem cells (BMSCs) loaded onto freeze-dried mineral bone block (FDMBB) in the presence or absence of recombinant platelet-derived growth factor-BB (rh PDGF-BB). Eight weeks after the bilateral extraction of premolars of four mongrel dogs, 25 × 10 mm defects were created at both sides of the mandible. The right mandible received autogenous-BMSC loaded on FDMBB (MSC group), whereas the left mandible received cellular blocks impregnated with rhPDGF-BB (MSC + PDGF Group). Animals were euthanized 8 weeks after grafting. Micro-computed tomography (micro-CT) and histomorphometric analysis demonstrated higher levels of bone formation for the test group (10.34% ± 0.20 and 26.63% ± 3.14, respectively) when compared to the control group (8.20% ± 0.20 and 21.38% ± 5.11). The differences were not statistically significant (P > 0.05). According to the performed micro-CT and histomorphometric analysis, adding 0.5 mg rhPDGF-BB (0.3 mg/mL) to the combination of BMSC/FDMBB did not significantly increase bone formation in supracrestal defect in dog mandible.

RhPDGF-BB Promotes Proliferation and Osteogenic Differentiation of Bone Marrow Stromal Cells from Streptozotocin-Induced Diabetic Rats through ERK Pathway

Management of nonunion fracture and massive segmental bone defects in diabetes remains a challenging clinical problem. Bone marrow stromal cells (BMSCs) are crucial for bone remodeling and hold promise for bone regeneration. However, we have showed previously that diabetes can affect the proliferation and osteogenic potential of BMSCs adversely and a strategy to attenuate the impaired functions of BMSCs is required. Platelet-derived growth factor-BB (PDGF-BB) plays an important role in bone formation. However, little information is available about its effect on diabetic BMSCs. In this study, BMSCs were isolated from streptozotocin-induced diabetic rats. After treatment with recombinant human PDGF-BB (rhPDGF-BB), diabetic BMSCs demonstrated enhanced cell proliferation and osteogenic differentiation based on increased expressions of osteogenic genes (Runx2, alkaline phosphatase, and osteocalcin) and Runx2 protein, as well as upregulated alkaline phosphatase activity and mineralization. Furthermore, blocking extracellular signal regulated kinase (ERK) pathway by inhibitor PD98059 repressed the enhanced proliferation and osteogenic differentiation in diabetic BMSCs induced by rhPDGF-BB. Together, these results indicated that rhPDGF-BB stimulates proliferation and osteogenic differentiation partially through ERK pathway in diabetic BMSCs. Therefore, modulation of diabetic BMSCs could augment BMSCs function affected by diabetes and holds significance for future strategies to treat diabetic bone complications.

Platelet-derived growth factor BB mediates the glioma-induced migration of bone marrow-derived mesenchymal stem cells by promoting the expression of vascular cell adhesion molecule-1 through the PI3K, P38 MAPK and NF-κB pathways

Platelet-derived growth factor BB (PDGFBB) has been shown to activate the migration of bone marrow-derived mesenchymal stem cells (BM-MSCs), and to contribute to mediating the tropism of BM-MSCs towards gliomas. However, the exact mechanism of this migratory behavior remains to be elucidated. The present study investigated the role of vascular cell adhesion molecule-1 (VCAM-1) in the PDGFBB-induced migration of BM-MSCs, the effect of PDGFBB on VCAM-1 expression of BM-MSCs and related signaling pathways involved in this process. Rat BM-MSCs were isolated and cultured by their characteristics of adherence to plastics. The concentrations of PDGFBB in the conditioned medium of C6 and U87 cells were measured using the ELISA method. In vitro migration assays using a VCAM-1 blocking antibody were performed to evaluate the role of VCAM-1 in PDGFBB-induced migration of BM-MSCs. The effect of rat recombinant PDGFBB on VCAM-1 expression of BM-MSCs was studied by RT-PCR and western blotting. LY294002, SB203580, PD98059, SP600125 and BAY11-7082 were used to explore the role of PI3K, p38 MAPK, MEK, JNK and NF-κB in the related intracellular signal transduction of PDGFBB stimulation on VCAM-1 expression of BM-MSCs. The data demonstrated that the neutralization of VCAM-1 inhibited the migration of BM-MSCs induced by PDGFBB. Additionally, PDGFBB stimulation increased VCAM-1 expression of BM-MSCs, which could be inhibited by LY294002, SB203580 and BAY11-7082. It is reasonable to conclude that PDGFBB significantly enhanced the expression of VCAM-1 in BM-MSCs, which facilitated the migration of BM-MSCs towards PDGFBB. PI3K, p38 MAPK and NF-κB were involved in the signal transduction of this process.

Postextraction ridge preservation and augmentation with mineralized allograft with or without recombinant human platelet-derived growth factor BB (rhPDGF-BB): a consecutive case series.

In an attempt to reduce postextraction alveolar bone resorption, ridge preservation and augmentation procedures have become standard-of-care treatment following tooth removal. This consecutive case series compares histologic and histomorphometric bone regenerative findings at 4 months following grafting for ridge preservation and augmentation in intact sockets and sockets with buccal wall defects. Sites were treated with mineralized allograft alone (control) or in combination with 0.3 mg/mL recombinant human platelet-derived growth factor BB (rhPDGF-BB) (test). Sites were allowed to heal for 4 months and then re-entered for trephine core biopsy and implant placement. At the end of 4 months, the mean percent remaining mineralized allograft was statistically significantly less in the test group than in the control group. The difference in mean percent vital bone between the groups showed a strong trend toward greater bone formation for the test group (41.8%) compared to the control group (32.5%) at the end of 4 months. Addition of growth factor signaling molecules to current grafting procedures may lead to accelerated bone regeneration, making it possible to successfully place implants at earlier time points.

Human periodontal ligament fibroblast response to rhPDGF-BB application on periodontally diseased root surfaces-in vitro

The aim of the present study was to evaluate the influence of recombinant Platelet Derived Growth Factor-BB (rhPDGF-BB) on the cellular proliferation, morphology and adherence of human periodontal ligament fibroblasts (PLFs) to the root surfaces of periodontally diseased and healthy human teeth roots at two different time periods. Primary cell cultures of PLFs were obtained from clinically healthy premolar or mandibular third molar teeth. 11 scaffolds were prepared from healthy teeth for group-1 (Healthy Substrates), 33 scaffolds were prepared from periodontally diseased teeth, which were further divided in Group-2 – Periodontally diseased substrates, Group-3 – Scaled and Root planed (SRP) substrates, Group-4 – SRP + rhPDGF-BB (50 ng/ml). Groups were further subdivided into two groups (n = 5 scaffolds per subgroup) and PLFs were incubated on the scaffolds for three and seven days, topographical assessment was done on the remaining substrate. Cell morphology and counting was assessed under a scanning electron microscope at 350× on day three and seven and statistically compared with the Mann–Whitney U test and the Kruskal–Wallis test. On day three, Group 1 showed least number of cells attached, whereas maximum number of cells were attached on Group 3 (SRP only) substrates. For day 7, Group 1 and Group 4 showed increase in the number of cells from day 3 to 7, while number of cells attached/substrate reduced drastically for Group 2 and 3 substrates. Group 3 and 4 showed better adhesion and proliferation of PLFs as compared to Group 1 and 2. Group-1 and Group-4 showed predominantly spindle cells with flat appearance, Group-3 showed stellate cells and Group-2 showed predominantly distorted spindle shaped cells. The results of this in-vitro study indicates that rhPDGF-BB plays a significant role as an adjunct to periodontal therapy in influencing maturity, attachment and proliferation of PLFs.