Effect of Curcumin and Bromelian on Osteogenesis of Gingiva Derived Stem Cells

Background: Mesenchymal stem cells (MSCs) are attractive cell sources for tissue engineering application because of their ability to proliferate and differentiate into mesenchymal tissues. MSCs derived from human gingiva are readily accessible and highly proliferative with the ability to differentiate into an osteogenic lineage, making them ideal sources for tissue regeneration of craniofacial defects. While dexamethasone is a traditional inducer of osteogenic differentiation, studies have shown that its prolonged presence in culture medium may have toxic effects on osteoblasts. Many antioxidants play a vital role in promoting osteogenic differentiation and offer a potential alternative to dexamethasone. It has been demonstrated that curcumin can promote osteogenesis of rat derived bone marrow mesenchymal cells, suggesting that curcumin can be used in the treatment of bone lesions. Previous studies reported that bromelain treatment relieved osteoarthritis, indicating that bromelain may be able to promote bone health, which gives a cue that bromelain can induce osteogenic differentiation in MSCs. Objective: The aim of this study was to investigate the effects of curcumin and bromelain on osteogenic differentiation of human gingiva derived mesenchymal stem cells (HGMSCs) and to compare their differentiation potential to dexamethasone. Methodology: Stem cells were isolated from human gingival tissue samples. Surface markers were detected using flow cytometry. Guided osteogenic differentiation assay was conducted to confirm mineralization. The effects of curcumin and bromelain on HGMSCs proliferation on day1, 3 and 5 was examined using a MTT assay. Cells were treated with various concentrations of curcumin (2, 5 and 10µM) and bromelain (1, 2.5 and 5µg/ml) for two weeks and gene expression was investigated using quantitative PCR. Results: Our findings demonstrated that curcumin and bromelain induced osteogenic differentiation in a dose dependent manner. At 2.5µg/ml the peak up-regulation could be seen for genes Collagen, ALP, and OPG for bromelain treated cells. In curcumin the ideal concentration found was to 2µM. The maximum enhancement has been observed at 2µM for all genes. Conclusion: Cells treated with curcumin and bromelain induced the osteogenic differentiation, however, future in vivo studies need to be conducted to confirm findings

Aqueductal developmental venous anomaly as an unusual cause of congenital hydrocephalus: a case report and review of the literature

<p>Abstract</p> <p>Introduction</p> <p>Aqueductal stenosis may be caused by a number of etiologies including congenital stenosis, tumor, inflammation, and, very rarely, vascular malformation. However, aqueductal stenosis caused by a developmental venous anomaly presenting as congenital hydrocephalus is even more rare, and, to the best of our knowledge, has not yet been reported in the literature. In this study, we review the literature and report the first case of congenital hydrocephalus associated with aqueductal stenosis from a developmental venous anomaly.</p> <p>Case presentation</p> <p>The patient is a three-day-old, African-American baby girl with a prenatal diagnosis of hydrocephalus. She presented with a full fontanelle, splayed sutures, and macrocephaly. Postnatal magnetic resonance imaging showed triventricular hydrocephalus, suggesting aqueductal stenosis. Examination of the T1-weighted sagittal magnetic resonance imaging enhanced with gadolinium revealed a developmental venous anomaly passing through the orifice of the aqueduct. We treated the patient with a ventriculoperitoneal shunt.</p> <p>Conclusions</p> <p>Ten cases of aqueductal stenosis due to venous lesions have been reported and, although these venous angiomas and developmental venous anomalies are usually considered congenital lesions, all 10 cases became symptomatic as older children and adults. Our case is the first in which aqueductal stenosis caused by a developmental venous anomaly presents as congenital hydrocephalus. We hope adding to the literature will improve understanding of this very uncommon cause of hydrocephalus and, therefore, will aid in treatment.</p

EFFECT OF BROMELAIN ON OSTEOGENIC DIFFERENTIATION OF HUMAN GINGIVA DERIVED STEM CELLS

Objective. The aim of this study was to investigate the effect of bromelain on cell proliferation and osteogenic differentiation of human gingiva derived mesenchymal stem cells (HGMSCs). Background. HGMSCs are highly proliferative with the ability to differentiate into osteogenic precursor cells. While dexamethasone is a traditional inducer of osteogenic differentiation, many antioxidants play a vital role in enhancing osteogenic differentiation and offer a potential alternative to dexamethasone. Bromelain, an antioxidant derived from pineapple extract, is known to modulate NF kappa B signaling. Previous studies showed that the inhibition n of NF-kappa B significantly enhanced MSC-mediated bone formation. Methods. Cryopreserved HGMSCs were used for the study. The cells obtained from passage 4 were seeded onto a 96- well plate and treated with different concentrations of bromelain (1, 2.5, 5, 7.5, 10 and 15 mg/mL) and cell viability was determined by MTT assay. Osteogenic differentiation was examined by osteogenic marker gene expression at 2 weeks. Results: While cells exposed to bromelain on day 1 showed significant increase in the proliferation at 1 μg/ml concentration, on day 3, there was significant increase in cell proliferation (P\u3c0.05) at all concentrations. The cells induced with bromelain for 2 weeks showed significant upregulation of several genes such as alkaline phosphatase, osteonectin, osteoprotegerin. Results. While cells exposed to bromelain on day 1 showed significant increase in the proliferation at 1 μg/ml concentration, on day 3, there was significant increase in cell proliferation (P\u3c0.05) at all concentrations. The cells induced with bromelain for 2 weeks showed significant upregulation of several genes such as alkaline phosphatase, osteonectin, osteoprotegerin. Conclusion. The findings from the study demonstrated that bromelain treatment induced cell proliferation. Nevertheless, the survival of HGMSCs is concentration dependent. The results of the study suggests that bromelain can induce osteogenic differentiation of HGMSCs. Grants. HPD grant

SMALL MOLECULES IN OSTEOGENIC DIFFERENTIATION

Objective. The objective of this study was to investigate the osteogenic potential of curcumin induced human gingiva derived mesenchymal stem cells (HGMSCs) Background. Mesenchymal stem cells have multilineage potential thus being an attractive source in regenerative medicine. Stem cell differentiation and modulation of functional activities are generally modulated by growth factors and small regulatory molecules. Dexamethasone (Dex) and curcumin are such well-known small molecules. While Dex is a traditional osteogenic inducer, the role of curcumin in inducing osteogenic differentiation needs to be determined. It has been proved that curcumin has anti-inflammatory property and inhibits osteoclastic activity. Methods. The HGMSCs obtained from human gingival tissue were cultured under standard culture conditions. Cells were treated with curcumin (2, 5 and 10uM) for 2 days, and then transferred to osteogenic medium. Cells in medium devoid of curcumin was used as control. Osteogenesis was assessed by alkaline phosphatase (ALP) gene expression and the ALP enzyme activity was measured by pNPP assay. Results. Compared to controls, curcumin enhanced ALP gene expression in a dose dependent manner. A significant increase in ALP activity was observed on day 7 and 14. The upregulation was comparable with Dex treated cells. Conclusion. The results of this study suggested that curcumin induces osteogenic differentiation of HGMSCs. Grants. NSU-HPD Grant