In vitro biomineralization and osteogenesis of Cissus quadrangularis stem extracts: An osteogenic regulator for bone tissue engineering.

Contemporary demand calls for a high restorative index as an indispensable requirement for bone tissue engineering scaffolds, where therapeutic agents of natural origin function as a modulator for new bone formation become of utmost importance. The study presents a systematic investigation of the edible stem part of Cissus quadrangularis (CQ) as a natural resource of bioactive metabolites capable of invoking early biomineralization and osteogenesis in vitro. Phytochemical screening of CQ stem extracts (sequential solvent extraction: polarity hexane

Radiopacity endowed magnetic nanocomposite with hyperthermia andin vitromineralization potential: a combinatorial therapeutic system for osteosarcoma.

The development of clinically advanced multifaceted therapeutic materials for osteosarcoma is at the forefront of cancer research. Accordingly, this work presents the design of a multifunctional magnetic nanocomposite composed of maghemite, strontium doped hydroxyapatite and silica nanoparticles prospectively holding indispensable therapeutic features such as magnetic hyperthermia,in vitrobiomineralization, sustained drug release and intrinsic radiopacity for the treatment of osteosarcoma. The optimal composition has been identified by sequentially modulating the ratio of precursors of the magnetic nanocomposite synthesized by sol-gel technique. Structural and morphological characterization by x-ray diffraction, fourier transform infrared spectrum, Brunauer-Emmet-Teller and transmission electron microscopy analyses followed by VSM, hyperthermia and micro-CT analyses essentially assisted in the selective configuration of biofunctional properties. Results exemplify that MSHSr1 has a saturation magnetization of 47.4 emu g-1and attained hyperthermia temperature (42 °C) at a very low exposure time of 4 min. MSHSr1 is further unique with respect to its exceptional x-ray attenuation ability (contrast enhancement 154.5% in digital radiography; CT number 3100 HU), early biomimetic mineralization (in vitro) evident by the formation of spheroidal apatite layer (Ca/P ratio 1.33) harvested from FESEM-EDX analysis and controlled release of Doxorubicin, the clinically used chemotherapeutic drug: 87.7% at 120 h in tumour analogous pH (6.5) when compared to physiological pH (71.3% at 7.4). MTT assay complemented with cytoskeleton (F-actin) staining of human osteosarcoma (HOS) cells affirm biocompatibility of MSHSr1.In vitrobiomineralization authenticated by Alizarin red S and von Kossa staining has been further corroborated by semi-quantitative calcium estimation of HOS cells cultured with MSHSr1 for two weeks. The results therefore validate the multifunctionality of MSHSr1, and hence could be proposed as a combinatorial therapeutic nanocomposite for osteosarcoma treatment.

Melatonin rescues glucocorticoid-induced inhibition of osteoblast differentiation in MC3T3-E1 cells via the PI3K/AKT and BMP/Smad signalling pathways.

AIMS: High-dose glucocorticoid (GC) administration causes osteoporosis. Many previous studies from our group and other groups have shown that melatonin participates in the regulation of osteoblast proliferation and differentiation, especially low concentrations of melatonin, which enhance osteoblast osteogenesis. However, the role of melatonin in glucocorticoid-induced osteoblast differentiation remains unknown. MATERIALS AND METHODS: An examination of the expression of osteoblast differentiation markers (ALP, OCN, COLL-1), as well as alkaline phosphatase staining and alkaline phosphatase enzymatic activity assay to measure osteoblast differentiation and quantifying Alizarin red S staining to measure mineralization, were performed to determine the effects of dexamethasone (Dex) and melatonin on the differentiation of MC3T3-E1 cells. We used immunofluorescence staining to detect the expression of Runx2 in melatonin-treated MC3T3-E1 cells. The expression of mRNA was determined by qRT-PCR, and protein levels were measured by western blotting. KEY FINDINGS: In the present study, we found that 100 µM Dex significantly reduced osteoblast differentiation and mineralization in MC3T3-E1 cells and that 1 µM melatonin attenuated these inhibitory effects. We found that only inhibition of PI3K/AKT (MK2206) and BMP/Smad (LDN193189) signalling abolished melatonin-induced differentiation and mineralization. Meanwhile, MK2206 decreased the expression of P-AKT and P-Smad1/5/9 and LDN193189 decreased the expression of P-Smad1/5/9 but had no obvious effect on P-AKT expression in melatonin-treated and Dex-induced MC3T3-E1 cells. SIGNIFICANCE: These findings suggest that melatonin rescues Dex-induced inhibition of osteoblast differentiation in MC3T3-E1 cells via the PI3K/AKT and BMP/Smad signalling pathways and that PI3K/AKT signalling may be the upstream signal of BMP/Smad signalling.

Biomineralization Precursor Carrier System Based on Carboxyl-Functionalized Large Pore Mesoporous Silica Nanoparticles.

Bone and teeth are derived from intrafibrillarly mineralized collagen fibrils as the second level of hierarchy. According to polymer-induced liquid-precursor process, using amorphous calcium phosphate precursor (ACP) is able to achieve intrafibrillar mineralization in the case of bone biomineral in vitro. Therefore, ACP precursors might be blended with any osteoconductive scaffold as a promising bone formation supplement for in-situ remineralization of collagens in bone. In this study, mesoporous silica nanoparticles with carboxyl-functionalized groups and ultra large-pores have been synthesized and used for the delivery of liquid like biomimetic precursors (ACP). The precursor delivery capacity of the nanoparticles was verified by the precursor release profile and successful mineralization of 2D and 3D collagen models. The nanoparticles could be completely degraded in 60 days and exhibited good biocompatibility as well. The successful translational strategy for biomineralization precursors showed that biomineralization precursor laden ultra large pore mesoporous silica possessed the potential as a versatile supplement in demineralized bone formation through the induction of intrafibrillar collagen mineralization.

Improved Multicellular Response, Biomimetic Mineralization, Angiogenesis, and Reduced Foreign Body Response of Modified Polydioxanone Scaffolds for Skeletal Tissue Regeneration.

The potential of electrospun polydioxanone (PDX) mats as scaffolds for skeletal tissue regeneration was significantly enhanced through improvement of the cell-mediated biomimetic mineralization and multicellular response. This was achieved by blending PDX ( i) with poly(hydroxybutyrate- co-valerate) (PHBV) in the presence of hydroxyapatite (HA) and ( ii) with aloe vera (AV) extract containing a mixture of acemannan/glucomannan. In an exhaustive study, the behavior of the most relevant cell lines involved in the skeletal tissue healing cascade, i.e. fibroblasts, macrophages, endothelial cells and preosteoblasts, on the scaffolds was investigated. The scaffolds were shown to be nontoxic, to exhibit insignificant inflammatory responses in macrophages, and to be degradable by macrophage-secreted enzymes. As a result of different phase separation in PDX/PHBV/HA and PDX/AV blend mats, cells interacted differentially. Presumably due to varying tension states of cell-matrix interactions, thinner microtubules and significantly more cell adhesion sites and filopodia were formed on PDX/AV compared to PDX/PHBV/HA. While PDX/PHBV/HA supported micrometer-sized spherical particles, nanosized rod-like HA was observed to nucleate and grow on PDX/AV fibers, allowing the mineralized PDX/AV scaffold to retain its porosity over a longer time for cellular infiltration. Finally, PDX/AV exhibited better in vivo biocompatibility compared to PDX/PHBV/HA, as indicated by the reduced fibrous capsule thickness and enhanced blood vessel formation. Overall, PDX/AV blend mats showed a significantly enhanced potential for skeletal tissue regeneration compared to the already promising PDX/PHBV/HA blends.