Proteoliposomes as matrix vesicles' biomimetics to study the initiation of skeletal mineralization

During the process of endochondral bone formation, chondrocytes and osteoblasts mineralize their extracellular matrix by promoting the formation of hydroxyapatite (HA) seed crystals in the sheltered interior of membrane-limited matrix vesicles (MVs). Ion transporters control the availability of phosphate and calcium needed for HA deposition. The lipidic microenvironment in which MV-associated enzymes and transporters function plays a crucial physiological role and must be taken into account when attempting to elucidate their interplay during the initiation of biomineralization. In this short mini-review, we discuss the potential use of proteoliposome systems as chondrocyte- and osteoblast-derived MVs biomimetics, as a means of reconstituting a phospholipid microenvironment in a manner that recapitulates the native functional MV microenvironment. Such a system can be used to elucidate the interplay of MV enzymes during catalysis of biomineralization substrates and in modulating in vitro calcification. As such, the enzymatic defects associated with disease-causing mutations in MV enzymes could be studied in an artificial vesicular environment that better mimics their in vivo biological milieu. These artificial systems could also be used for the screening of small molecule compounds able to modulate the activity of MV enzymes for potential therapeutic uses. Such a nanovesicular system could also prove useful for the repair/treatment of craniofacial and other skeletal defects and to facilitate the mineralization of titanium-based tooth implants

Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation

Endochondral calcification involves the participation of matrix vesicles (MVs), but it remains unclear whether calcification ectopically induced by implants of demineralized bone matrix also proceeds via MVs. Ectopic bone formation was induced by implanting rat demineralized diaphyseal bone matrix into the dorsal subcutaneous tissue of Wistar rats and was examined histologically and biochemically. Budding of MVs from chondrocytes was observed to serve as nucleation sites for mineralization during induced ectopic osteogenesis, presenting a diameter with Gaussian distribution with a median of 306 ± 103 nm. While the role of tissue-nonspecific alkaline phosphatase (TNAP) during mineralization involves hydrolysis of inorganic pyrophosphate (PPi), it is unclear how the microenvironment of MV may affect the ability of TNAP to hydrolyze the variety of substrates present at sites of mineralization. We show that the implants contain high levels of TNAP capable of hydrolyzing p-nitrophenylphosphate (pNPP), ATP and PPi. The catalytic properties of glycosyl phosphatidylinositol-anchored, polidocanol-solubilized and phosphatidylinositol-specific phospholipase C-released TNAP were compared using pNPP, ATP and PPi as substrates. While the enzymatic efficiency (k cat/Km) remained comparable between polidocanol-solubilized and membrane-bound TNAP for all three substrates, the k cat/Km for the phosphatidylinositol-specific phospholipase C-solubilized enzyme increased approximately 108-, 56-, and 556-fold for pNPP, ATP and PPi, respectively, compared to the membrane-bound enzyme. Our data are consistent with the involvement of MVs during ectopic calcification and also suggest that the location of TNAP on the membrane of MVs may play a role in determining substrate selectivity in this micro-compartment

Solos urbanos Urban soils

A forte pressão provocada pela expansão urbana desordenada sobre os recursos naturais, principalmente os solos, tem provocado danos, muitas vezes de difícil reparo. A grande concentração populacional em centros urbanos cada vez maiores tem dirigido a atenção de diferentes profissionais para o recurso solo, no sentido de entender sua dinâmica para minimizar sua degradação. No entanto, a falta de conhecimento sobre as propriedades, bem como sobre a aptidão dos solos sob uso urbano tem provocado o seu mau uso, resultando em processos como compactação, erosão, deslizamentos e inundações, assim como poluição com substâncias orgânicas, inorgânicas e patógenos, aumentando os custos do desenvolvimento afetando toda a sociedade. Neste sentido, este texto discute como o conhecimento pedológico pode diminuir os efeitos negativos provocados pelo processo de urbanização.<br>The strong pressure caused by the disordered urban expansion over the natural resources, mainly the soils, has caused damages, many times difficult to repair. The great population concentration in urban centers getting larger and larger has been driving the attention of different professionals to soil resource, in the sense of understanding its dynamics to minimize its degradation. The lack of knowledge related to the soils properties and capability promote their inappropriate use, resultig in degrading processes as compaction, erosion, sliding, floods, and organic, inorganic and patogenic pollution, increasing the cost of development and affecting the whole society. This text discusses how pedologic knowledge can reduce the negative effects caused by the urbanization process