Nanohybrid composed of graphene oxide functionalized with sodium hyaluronate accelerates bone healing in the tibia of rats.

This study synthesized and characterized a nanohybrid composed of graphene oxide (GO) functionalized with sodium hyaluronate (HY) (GO-HY), evaluated its effect in vitro and determined its osteogenic potential in vivo. The synthesized nanohybrid was analyzed by Scanning electron microscopy (SEM), Raman spectrometry, Thermogravimetry, Fourier-transform infrared (FTIR) spectroscopy and X-ray diffraction. MC3T3-E1 cell viability was assessed by MTT assay in 48 and 72 h. Bone defects were created in tibia of 40 Wistar rats and filled with blood clot (control), 1% HY, GO (50, 100 and 200 µg/mL) and the nanohybrid (50, 100 and 200 µg/mL). After 7 and 14 days, histomorphometric analysis was carried out to assess osteogenic potential of the nanohybrid. Immunohistochemical analysis evaluated the expression of vascular endothelial growth factor (VEGF) in bone defects. Thermogravimetric analysis, Raman and FTIR spectrometry confirmed the functionalization of GO with HY by covalent bonds. Five µg/mL concentrations of the nanohybrid did not alter the viability of the MC3T3-E1 cells. Histomorphometric analysis demonstrated that the nanohybrid at 100 µg/mL significantly accelerated the bone repair in tibia of rats when compared to controls (p < 0.01). Immunohistochemical analysis showed a significantly less intense VEGF expression in tibia treated with the nanohybrid when compared to controls (p < 0.05). The nanohybrid composed of GO functionalized with HY was able to induce the acceleration of the tissue regeneration process in bone defects created in the tibia of rats. This novel nanohybrid is a promising material for the field of bone tissue engineering.

Cobalt-containing bioactive glass mimics vascular endothelial growth factor A and hypoxia inducible factor 1 function.

Bioactive glasses (BGs) have shown great potential for tissue regeneration and their composition flexibility allows the incorporation of different ions with physiological activities and therapeutic properties in the glass network. Among the many ions that could be incorporated, cobalt (Co) is a significant one, as it mimics hypoxia, triggering the formation of new blood vessels by the vascular endothelial growth factor A (VEGFA), due to the stabilizing effect on the hypoxia inducible factor 1 subunit alpha (HIF1A), an activator of angiogenesis-related genes, and is therefore of great interest for tissue engineering applications. However, despite its promising properties, the effects of glasses incorporated with Co on angiogenesis, through human umbilical cord vein endothelial cells (HUVECs) studies, need to be further investigated. Therefore, this work aimed to evaluate the biocompatibility and angiogenic potential of a new sol-gel BG, derived from the SiO2 -CaO-P2 O5 -CoO system. The structural evaluation showed the predominance of an amorphous glass structure, and the homogeneous presence of cobalt in the samples was confirmed. in vitro experiments showed that Co-containing glasses did not affect the viability of HUVECs, stimulated the formation of tubes and the gene expression of HIF1A and VEGFA. in vivo experiments showed that Co-containing glasses stimulated VEGFA and HIF1A expression in blood vessels and cell nuclei, respectively, in the deep dermis layer of the dorsal region of rats, featuring considerable local stimulation of the angiogenesis process due to Co-release. Co-containing glasses showed therapeutic effect, and Co incorporation is a promising strategy for obtaining materials with superior angiogenesis properties for tissue engineering applications.

The angiotensin converting enzyme 2/angiotensin-(1-7)/Mas Receptor axis as a key player in alveolar bone remodeling.

The renin-angiotensin system (RAS), aside its classical hormonal properties, has been implicated in the pathogenesis of inflammatory disorders. The angiotensin converting enzyme 2/angiotensin-(1-7)/Mas Receptor (ACE2/Ang-(1-7)/MasR) axis owns anti-inflammatory properties and was recently associated with bone remodeling in osteoporosis. Thus, the aim of this study was to characterize the presence and effects of the ACE2/Ang-(1-7)/MasR axis in osteoblasts and osteoclasts in vitro and in vivo. ACE2 and MasR were detected by qPCR and western blotting in primary osteoblast and osteoclast cell cultures. Cells were incubated with different concentrations of Ang-(1-7), diminazene aceturate (DIZE - an ACE2 activator), A-779 (MasR antagonist) and/or LPS in order to evaluate osteoblast alkaline phosphatase and mineralized matrix, osteoclast differentiation and cytokine expression, and mRNA levels of osteoblasts and osteoclasts markers. An experimental model of alveolar bone resorption triggered by dysbiosis in rats was used to evaluate bone remodeling in vivo. Rats were treated with Ang-(1-7), DIZE and/or A-779 and periodontal samples were collected for immunohistochemistry, morphometric analysis, osteoblast and osteoclast count and cytokine evaluation. Human gingival samples from healthy and periodontitis patients were also evaluated for detection of ACE2 and MasR expression. Osteoblasts and osteoclasts expressed ACE2 and MasR in vitro and in vivo. LPS stimulation or alveolar bone loss induction reduced ACE2 expression. Treatment of bone cells with Ang-(1-7) or DIZE stimulated osteoblast ALP, matrix synthesis, upregulated osterix, osteocalcin and collagen type 1 transcription, reduced IL-6 expression, and decreased osteoclast differentiation, RANK and IL-1ß mRNA transcripts, and IL-6 and IL-1ß levels, in a MasR-dependent manner. In vivo, Ang-(1-7) and DIZE decreased alveolar bone loss through improvement of osteoblast/osteoclast ratio. A-779 reversed such phenotype. ACE2/Ang-(1-7)/MasR axis activation reduced IL-6 expression, but not IL-1ß. ACE2 and MasR were also detected in human gingival samples, with higher expression in the healthy than in the inflamed tissues. These findings show that the ACE2/Ang-(1-7)/MasR is an active player in alveolar bone remodeling.

Avaliação topográfica e in vitro de superfícies de titânio revestidas com vidro bioativo / Topographic and in vitro evaluation of titanium surfaces coated with bioative glass

Objetivo: Avaliar e comparar a rugosidade superficial e a atividade dos osteoblastos em contato com uma nova superfície bioativa e nanoestruturada de titânio grau 4 revestida com vidro bioativo contendo fosfato de cálcio, sintetizada pelo método sol-gel. Material e método: Sessenta e três discos de titânio, medindo 4 mm de diâmetro por 2 mm de altura, foram preparados e divididos em três grupos: microtexturizado (Ticp - controle); revestido com vidro bioativo e seco a vácuo a 37 °C por 10 dias (BGTi37), e revestido com vidro bioativo e aquecido a 600 °C por cinco horas (BGTi600). Três espécimes de cada grupo foram utilizados para avaliação da topografia superficial e 18 espécimes, para cultura celular. Resultado: O revestimento de vidro bioativo diminuiu a rugosidade média quando comparado ao titânio microtexturizado. A proporção de células viáveis, a produção de fosfatase alcalina e o grau de mineralização da matriz óssea em contato com os espécimes de titânio do grupo BGTi600 foram significativamente menores em relação aos grupos controle e do titânio microtexturizado. Conclusão: Apesar de sua marcante menor rugosidade, a superfície BGTi37 apresentou comportamento biológico semelhante a uma superfície de titânio microtexturizada e moderadamente rugosa. A outra superfície experimental (BGTi600), a de menor rugosidade entre todas as testadas, apresentou os piores resultados de ativação dos osteoblastos.

Objective: To evaluate and compare the surface roughness and the activity of the osteoblasts in contact with a new bioactive and nanostructured surface of grade 4 titanium coated with bioactive glass containing calcium phosphate synthesized by the sol-gel method. Material and method: Sixty-three titanium disks, measuring 4 × 2 mm, were prepared and divided into three groups: rough surface, obtained by sandblasted, large-grit, acid-etched (SLA) treatment (Ticp); SLA surface coated with bioglass and dried in a vacuum at 37 °C for 10 days (BGTi37) and SLA surface coated with bioglass and dried in air at 600 °C for 5 hours (BGTi600). Three specimens of each group were used for evaluation of surface topography and 18 for cell cultures. Result: The bioactive glass coating decreased the average roughness when compared to rough titanium surface. The proportion of viable cells, the production of alkaline phosphatase and the degree of mineralization of the bone matrix in contact with the titanium specimens of the BGTi600 group was significantly lower in relation to the control and rough titanium surface groups. Conclusion: Despite its marked lower roughness, BGTi37 surface presented a similar biological behavior to a titanium rough surface obtained by SLA treatment. The other experimental surface (BGTi600), the one with the least roughness among all tested, presented the worst results of osteoblast activation.