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Topical formulations that include natural active compounds to treat inflamed and aching skin are favored by patients. Small local producers use commercial creams and gels to which they add compounds, often with criteria based more on trends than scientific evidence, to obtain products to treat inflamed joints or skin. Based on our previous studies that demonstrated the anti-inflammatory and analgesic effects of Harpagophytum procumbens extracts and N-acetylglucosamine, we were interested in preparing formulations with these natural substances for topical applications. In the present study, we compared the rheological properties of various creams and gels with bulk and nanoparticle forms of N-acetylglucosamine added, focusing on the flow properties of multiphase emulsion. Moreover, the ability of nanoparticles to reduce inflammation and stimulate collagen production was demonstrated in both chondrocyte and fibroblast spheroids. The results showed the activity of nano-N-acetylglucosamine in our cell models, suggesting its suitability for use as a component in creams and gels. Furthermore, its ability to affect the flow properties of commercial creams and gels makes it an ideal component of topical formulations for the treatment of inflamed skin and joints. [ABSTRACT FROM AUTHOR]

The intron retention (IR) is a phenomenon utilized by cells to allow diverse fates at the same mRNA, leading to a different pattern of synthesis of the same protein. In this study, we analyzed the modulation of phosphoinositide‐specific phospholipase C (PI‐PLC) enzymes by Harpagophytum procumbens extract (HPE) in synoviocytes from joins of osteoarthritis (OA) patients. In some samples, the PI‐PLC γ1 isoform mature mRNA showed the IR and, in these synoviocytes, the HPE treatment increased the phenomenon. Moreover, we highlighted that as a consequence of IR, a lower amount of PI‐PLC γ1 was produced. The decrease of PI‐PLC γ1 was associated with the decrease of metalloprotease‐3 (MMP‐3), and MMP‐13, and ADAMTS‐5 after HPE treatment. The altered expression of MMPs is a hallmark of the onset and progression of OA, thus substances able to decrease their expression are very desirable. The interesting outcomes of this study are that 35% of analyzed synovial tissues showed the IR phenomenon in the PI‐PLC γ1 mRNA and that the HPE treatment increased this phenomenon. For the first time, we found that the decrease of PI‐PLC γ1 protein in synoviocytes interferes with MMP production, thus affecting the pathways involved in the MMP expression. This finding was validated by the silencing of PI‐PLC γ1 in synoviocytes where the IR phenomenon was not present. Our results shed new light on the biochemical mechanisms involved in the degrading enzyme production in the joint of OA patients, suggesting a new therapeutic target and highlighting the importance of personalized medicine. Significant Statement: This manuscript focuses on the modulation of phosphoinositide‐specific phospholipase C (PI‐PLC) γ1 in human primary fibroblast‐like synoviocytes, which leads to the modulation of metalloprotease‐3 (MMP‐3), MMP‐13, and ADAMTS‐5, involved in the degradation of extracellular matrix in degenerative joint diseases. We found that in some synoviocyte samples, the PI‐PLC γ1 mRNA incurs in the intron retention (IR) phenomenon, which results in an unsuccessful translation of the proteins associated with the decrease of MMP enzymes. Moreover, IR is increased after cell treatment with Harpagophytum procumbens extract (HPE), traditionally used to treat osteoarthritis. Considering that HPE results are effective in modulating MMPs only in synoviocyte samples where the IR phenomenon is present, it is necessary to evaluate individual variability in the personalized medicine field. [ABSTRACT FROM AUTHOR]

Over the last decades, a variety of biomaterials, ranging from synthetic products to autologous and heterologous grafts, have been recommended to conserve and regenerate bone tissue after tooth extraction. We conducted a biochemical study on ground extracted teeth that aimed to evaluate the osteoinductive and osteoconductive potential of dentin by assessing the releases of bone morphogenetic protein (BMP-2), osteocalcin (OC) and osteonectin (ON) over time (24 h, 10 days and 28 days). Twenty-six patients, who required the extraction of nonrestorable teeth, were enrolled in the study according to the inclusion criteria, as follows: thirteen young patients 18 to 49 years of age (UNDER 50), and thirteen patients of 50 to 70 years (OVER 50); a total of twenty-six teeth were extracted, ground and analyzed by enzyme-linked immunosorbent assays (ELISA). All ground teeth released BMP-2, OC and ON at each time point; no differences were observed between the UNDER-50 and OVER-50 patients. The results of the study support the use of autologous dentin as osteoinductive material for bone regeneration procedures, irrespective of patients' ages. [ABSTRACT FROM AUTHOR]

Inteins are proteins involved in the protein splicing mechanism, an autoprocessing event, where sequences (exteins) separated by inteins become ligated each other after recombination. Two kinds of inteins have been described, contiguous inteins and split inteins. The former ones are transcribed and translated as a single peptide along with their exteins, while the latter are fragmented between two different genes and are transcribed and translated separately. The aim of this study is to establish a method to obtain a fluorescent eukaryotic protein to analyze its cellular localization, using the natural split gp41‐1 inteins. We chose natural split inteins due to their distribution in all three domains of life. Two constructs were prepared, one containing the N‐terminal split intein along with the N‐moiety of the Red Fluorescent Protein (RFP) and a second construct containing the C‐terminal of split intein, the C‐moiety of RFP and the gene coding for Maspin, a tumor suppressor protein. The trans‐splicing was verified by transfecting both N‐terminal and C‐terminal constructs into mammalian cells. The success of the recombination event was highlighted through the fluorescence produced by reconstituted RFP after recombination, along with the overlap of the red fluorescence produced by recombined RFP and the green fluorescence produced by the hybridization of the recombinant Maspin with a specific antibody. In conclusion, we opted to use this mechanism of recombination to obtain a fluorescent Maspin instead to express a large fusion protein, considering that it could interfere with Maspin's structure and function. Significance statement: This manuscript focuses on the use of Ultra‐Fast Split Gp41‐1 intein to obtain a fluorescent eukaryotic protein, Maspin, a tumor suppressor protein, and to analyze its cellular localization. This method is based on protein recombination, using split intein and Red Fluorescent Protein. The protein recombination produces a fluorescent Maspin to be localized into cell compartments. The advantage of this method to obtain a fluorescent protein instead to express a large fusion protein is the ability to avoiding the interference that a large protein could have on Maspin structure and function. [ABSTRACT FROM AUTHOR]

The ultraviolet (UV) component of solar radiation is the driving force of life on earth, but it can cause photoaging and skin cancer. In this study, we investigated the effects of the glucosamine‐derivative 2‐(N‐Acetyl)‐L‐phenylalanylamido‐2‐deoxy‐β‐D‐glucose (NAPA) on human primary fibroblasts (FBs) stimulated in vitro with environmental levels of UVB radiation. FBs were irradiated with 0.04 J cm−2 UVB dose, which resulted a mild dosage as shown by the cell viability and ROS production measurement. This environmental UVB dose induced activation of MAP kinase ERK 1/2, the stimulation of c‐fos and at lower extent of c‐jun, and in turn AP‐1‐dependent up‐regulation of pro‐inflammatory factors IL‐6 and IL‐8 and suppression of collagen type I expression. On the contrary, 0.04 J cm−2 UVB dose was not able to stimulate metalloprotease production. NAPA treatment was able to suppress the up‐regulation of IL‐6 and IL‐8 via the inhibition of MAP kinase ERK phosphorylation and the following AP‐1 activation, and was able to attenuate the collagen type I down‐regulation induced by the UVBs. Taken together, our results show that NAPA, considering its dual action on suppression of inflammation and stimulation of collagen type I production, represents an interesting candidate as a new photoprotective and photorepairing agents. [ABSTRACT FROM AUTHOR]