Advanced Regenerative Techniques Based on Dental Pulp Stem Cells for the Treatment of Periodontal Disease

Recent progress in periodontology intended to reduce the risk represented by periodontal disease for systemic disorders and general human health condition. In this chapter, we overview the advantages and limitations of current techniques based on occlusive membranes for periodontal regeneration. Special emphasis is paid to advanced techniques using stem cells from dental pulp for the regeneration of bone defects caused by the chronic periodontal disease. Stem cells isolation, in vitro expansion and characterization techniques are presented. Therapeutic strategies of stem cells delivery using natural polymeric carriers are discussed. Stem cell-scaffold constructs application in bone tissue engineering is proposed, taking into account the marked decline of healing, and regenerative processes in elderly individuals. Future researchers envisage multiple effects of engineered constructs with antimicrobial, anti-inflammatory, and regenerative activity for periodontal treatment

Uklanjanje slobodnih radikala, postizanje redoks ravnoteže i cijeljenje rana pomoću bioaktivnih peptida dobivenih hidrolizom kolagena iz kože bijelog glavaša (Hypophthalmichthys molitrix) potpomognutom proteinazom K

Research background. Various protocols for enzymatic hydrolysis of fish by-products are increasingly tested to ensure value-added products with functional and biological properties important for food, cosmetic and medical applications. In addition, they attempt to minimize waste from industrial processing and environmental requirements. This study aims to establish an efficient protocol based on two-step enzymatic hydrolysis of freshwater fish skin and to evaluate the effect of resulting bioactive peptides on free radical scavenging, redox balance and regulation of fibroblast proliferation and migration. Experimental approach. Pepsin-soluble collagen extracted from silver carp (Hypophthalmichthys molitrix) skin was hydrolyzed by proteinase K at specific sites under controlled conditions. The molecular mass of ultrafiltration permeate was determined by gradient electrophoresis and gel filtration chromatography. The biological activity of intermediate and small size bioactive peptides was evaluated in experimental models in vitro mimicking oxidative stress and skin wound conditions. Results and conclusions. Extracted fish collagen was hydrolysed using proteinase K, the most efficient enzyme for the cleavage of the primary structure of the molecule, as previously found in silico. Established optimal conditions increased the enzyme specificity and the process yield. Bioactive peptides exerted significantly higher scavenging activity on free stable radicals and hydroxyl radicals often found in vivo, compared to fish collagen. They stimulated fibroblast metabolism in a dose-dependent manner and up-regulated cell migration in a scratch wound model. Pretreatment of fibroblasts with induced oxidative stress using optimal concentrations of fish peptides prevented the increase of reactive oxygen species production. In conclusion, bioactive peptides from carp skin demonstrated valuable properties of maintaining redox balance and skin wound healing process improvement, which indicated further potential applications in the development of pharmaceutical and nutraceutical formulations. Novelty and scientific contribution. In this study the enzymatic hydrolysis was applied to isolated protein, in contrast to previous studies using waste tissue with variable composition. Recovered bioactive peptides acted not only as antioxidant agents, but also as regulators of oxidative stress and wound healing processes in skin cell models. Their nutritional and cosmetic application is recommended in novel formulations fighting skin ageing phenomena.Pozadina istraživanja. U posljednje se vrijeme sve više ispituju postupci enzimske hidrolize nusproizvoda obrade ribe radi dobivanja proizvoda obogaćenih funkcionalnim i biološkim svojstvima važnim u proizvodnji hrane, kozmetike i medicinskih preparata. Osim toga, na taj se način smanjuju količina otpada industrijske proizvodnje i pritisak na okoliš. Svrha je ovoga rada bila osmisliti učinkoviti postupak dvostupanjske enzimske hidrolize kože slatkovodne ribe i ispitati učinak dobivenih bioaktivnih peptida na uklanjanje slobodnih radikala, redoks ravnotežu, te proliferaciju i migraciju fibroblasta. Eksperimentalni pristup. Kolagen razgradljiv pomoću pepsina izoliran je iz kože bijelog glavaša (Hypoph¬thalmichthys molitrix) i hidroliziran u kontroliranim uvjetima pomoću proteinaze K. Molekulska masa permeata dobivenog ultrafiltracijom određena je pomoću elektroforeze s gradijentom i gel-filtracijske kromatografije. Biološka aktivnost peptida srednje i male veličine ispitana je in vitro simulacijom oksidacijskog stresa i oštećenja kože. Rezultati i zaključci. Riblji kolagen je hidroliziran pomoću proteinaze K, najučinkovitijeg enzima koji cijepa primarnu strukturu molekule, što je potvrđeno prethodnim ispitivanjem in silico. Pri optimalnim uvjetima povećali su se specifičnost enzima i prinos reakcije. U usporedbi s riblijm kolagenom, bioaktivni su peptidi imali bitno veću sposobnost uklanjanja slobodnih i hidroksilnih radikala, često prisutnih in vivo. Ovisno o koncentraciji, stimulirali su metabolizam fibroblasta i migraciju stanica pri cijeljenju modelne rane. Prethodna obrada fibroblasta optimalnim koncentracijama ribljih peptida spriječila je porast sinteze reaktivnih spojeva kisika pri induciranju oksidacijskog stresa. Možemo zaključiti da bioaktivni peptidi kože bijelog glavaša imaju važna svojstva, kao što su održavanje redoks ravnoteže i pospješivanje cijeljenja rana, što upućuje na njihovu moguću primjenu u razvoju farmaceutika i nutraceutika. Novina i znanstveni doprinos. U ovom je radu provedena enzimska hidroliza izoliranog proteina, za razliku od dosadašnjih istraživanja u kojima je korišten otpad različitog sastava. Dobiveni bioaktivni peptidi nisu djelovali samo kao antioksidacijski agensi, već i kao regulatori oksidacijskog stresa i cijeljenja kože u staničnom modelu. Zbog svojih svojstava mogu se preporučiti za primjenu u proizvodnji kozmetičkih pripravaka koji suzbijaju starenje kože

Anti-Inflammatory Activity of Biomaterials Intended for Periodontal Disease Treatment

Periodontal disease is associated with chronic tissue inflammation, which besides bacterial [...

Nanoplatforms for Irinotecan Delivery Based on Mesoporous Silica Modified with a Natural Polysaccharide

Natural compounds are an important source of beneficial components that could be used in cancer therapy along with well-known cytostatic agents to enhance the therapeutic effect while targeting tumoral tissues. Therefore, nanoplatforms containing mesoporous silica and a natural polysaccharide, ulvan, extracted from Ulva Lactuca seaweed, were developed for irinotecan. Either mesoporous silica-ulvan nanoplatforms or irinotecan-loaded materials were structurally and morphologically characterized. In vitro drug release experiments in phosphate buffer solution with a pH of 7.6 emphasized the complete recovery of irinotecan in 8 h. Slower kinetics were obtained for the nanoplatforms with a higher amount of natural polysaccharide. Ulvan extract proved to be biocompatible up to 2 mg/mL on fibroblasts L929 cell line. The irinotecan-loaded nanoplatforms exhibited better anticancer activity than that of the drug alone on human colorectal adenocarcinoma cells (HT-29), reducing their viability to 60% after 24 h. Moreover, the cell cycle analysis proved that the irinotecan loading onto developed nanoplatforms caused an increase in the cell number trapped at G0/G1 phase and influenced the development of the tumoral cells

New Hydrogel Formulations Based on Natural and Synthetic Polymers for Skin Regeneration

The skin, which represents about 16% of the total body mass, acts as a protective barrier against external microbial factors [1]. Therefore, damaged tissues, especially burns, require rapid local coverage to avoid infections and to ensure the protective barrier function of the skin [2]. The aim of this study was to design and characterize new hydrogel formulations based on natural and synthetic polymers and that were biodegradable and cytocompatible to serve as temporary dressings with regenerative properties for skin wound healing. The proposed experimental variants of the hydrogels are based on mixtures of gelatin (Gel), sodium alginate (Alg), polyvinyl alcohol (PVA), and methylcellulose (MC1500) in different weight ratios: Gel-Alg (1:0.75, g/g), Gel-Alg-PVA (1:0.27:0.18, g/g/g) and Gel-Alg-MC1500 (1:0.26:0.35, g/g/g). Physicochemical and biochemical characterizations were performed to determine the swelling degree, biodegradation in physiological conditions (pH 7.4, 37 °C) and in the presence of collagenase (mimicking the inflamed wounded milieu), viscosity, and syneresis, while their ultrastructure was investigated by SEM analysis [3]. The L929 murine fibroblast culture was used to assess the in vitro cytocompatibility of the hydrogels after 24 h and 48 h of cultivation using quantitative MTT and LDH assays [4]. Cell morphology was observed in treated cultures by light microscopy after Giemsa staining. The physicochemical and biochemical analyses indicated that the novel polymeric hydrogels variants had a good swelling capacity due to the presence of Alg, had an adjustable viscosity, and controlled biodegradation over time in both physiological and inflamed conditions. Two mixture variants were outlined: Gel-Alg-PVA with reduced porosity and low biodegradability over time and Gel-Alg-MC1500 with increased porosity and higher biodegradation over time, even in the physiological environment. The SEM morphology observations showed that the hydrogels had a dense and microporous structure, with pores of irregular shapes and sizes, which could ensure skin protection against external microbial agents while also maintaining the required degree of humidity and oxygen exchange with the external environment. In vitro quantitative tests indicated a high degree of cytocompatibility for all of the tested hydrogels, with cell viability percentages higher than 90%. The cell morphology observations revealed that in the presence of hydrogel samples, the L929 murine fibroblasts maintained their normal phenotype, and the cell density was similar to that of the negative control (untreated cells). Overall, our findings indicated that the hydrogels containing synthetic polymers (Gel-Alg-PVA, Gel-Alg-MC1500) had adequate physicochemical, biochemical, and biological properties that should be further tested to determine their role as biomaterials for skin tissue engineering applications

Cerium-Containing Mesoporous Bioactive Glasses (MBGs)-Derived Scaffolds with Drug Delivery Capability for Potential Tissue Engineering Applications

Finding innovative solutions to improve the lives of people affected by trauma, bone disease, or aging continues to be a challenge worldwide. Tissue engineering is the most rapidly growing area in the domain of biomaterials. Cerium-containing MBG-derived biomaterials scaffolds were synthesized using polymethyl methacrylate (PMMA) as a sacrificial template. The obtained scaffolds were characterized by X-ray powder diffraction (XRPD), infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The Ce4+/Ce3+ ratio in the scaffolds was estimated. In vitro testing revealed good cytocompatibility of the investigated scaffolds in mouse fibroblast cell line (NCTC clone L929). The results obtained regarding bioactivity, antibacterial activity, and controlled drug delivery functions recommend these scaffolds as potential candidates for bone tissue engineering applications

Chemopreventive functional food through selenium biofortification of cauliflower plants

The aim of this work was to develop a biotechnological approach for production of cauliflower as safe functional food, with an optimal content of chemopreventive compounds, by a protective biofortification, through selenium application together with betaine and spraying adjuvants. In the control and treated cauliflower plants we determined the amount of total selenium, glucosinolates (sulforaphane) and SAH (S-Adenosyl-homocysteine). We also assayed the chemopreventive effects of compounds formed in the treated cruciferous plants through in vitro tests, using human colorectal tumor cell line (CaCo2). Extracts of plants treated with selenium applied together with betaine and spraying adjuvant were significantly more active on reduction of tumoral cell viability than the extract of control plants. Cauliflower plants, obtained after our treatments for protective biofortification, were used to feed rabbits, for 10 days. The ingestion of biofortified cauliflower did not modify the hematological and biochemical parameters on the laboratory animals

Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyvalerate) (P(3HB-<i>co</i>-3HV))/Bacterial Cellulose (BC) Biocomposites for Potential Use in Biomedical Applications

The aim of this study was to obtain biocomposites consisting of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), bacterial cellulose (BC) and α-tocopherol by a melt processing technique for potential use in biomedical applications. The melt processing and roughness of biocomposites were evaluated and compared to sample without BC. The degradation rate of PHBV/BC biocomposites was measured in phosphate buffer saline (PBS) by determining the mass variation and evidencing of thermal and structural changes by differential scanning calorimetry (DSC) and attenuated total reflectance-Fourier transformed infrared spectrometry (ATR-FTIR). The cell viability, cell morphology, cell cycle distribution and total collagen content were investigated on murine NCTC fibroblasts. Overall, the adding of BC to polyester matrix led to an adequate melt processing of biocomposites and increased surface roughness and cytocompatibility, allowing the cells to secrete the extracellular matrix (collagen) and stimulate cell proliferation. Results showed that the PHBV/BC biocomposites were favorable for long-term degradation and could be used for the design of medical devices with controlled degradability

Preparation and Biocompatibility of Poly Methyl Methacrylate (PMMA)-Mesoporous Bioactive Glass (MBG) Composite Scaffolds

In recent years, the rising number of bone diseases which affect millions of people worldwide has led to an increased demand for materials with restoring and augmentation properties that can be used in therapies for bone pathologies. In this work, PMMA- MBG composite scaffolds containing ceria (0, 1, 3 mol%) were obtained by the phase separation method. The obtained composite scaffolds were characterized by X-ray diffraction, infrared spectroscopy, and scanning electron microscopy. UV–Vis measurement and EDX analysis confirmed the presence of cerium ions in the composite scaffolds. Evaluation of the in-vitro biocompatibility using MTT assay showed that composite scaffold containing 1 mol% of ceria presented higher viability than control cells (100%) for concentrations ranging between 5 and 50% after 96 h of incubation