Intra-Articular Hyaluronic Acid in Osteoarthritis and Tendinopathies: Molecular and Clinical Approaches.

Musculoskeletal diseases continue to rise on a global scale, causing significant socioeconomic impact and decreased quality of life. The most common disorders affecting musculoskeletal structures are osteoarthritis and tendinopathies, complicated orthopedic conditions responsible for major pain and debilitation. Intra-articular hyaluronic acid (HA) has been a safe, effective, and minimally invasive therapeutic tool for treating these diseases. Several studies from bedside to clinical practice reveal the multiple benefits of HA such as lubrication, anti-inflammation, and stimulation of cellular activity associated with proliferation, differentiation, migration, and secretion of additional molecules. Collectively, these effects have demonstrated positive outcomes that assist in the regeneration of chondral and tendinous tissues which are otherwise destroyed by the predominant catabolic and inflammatory conditions seen in tissue injury. The literature describes the physicochemical, mechanical, and biological properties of HA, their commercial product types, and clinical applications individually, while their interfaces are seldom reported. Our review addresses the frontiers of basic sciences, products, and clinical approaches. It provides physicians with a better understanding of the boundaries between the processes that lead to diseases, the molecular mechanisms that contribute to tissue repair, and the benefits of the HA types for a conscientious choice. In addition, it points out the current needs for the treatments.

Violência Obstétrica na Perspetiva dos Enfermeiros Especialistas em Enfermagem de Saúde Materna e Obstétrica / Obstetric Violence from the Perspective of Maternal and Obstetric Health Nurse Specialists

No âmbito do Mestrado em Enfermagem de Saúde Materna e Obstétrica, foi desenvolvido um estágio de natureza profissional onde se procurou adquirir e desenvolver competências necessárias para a prática de enfermagem especializada. Assim, com o presente relatório pretende-se descrever e refletir sobre as atividades desenvolvidas ao longo do 2º ano deste ciclo de estudos no âmbito dos cuidados especializados à mulher, ao recém-nascido e à família. Durante a trajetória da experiência prática desenvolvida nos vários locais de ensino clínico, surgiu a inquietação que deu o mote para o tema de investigac?a?o ?Violência obstétrica na perspetiva dos Enfermeiros Especialistas em Enfermagem de Saúde Materna e Obstetri?cia?. Ao longo deste percurso, foi possível o contacto com diversas narrativas, não só de mulheres, como também de profissionais de saúde. As primeiras, falando de violência na primeira pessoa. Os segundos, considerando que têm pouca voz relativamente a este tema. Tendo em conta a abrangência e complexidade do tema, optou-se por direcionar o estudo para a perspetiva dos profissionais de saúde, nomeadamente os Enfermeiros Especialistas em Enfermagem de saúde Materna e Obstétrica. Neste sentido, foi realizado um estudo qualitativo, exploratório e descritivo. A amostra foi constituída por nove Enfermeiras Especialistas em Enfermagem de Saúde Materna e Obstétrica, que desenvolvem a sua atividade em contexto de prestação de cuidados a mulheres ao longo de todo o seu ciclo gravídico. Como resultados encontrados podemos referir que os participantes do estudo reconhecem a existência de práticas que se enquadram no fenómeno ?violência obstétrica?, sendo que estas práticas poderão estar associadas a fatores estruturais, sociais ou individuais. Existe, no entanto, alguma consciencialização para a mudança de práticas, que implica a formação dos profissionais de saúde, o aumento da literacia em saúde das grávidas/sociedade, bem como um envolvimento mais ativo das instituições que regulam a profissão, nomeadamente a Ordem dos Enfermeiros.

Platelet-Rich Plasma Gel Matrix (PRP-GM): Description of a New Technique.

Several musculoskeletal conditions are triggered by inflammatory processes that occur along with imbalances between anabolic and catabolic events. Platelet-rich plasma (PRP) is an autologous product derived from peripheral blood with inherent immunomodulatory and anabolic properties. The clinical efficacy of PRP has been evaluated in several musculoskeletal conditions, including osteoarthritis, tendinopathy, and osteonecrosis. When used in combination with hyaluronic acid (HA), a common treatment alternative, the regenerative properties of PRP are significantly enhanced and may provide additional benefits in terms of clinical outcomes. Recently, a new PRP-derived product has been reported in the literature and is being referred to as "plasma gel". Plasma gels are obtained by polymerizing plasmatic proteins, which form solid thermal aggregates cross-linked with fibrin networks. Plasma gels are considered to be a rich source of growth factors and provide chemotactic, migratory, and proliferative properties. Additionally, clot formation and the associated fibrinolytic reactions play an additional role in tissue repair. There are only a few scientific articles focusing on plasma gels. Historically, they have been utilized in the fields of aesthetics and dentistry. Given that the combination of three products (PRP, HA, and plasma gel) could enhance tissue repair and wound healing, in this technical note, we propose a novel regenerative approach, named "PRP-HA cellular gel matrix" (PRP-GM), in which leukocyte-rich PRP (LR-PRP) is mixed with a plasma gel (obtained by heating the plasma up) and HA in one syringe using a three-way stopcock. The final product contains a fibrin-albumin network entangled with HA's polymers, in which the cells and biomolecules derived from PRP are attached and released gradually as fibrinolytic reactions and hyaluronic acid degradation occur. The presence of leukocytes, especially monocytes and macrophages, promotes tissue regeneration, as type 2 macrophages (M2) possess an anti-inflammatory feature. In addition, HA promotes the viscosuplementation of the joint and induces an anti-inflammatory response, resulting in pain relief. This unique combination of biological molecules may contribute to the optimization of regenerative protocols suitable for the treatment of degenerative musculoskeletal diseases.

How centrifugation influences the recovery and soy peptone incorporation in hyaluronic acid coils from fermentation.

Hyaluronic acid from fermentation (bio-HA) is a high-added-value biopolymer with a wide range of applications in aesthetic and medical areas. Bio-HA is a capsular exopolysaccharide highly hydrophilic with a coiled structure and high negative charge density at physiological pH. Centrifugation, currently used to separate microbial cells after fermentation, should significantly affect the bio-HA interactions with the medium components. Although the literature shows various centrifugation conditions for bio-HA, there is no concern about its recovery and characterization. We examined how centrifugation influences its recovery and characterized interactions with soy peptone (SP), used as a nitrogen source. We collected experimental data according to a statistical central composite design (CCD) planning with replicates at the central point with time (8-22 min) and spin (582xg to 12,402xg) as independent variables. The obtained results show bio-HA recoveries ranged from 45 ± 0.5-100% as dosed in the supernatant. The incorporation of amino acids and peptides from SP increased HA coils' density, causing losses due to sedimentation at higher spins and times. Furthermore, SP incorporation also changed the coils' size regarding HA standards of molar mass 106,105 and 104 Da. We conclude centrifugation should significantly impact downstream processing and the total cost of bio-HA production.

Spinal Reflex Recovery after Dorsal Rhizotomy and Repair with Platelet-Rich Plasma (PRP) Gel Combined with Bioengineered Human Embryonic Stem Cells (hESCs).

Dorsal root rhizotomy (DRZ) is currently considered an untreatable injury, resulting in the loss of sensitive function and usually leading to neuropathic pain. In this context, we recently proposed a new surgical approach to treat DRZ that uses platelet-rich plasma (PRP) gel to restore the spinal reflex. Success was correlated with the reentry of primary afferents into the spinal cord. Here, aiming to enhance previous results, cell therapy with bioengineered human embryonic stem cells (hESCs) to overexpress fibroblast growth factor 2 (FGF2) was combined with PRP. For these experiments, adult female rats were submitted to a unilateral rhizotomy of the lumbar spinal dorsal roots, which was followed by root repair with PRP gel with or without bioengineered hESCs. One week after DRZ, the spinal cords were processed to evaluate changes in the glial response (GFAP and Iba-1) and excitatory synaptic circuits (VGLUT1) by immunofluorescence. Eight weeks postsurgery, the lumbar intumescences were processed for analysis of the repaired microenvironment by transmission electron microscopy. Spinal reflex recovery was evaluated by the electronic Von Frey method for eight weeks. The transcript levels for human FGF2 were over 37-fold higher in the induced hESCs than in the noninduced and the wildtype counterparts. Altogether, the results indicate that the combination of hESCs with PRP gel promoted substantial and prominent axonal regeneration processes after DRZ. Thus, the repair of dorsal roots, if done appropriately, may be considered an approach to regain sensory-motor function after dorsal root axotomy.

Hyaluronic Acid in the Intestinal Tract: Influence of Structure, Rheology, and Mucoadhesion on the Intestinal Uptake in Rats.

Oral hyaluronic acid (HA) is a ubiquitous biopolymer that has gained attention as a treatment for local or systemic diseases. Here, we prepared and characterized structures of free HA (f-HA) with a high (>105 Da), intermediate (≤105 Da), and low (≤104 Da) average molar mass (MM); nanoparticles crosslinked with adipic dihydrazide (n-HA); and mixed formulations (mixed-HA) containing f-HA and n-HA. MM distribution determined the structure, hydrodynamic diameter, and zeta potential of the f-HAs. Crosslinking changed the physicochemical properties in n-HA. In vitro tack adhesion assays, using mucin tablets or a viable rat intestinal mucosa, showed better mucoadhesion with f-HA (intermediate MM) and mixed-HA (25% n-HA), especially in the jejunum segment. High MM f-HA presented negligible mucoadhesion. n-HA showed the deepest diffusion into the porous of the membranes. In vivo results showed that, except for high MM f-HA, there is an inverse relationship between rheological changes in the intestinal membrane macerates resulting from mucoadhesion and the effective intestinal permeability that led to blood clearance of the structures. We conclude that the n-HA formulations are promising for targeting other tissues, while formulations of f-HA (intermediate MM) and mixed-HA are better for treating dysbiosis.

Hyaluronic acid and fibrin from L-PRP form semi-IPNs with tunable properties suitable for use in regenerative medicine.

Autologous leukocyte- and platelet-rich plasma (L-PRP) combined with hyaluronic acid (HA) has been widely used in local applications for cartilage and bone regeneration. The association between L-PRP and HA confers structural and rheological changes that differ among individual biomaterials but has not been investigated. Therefore, the standardization and characterization of L-PRP-HA are important to consider when comparing performance results to improve future clinical applications. To this end, we prepared semi-interpenetrating polymer networks (semi-IPNs) of L-PRP and HA and characterized their polymerization kinetics, morphology, swelling ratio, stability and rheological behavior, which we found to be tunable according to the HA molar mass (MM). Mesenchymal stem cells derived from human adipose tissue (h-AdMSCs) seeded in the semi-IPNs had superior viability and chondrogenesis and osteogenesis capabilities compared to the viability and capabilities of fibrin. We have demonstrated that the preparation of the semi-IPNs under controlled mixing ensured the formation of cell-friendly hydrogels rich in soluble factors and with tunable properties according to the HA MM, rendering them suitable for clinical applications in regenerative medicine.

Leukocyte-rich PRP for knee osteoarthritis: Current concepts.

Knee osteoarthritis is a major painful and debilitating orthopaedic disease affecting a large number of adult individuals on a global scale. Over the years, this severe condition has been widely studied and while many alternatives have been utilized, platelet-rich plasma (PRP) remains one of the most popular solutions among researchers and clinicians alike. While there are different formulations and techniques involved in the preparation of PRP, produced either manually or via the use of commercial kits, the presence of leukocytes in a PRP mixture is a factor that raises concern due to their well-known pro-inflammatory activity. Although it is reasonable to worry about this, it should be taken into consideration that in order for the healing process to occur, the inflammatory phase is necessary. Leukocytes present in the inflammatory phase release both pro and anti-inflammatory molecules and, when combined with activated platelets, their potential increases. Additionally, due to the macrophage's plasticity to switch from the subtype 1 to subtype 2, it is suggested that the inclusion of the components from the buffy coat layer in a PRP mixture, classifying it as leukocyte-rich platelet-rich plasma or L-PRP, may provide benefits instead of detriments, from a standpoint of the regenerative potential of PRP.

Production of humic acids by solid-state fermentation of Trichoderma reesei in raw oil palm empty fruit bunch fibers.

Humic acids (HA) are organic macromolecules of high structural complexity and are primarily obtained from non-renewable carbon sources such as peat and coal. HA is widely used in agriculture but is known to have therapeutic properties, which are still poorly explored. Previous studies have shown the potential of biotechnological processes in the production of HA in submerged fermentation (SF) and solid-state fermentation (SSF) using pre-treated fibers of oil palm of empty fruit bunch (EFB) for the cultivation of Trichoderma strains. EFB is an agro-industrial residue that is readily available at a low cost. The present study aimed to study the production of HA by Trichoderma reesei in the SSF of raw fibers of EFBs from two different palm oil producers. Profiles of HA production, cellular protein, pH, glucose, moisture, and oxygen transfer were obtained during SSF by EFB with and without lipids, as well as, images by electron microscopy of fibers. Results showed efficient HA production in the raw fibers of the EFBs. HA production followed the cellular protein (6 g HA per 100 g of fibers) evolution of the fermentations in the absence of lipids, while the composition of lipids greatly affected its production. The best HA production (350 mg HA per 100 g of fibers) was obtained from EFB that was richer in lignocellulosics and the residual lipids were similar to the fractions of palm and kernel oils, while EFB with lower lignocellulosic presented a production 110 mg HA per 100 g of fibers.

Association of Platelet-Rich Plasma and Auto-Crosslinked Hyaluronic Acid Microparticles: Approach for Orthopedic Application.

Platelet-rich plasma (PRP) associated with high molecular weight hyaluronic acid (HA) has been clinically used for tissue regeneration in orthopedics. Despite the recognized beneficial clinical outcomes (e.g., early pain control, improvement of patients' functional limitation and longer-term effectiveness compared to PRP and HA alone in mild and moderate osteoarthritis treatments), its use is still challenging and controversial due to lack of standardization of association practical protocols. Moreover, most studies neglect the matrix structure, that generates the ultimate properties of the association among platelets, fibrin network and the microparticles. In the present work, we aimed to analyze the influence of the PRP/HA association with a controlled matrix structure on the stability, rheological behavior, release of growth factors and in vitro proliferation of human adipose-derived mesenchymal cells (h-AdMSCs). The attenuation of the negative charge of HA was also evaluated. Pure PRP (P-PRP) (i.e., plasma enriched with platelets and poor in leukocytes) was prepared by centrifugation and activated with serum and calcium chloride (AP-PRP). Autocrosslinked hyaluronic acid (AHA) was prepared by organocatalyzed auto-esterification and structured in microparticles (MPAHA) by shearing. The attenuation of the negative charge of MPAHA was performed with chitosan (CHT) by polyelectrolyte complexation yielding MPAHA-CHT. The results showed that microparticles (MPs) have viscoelastic properties, extrusion force and swelling ratio appropriate for injectable applications. The association of AP-PRP with the controlled structure of MPAHA and MPAHA-CHT formed a matrix composed of platelets and of a fibrin network with fibers around 160 nm located preferably on the surface of the MPs with an average diameter of 250 µm. Moreover, AP-PRP/MPAHA and AP-PRP/MPAHA-CHT associations were non-toxic and supported controlled growth factor (PDGF-AB and TGF-ß1) release and in vitro proliferation of h-AdMSC with a similar pattern to that of AP-PRP alone. The best h-AdMSC proliferation was obtained with the AP-PRP/MPAHA-CHT75:25 indicating that the charge attenuation improved the cell proliferation. Thus, the association of AP-PRP with the controlled structure of HA can be a valuable approach for orthopedic applications.

Hyaluronic acid behavior in oral administration and perspectives for nanotechnology-based formulations: A review.

Hyaluronic acid (HA) is a ubiquitous polysaccharide with diverse biological functions. Is known that in the intestinal epithelium, the exogenous HA of molar mass ≥105 Da orally administered antagonizes TLR4 overexpression resulting from dysbiosis and promotes immunomodulation in multifactorial crosstalk, thus helping to treat or to prevent injuries. As macromolecules mediate cell signaling, the three-dimensional structure of HA plays a vital role in those functions. Introducing HA in terms of its molecular structure, its spatial architecture as dependent on pH, concentration and molar mass, occurrence, biological functions and turnover in the tissues, this review addresses the HA in the gastrointestinal system, the molecular dynamics of intestinal uptake and signaling, immunomodulation at intestinal and systemic levels and HA fate to other tissues. Finally, at the light of these behaviors, a nanotechnological approach is presented as progress in the field of the oral HA administration and discussed with perspectives for future developments.

Centrifugation Conditions in the L-PRP Preparation Affect Soluble Factors Release and Mesenchymal Stem Cell Proliferation in Fibrin Nanofibers.

Leukocyte and platelet-rich plasma (L-PRP) is an autologous product that when activated forms fibrin nanofibers, which are useful in regenerative medicine. As an important part of the preparation of L-PRP, the centrifugation parameters may affect the release of soluble factors that modulate the behavior of the cells in the nanofibers. In this study, we evaluated the influences of four different centrifugation conditions on the concentration of platelets and leukocytes in L-PRP and on the anabolic/catabolic balance of the nanofiber microenvironment. Human adipose-derived mesenchymal stem cells (h-AdMSCs) were seeded in the nanofibers, and their viability and growth were evaluated. L-PRPs prepared at 100× g and 100 + 400× g released higher levels of transforming growth factor (TGF)-ß1 and platelet-derived growth factor (PDGF)-BB due to the increased platelet concentration, while inflammatory cytokines interleukin (IL)-8 and tumor necrosis factor (TNF)-α were more significantly released from L-PRPs prepared via two centrifugation steps (100 + 400× g and 800 + 400× g) due to the increased concentration of leukocytes. Our results showed that with the exception of nanofibers formed from L-PRP prepared at 800 + 400× g, all other microenvironments were favorable for h-AdMSC proliferation. Here, we present a reproducible protocol for the standardization of L-PRP and fibrin nanofibers useful in clinical practices with known platelet/leukocyte ratios and in vitro evaluations that may predict in vivo results.

Reflex arc recovery after spinal cord dorsal root repair with platelet rich plasma (PRP).

Spinal dorsal roots can be affected by a wide range of lesions, leading to a significant loss of proprioceptive information transmission and greatly affecting motor behavior. In this context, the reimplantation of lesioned roots with platelet-rich plasma (PRP) may allow nerve regeneration. Therefore, the present study evaluated sensorimotor improvement following dorsal root rhizotomy and repair with PRP. For this purpose, female Lewis rats were subjected to unilateral rhizotomy (RZ) of the L4-L6 dorsal roots and divided into the following groups: (1) the unlesioned control group; (2) the group that underwent rhizotomy (RZ) without repair; and (3) the group that underwent RZ followed by root repair with PRP. PRP was obtained from human blood and characterized regarding platelet concentration, integrity, and viability. Reflex arc recovery was evaluated weekly for eight weeks by the electronic von Frey method. The spinal cords were processed 1 week postlesion to evaluate the in vivo gene expression of TNFα, TGF-ß, BDNF, GDNF, VEGF, NGF, IL-4, IL-6, IL-13 by qRT-PCR and eight weeks postlesion to evaluate changes in the glial response (GFAP and Iba-1) and excitatory synaptic circuits (VGLUT1) by immunofluorescence. The results indicated that PRP therapy partially restores the paw withdrawal reflex over time, indicating the reentry of primary afferents from the dorsal root ganglia into the spinal cord without exacerbating glial reactivity. Additionally, the analysis of mRNA levels showed that PRP therapy has immunomodulatory properties. Overall, the present data suggest that the repair of dorsal roots with PRP may be considered a promising approach to improve sensorimotor recovery following dorsal rhizotomy.

Nanostructured lipid carriers loaded with free phytosterols for food applications.

The objective of this study was to develop nanostructured lipid carriers (NLCs) with free phytosterols (FP) using conventional fats and oils. Lipid matrices (LMs) and NLCs were produced with high oleic sunflower oil, fully hydrogenated canola (CA) and crambe (CR) oils by high-pressure homogenization (HPH). The NLCs were evaluated for hydrodynamic diameter (Z-ave), polydispersity index (PDI), and zeta potential (ZP). The melting behavior and polymorphism were investigated for both, the LMs and NLCs. The NLCs presented particle sizes ranging from 148.23 to 342.10 nm, PDI from 0.275 to 0.481, and ZP between -22.27 and -29.70 mV. The NLCs presented higher thermal resistance than that of the LMs. The use of CA and CR separately in the NLC formulations favored the incorporation of FP. The LMs and NLCs presented crystals in ß-form and in mixtures of ß' and ß forms. The developed NLCs can be used for food enrichment, such as spreads, margarine, and beverages.

Solid lipid nanoparticles as carriers for lipophilic compounds for applications in foods.

Nanotechnology is a new subject of interest in the field of food industry. Therefore, scientific and technological studies have been intensified in the last 10 years because of the promising results associated with the potential application of functional properties in food products, such as physical and chemical stability, protection and controlled release of bioactive compounds, and facilitated solubility of lipophilic compounds. Lipids have been used as raw material for the preparation of nanostructures, mainly owing to the solubilization capacity of lipophilic bioactive compounds, as well as because of the advantage of potentially using natural ingredients for production on an industrial scale. Thus, in this review, we describe the information reported in scientific literature on the chemical, physical, and structural properties of lipids used in the preparation of solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs). We reviewed the production methods; structural lipid components; emulsifying systems; bioactive lipophilic compounds; and the physical, thermal, and oxidative properties of SLN and NLC. In addition, important methods for characterizing these systems with regard to particle size, polydispersity index, zeta potential, morphology, crystallization behavior, and polymorphism are discussed with examples, in order to support studies that consider physical stability during processing and storage. Furthermore, studies on the applications of SLNs and NLCs in foods are only found for model systems, justifying the compilation of a series of studies on the potential applications to encourage future works. In addition, we have described the aspects still under discussion, related to the possible risks and regulatory aspects of nanotechnology in food.

Structural Modifications and Solution Behavior of Hyaluronic Acid Degraded with High pH and Temperature.

Hyaluronic acid (HA) is a macromolecule with valuable benefits over its range of molar masses (MM). Degradation studies are relevant to maintain the same purity level in biomedical studies when using HA of different MM. We degraded HA via high pH and temperature and evaluated its MM, solution behavior, and structure over time. After 24 h, low MM HA was predominant, and the MM decreased from 753 to 36.2 kDa. Dynamic light scattering (DLS) showed a decrease in the number of HA populations, and the solution tended to be less polydispersed. The zeta potential varied from - 10 to - 30 mV, close to the stable range. FTIR showed that the primary structure of HA was affected after only 48 h of reaction. These results are relevant for the production of low MM HA to be used or mixed with high MM HA, generating structured biomaterials for biomedical applications.

Liposomal-based lidocaine formulation for the improvement of infiltrative buccal anaesthesia.

This study describes the encapsulation of the local anaesthetic lidocaine (LDC) in large unilamellar liposomes (LUV) prepared in a scalable procedure, with hydrogenated soybean phosphatidylcholine, cholesterol and mannitol. Structural properties of the liposomes were assessed by dynamic light scattering, nanoparticle tracking analysis and transmission electron microscopy. A modified, two-compartment Franz-cell system was used to evaluate the release kinetics of LDC from the liposomes. The in vivo anaesthetic effect of liposomal LDC 2% (LUVLDC) was compared to LDC 2% solution without (LDCPLAIN) or with the vasoconstrictor epinephrine (1:100 000) (LDCVASO), in rat infraorbital nerve blockade model. The structural characterization revealed liposomes with spherical shape, average size distribution of 250 nm and low polydispersity even after LDC incorporation. Zeta potential laid around -30 mV and the number of suspended liposomal particles was in the range of 1012 vesicles/mL. Also the addition of cryoprotectant (mannitol) did not provoke structural changes in liposomes properties. In vitro release profile of LDC from LUV fits well with a biexponential model, in which the LDC encapsulated (EE% = 24%) was responsible for an increase of 67% in the release time in relation to LDCPLAIN (p < 0.05). Also, the liposomal formulation prolonged the sensorial nervous blockade duration (∼70 min), in comparison with LDCPLAIN (45 min), but less than LDCVASO (130 min). In this context, this study showed that the liposomal formulations prepared by scalable procedure were suitable to promote longer and safer buccal anaesthesia, avoiding side effects of the use of vasoconstrictors.

Distribution, recovery and concentration of platelets and leukocytes in L-PRP prepared by centrifugation.

Platelet-rich plasma (PRP) is an autologous product prepared from whole blood (WB) that is widely used in regenerative medicine. In clinical practice, discontinuous centrifugation is used for both hand- and machine-prepared PRP. However, separation of WB fractions via centrifugation is a complex process, and the lack of clear mechanisms limits the understanding and evaluation of PRP preparation methods This paper focuses on the distribution, recovery and concentration factor of platelets and leukocytes in L-PRP (leukocyte and platelet-rich plasma) to define a concentration pattern for these blood components due to centrifugation conditions. WB collected from three healthy donors was centrifuged for 10min at 50-800 xg in a first step and then at 400 xg in a second step. The results from the first centrifugation step showed most platelets to be distributed in the upper layer (UL) and the buffy coat (BC), with approximately 14.5±5.2% retained in the bottom layer (BL). Most leukocytes were present in the BL. The greatest platelet recoveries from L-PRP were obtained at up to 150 xg (88.5±16.9%). The cumulative concentration factors with respect to the WB from the second centrifugation step were 6 and 1.2 for platelets and leukocytes, respectively. Thus, the concentration patterns delineated three centrifugation ranges with platelet/leukocyte ratios of 205±18, 325±15 and 107±4 and lymphocyte/granulocyte ratios of 1.54±0.74, 0.90±0.08 and 0.42±0.07. These findings contribute to a scientifically based standardization of L-PRP preparations.

Contributions for classification of platelet rich plasma - proposal of a new classification: MARSPILL.

Platelet-rich plasma (PRP) has emerged as a significant therapy used in medical conditions with heterogeneous results. There are some important classifications to try to standardize the PRP procedure. The aim of this report is to describe PRP contents studying celular and molecular components, and also propose a new classification for PRP. The main focus is on mononuclear cells, which comprise progenitor cells and monocytes. In addition, there are important variables related to PRP application incorporated in this study, which are the harvest method, activation, red blood cells, number of spins, image guidance, leukocytes number and light activation. The other focus is the discussion about progenitor cells presence on peripherial blood which are interesting due to neovasculogenesis and proliferation. The function of monocytes (in tissue-macrophages) are discussed here and also its plasticity, a potential property for regenerative medicine treatments.

Antimicrobial activity of polymyxin-loaded solid lipid nanoparticles (PLX-SLN): Characterization of physicochemical properties and in vitro efficacy.

Antimicrobial resistance is a current public health concern, limiting the available therapeutic options used for the treatment of common bacterial infections. The development of new drug entities via biotechnological processes is however expensive and time-consuming. Therefore, old antimicrobial agents have been recovered for clinical use. An example of these drugs is polymyxin, which is known for its serious adverse side effects, such as nephrotoxicity, neurotoxicity and promotion of skin pigmentation. To overcome these limitations, the use of biodegradable nanoparticles has been proposed to allow site-specific targeting, increasing the drug's bioavailability and decreasing its side effects. The aim of this work was the development of an optimized pharmaceutical formulation composed of solid lipid nanoparticles (SLN) loading polymyxin B sulphate (PLX) for the treatment of bacterial infections. The PLX-loaded SLN were produced by a double emulsion method (w/o/w), obtaining particles with a mean size of approximately 200nm, polydispersity of 0.3 and zeta potential of -30mV. The encapsulation efficiency reached values above 90% for all developed formulations. SLN remained stable for a period of 6months of storage at room temperature. The occlusive properties of the SLN was shown to be dependent on the type of lipid, while the antimicrobial properties of PLX-loaded SLN were effective against resistant strains of Pseudomonas aeruginosa. Results from the differential scanning calorimetry (DSC), wide angle X-ray diffraction (WAXD) and small angle X-ray scattering (SAXS) analyses confirmed the crystallinity of the inner SLN matrices, suggesting the capacity of these particles to modify the release profile of the loaded drug.