Maria Helena Andrade Santana

Hyaluronan-based materials are widely described as hydrogels, yet the term is applied without distinction to preparations that differ fundamentally in structure. This review addresses that imprecision by separating three material states of hyaluronan and by stating the criteria under which each may be called a gel: native linear hyaluronan, which forms a viscoelastic solution; the hexadecylamide derivative (HYADD-4), which self-assembles into a transient physical network through hydrophobic interactions; and hyaluronan cross-linked with 1,4-butanediol diglycidyl ether (BDDE), which forms a covalent three-dimensional network. A material is treated as a gel here only where both a percolated network and a solid-like mechanical response are documented. On that basis the covalent network qualifies; the derivative is elastic-dominated across the walking-to-running frequency band of the knee but has not been subjected to a formal Winter–Chambon analysis and is therefore described as gel-like; and the linear solution does not qualify at the molecular weights and concentrations used in marketed viscosupplements. We review the chemistry of network formation, the comparative physicochemical and rheological behavior, including viscosity, the storage and loss moduli, boundary lubrication, and resistance to enzymatic and oxidative degradation, and the receptor-mediated and anti-inflammatory mechanisms of action involving CD44 and molecular-weight-dependent signaling. Clinical and regenerative applications in osteoarthritis viscosupplementation and in scaffolds and carriers for cells and growth factors are summarized for each state. The networked materials show higher viscoelasticity and greater resistance to degradation than the linear solution, which provides a rationale for longer residence; the evidence for a clinical advantage over conventional hyaluronan, however, rests largely on preclinical and rheological data. A three-state framework is proposed, pending validation, to support clearer interpretation of the literature and more rational material selection.

To mimic physiological microenvironments in organ-on-a-chip systems, physiologically relevant parameters are required to precisely access drug metabolism. Oxygen level is a critical microenvironmental parameter to maintain cellular or tissue functions and modulate their behaviors. Current organ-on-a-chip setups are oftentimes subjected to the ambient incubator oxygen level at 21%, which is higher than most if not all physiological oxygen concentrations. Additionally, the physiological oxygen level in each tissue is different ranging from 0.5 to 13%. Here, a closed-loop modular multiorgan-on-chips platform is developed to enable not only real-time monitoring of the oxygen levels but, more importantly, tight control of them in the range of 4 to 20% across each connected microtissue-on-a-chip in the circulatory culture medium. This platform, which consists of microfluidic oxygen scavenger(s), an oxygen generator, a monitoring/controller system, and bioreactor(s), allows for independent, precise upregulation and downregulation of dissolved oxygen in the perfused culture medium to meet the physiological oxygen level in each modular microtissue compartment, as needed. Furthermore, drug studies using the platform demonstrate that the oxygen level affects drug metabolism in the parallelly connected liver, kidney, and arterial vessel microtissues without organ–organ interactions factored in. Overall, this platform can promote the performances of organ-on-a-chip devices in drug screening by providing more physiologically relevant and independently adjustable oxygen microenvironments for desired organ types on a single- or a multiorgan-on-chip(s) configuration.

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.

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.

Hyaluronic acid (HA) is a natural polyelectrolyte abundant in mammalian connective tissues, such as cartilage and skin. Both endogenous and exogenous HA produced by fermentation have similar physicochemical, rheological, and biological properties, leading to medical and dermo-cosmetic products. Chemical modifications such as cross-linking or conjugation in target groups of the HA molecule improve its properties and in vivo stability, expanding its applications. Currently, HA-based scaffolds and matrices are of great interest in tissue engineering and regenerative medicine. However, the partial oxidation of the proximal hydroxyl groups in HA to electrophilic aldehydes mediated by periodate is still rarely investigated. The introduced aldehyde groups in the HA backbone allow spontaneous cross-linking with adipic dihydrazide (ADH), thermosensitivity, and noncytotoxicity to the hydrogels, which are advantageous for medical applications. This review provides an overview of the physicochemical properties of HA and its usual chemical modifications to better understand oxi-HA/ADH hydrogels, their functional properties modulated by the oxidation degree and ADH concentration, and the current clinical research. Finally, it discusses the development of biomaterials based on oxi-HA/ADH as a novel approach in tissue engineering and regenerative medicine.

Hyaluronic acid (HA) is a polysaccharide of alternating d-glucuronic acid and N-acetyl-d-glucosamine residues present in the extracellular matrix of connective, epithelial, and nervous tissues. Due to its singular hydrating, rheological and adhesive properties, HA has found numerous cosmetic and medical applications. However, techno-economic analyses of high value-added bioproducts such as HA are scarce in the literature. Here, we present a techno-economic analysis of a process for producing HA using Streptococcus zooepidemicus, simulated in SuperPro Designer. In the baseline scenario, HA is produced by batch fermentation, reaching 2.5 g/L after 24 h. It is then centrifuged, diafiltered, treated with activated carbon and precipitated with isopropanol. The product is suitable for topical formulations and its production cost was estimated as 1115 $/kg. A similar scenario, based on fed-batch culture and assuming a titer of 5.0 g/L, led to a lower cost of 946 $/kg. Moreover, in two additional scenarios, 10% of the precipitated HA is diverted to the production of a highly pure and high-molecular weight HA, suitable for injectable applications. These scenarios resulted in higher capital and operating costs, but also in higher profits, because HA for injectable use has a higher selling price that more than compensates for its higher production costs.

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.