Bacterial Sources Research Articles

The secondary metabolism collaboratory: a database and web discussion portal for secondary metabolite biosynthetic gene clusters.

Secondary metabolites are small molecules produced by all corners of life, often with specialized bioactive functions with clinical and environmental relevance. Secondary metabolite biosynthetic gene clusters (BGCs) can often be identified within DNA sequences by various sequence similarity tools, but determining the exact functions of genes in the pathway and predicting their chemical products can often only be done by careful, manual comparative analysis. To facilitate this, we report the first release of the secondary metabolism collaboratory (SMC), which aims to provide a comprehensive, tool-agnostic repository of BGC sequence data drawn from all publicly available and user-submitted bacterial and archaeal genome and contig sources. On the website, users are provided a searchable catalog of putative BGCs identified from each source, along with visualizations of gene and domain annotations derived from multiple sequence analysis tools. SMC's data is also available through publicly-accessible application programming interface (API) endpoints to facilitate programmatic access. Users are encouraged to share their findings (and search for others') through comment posts on BGC and source pages. At the time of writing, SMC is the largest repository of BGC information, holding 13.1M BGC regions from 1.3M source sequences and growing, and can be found at https://smc.jgi.doe.gov.

Bioengineered therapeutic systems for improving antitumor immunity

Abstract Immunotherapy, a monumental advancement in antitumor therapy, still yields limited clinical benefits owing to its unguaranteed efficacy and safety. Therapeutic systems derived from cellular, bacterial, and viral sources possess inherent properties that are conducive to antitumor immunotherapy. However, crude biomimetic systems have restricted functionality and may produce undesired toxicity. With the advances in biotechnology, various toolkits are available to add or subtract certain properties of living organisms to create flexible therapeutic platforms. This review elaborates on the creation of bioengineered systems by gene editing, synthetic biology, and surface engineering to enhance immunotherapy. The modifying strategies on the systems are discussed, including equipment of navigation and recognition systems to improve therapeutic precision, introduction of controllable components to control the duration and intensity of treatment, addition of immunomodulatory components to amplify immune activation, and removal of toxicity factors to ensure biosafety. Finally, we summarize the advantages of bioengineered immunotherapeutic systems and possible directions for their clinical translation.

The effect of clinically relevant antibiotics on bacterial extracellular vesicle release from E. coli.

Sepsis, a leading cause of death in hospitals, can be defined as a dysregulated host inflammatory response to infection, which can lead to tissue damage, organ failure, and cardiovascular complications. Although there is no cure for sepsis, the condition is typically managed with broad-spectrum antibiotics to eliminate any potential bacterial source of infection. However, a potential side-effect of antibiotic treatment is the enhanced release of bacterial extracellular vesicles (BEVs), membrane-bound nanoparticles containing proteins and other biological molecules from their parent bacterium. Some of the Gram-negative EV cargo, including Peptidoglycan associated lipoprotein (Pal) and Outer membrane protein A (OmpA), have been shown to induce both acute and chronic inflammation in host tissue. We hypothesize that antibiotic concentration and its mechanism of action can have an effect on the amount of released BEVs, which could potentially exacerbate the host inflammatory response. In this study, we evaluated nine clinically relevant antibiotics for their effect on EV release from Escherichia coli. Several beta-lactam antibiotics caused significantly more EV release, while quinolone and aminoglycosides caused relatively less vesiculation. Further study is warranted to corroborate the correlation between an antibiotic's mechanism of action and its effect on EV release, but these results underline the importance of antibiotic choice when treating sepsis patients.

Bacterial Communities of House Flies from Beef and Dairy Cattle Operations are Diverse and Contain Pathogens of Medical and Veterinary Importance.

Adult house flies (Musca domestica L.) are important reservoirs and mechanical vectors of bacteria in livestock operations. House fly bacterial communities are influenced by their local environment, yet a comprehensive understanding of bacterial diversity, pathogen prevalence, and bacterial source is not fully understood. We characterized bacterial communities from adult female house flies and associated manure samples from beef and dairy cattle farms in Kansas, Oklahoma, and Texas over four months (July-October). Bacterial community composition in flies and manure reflected the local environment, and house flies shared the majority (≥ 99%) of bacterial taxa with manure. The variability of bacterial diversity was greater among individual fly (species richness range: 48-1747) samples than manure (species richness range: 345-1162). Temporal variability of fly bacterial diversity was observed within each farm type. Bacterial taxa of veterinary and medical importance such as Corynebacterium, Turicibacter, Staphylococcus, Streptococcus, and Acinetobacter were highly prevalent in flies, constituting core bacterial communities. The prevalence of bacterial taxa associated with bovine keratoconjunctivitis (IBK) and bovine respiratory disease (BRD) was higher in flies than in manure and prevalence varied monthly. This study underscores the crucial role house flies play as carriers of cattle pathogens, contributing to their dissemination among animals and to off-site locations, where they pose a threat to surrounding communities and agricultural operations.

Enzymatic Glycosylation of 4'-Hydroxychalcones: Expanding the Scope of Nature's Catalytic Potential.

Chalcones, including 4'-hydroxychalcones, have garnered significant attention in the area of drug discovery due to their diverse pharmacological properties, such as anti-inflammatory, antioxidative, and anticancer effects. However, their low water solubility and bioavailability limit their efficacy in vivo. Glycosylation presents a promising approach to enhance the water solubility, stability, and metabolic properties of chalcones. This study investigates the enzymatic glycosylation of eight chemically synthesized 4'-hydroxychalcones using a diverse set of sugar glucosyltransferases from bacterial, plant, and fungal sources, alongside Glycine max sucrose synthase (GmSuSy) in a cascade reaction. Among the tested enzymes, five exhibited a remarkable versatility for glycoside production, and for large-scale biotransformation, flavonoid 7-O-glucosyltransferase Sbaic7OGT from Scutellaria baicalensis was selected as the most effective. As a result of the experiments conducted, eight trans-chalcone glycosides were obtained. During the purification of the reaction products, we also observed the isomerization of the products by simple sunlight exposure, which resulted in eight additional cis-chalcone glycosides. This study highlights the novel use of a cascade reaction involving Glycine max sucrose synthase (GmSuSy) for the efficient glycosylation of trans-4'-hydroxychalcones, alongside the unexpected discovery of cis-chalcone glycosides during the purification process.

Molecular Characterization of a New Bacterial Source of Naringinase as Paenibacillus stellifer RAMCM-44 and Fermentative Production of Naringinase at Shake-Flask Level

Molecular Characterization of a New Bacterial Source of Naringinase as Paenibacillus stellifer RAMCM-44 and Fermentative Production of Naringinase at Shake-Flask Level

B-019 Reduction in Culture Media Costs, without Impacting Turnaround Time, Through Implementation of an Automated Microbiology Platform and Gram Stain Interpretation

Abstract Background Automated plating instruments have been shown to improve quality in Microbiology laboratories in a number of ways including consistent streaking patterns that provide better isolation of bacterial colonies compared to manual methods. The time per culture set up and automated camera imaging is dependent upon the amount of media set, such that more individual plates slow down turn around time. Conversely, smaller media surface area and the use of non-selective media may result in mixed colonies that require sub-culture for isolation before identification and susceptibilities can be performed. Based upon these findings, we attempted to reduce our media usage per culture with the implementation of automation, with the goal of using as few plates as possible while at the same time not negatively impacting the turn around time of the culture via the need to subculture poorly isolated colonies. We conducted a pre-and post-implementation study evaluating the costs per culture and turn around time delays related to subculturing in a subset. Methods In the pre-implementation period, from April-January 2023, three individual media plates (sheep blood agar, chocolate agar and CNA/MacConkey bi-plate) were routinely set for general aerobic bacterial culture sources including swabs and body fluids. In the post implementation period, from April-January 2024, a single sheep blood/chocolate agar bi-plate was set unless the gram stain showed evidence of mixed organisms (defined as more than one morphology) at which time a secondary selective media (CNA/MacConkey) was manually set. For urine cultures, in the pre-implementation period 2 plates were set (sheep blood and CNA/MacConkey bi-plate) and in the post-implementation period a single chromogenic agar/sheep blood bi-plate was set. Total numbers of cultures in both periods were collected as well as the number of cultures for which additional media were set based on gram stain results. Orders of subculture media in the laboratory information system were compared per positive cultures. Statistical analysis was performed in GraphPad Prism. Results In the pre- versus post-implementation periods, a total of 16,454 vs 17,782 general aerobic cultures and 50,680 vs 53,916 urine cultures were set, respectively. Only 3.6% of general cultures required additional media set based on gram stain. The difference in cost between pre- and post-implementation was approximately a two-thirds reduction in cost for general cultures and a 25% reduction for urine cultures, with a total savings of approximately $34,238 in media avoidance in the post-implementation period. The frequency of positive cultures requiring subculture was not significantly different between the pre- (average 36% of positive growth) and post-implementation (average 22% of positive growth) periods for both culture types and may actually have been reduced due to automation. Conclusions The implementation of reduced media concomitant with streaking via automation resulted in significant cost savings while not negatively impacting isolation, which in turn impacts culture turn around times. The automation may have reduced need for subculture, perhaps due to better isolation, thus subsequent analyses will explore this additional finding.

Isolation and characterization of alginate producing Azotobacter sp, isolated from rhizospheric soil of Oryza sativa (L.) and Triticum aestivum (L.)

Alginate as biopolymer consisting of β-D Mannuronic acid and α-L Guluronic acid monomer residues linked by (1-4) glycosidic bond. Brown seaweeds mostly Laminaria sp, Ascophyllum nodosum, Eclonia maxima, Sargassum sp. and Macrocystis pyrifera were earlier used for commercial alginate production. Major drawback of using sea weeds for alginate production were high production cost, impurity issue and safety concerns includes the reasons behind discovery of novel bacterial sources to make an efficient alginate production system. Later two bacterial genera Pseudomonas and Azotobacter were reported for alginate production. However, due to pathogenicity, and quality issue associated with some Pseudomonas strains, Azotobacter sp. was found to be most reliable and prioritized bacterial sources for alginate production with appreciable qualities which will be ready to be used for industrial applications. Therefore, current research aims to isolate and characterize a new strain of Azotobacter sp. with alginate-producing ability. Two different soil samples were collected from two places Deokali and Parsurampur, Jaunpur district in the spring (06 August 2022) and winter (06 February, 2023) seasons from Oryza sativa and Triticum aestivum rhizospheric region respectively. Azotobacter sp. was isolated by using Nitrogen Free Burk’s medium and alginate production was confirmed by gel formation assay method in the presence of CuSO4 and CaCl2 solutions. The carbohydrate qualitative test was done by using Molisch’s Test. Two novel strains Azotobacter chroococcum PU-RSD01 and Azotobacter chroococcum PU-RSD02 were isolated on the basis of high alginate producing ability. Morphological, biochemical and molecular techniques further characterized the isolates.

Molecular phylogenetic characterization of L-asparaginase-producing endophytic fungi inhabiting Prunus africana and Periploca linearifolia: Effect of incubation time and pH on enzyme production

The therapeutic use of L-asparaginase derived from bacterial sources has been constrained by various challenges, including toxicity and repression. This has prompted the exploration of alternative sources, particularly eukaryotic microorganisms like fungi, to enhance the safety and effectiveness of the L-asparaginase enzyme. This study aims to investigate the fungal endophytes inhabiting Periploca linearifolia and Prunus africana as a potential source of novel L-asparaginase. Additionally, the study seeks to examine the impact of incubation time and pH on L-asparaginase production. Standard surface sterilization techniques were used for isolation of endophytic fungi. Based on the research findings, 24 % of the fungal endophytes demonstrated a positive reaction for L-ASNase activity and were identified as Colletotrichum sydowii, Fusarium sporotrichioides, Fusarium solani, Cercospora canescens, Penicillium crustosum, Penicillium pancosmium, Phoma sp, Penicillium ubiquetum, septoria sp and Penicillium commune. A significant variation in the production of L-asparaginase based on the time of incubation and pH was observed. Most endophytic fungal isolates exhibit optimal enzyme activity on the 6th day of incubation. However, both Septoria sp and Colletotrichum sydowii recorded the highest L-asparaginase activity on the 9th day of incubation. Fusarium solani demonstrated peak activity of 12.4 ± 1.12 UI/mL on the 12th day of incubation. The optimal pH for L-asparaginase production by fungal endophytes was found to be between 5 and 6. The fungal endophytes isolated from medicinal plants have the potential to serve as sources of novel L-asparaginase. Furthermore, it is evident that fungal endophytes display considerable variation in L-asparaginase production, influenced by the duration of incubation and pH conditions.

Bacterial Communities of House Flies from Dairy Farms Highlight Their Role as Reservoirs, Disseminators, and Sentinels of Microbial Threats to Human and Animal Health.

Adult house flies (Musca domestica L.) inhabiting dairy farms not only are nuisance pests but also harbor and disseminate bacteria. We examined the bacterial community composition, diversity, environmental sources, and prevalence in individual adult female house flies and cattle manure samples collected monthly from Florida, North Carolina, and Tennessee dairy farms between May and August 2021. Individual house flies carried diverse bacterial communities, encompassing all bacterial taxa (100%) identified across manure samples, and additional species likely acquired from the animals. Bacterial community assemblage in house flies and manure samples within farms varied by month. Some taxa were differentially associated with either house flies (Corynebacterium, Acinetobacter, and Staphylococcus) or manure samples (Treponema, Succinivibrio, and Clostridia). House fly bacterial communities mostly contained specialist species originating from manure, with several taxa (Escherichia, Corynebacterium, Turicibacter) being potential pathogens of livestock and humans. These findings further support the role of house flies as carriers of cattle-associated bacteria, including pathogens, and their potential for disseminating these microbes among cattle and to neighboring environments. Since their bacterial communities provide a snapshot of their surrounding environment, house flies also serve as effective sentinels in xenosurveillance strategies.

Structural identification and immunostimulatory effect of Bacillus velezensis GV1 polysaccharides via TLR4/NF-κB signaling pathway in RAW264.7 macrophages

Microbial polysaccharides derived from bacterial sources possess unique properties because of their structural complexity contributing to exceptional characteristics, including potent immunostimulatory effects. In this study, we extracted crude polysaccharide from Bacillus velezensis GV1 (BPS) which was isolated from Korean ginseng vinegar, and subsequently characterized for sugar composition and functional groups using FT-IR and methylation method. Structural analysis indicated that BPS was composed of mannan and glucan in a ratio of 7.5:2.5. The immunostimulatory effect of BPS was investigated in RAW264.7 macrophages. The results revealed that BPS significantly increased NO production, as well as the secretion and expression of key cytokines, such as IL-6, TNF-α, and IL-1β. These effects were confirmed using a TLR4 antagonist (TAK-242). Moreover, BPS exhibited immunostimulatory potential by promoting the NF-κB signaling pathway. In conclusion, this study establishes a foundation for the potential application of BPS as an immunostimulatory adjuvant or alternative component in functional foods, particularly for enhancing innate immune responses.

Advances in Microbial Exopolysaccharides: Present and Future Applications.

Microbial exopolysaccharides (EPSs) are receiving growing interest today, owing to their diversity in chemical structure and source, multiple functions, and immense potential applications in many food and non-food industries. Their health-promoting benefits for humans deserve particular attention because of their various biological activities and physiological functions. The aim of this paper is to provide a comprehensive review of microbial EPSs, covering (1) their chemical and biochemical diversity, including composition, biosynthesis, and bacterial sources belonging mainly to lactic acid bacteria (LAB) or probiotics; (2) their technological and analytical aspects, especially their production mode and characterization; (3) their biological and physiological aspects based on their activities and functions; and (4) their current and future uses in medical and pharmaceutical fields, particularly for their prebiotic, anticancer, and immunobiotic properties, as well as their applications in other industrial and agricultural sectors.

Bacterial Pigments as a Promising Alternative to Synthetic Colorants: From Fundamentals to Applications.

Pigments find widespread application in the fields of food, medicine, textiles, and cosmetics. At present, synthetic colorants dominate the global pigment market. However, the environmental and health hazards associated with synthetic colorants have spurred extensive research on eco-friendly and safe alternatives. Natural pigments are particularly intriguing for meeting consumer demands and sustainable development, as they not only exhibit various vibrant color shades without discernible toxic side effects but also offer additional healthful features such as antibacterial, antioxidant, anticancer, and antiviral properties compared with their synthetic counterparts. Among natural sources, bacterial strains share distinct advantages for large-scale pigment production because of their intrinsic robustness of cellular metabolic systems. This review comprehensively outlines the bacterial sources, extraction and purification methods, structural characteristics, biological activities, and potential applications of typical pigments, including but not limited to violacein, indigoidine, melanin, carotenoids, prodigiosin, and rhodopsin. Additionally, it underscores the primary obstacles to the development and production of bacterial pigments for commercial applications, discussing feasible strategies for overcoming production bottlenecks. This work also provides valuable insights for the scientific and rational advancement of bacterial pigment development.

How carbon sources drive cellulose synthesis in two Komagataeibacter xylinus strains

Bacterial cellulose synthesis from defined media and waste products has attracted increasing interest in the circular economy context for sustainable productions. In this study, a glucose dehydrogenase-deficient Δgdh K2G30 strain of Komagataeibacter xylinus was obtained from the parental wild type through homologous recombination. Both strains were grown in defined substrates and cheese whey as an agri-food waste to assess the effect of gene silencing on bacterial cellulose synthesis and carbon source metabolism. Wild type K2G30 boasted higher bacterial cellulose yields when grown in ethanol-based medium and cheese whey, although showing an overall higher d-gluconic acid synthesis. Conversely, the mutant Δgdh strain preferred d-fructose, d-mannitol, and glycerol to boost bacterial cellulose production, while displaying higher substrate consumption rates and a lower d-gluconic acid synthesis. This study provides an in-depth investigation of two K. xylinus strains, unravelling their suitability for scale-up BC production.

Potential antimicrobial and fruit juice clarification activity of amylase enzyme from Bacillus strains

The hydrolytic enzyme, amylase possesses wide industrial applications and its production from bacterial sources by submerged fermentation is much simplified and economical. The research aimed to characterize amylase-producing bacteria and evaluate their potential for amylase activity regarding antimicrobial and fruit juice clarification. In current study, Bacillus licheniformis, Bacillus amyloliquifaciens, Bacillus cereus, Bacillus subtilis and Bacillus paramycoides was identified by 16S rRNA sequencing. After submerged fermentation, amylase activity of bacteria was measured by 3, 5-dinitro salicylic acid (DNS) assay. A substantial amount of amylase (423.47 mg/ml) in crude extract was measured by Bradford protein assay. Later, ammonium sulfate (80 %) precipitated partially purified amylase showed 1.6 times enhanced amylase activity (1484.94 U/ml) compared to crude amylase (973.23 U/ml). For highest amylase production, 72 h of optimum fermentation period was recorded at pH 7 with 2 % starch as substrate. Potent thermophilic amylase activity was observed at 65 °C. In apple juice clarification activity of amylase, turbidity of juice was reduced to 54.18 %. Potential antimicrobial property of amylase was detected with largest zone of inhibition against Escherichia coli ATCC 25922 (22.36 mm) and Mucor sp. ATCC 48559 (22.45 mm). Considering promising amylase properties, amylase-producing Bacillus strains from rice mill soil can be fermented for large scale amylase production providing application for industrial purposes including fruit juice clarification and antimicrobial activities. It will also overthrow the requirement of employing expensive and harmful chemicals in fruit juice clarification and combating pathogens.

Activating UCHL1 through the CRISPR activation system promotes cartilage differentiation mediated by HIF-1α/SOX9.

Developing strategies to enhance cartilage differentiation in mesenchymal stem cells and preserve the extracellular matrix is crucial for successful cartilage tissue reconstruction. Hypoxia-inducible factor-1α (HIF-1α) plays a pivotal role in maintaining the extracellular matrix and chondrocyte phenotype, thus serving as a key regulator in chondral tissue engineering strategies. Recent studies have shown that Ubiquitin C-terminal hydrolase L1 (UCHL1) is involved in the deubiquitylation of HIF-1α. However, the regulatory role of UCHL1 in chondrogenic differentiation has not been investigated. In the present study, we initially validated the promotive effect of UCHL1 expression on chondrogenesis in adipose-derived stem cells (ADSCs). Subsequently, a hybrid baculovirus system was designed and employed to utilize three CRISPR activation (CRISPRa) systems, employing dead Cas9 (dCas9) from three distinct bacterial sources to target UCHL1. Then UCHL1 and HIF-1α inhibitor and siRNA targeting SRY-box transcription factor 9 (SOX9) were used to block UCHL1, HIF-1α and SOX9, respectively. Cartilage differentiation and chondrogenesis were measured by qRT-PCR, immunofluorescence and histological staining. We observed that the CRISPRa system derived from Staphylococcus aureus exhibited superior efficiency in activating UCHL1 compared to the commonly used the CRISPRa system derived from Streptococcus pyogenes. Furthermore, the duration of activation was extended by utilizing the Cre/loxP-based hybrid baculovirus. Moreover, our findings show that UCHL1 enhances SOX9 expression by regulating the stability and localization of HIF-1α, which promotes cartilage production in ADSCs. These findings suggest that activating UCHL1 using the CRISPRa system holds significant potential for applications in cartilage regeneration.

Co-exposure to different bacterial species’ lipopolysaccharides with the NASH diet exacerbates NASH and liver fibrosis progression in mice

Background and aimWith the obesity epidemic, nonalcoholic fatty liver disease (NAFLD) has become a public health concern, but its progression mechanism remains unclear. Experimental models mimicking human NAFLD/steatohepatitis (NASH) are crucial. This study simulates gut microbiota imbalance effects on NASH and liver fibrosis. MethodsWe used different bacterial sources of lipopolysaccharide (LPS), including Escherichia coli (GEC) and Salmonella abortus equi (GSE), combined with a Gubra Amylin NASH (GAN) diet to induce NASH and liver fibrosis. ResultsThe GSE group showed significantly higher serum alanine aminotransferase, hydroxyproline, CD68-positive cells, α-smooth muscle actin, glial fibrillary acidic protein, and TNF-α, COL1A1, TGF-β, and NLRP3 expressions compared to the the GAN group. The GSE group also had higher Erysipelotrichaceae, Akkermansiaceae, and Bacteroidaceae family numbers. ConclusionsThe GAN diet with LPS treatment successfully induced NASH and fibrosis making this model useful for preclinical NASH drug testing.

Bacterial Purine Nucleoside Phosphorylases from Mesophilic and Thermophilic Sources: Characterization of Their Interaction with Natural Nucleosides and Modified Arabinofuranoside Analogues.

The enzymatic synthesis of nucleoside derivatives is an important alternative to multi-step chemical methods traditionally used for this purpose. Despite several undeniable advantages of the enzymatic approach, there are a number of factors limiting its application, such as the limited substrate specificity of enzymes, the need to work at fairly low concentrations, and the physicochemical properties of substrates-for example, low solubility. This research conducted by our group is dedicated to the advantages and limitations of using purine nucleoside phosphorylases (PNPs), the main enzymes for the metabolic reutilization of purines, in the synthesis of modified nucleoside analogues. In our work, the substrate specificity of PNP from various bacterial sources (mesophilic and thermophilic) was studied, and the effect of substrate, increased temperature, and the presence of organic solvents on the conversion rate was investigated.

Cerebellar abscess secondary to metastatic lung adenocarcinoma: a case report

BackgroundCerebellar abscesses are rare, life-threatening infections often originating from bacterial sources, while metastatic brain lesions from lung adenocarcinoma are relatively common. However, the coexistence of a cerebellar abscess secondary to metastatic lung adenocarcinoma is exceedingly rare and presents unique diagnostic and management challenges.Case PresentationWe report a case of a 35 year-old Pakistani female patient with persistent headaches, nausea, and vertigo, who was found to have a large cerebellar mass with features suggestive of metastatic lung adenocarcinoma. Further investigation revealed a concomitant cerebellar abscess. Surgical excision and broad-spectrum antibiotics were initiated, resulting in a favorable outcome.ConclusionThis case showcases the rarity and complexity of cerebellar abscesses due to metastatic lung adenocarcinoma. Timely intervention, including surgery and targeted therapy, is crucial for successful management. Further research is needed to enhance treatment strategies.

ViSCNOVAS: A Novel Classification System for Hyaluronic Acid-Based Gels in Orthobiologic Products and Regenerative Medicine.

Hyaluronic acid (HA), a naturally occurring polysaccharide, holds immense potential in regenerative medicine due to its diverse biological functions and clinical applications, particularly in gel formulations. This paper presents a comprehensive exploration of HA, encompassing its origins, molecular characteristics, and therapeutic roles in gel-based interventions. Initially identified in bovine vitreous humor, HA has since been found in various tissues and fluids across vertebrate organisms and bacterial sources, exhibiting consistent physicochemical properties. The synthesis of HA by diverse cell types underscores its integral role in the extracellular matrix and its relevance to tissue homeostasis and repair. Clinical applications of HA, particularly in addressing musculoskeletal ailments such as osteoarthritis, are examined, highlighting its efficacy and safety in promoting tissue regeneration and pain relief. Building upon this foundation, a novel classification system for HA-based interventions is proposed, aiming to standardize treatment protocols and optimize patient outcomes. The ViSCNOVAS classification system refers to viscosity, storage, chain, number, origin, volume, amount, and size. This classification is specifically designed for HA-based orthobiologic products used in regenerative medicine, including orthopedics, sports medicine, aesthetics, cosmetic dermatology, and wound healing. It aims to provide clinicians with a structured framework for personalized treatment strategies. Future directions in HA research are also discussed, emphasizing the need for further validation and refinement of the proposed classification system to advance the field of regenerative medicine. Overall, this manuscript elucidates the biological functions of hyaluronic acid and its potential in clinical practice while advocating for standardization to enhance patient care in various regenerative applications.