Flatworms are widespread in the marine realm, but only a few species parasitize corals. Several of these are considered pests in the aquarium trade, and the invasive potential of these species is a concern. In the Caribbean there are no records to date of flatworms naturally occurring in coral nurseries and/or parasitizing corals in the wild. Here we report, for the first time, a flatworm identified as Convolutriloba longifissura affecting a Caribbean coral and include a phylogenetic and histological analyses. Flatworms were collected from nursery corals and artificial recruitment units (bioballs) suspended in the water column at Punta Cana reef. Sequencing and morphology of individuals from both indicate that C. longifissura is present in the wild as well as the nursery. Experiments were conducted to investigate the effect of flatworm exposure on signs of macroscopic stress (i.e. mucus production, paleness, bleaching, and tissue loss) on microfragments of Pseudodiploria clivosa. Standard histopathological techniques were applied to describe microscopic alterations of the coral exposed to C. longifissura. We found corals exposed to 30 individuals of C. longifissura had a 2.5-fold increased probability of developing macroscopic stress signs compared to controls after 14 d of exposure to different flatworm densities. While the actual impacts of these flatworms on coral restoration and microfragmentation programs or wild corals in the Caribbean remain to be determined, our results indicate that flatworms are no longer an exclusive problem of Indo-Pacific corals.
The gain-of-function allele rol-6 ( su1006 ) has been used as a co-injection marker to visually identify transgenic progeny following microinjection in Caenorhabditis elegans . The rol-6 (gf) allele yields a clear, visual roller phenotype; however, affected worms are twisted along the anterior-posterior axis, obscuring tissues and complicating visual analyses. We deployed CRISPR/Cas9 to "unroll" transgenic worm strains harboring rol-6 (gf) . We discovered that our successfully unrolled strains introduced additional nucleotides into the rol-6 (gf) loci, rendering the rol-6 (gf) copies inactive. Our results indicate that genome engineering can be easily deployed to modify existing transgenic worm strains and could be applied to other gain-of-function co-injection markers.
Researches are being frequently conducted on the utility aspects of helminth parasites worldwide. The evolution is driven by the need for survival against the toxicity of biometals that emerged in zoonotic roundworms of Anisakidae. This study is aimed at investigating whether the sequestration of biometals from the environment into the body tissues of parasites serves a dual purpose: firstly, the removal of toxic metals and other biotoxins from the immediate environment of worms and, secondly, the utilization of extra available biometals, like sulfur and cysteine, in the surrounding environment to strengthen the cuticularization for the body's defense. Additionally, it examines whether the toxins absorbed by the tissues of worms thus protect tissues of the fish host from damage caused by toxicity; the potential "environmental sink" thus came into operation. These worms exploited the alterations in chemical composition to adapt and evolve in the newer environment. An assessment of biotoxic influence generated through heavy metal toxicity was conducted during this investigation. Scanning electron microscopy and energy-dispersive x-ray micrographic analysis were the techniques employed. The studies were conducted in anisakid nematodes parasitizing reef-associated fishes at Ilha "Grande" Island, 19 km from Panaji, Goa, in India. The first record of Anisakis typica third-stage larvae has recently appeared in Indian fish. The survival of worms of Anisakidae under the influence of a hostile environment comprising toxic biometals, like Hg, Pb, and Cd, was analyzed. The hypothesis of the metal filter being functional at the worm-iron influx interface was propounded. The typical anisakids of the family Anisakidae trigger a zoonotic mechanism that involves damage to human health. The superfluous iron influx within the intestinal environment apparently brought about physiological changes in the host's body tissues as well. The excessive iron content from the tissues of nematodes first crossed the worm's membrane barrier. Next, the iron content was stored in different body organs of roundworms to avoid the influence of toxicity on the fish host's body. Apparently, the parasitic tissues of the nematode functionally operated as an environmental sink which could relieve body tissues of the host fish due to unwarranted duress under biometallic stress.
Among parasites, roundworms release substances that quietly reshape how the body's defences respond - a dance shaped by long evolution. One player, called p43 - also known as Tm-DLP-1 - (Trichuris muris Dorylaimia Lipid-carrying Protein-1), makes up nearly all of it. Scientists now see this protein not just as a worm tool but as something that can steer immune responses, possibly helping fight tumours. Here lies an exploration: how p43 alters immunity, teams up with gut microbes, and shows promise in early tests and trials against cancer. Beyond its role inside the parasite hauling lipids around, p43 works outside by grabbing tightly onto IL-13, a signal involved in fighting worms - and oddly enough - in feeding some cancers. Thanks to parts resembling the IL-13 receptor alpha-2 and sections like those found in thrombospondin, this protein binds IL-13 strongly while also attaching to sugar chains in the tissue scaffold, leaving pockets where immune behaviour shifts subtly. Despite strong results across several animal cancer models - slowed tumours, stronger CD8+ T cell attacks, weaker regulatory T cell activity - the path forward stays uncertain. Safety data in healthy humans come from trials using Trichuris suis ova, yet those with weakened immune systems face lingering risks. A molecule called p43/Tm-DLP-1 emerges here, not by chance, due to its ability to block IL-13 while shifting gut microbes. Still, progress demands more lab refinement, larger production methods, and human studies built with precision before any real impact can take hold.
Loiasis is a parasitic infection caused by the filarial nematode Loa loa, endemic to Central and West Africa. Its occurrence in non-endemic regions such as Morocco is rare and may pose a diagnostic challenge. We report the case of a Moroccan patient presenting with an intermittent sensation of a mobile foreign body in the eye, associated with redness and photophobia. The patient reported no recent travel to endemic regions but had close contact with individuals originating from Cameroon, a loiasis-endemic area, raising questions regarding possible indirect or remote exposure. Following several consultations, spontaneous exteriorization of a subconjunctival worm was observed. The diagnosis was supported by clinical observation of the subconjunctival worm, blood parasitological testing demonstrating microfilaremia, and the presence of Calabar swellings. The patient was treated with ivermectin followed by albendazole, resulting in complete clinical recovery without recurrence during follow-up. This case highlights the importance of considering loiasis in the differential diagnosis of atypical ocular symptoms, even in non-endemic settings, and underscores the value of multidisciplinary collaboration for timely diagnosis and appropriate management.
Accurate quantification of egg-laying in Caenorhabditis elegans from 4K video recordings remains a bottleneck due to the cost of manual annotation and, above all, the risk of missing events. We present a human-in-the-loop assistance method explicitly designed under a recall-first criterion: to avoid missed egg-laying events (recall close to 1.0) while reducing operator review time. The workflow combines two complementary, deliberately over-detecting approaches - appearance by inter-frame difference restricted to a worm-centred local mask, and trajectory-based sustained-change detection - with a minimalist GUI that allows events to be confirmed or discarded within seconds and exports traceable results. The method was validated on six videos (4K, 5 fps; reviewed at 25 fps; one worm per video). Using manual annotation as the ground truth, the system achieved a recall of 0.992 (95% CI: 0.955-0.999) with moderate precision (0.77), consistent with the recall-first strategy. The approach shifts operator effort from exhaustive frame-by-frame searching to rapidly confirming pre-filtered candidates, maintaining full traceability and reproducibility.
Oviposition-site selection is a key maternal decision in oviparous animals, involving trade-offs among incubation conditions, offspring performance, and maternal constraints. In amphisbaenians (worm lizards), a clade of highly specialized fossorial reptiles, oviposition has long been assumed to occur obligatorily in ant and termite nests, implying strong ecological specialization. Here, we re-evaluate this assumption using new field observations and a critical synthesis of published records. We describe a natural oviposition site of the smallhead worm lizard (Leposternon microcephalum) located in a soil cavity with no evidence of ants or termites and review 18 records from nine amphisbaenian species. Across species, eggs occur not only in ant and termite nests but also in subterranean cavities and decaying logs, indicating facultative rather than exclusive use of social-insect nests. A field inspection of 31 active ant nests during the reported oviposition season yielded no eggs or amphisbaenians. Additional evidence suggests some species excavate or modify underground chambers and that at least one species oviposits communally. These findings challenge the view of strict dependence on ant and termite nests and instead support oviposition-site choice in amphisbaenians as a flexible, context-dependent maternal behavior shaped by ecological trade-offs rather than rigid specialization. Standardized surveys across microhabitats and experimental tests of incubation environments are needed to clarify how availability, costs, and benefits interact to shape nesting decisions in fossorial reptiles.
Owing to their overlapping geographical distribution and the consequent occurrence of coinfections, several studies have evaluated the impact of helminth infections on malaria immune responses and clinical outcomes. However, little has been reported on how malaria coinfection affects anthelmintic responses in children harbouring worm infections. We therefore aimed to assess the impact of malaria coinfection on helminth-related clinical and immune outcomes in children. Here, we measured in plasma the levels of 30 cytokines, total IgE, and helminth-specific IgM, IgA, IgG, IgG1-4, and IgE antibodies to a panel of 11 helminth antigens by Luminex. Samples were obtained from 441 children aged 2-10 years with diverse symptomatology, recruited from two hospitals in the Manhiça District (Mozambique). Parasite infections were diagnosed by rapid antigen diagnostic test, microscopy, and/or quantitative PCR. Among the recruited children, 96 were diagnosed with helminth infection, of whom 16 (17%) were coinfected with Plasmodium falciparum, and 80 (83%) had helminth infection alone. Coinfection was associated with an increased prevalence of anaemia (33% [5/15], p = 0.006) compared to helminth infection alone. Coinfected children also exhibited higher concentrations of pro-inflammatory cytokines (IL-2R and IL-6), cell-recruiting chemokines (MCP-1 and MIG), anti-inflammatory proteins (IL-1RA and IL-10), and the growth factor HGF (p < 0.05). Additionally, helminth-specific antibody responses were significantly enhanced (p < 0.05) in coinfected participants, with total IgG and IgM responses showing the strongest associations with coinfection. Notably, antibodies to several antigens, including Sh-SERPIN, Sm-SERPIN, As-37, Na-SAA-2, Na-GST-1, and Tm-16, were consistently increased across multiple isotypes (IgG, IgG cytophilic subclasses, IgM, and IgE) among coinfected children. In summary, malaria coinfection was associated with an increased risk of anaemia and enhanced inflammation and specific antibody responses in helminth-infected children, which could potentially influence worm expulsion or survival, as well as helminth vaccine efficacy.
Schistosomiasis, or bilharzia, is a highly prevalent water-borne helminth infection that accounts for an estimated 1.5-1.7 billion disability-adjusted life years lost annually. Pathology driven by adult worms and eggs is accompanied by robust immunomodulation in the human host. Protective responses may promote worm elimination, limit bystander tissue injury caused by parasite migration and contain eggs within granulomatous lesions. However, evidence also suggests that schistosomes can actively or indirectly influence haematopoiesis, thereby reshaping downstream mature immune cell responses. This perspective reviews current literature on schistosome-dependent and host-derived regulation of haematopoiesis, the importance of preclinical modelling and the emerging relevance of controlled human challenge models. It also discusses bone marrow organoid models and their utility for recapitulating complex infection dynamics, with the aim of informing clinically relevant studies and therapeutic strategies.
Northern bobwhite (Colinus virginianus) populations have experienced sustained declines across their native range, with parasitic helminths, particularly the eyeworm (Oxyspirura petrowi) and cecal worm (Aulonocephalus pennula), emerging as contributing stressors. This study assessed the long-term efficacy of a biannually administered anthelmintic feed treatment (QuailGuard®) and characterized seasonal and annual reinfection patterns in wild bobwhite populations at 2 sites (Oklahoma and Texas) from 2023 to 2025. Fecal samples were collected from wild bobwhite from March to October and analyzed via quantitative polymerase chain reaction (qPCR) to estimate infection levels. Aulonocephalus pennula infection levels were consistently reduced following spring and fall treatments, with efficacy ranging from 61 to 92%, except for 1 unsuccessful treatment in the fall of 2024 at the Texas site. Reinfection was rapid during the summer, and overwinter reinfection was more severe in 2024-2025 than the previous year. Oxyspirura petrowi prevalence was lower overall but showed similar seasonal trends. Birds under approximately 10 wk of age were less likely to have high infection levels, and the population's age structure influenced infection dynamics in late summer. A significant co-infection association between A. pennula and O. petrowi suggests overlapping parasite incidence, which is expected given their shared intermediate hosts. Although the medicated feed demonstrated sustained effectiveness, treatment outcomes varied, likely due to climate conditions and bird behavior. These results support continued biannual treatments but emphasize the need for site-specific adjustments, such as extending pre-baiting periods, to enhance efficacy. Continued monitoring will refine treatment timing, enhance cost-effectiveness, and mitigate the impacts of helminths on bobwhite populations.
In the past decade, the emergence of feline metastrongyloid lungworms has been increasingly recognized across Europe. The parasite is now being reported in previously non-endemic areas, with severe clinical cases described more frequently. This study describes the first molecular identification of Aelurostrongylus abstrusus in a naturally infected domestic cat in Serbia and treatment efficacy clinical and parasitological follow up. Nested PCR targeting specific 233-bp long fragment of the internal transcribed spacer 2 region was used for species identification, which was further confirmed by sequence analysis. The animal was treated with 50 mg/kg fenbendazole orally during 15 days. The efficacy of treatment was monitored by examining the presence of larvae in faecal samples each day throughout the course of therapy by using Baerman technique. Fenbendazole has shown to be effective in eliminating parasites, with larvae being completely absent from faecal samples by the 13th day of treatment. A worsening of clinical signs was observed following the initiation of therapy. The period of the highest larval burden in faecal samples correlated with the most intense coughing, which was also consistent with radiographic findings in the form of alveolar patterns. An additional experiment was done, showing that larvae can survive fluctuating daily temperatures in liquid environment for up to 51 days. The findings in this study indicate the emergence of feline lungworms and underscore its inclusion in differential diagnoses of feline respiratory syndrome. The importance of close monitoring during therapy highlights the need for further research, particularly in relation to treatment and prevention.
Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) enables spatial mapping of biomolecules within biological tissues, where conventional MS1-based workflows often result in ambiguous compound annotations. Data-dependent acquisition (DDA) can improve annotation specificity but is biased toward high-abundant ions and lacks reproducibility. Here, we present a novel MALDI MSI workflow integrating data-independent acquisition (DIA) to obtain both spatial and fragmentation information. In this approach, MS1 and DIA MS2 spectra are acquired alternately across the sample without prior knowledge of compound localization. Each image pixel consists of one MS1 and several DIA MS2 subpixels, providing both high spatial resolution for MS1 data and broad m/z coverage. Using small, randomized m/z isolation windows reduces spectral overlap and improves fragment-ion specificity. Data were processed using an extended Compound Discoverer, integrating mzCloud and LipidSearch for compound annotation. Applied to tissue of the parasitic worm Fasciola hepatica, this workflow produced detailed lipid maps and improved annotation confidence by combining precursor-mass, DIA MS2, and spatial-correlation information. Our results demonstrate that DIA offers a flexible strategy to integrate fragmentation information into MALDI MSI, expanding its capabilities for spatial metabolomics.
Parkinson's disease is associated with amyloid fibrillation of alpha-synuclein (α-syn) that involves the aggregation of protein into insoluble fibrils. In this study, we investigate the effect of luteolin (LUT) on α-syn aggregation in vitro, in silico, and in Caenorhabditis elegans. The decrements in light scattering and ThT fluorescence reveal that there is less formation of α-syn aggregates with an increase in the concentration of LUT. Also, a decrease in the ANS fluorescence depicts less exposure of hydrophobic patches in α-syn in the presence of LUT. Far UV CD data shows that LUT resists conversion structure into beta sheets. Dynamic light scattering and transmission electron microscopy revealed that LUT decreases the size and number of fibrils formed. Additionally, LUT reduced RBC hemolysis, indicating its potential therapeutic value in preventing the formation of toxic aggregates. In vivo studies in C. elegans also show that LUT improves the mitochondrial health, reduces reactive oxygen species (ROS), and enhances worm motility. Computational studies provide probable mechanistic insight that LUT interacted with α-syn via hydrophobic and hydrogen interactions, thus limiting the formation of fibrils by masking the sites that might be involved in aggregation. Consequently, LUT is an inhibitor of α-syn aggregation and may be a potential therapeutic agent against Parkinson's disease.
We report a pediatric case in which a retrolenticular Armillifer armillatus infestation mimicked a white cataract. A ten-year-old girl presented with a white cataract of her right eye during a humanitarian mission in the Democratic Republic of Congo. Fundoscopy was inaccessible. During surgery, an annulated worm-like parasite was found in the retrolenticular space. Further anamnesis was remarkable for regular snake and crocodile meat consumption, and a clinical diagnosis of ocular pentastomiasis was made. The parasite was morphologically and molecularly identified as A. armillatus. After surgery, fundoscopy showed diffuse retinal atrophy and the vision did unfortunately not improve. Human pentastomiasis is a rare, yet emerging zoonosis in rural tropical areas. Consumption of undercooked, infected snake (or rarely other reptile) meat may lead to visceral pentastomiasis, with uncommon ophthalmological involvement. Ocular pentastomiasis can be a devastating cause of vision loss in endemic regions. This case illustrates that a retrolenticular location of the parasite was likely responsible for a white cataract in an otherwise quiet, white eye, highlighting intraocular pentastomiasis as a potentially underdiagnosed cause of vision loss in endemic regions.
Lavandula angustifolia (lavender) extract displayed antioxidant properties in mammalian studies and has been promoted as a candidate neurotherapeutic for Alzheimer's disease (AD). To better inform its clinical utility, we exposed wildtype ( N2 ) and spr-4 mutant strains of C. elegans to extracts from this lavender species and examined animals for neurobehavioral changes in a mechanosensory phenotype. Importantly, spr-4 encodes the worm ortholog of repressor element 1-silencing transcription factor (REST), an established genetic modifier of AD. While low concentrations of lavender did not alter behavioral responses, spr-4 mutants selectively displayed neuronal vulnerability at the highest concentration tested, thereby revealing dose-responsive, lavender-associated neurotoxicity.
Dioctophymosis is a zoonotic disease caused by Dioctophyme renale, a nematode that can reach up to 100 cm in length and is commonly known as the "giant kidney worm." The parasite shows a marked predilection for the right kidney, and ultrasonography is the diagnostic method of choice, allowing visualization of characteristic tubular structures within the renal parenchyma. Surgical intervention remains the only established treatment, with nephrectomy being the most commonly performed procedure, although nephrotomy may be considered in cases with residual renal parenchyma. This study aimed to retrospectively describe the surgical technique, patient selection criteria, and postoperative outcomes of open nephrotomy in dogs with renal D. renale infection. Eighteen dogs diagnosed with right renal dioctophymosis between 2017 and 2024 were included. Patient selection was based on ultrasonographic evidence of preserved renal parenchyma. A paracostal approach was used in most cases to optimize renal exposure, and vascular occlusion was achieved using a Rummel tourniquet, with ischemia time maintained below 20 min. Parasite removal was successfully achieved in all patients. Postoperative monitoring showed stable or improved serum creatinine concentrations in the evaluated cases. Nephrotomy was a feasible surgical option in selected patients with renal dioctophymosis. Early diagnosis, before complete parenchymal destruction occurs, may increase the number of candidates for this approach, although further prospective studies are needed to better characterize long-term outcomes and refine patient selection criteria.
Chromatographic isolation of Crotalaria madurensis Wight & Arn leaves aqueous methanol extract availed a new potassium sulfonate acylated flavonol glycosides named quercetin 8-potassium sulfonate 3-O-[2-O-sulfonyl]-β-D-4C1-glucopyranosyl-(1''''→3''')-4-O-[E- caffeoyl]-β-D-4C1-glucopyranosyl-(1'''→2'')-3-O-[E-caffeoyl]-β-D-4C1-glucopyranoside (Crotalamadoside A) (1), along with three known flavonoids identified as quercetin 7-O-neohispredoside (2), 3',4'-dimethoxy quercetin 3-O-neohispredoside (3) and isoquercetin (6) besides two triterpene saponin compounds named as hedragenin 3-O-β-D-4C1-glucopyranoside (4) and hedragenin 3-O-α-L-1C4-rhamnopyranoside (5). This was in addition, to two cinnamic acid derivatives named as E-caffeic acid 4-O-β-D-4C1-glucopranoside (7) and E-caffeic acid (8). Identification of the isolated compounds relied on their chromatographic properties and spectral data (UV, ESI-MS, 1H NMR, 13C NMR, 1H-1H COSY, HSQC, and HMBC). The antischistosomal activity of the C. madurensis different extracts were assessed in vitro using schistosome worm killing, findings revealed that the aqueous methanol extract was the most effective extract, chromatographic fractionation of this extract revealed significant antischistosomal activity for fractions number V and VI rich in flavonoids and triterpene saponins, respectively.
Schistosomiasis, caused by parasitic worms, affects over 250 million people globally, with limited treatment options due to praziquantel's inability to prevent reinfection or reduce immunopathology. In Schistosoma mansoni (S. mansoni) infection, egg-induced granulomatous inflammation in the liver and intestines is driven by CD4 T helper (Th) cell responses, with severe pathology in some individuals mediated by Th17 cell activation. We previously demonstrated that the stimulator of interferon genes (STING) mitigates schistosome egg-induced immunopathology by promoting type I Interferon (IFN-I) production and suppressing Th17 responses. Here, we investigate the therapeutic potential of the STING agonist diABZI-3 in a high-pathology CBA mouse model. In vitro, diABZI-3 pretreatment of bone marrow-derived dendritic cells (BMDCs) significantly enhanced IFNβ production while abolishing IL-1β and IL-17 expression in response to schistosome egg stimulation, an effect dependent on early administration. In vivo, a single dose of diABZI-3 administered to S. mansoni-infected CBA mice reduced liver and intestine granuloma size, lowered IL-1β, IL-17, and CD209a levels, promoted the Foxp3⁺ regulatory T cells, reduced Th17 recruitment, and mitigated inflammation-associated shifts in gut microbiota populations. Furthermore, blocking STING degradation with bafilomycin A1 sustained STING signaling, leading to pronounced IL-1β suppression. These findings highlight diABZI-3 as a promising therapeutic agent for reducing schistosome-induced immunopathology.
Pyruvate dehydrogenase complex (PDHc) deficiency (PDCD) is a primary mitochondrial disorder characterized by neurodevelopmental disability, altered intermediary metabolism and early mortality. Dichloroacetate (DCA), a pyruvate analogue, is a well-described PDHc activator that remains under clinical investigation for treatment of PDCD. Here, we studied the in vivo efficacy of a 5-point log concentration range of DCA on animal health and metabolism in C. elegans with feeding RNA interference (RNAi) expression knockdown of either PDHA-1 or DLD-1 homologues at graded degrees to model variable disease severity. These worm models recapitulate phenotypic features of PDCD observed in human patients, including reduced survival, delayed growth, locomotor impairment, and elevated lactate and/or pyruvate tissue levels. DCA treatment appeared well-tolerated, with no gross morphologic toxicity seen at doses up to 25 mM. Significantly improved health, survival, tissue lactate levels, and mitochondrial physiology were observed at 25 mM in pdha-1(RNAi) knockdown animals. DCA treatment in dld-1(RNAi) C. elegans models (undiluted, 1:20 dilution, and 1:100 dilution) showed significant therapeutic benefits on survival, neuromuscular function and metabolic phenotypes primarily in the moderate (1:20) and/or mild (1:100) dld-1(RNAi) deficiency strains, but not in full-dose dld-1(RNAi) . Importantly, linear growth, neuromuscular activity, and mitochondrial physiology were significantly improved with DCA treatment even in the most severe dld-1(RNAi) undiluted model. Overall, preclinical modeling provides objective evidence of DCA therapeutic efficacy in C. elegans expression knockdown strains for two well-conserved homologues of PDHA1 and DLD that represent distinct genetic etiologies of PDHc deficiency, with demonstrated beneficial effects on survival, healthspan, tissue lactate, and mitochondrial physiology. These data further confirm that DCA's therapeutic effect correlates with PDHc disease phenotype severity in dld-1(RNAi) animals. SYNOPSIS: Dichloroacetate (DCA) treatment demonstrated significant preclinical beneficial effect on survival, neuromuscular function, linear growth, tissue lactate, and mitochondrial metabolism in two C. elegans models with variable degrees of pyruvate dehydrogenase complex (PDHc) deficiency (PDCD), providing confirmatory evidence to support its therapeutic potential in human PDCD patients.
Knee osteoarthritis (KOA) is a leading global cause of pain and functional impairment, with a high prevalence in India driven by aging and obesity. Traditional treatments often provide only symptomatic relief without addressing underlying structural degeneration. This study evaluates the efficacy of intra-articular growth factor concentrate (GFC) as a regenerative and disease-modifying therapy for moderate KOA. In this randomized controlled trial, 72 patients with Kellgren-Lawrence (KL) Grade II or III KOA were randomized into two groups. Group A (n = 36) received two ultrasound-guided intra-articular injections of autologous GFC spaced four weeks apart, while Group B (n = 36) received normal saline. Clinical outcomes were assessed using the Visual Analog Scale (VAS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Structural and biochemical changes were monitored via magnetic resonance imaging (MRI) (Whole-Organ Magnetic Resonance Imaging Score) and serum Coll2-1NO2 levels at baseline, 6 months, and 9 months. A total of 66 patients completed the 9-month follow-up. Group A demonstrated a significant reduction in mean VAS scores (6.48-2.79; P < 0.0001) and WOMAC scores (62.58-47.40; P < 0.0001), indicating superior pain relief and functional improvement compared to Group B, which showed mild worsening. Biochemically, Group A showed a significant decrease in serum Coll2-1 levels (P = 0.03), suggesting reduced cartilage breakdown. MRI analysis revealed increased WORM scores in Group A (130.03-136.91; P = 0.03), indicating cartilage preservation and potential regeneration, whereas Group B scores decreased (P = 0.027). Intra-articular GFC injection is a safe and effective intervention for KL Grade II and III KOA. It provides sustained symptomatic relief and demonstrates potential disease-modifying effects through cartilage preservation and reduced degradation biomarkers. GFC represents a viable biological therapy to bridge the gap between conservative management and invasive surgical procedures.