Biomedical Sciences and Clinical Medicine

ANGPT2 as a Therapeutic Target in Endometriosis: Evolving Perspectives in Angiogenesis - ANGPT2, an Emerging Target in Endometriosis Therapy

Sheeja M J — 2025-11-24

Endometriosis is a chronic, estrogen-dependent inflammatory condition marked by the ectopic implantation of endometrial-like tissue, most commonly involving the ovaries, peritoneum, and pelvic structures. This ectopic tissue responds to hormonal cycles, leading to symptoms such as
pelvic pain, dysmenorrhea, dyspareunia, and infertility. A critical process in the pathophysiology of endometriosis is angiogenesis, which sustains
lesion growth and survival. Angiopoietin-2 (Ang-2), encoded by the ANGPT2 gene, is a key regulator of abnormal vascular remodeling in endometriosis, acting as a vascular destabilizer and enhancing angiogenic responses, parti-
cularly in conjunction with vascular endothelial growth factor (VEGF).
This review synthesizes current evidence on the pathological role of Ang-2
in endometriosis-associated angiogenesis and evaluates its potential as a molecular target for therapy. A comprehensive literature search was performed using PubMed, Scopus, Google Scholar, and Web of Science,
covering articles published from 1997 to 2025. Relevant studies were selected
based on Ang-2’s involvement in angiogenesis, its expression patterns in endometriotic tissue, and the outcomes of therapeutic interventions.
The findings reveal that ANGPT2 expression is upregulated in endometriotic lesions and is modulated by hypoxia, estrogen, and inflammatory factors. Elevated levels of Ang-2 are associated with increased disease severity and vascular immaturity in lesions. Preclinical studies targeting Ang-2, either alone or in combination with VEGF inhibitors, have demonstrated reductions in lesion vascularization and growth, highlighting its therapeutic promise.
In conclusion, Ang-2 serves as a critical mediator of pathological angiogenesis in endometriosis and presents a promising target for novel anti- angiogenic therapies. Further translational research and clinical trials are warranted to explore its full therapeutic potential.

MC4R: A Genetic Key to Metabolic and Cardiovascular Disorders

Joby P Jose — 2025-12-08

The melanocortin-4 receptor (MC4R) is a critical regulator of energy homeo-stasis, lipid metabolism, and cardiovascular risk factors. First cloned in 1993, MC4R’s role in appetite regulation was confirmed through animal studies, and its genetic mutations were later identified as major contribu-tors to monogenic obesity. This review synthesizes current research on MC4R’s genetic variations and their implications for obesity, metabolic disorders, and cardiovascular health. A systematic literature review was conducted, adhering to PRISMA guidelines, with data extracted from PubMed, Scopus, Web of Science, and Google Scholar. Studies from 1992 to 2025 focusing on MC4R mutations, their effects on obesity, type 2 diabetes, dyslipidemia, and hypertension, as well as their contribution to cardiovascular risk were included. Recent insights highlight rare MC4R variants, such as p.Ser36Thr and p.Ala175Thr which are associated with severe obesity, and novel structural variants, e.g., p.Ser85Gly, which destabilize signaling pathways. Loss-of-function MC4R variants cause hyperphagia and early-onset obesity, whereas gain-of-function variants enhance satiety, providing protection against obesity. MC4R dysfunction also impacts glucose homeostasis and lipid metabolism, thereby increasing the risk of type 2 diabetes, dyslipidemia, and hypertension. Additionally, alterations in MC4R increase cardiovascular risk via pathways involving energy imbalance, sympathetic activation, and endothelial dysfunction. This review highlights the genetic underpinnings of MC4R in metabolic and cardiovascular diseases and underscores the need for developing targeted therapeutic strategies.

Phenethylamine Does Not Accelerate Healing in Staphylococcus aureus-infected Wounds in Mice

Hana Dian Mufina — 2025-12-08

Objective Bacterial infections can lead to a delay in the wound healing process. Recent studies have reported that trace amines accelerate wound healing by enhancing keratinocyte migration thus promoting a faster wound re-epithelization process. This study aimed to investigate the effects of phenethylamine on Staphylococcus aureus-infected wounds by using a mouse model.

Methods This study used mice as a model for in vivo skin wound experi-ments. S. aureus was applied on skin wounds on the backs of the mice. Phenethylamine in different concentrations was also applied either once daily or every two days. The wound diameter, abscess formation, and swollen area were observed every two days for 14 days.

Results Different concentrations and frequencies of phenethylamine applications on the wound exhibited no significant wound healing acceleration. Phenethylamine applications also did not show significant effects on abscess formation or swelling.

Conclusions S. aureus infection may overwhelm the wound healing acceleration effects of phenethylamine resulting in no significant improvement in healing.

Nanofibrous Scaffolds Functionalized with Serine for Dentin Mineralization: A Characterization Study

Aruna Krishnan — 2025-11-24

Objective Dentin repair and regeneration pose significant challenges in restorative dentistry due to its hierarchical structure and susceptibility to demineralization. Traditional approaches often fail to restore the naturalarchitecture and functionality of dentin. The aim of this study was to develop and characterize a biomimetic polycaprolactone (PCL)/nanohydroxyapatite (nHA)/serine (Ser) scaffold to enhance dental mineralization and support tissue regeneration. The inclusion of serine, known for its ability to bind calcium and phosphate ions and guide hydroxyapatite crystal formation, could enhance nucleation and cell attachment, making the scaffold a promising biomaterial for dentin repair.

Methods Nanofibrous scaffolds were fabricated using PCL, nHA, and Ser by the electrospinning method. The structural and functional properties of the scaffold were characterized through scanning electron microscopy (SEM), energy-dispersive x-ray spectroscopy (EDX), x-ray diffraction (XRD), and fourier transform infrared spectroscopy (FTIR). SEM and EDX were used to evaluate the morphology, porosity, and elemental composition. XRD analyzed the crystalline and amorphous phases, while FTIR analyzed chemical interactions among the components.

Results SEM analysis revealed a uniform fibrous structure with interconnected porosity, mimicking the extracellular matrix. EDX confirmed the successful incorporation of calcium and phosphorus, indicating the presence of nHA. XRD analysis demonstrated a combination of crystalline and amorphous phases, highlighting the scaffold’s structural stability and bioactivity. FTIR spectroscopy identified characteristic peaks corresponding to PCL, nHA, and Ser, validating their successful integration and interaction within the scaffold matrix.

Conclusions The nanofibrous scaffold exhibited structural and compositional properties, confirming the presence of PCL, nHA and Ser. The properties analyzed support its potential for biomineralization and tissue regeneration. The inclusion of Ser could enhance hydroxyapatite nucleation and cell attachment, making the scaffold a promising biomaterial for dentin repair. Further in-vitro and in-vivo studies are recommended to validate its clinical application.

Prevalence of Vitamin D Deficiency in Diabetic Neuropathy Patients in the South Indian Population

Vadivel Mani — 2025-12-08

Objective This research explores the prevalence of vitamin D deficiency among diabetic neuropathy patients in South India, highlighting the significant impact of this deficiency on health outcomes, particularly of the sensory-motor type.

Methods A cross-sectional study at Koonaseema Institute of Medical Science and Research Foundation (KIMS&RF) found a significant association between vitamin D deficiency and diabetic neuropathy, with factors such as sociodemographic, risk factors, treatment duration, treatment modality, DPN score, hyperalgesia, and neuropathic pain.

Results A study of 480 diabetic neuropathy patients found that 88.54% of them had a deficiency in vitamin D. Of those patients, 51.43% had insufficiency, while 37.11% had a more severe deficiency. The statistical analy-sis was conducted using Excel and the 16^th edition of SPSS (Statistical Package for the Social Sciences). Literate individuals had lower vitamin D levels, while middle-class individuals were more likely. Obesity was linked to vitamin D insufficiency, with 65.60% of obese patients having insufficient vitamin D. Treatment modalities, sensory thresholds, and neuropathic pain symptoms were also associated with vitamin D deficiency.

Conclusions This study in South India found that 88.54% of individuals, particularly those with diabetic neuropathy, were deficient in vitamin D. This is lower than in Egypt, but higher than in the UAE and China. Factors affecting vitamin D levels affect both males and females, with a higher impact on males. Complications from diabetic neuropathy are linked tovitamin D insufficiency. Further research is needed to validate these findings.

Effects of Inflammatory Biomarkers (Fetuin-A and High Sensitivity C-reactive Protein) on Glycemic Control in Type 2 Diabetic Women

Ali Al-bayati — 2025-11-24

Objective Inflammation plays an essential role in the development of insulin resistance and type 2 diabetes. The objective of this study was to evaluate the effects of inflammatory markers (hs-CRP and fetuin-A) on glycemic control in Iraqi women patients with type 2 diabetes, and to examine the correlation of these markers with indices of glycemic control.

Methods This case-control study included 45 Iraqi type 2 diabetic female patients and 44 non-diabetic Iraqi female patients matched for age and sex as control subjects. For both patients and controls hs-CRP and fetuin-A were measured using ELISA, HbA1c by the turbidimetric immunoassay, FBG and lipid profile measures using the spectrophotometer method.

Results High levels of both fetuin-A and hs-CRP were observed in diabetic patients compared to the healthy controls. hs-CRP was significantly higher in the diabetic group 3.1 ± 0.59 mg/dL than the control 1.22 ± 0.66 mg/dL and was statistically significantly correlated with the marker of glycemic control (p < 0.001). The serum level of fetuin-A in the diabetic group (768.7 ± 173.03 µg/mL) was also significantly higher than that of control (311.95 ± 94.13 µg/mL) (p < 0.001). A strong positive correlation was observed between fetuin-A concentrations with serum hs-CRP concentrations in the diabetic individuals but not in the controls.

Conclusions Diabetic Iraqi women have higher inflammatory markers (fetuin-A and hs-CRP) than non-diabetic women. A strong association was observed for these inflammatory biomarkers and glycemic control in diabetic subjects.

Development of Zinc Oxide Nanoparticle–Infused Carrageenan Membranes with Enhanced Structural and Antimicrobial Properties for Guided Tissue Regeneration

PRIYANGHA P T — 2025-11-24

Objective The fundamental aim of periodontal therapy is to promote the regeneration of tissues affected by the disease. In recent times, there has been a rising trend in using guided tissue regeneration (GTR) membranes. These membranes are increasingly relied upon to direct the growth of gingival tissue away from the root surface. Both resorbable and non-resorbable membranes presently in use serve as physical barriers, hindering the infiltration of connective and epithelial tissues into the defect area, thus fostering periodontal tissue regeneration. This study aims to develop a polymeric membrane reinforced with carrageenan and zinc oxide nanoparticles and assess its potential for periodontal regeneration.

Methods 3g of dried Kappaphycus was mixed with 100 mL of distilled water in a container and autoclaved for 15 minutes at 121 degrees Celsius. After cooling, the mixture was blended thoroughly for carrageenan extraction. Then, 1.5 g of PEG 6000 was added as a plasticizer. Zinc oxide nanoparticles (ZnO) were added and homogenized into the carrageenan extract. The mixture was microwave-boiled for 2 minutes, poured onto a petri dish, and air-dried for 24 hours. Further drying at 50 degrees Celsius for 2 hours ensured complete moisture removal. Finally, the membrane was carefully peeled from the petri dish for testing and use. The membrane underwent SEM, tensile strength, Fourier transform infrared, and antimicrobial activity analysis.

Results Scanning electron microscopy analysis revealed a densely packed and moderately rough surface in the ZnO-incorporated membrane, with a broader particle size distribution (1.28 ± 3.67 µm) compared to the smoother carrageenan-only membrane (1.15 ± 1.68 µm). Tensile testing showed improved mechanical strength in the ZnO composite (6.89 MPa) relative to the plain carrageenan membrane (5.39 MPa). FTIR spectra confirmed successful integration of ZnO nanoparticles through characteristic Zn–O peaks, along with preserved polysaccharide functional groups. Antimicrobial activity, evaluated via OD₆₀₀ measurements over 4 hours, demonstrated time- and size-dependent bacterial inhibition against S. mutans, with the 4 cm² membrane showing superior performance, comparable or superior to chloramphenicol at later time points.

Conclusions The developed membrane reinforced with carrageenan and zinc oxide nanoparticles exhibited adequate tensile strength and sufficient antimicrobial properties, suggesting its suitability for utilization in periodontal therapy as an effective regenerative material.

Automated Immunohistochemistry as an Alternative to Conventional Immunohistochemistry for Analyzing the Expression of BCL2 and Ki-67 in Diffuse Large B-Cell Lymphoma Patients

Phuttirak Yimpak — 2025-11-27

Objective Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma. Immunohistochemistry (IHC) is widely used to assess prognostic markers such as BCL2 and Ki-67. Conventional IHC relies on manual staining and visual estimation by pathologists, while automated IHC incorporates computer-based image analysis. This study aimed to evaluate the level of agreement between these two IHC workflows in assessing BCL2 and Ki-67 expression in DLBCL patients.

Methods Formalin-fixed, paraffin-embedded tissue samples from nodal and extranodal sites of 42 and 60 DLBCL cases were analyzed for BCL2 and Ki-67, respectively. Conventional IHC involved manual staining and evaluation by a pathologist in 10.00% increments. Automated IHC used the Ventana BenchMark ULTRA system with digital image analysis. Expression levels of the two systems were compared using paired t-tests, intraclass correlation coefficients (ICC), Pearson’s correlation, and Bland–Altman plots.

Results For BCL2, no significant difference was found between the two methods (mean difference = 5.79%, p = 0.103), with moderate to good agreement (ICC = 0.739) and strong correlation (r = 0.772). For Ki-67, patho-logist-based assessment yielded significantly higher values (mean difference = 13.91%, p < 0.001), with moderate agreement (ICC = 0.569) and strong correlation (r = 0.688). Bland–Altman analysis confirmed acceptable limits of agreement, although a slight underestimation in the automated Ki-67 results was observed.

Conclusions Automated IHC showed moderate to good agreement with conventional IHC, particularly for BCL2. Despite some differences in Ki-67 quantification, both methods demonstrated consistent trends. Auto-mated IHC offers reproducibility and scalability, making it a promising complementary tool in pathology.

Comparative Study of Guava Leaf Bioactive Extracts Using Ultrasonication Assisted Extraction

Somya Khanna — 2025-11-24

Objective Guava (Psidium guajava L.), a member of the Myrtaceae family, is grown worldwide and is widely consumed in India because of its good nutritional value as well as its medicinal properties. various parts of the guava tree, e.g., its root, bark, leaves, and fruit, have been found to be rich in pharmacological properties. Despite the health benefits, parts of the tree other than the guava fruit are not widely used by people. Guava leaves are generally considered as a non-conventional food with high nutritive value, medicinal properties, essential oils and good amounts of bioactive compounds.

Methods In this study, these bioactive compounds were extracted by ultrasound-assisted extraction (UAE) using three different solvents (water, ethanol and methanol) and the extracts were tested for finding the bioactive content in it.

Results This study found that dilute methanol (50 : 50) extracts contain significant amounts of flavonoids, alkaloids, saponins and terpenoids. Ethanol extracts showed good quantities of tannins and aqueous extract showed good extraction of total phenolic content.

Conclusions These findings show that guava leaves are rich in bioactive compounds which can be extracted using environmentally friendly methods which can be used in food and pharmaceutical industries

Can the Indonesian Version of the Work-Related Quality of Life Scale-2 (WRQoLS-2) Be Used to Measure Quality of Work Life Among Hospital Personnel?

Naesinee Chaiear — 2025-11-27

Objective This study aimed to construct and validate the Work-Related Quality of Life Scale-2 (WRQoLS-2) for use in Indonesia.

Methods A cross-cultural adaptation process, including forward and backward translations by multilingual experts, ensured linguistic and conceptual equivalence of the items. Six nursing experts assessed the content validity, and the reliability was tested using Cronbach’s alpha, Intraclass Correlation Coefficients (ICCs), and Spearman rank correlation coefficients among 38 registered nurses.

Results The Indonesian WRQoLS-2 showed strong content validity (S-CVI/Ave of 0.97, S-CVI/AU of 0.84) and high internal consistency (Cronbach’s alpha: 0.955-0.961). The ICC analysis yielded an overall ICC of 0.921, indicating excellent reliability. Most dimensions showed positive Spearman correlations, with the exception of three items in the Stress at Work dimension, which were revised and retested. The revised items confirmed that there were no comprehension issues vis-à-vis the questionnaire.

Conclusions Overall, the Indonesian version of the WRQoLS-2 proved to be a valid and reliable tool for assessing the quality of work life among hospital personnel in Indonesia.

Virgin Coconut Oil Inhibits Leukemic Cell Proliferation Via Apoptosis in K562 and MOLT-4 Cell Lines

Sawalee Saosathan — 2025-12-08

Objective Leukemia is a blood cancer which is expressed by an uncontrolled increase in abnormal white blood cells. To date, chemotherapy has been regularly used to treat leukemia. However, the major problem with chemotherapy is the side effects for leukemia patients. Nowadays, medicinal plants are of interest as an additional form of cancer treatment. Virgin coconut oil (VCO) is a source of antioxidants, anti-viral, anti-inflammatory, and anti-cancer properties. This study was designed to investigate the potential of VCO to inhibit the growth of leukemic cells by inducing cell cycle arrest and apoptosis.

Methods The VCO was analyzed using gas chromatography coupled with mass spectrometry (GC-MS). In the experiment, two leukemic cell lines, K562 and MOLT-4, were treated with various concentrations of VCO for 48 h. Cytotoxic effects were measured using the MTT assay and compared to commercial lauric acid. Cell cycle analysis was performed using propidium iodide (PI) staining, and apoptosis was assessed using an Annexin V-FITC kit.

Results The study found that VCO is primarily composed of lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, linoleic acid, oleic acid, and stearic acid. In addition, VCO showed cytotoxic effects in both K562 and MOLT-4 cells, with IC₅₀ values of 618.2 µg/mL and between 654 and 760 µg/mL, respectively. Cell cycle analysis demonstrated that VCO induced accumulation of cells in the G2/M phase. Additionally, the apoptosis assay demonstrated that VCO treatment induces apoptosis, particularly in K562 cells, in a cell-dependent manner.

Conclusions VCO is a promising source of bioactive compounds which exhibit cytotoxic, cell cycle arrest and apoptotic effects on leukemic cells in-vitro, suggesting its potential for further investigation as an anti- leukemic agent.

Sex-specific Differences in the Association between Insulin Resistance and Metabolic Syndrome and Ferritin in Korean Adults: A Nationwide Population-based Study

Mi Young Gi — 2025-11-28

Objective This study assessed the association between metabolic syndrome (MetS) and homeostasis model assessment of insulin resistance (HOMA-IR) and ferritin based on gender in Korean adults.

Methods This study was conducted using data from 5,730 adults (aged 20 or older) and the Korea National Health and Nutrition Examination Survey.

Results This study included some key findings First, HOMA-IR levels showed a positive correlation with quartiles (Q) of ferritin both in postmenopausal women (p = 0.001) and in men (p < 0.001). However, there was no statistically significant difference between the HOMA-IR levels and the Q of ferritin in premenopausal women (p = 0.149). Second, using the Q1 of ferritin as the reference, the ORs of MetS were positively significant in men (Q3 [OR, 1.770; 95% CI, 1.308–2.397] and Q4 [OR, 2.385; 95% CI, 1.775–3.20]) and postmenopausal women (Q4 [OR, 1.873; 95% CI, 1.351–2.596]). However, there was no significant difference between MetS and the quartiles of ferritin in premenopausal women.

Conclusions An increase in ferritin is associated with both metabolic syndrome and insulin resistance in both Korean postmenopausal women and men. However, there is no significant difference between ferritin and metabolic syndrome in premenopausal women.

Validity and Reliability of the Thai Version of the Barthel Index Self-Report (BI-SR-TH) for People with Spinal Cord Injury

Siam Tongprasert — 2025-11-28

Objective This study aimed to cross-culturally translate and evaluate the psychometric properties of the Thai version of the Barthel Index Self-Report (BI-SR-TH) for Thai people with spinal cord injury (SCI).

Methods A descriptive cross-sectional validation and reliability study was conducted at the Maharaj Nakorn Chiang Mai Hospital, involving 109 Thai people aged 18 or older with either traumatic or non-traumatic SCI. The English postal version of the Barthel Index was translated into Thai following standard cross-cultural procedures, including forward and backward translation. Participants completed both the BI-SR-TH and the Thai EuroQoL (EQ-5D-5L) questionnaires via self-report and clinician assessment using the Barthel Index. The BI-SR-TH was re-administered to the participants one week later. Psychometric evaluation included assessments of internal consistency (Cronbach’s alpha), concurrent validity (Spearman’s correlation with clinician-rated BI), construct validity (convergent and discriminant validity assessed by correlations with relevant and distinct EQ-5D-5L domains), and test-retest reliability (intraclass correlation coefficient, ICC). The agreement between the BI-SR-TH and BI scores was assessed using ICC and the Bland-Altmand Altman analysis. Item-level agreement was assessed using the Kappa statistic.

Results The BI-SR-TH demonstrated good internal consistency (Cronbach’s alpha = 0.836) and excellent concurrent validity with a clinician-rated BI (r = 0.884). In terms of construct validity, the BI-SR-TH showed strong negative correlations with relevant EQ-5D-5L domains, e.g., mobility, self-care, and usual activities (r = –0.67 to –0.81, p < 0.01), demonstrating convergent validity. In contrast, correlations with unrelated domains such as anxiety and pain were weak or nonsignificant, indicating discriminant validity. The total score agreement between BI-SR-TH and the clinician BI was high (ICC = 0.916), and Bland-Altman analysis indicated a small mean difference. Most BI-SR-TH items showed good agreement with clinician ratings (Kappa 0.61–0.72) with the exception of the bowels and bladder items which showed only fair agreement (Kappa = 0.34 and 0.35, respectively). Test-retest reliability was also high (ICC = 0.924).

Conclusions The BI-SR-TH is a valid and reliable tool for assessing functional independence in Thai people with SCI. However, the bowels and bladder items require further refinement for this population

Uncovering the Molecular Landscape of Secreted Phosphoprotein 1 and Its Expression Network in Kidney Renal Papillary Cell Carcinoma

Danang Prasetyaning Amukti — 2025-11-28

Objective This study aims to investigate the expression pattern of Secreted Phosphoprotein 1 (SPP1), its prognostic relevance, and the associated co-expression gene network in kidney renal papillary cell carcinoma (KIRP).

Methods Publicly available transcriptomic and clinical datasets were analyzed using GEPIA, UALCAN, and the GTEx Portal. SPP1 expression levels in KIRP versus normal kidney tissues were evaluated, followed by Kaplan–Meier survival analysis stratified by ethnicity, gender, and body weight. Co-expression analysis was performed to identify genes significantly correlated with SPP1 and to explore their potential biological functions.

Results SPP1 expression was significantly upregulated in KIRP tissues compared with normal kidney tissues, consistent with GTEx data showing naturally high expression in kidney cortex and medulla. High SPP1 expression was associated with poorer overall survival, particularly among Caucasian patients, while no significant survival differences were observed by gender or body weight. Ten genes strongly co-expressed with SPP1, MLLT3, AKIRIN2, YIPF1, SESTD1, LAMC1, FAS, ABCC3, MTHFD1L, ANXA2P2, and ARHGAP29 were identified, many of which are implicated in cell adhe-sion, migration, extracellular matrix remodeling, and apoptosis resistance.

Conclusions SPP1 is markedly overexpressed in KIRP and correlates with unfavorable prognosis, highlighting its potential as a diagnostic and prognostic biomarker. The identified SPP1-associated co-expression gene network may offer novel insights and therapeutic targets for improving molecular-based interventions in KIRP.

Next-Generation Dental Bone Implants − Sustainable 3D Nanostructures Derived from Animal By-Products: A Preliminary Study

Rethinam Senthil — 2025-12-08

Objective The development of next-generation dental bone implants focuses on sustainable, bioactive materials derived from animal by pro-ducts, offering a cost-effective and eco-friendly alternative for bone regeneration. This study explores the fabrication of a 3D nano-structured dental implant (3D-DI) using fish bone collagen (FBC) and fibrin nanoparticles (FN), chosen for their excellent biocompatibility, bioactivity, and availability.

Methods FBC and FN scaffolds were synthesized and incorporated into an implant framework. Fourier-transform infrared spectroscopy (FTIR) analysis confirmed the presence of bioactive functional groups. High-resolution scanning electron microscopy (HRSEM) characterized the porous microstructure (100–500 nm) as conducive to cell adhesion and proliferation. In-vitro cytotoxicity (MTT assay) and cell viability (live cell staining) were evaluated using MG63 osteoblast-like cells.

Results The 3D-DI demonstrated significant bioactivity after 14 days in simulated body fluid (SBF), with a mineralization rate of 68% (p < 0.01). MG63 cell viability increased to 85% after 72 hours (p < 0.05) (ANOVA using Microsoft Excel Office 2013), indicating excellent biocompatibility. Mechanical testing showed compressive strength of 83.41 ± 0.56 MPa and tensile strength of 76.83 ± 0.81 MPa suitable for load-bearing dental and orthopedic applications (p < 0.05).

Conclusions This study presents a novel approach for transforming animal waste into sustainable, bioactive dental bone implants, offering an eco-friendly alternative to conventional graft materials. Unlike previous reports that primarily focus on single-component or synthetic scaffolds, 3D-DI exhibits superior mechanical strength, enhanced mineralization, and excellent biocompatibility. These unique properties highlight its potential for load-bearing dental and orthopedic applications, addressing the current gap in development of cost-effective, high-performance biomaterials from bio-waste sources.